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Sample records for c2h2 zinc finger

  1. Herbivory responsive C2H2 zinc finger transcription factor protein StZFP2 from potato

    Science.gov (United States)

    While C2H2 zinc finger transcription factors are often regulated by abiotic stress, their role during insect infestation has been overlooked. This study demonstrates that the transcripts of the zinc finger transcription factors StZFP1 and StZFP2 are induced in potato (Solanum tuberosum) upon infesta...

  2. Conservation, diversification and expansion of C2H2 zinc finger proteins in the Arabidopsis thaliana genome

    Directory of Open Access Journals (Sweden)

    Böhm Siegfried

    2004-07-01

    Full Text Available Background The classical C2H2 zinc finger domain is involved in a wide range of functions and can bind to DNA, RNA and proteins. The comparison of zinc finger proteins in several eukaryotes has shown that there is a lot of lineage specific diversification and expansion. Although the number of characterized plant proteins that carry the classical C2H2 zinc finger motifs is growing, a systematic classification and analysis of a plant genome zinc finger gene set is lacking. Results We found through in silico analysis 176 zinc finger proteins in Arabidopsis thaliana that hence constitute the most abundant family of putative transcriptional regulators in this plant. Only a minority of 33 A. thaliana zinc finger proteins are conserved in other eukaryotes. In contrast, the majority of these proteins (81% are plant specific. They are derived from extensive duplication events and form expanded families. We assigned the proteins to different subgroups and families and focused specifically on the two largest and evolutionarily youngest families (A1 and C1 that are suggested to be primarily involved in transcriptional regulation. The newly defined family A1 (24 members comprises proteins with tandemly arranged zinc finger domains. Family C1 (64 members, earlier described as the EPF-family in Petunia, comprises proteins with one isolated or two to five dispersed fingers and a mostly invariant QALGGH motif in the zinc finger helices. Based on the amino acid pattern in these helices we could describe five different signature sequences prevalent in C1 zinc finger domains. We also found a number of non-finger domains that are conserved in these families. Conclusions Our analysis of the few evolutionarily conserved zinc finger proteins of A. thaliana suggests that most of them could be involved in ancient biological processes like RNA metabolism and chromatin-remodeling. In contrast, the majority of the unique A. thaliana zinc finger proteins are known or

  3. Involvement of C2H2 zinc finger proteins in the regulation of epidermal cell fate determination in Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    An Yan; Minjie Wu; Yongqin Zhao; Aidong Zhang; Bohan Liu; John Schiefelbein; Yinbo Gan

    2014-01-01

    Cell fate determination is a basic developmental process during the growth of multicellular organisms. Trichomes and root hairs of Arabidopsis are both readily accessible structures originating from the epidermal cells of the aerial tissues and roots respectively, and they serve as excellent models for understanding the molecular mecha-nisms controlling cell fate determination and cell morphogen-esis. The regulation of trichome and root hair formation is a complex program that consists of the integration of hormonal signals with a large number of transcriptional factors, including MYB and bHLH transcriptional factors. Studies during recent years have uncovered an important role of C2H2 type zinc finger proteins in the regulation of epidermal cell fate determination. Here in this minireview we briefly summarize the involvement of C2H2 zinc finger proteins in the control of trichome and root hair formation in Arabidopsis.

  4. Characterization of How DNA Modifications Affect DNA Binding by C2H2 Zinc Finger Proteins

    Science.gov (United States)

    Patel, A.; Hashimoto, H.; Zhang, X.; Cheng, X.

    2016-01-01

    Much is known about vertebrate DNA methylation and oxidation; however, much less is known about how modified cytosine residues within particular sequences are recognized. Among the known methylated DNA-binding domains, the Cys2-His2 zinc finger (ZnF) protein superfamily is the largest with hundreds of members, each containing tandem ZnFs ranging from 3 to >30 fingers. We have begun to biochemically and structurally characterize these ZnFs not only on their sequence specificity but also on their sensitivity to various DNA modifications. Rather than following published methods of refolding insoluble ZnF arrays, we have expressed and purified soluble forms of ZnFs, ranging in size from a tandem array of two to six ZnFs, from seven different proteins. We also describe a fluorescence polarization assay to measure ZnFs affinity with oligonucleotides containing various modifications and our approaches for cocrystallization of ZnFs with oligonucleotides. PMID:27372763

  5. A C2H2 zinc finger protein FEMU2 is required for fox1 expression in Chlamydomonas reinhardtii.

    Directory of Open Access Journals (Sweden)

    Xiaodong Deng

    Full Text Available Chlamydomonas reinhardtii fox1 gene encodes a ferroxidase that is involved in cellular Fe uptake and highly induced during Fe deficient conditions. In an effort to identify fox1 promoter regulatory elements, an insertional library was generated in a transgenic Chlamydomonas strain (2A38 harboring an arylsulfatase (ARS reporter gene driven by the fox1 promoter. Mutants with a defective response to low iron conditions were selected for further study. Among these, a strain containing a disrupted femu2 gene was identified. Activation of the fox1 promoter by the femu2 gene product was confirmed by silencing the femu2 gene using RNA interference. In three femu2 RNAi transgenic lines (IR3, IR6, and IR7, ARS reporter gene activities declined by 84.3%, 86.4%, and 88.8%, respectively under Fe deficient conditions. Furthermore, RT-PCR analysis of both the femu2 mutant and the RNAi transgenic lines showed significantly decreased transcript abundance of the endogenous fox1 gene under Fe deficient conditions. Amino acid sequence analysis of the femu2 gene product identified three potential C2H2 zinc finger (ZF motifs and a nuclear localization study suggests that FEMU2 is localized to the nucleus. In addition, a potential FEMU2 binding site ((G/TTTGG(G/T(G/TT was identified using PCR-mediated random binding site selection. Taken together, this evidence suggests that FEMU2 is involved in up-regulation of the fox1 gene in Fe deficient cells.

  6. Genome-Wide Analysis of C2H2 Zinc-Finger Family Transcription Factors and Their Responses to Abiotic Stresses in Poplar (Populus trichocarpa.

    Directory of Open Access Journals (Sweden)

    Quangang Liu

    Full Text Available C2H2 zinc-finger (C2H2-ZF proteins are a large gene family in plants that participate in various aspects of normal plant growth and development, as well as in biotic and abiotic stress responses. To date, no overall analysis incorporating evolutionary history and expression profiling of the C2H2-ZF gene family in model tree species poplar (Populus trichocarpa has been reported.Here, we identified 109 full-length C2H2-ZF genes in P. trichocarpa, and classified them into four groups, based on phylogenetic analysis. The 109 C2H2-ZF genes were distributed unequally on 19 P. trichocarpa linkage groups (LGs, with 39 segmental duplication events, indicating that segmental duplication has been important in the expansion of the C2H2-ZF gene family. Promoter cis-element analysis indicated that most of the C2H2-ZF genes contain phytohormone or abiotic stress-related cis-elements. The expression patterns of C2H2-ZF genes, based on heatmap analysis, suggested that C2H2-ZF genes are involved in tissue and organ development, especially root and floral development. Expression analysis based on quantitative real-time reverse transcription polymerase chain reaction indicated that C2H2-ZF genes are significantly involved in drought, heat and salt response, possibly via different mechanisms.This study provides a thorough overview of the P. trichocarpa C2H2-ZF gene family and presents a new perspective on the evolution of this gene family. In particular, some C2H2-ZF genes may be involved in environmental stress tolerance regulation. PtrZFP2, 19 and 95 showed high expression levels in leaves and/or roots under environmental stresses. Additionally, this study provided a solid foundation for studying the biological roles of C2H2-ZF genes in Populus growth and development. These results form the basis for further investigation of the roles of these candidate genes and for future genetic engineering and gene functional studies in Populus.

  7. Activation of the ustilagic acid biosynthesis gene cluster in Ustilago maydis by the C2H2 zinc finger transcription factor Rua1.

    Science.gov (United States)

    Teichmann, Beate; Liu, Lidan; Schink, Kay Oliver; Bölker, Michael

    2010-04-01

    The phytopathogenic basidiomycetous fungus Ustilago maydis secretes, under conditions of nitrogen starvation, large amounts of the biosurfactant ustilagic acid (UA). This secreted cellobiose glycolipid is toxic for many microorganisms and confers biocontrol activity to U. maydis. Recently, a large gene cluster that is responsible for UA biosynthesis was identified. Here, we show that expression of all cluster genes depends on Rua1, a nuclear protein of the C(2)H(2) zinc finger family, whose gene is located within the gene cluster. While deletion of rua1 results in complete loss of UA production, overexpression of rua1 promotes increased UA synthesis even in the presence of a good nitrogen source. Bioinformatic analysis allowed us to identify a conserved sequence element that is present in the promoters of all structural genes involved in UA biosynthesis. Deletion analysis of several promoters within the cluster revealed that this DNA element serves as an upstream activating sequence (UAS) and mediates Rua1-dependent expression. We used the yeast one-hybrid system to demonstrate specific recognition of this DNA element by Rua1. Introduction of nucleotide exchanges into the consensus sequence interfered with Rua1-dependent activation, suggesting that this sequence element acts as a direct binding site for Rua1. PMID:20173069

  8. C2H2 zinc finger proteins of the SP/KLF, Wilms tumor, EGR, Huckebein, and Klumpfuss families in metazoans and beyond.

    Science.gov (United States)

    Pei, Jimin; Grishin, Nick V

    2015-11-15

    Specificity proteins (SPs) and Krüppel-Like Factors (KLFs) are C2H2-type zinc finger transcription factors that play essential roles in differentiation, development, proliferation and cell death. SP/KLF proteins, similarly to Wilms tumor protein 1 (WT1), Early Growth Response (EGR), Huckebein, and Klumpfuss, prefer to bind GC-rich sequences such as GC-box and CACCC-box (GT-box). We searched various genomes and transcriptomes of metazoans and single-cell holozoans for members of these families. Seven groups of KLFs (KLFA-G) and three groups of SPs (SPA-C) were identified in the three lineages of Bilateria (Deuterostomia, Ecdysozoa, and Lophotrochozoa). The last ancestor of jawed vertebrates was inferred to have at least 18 KLFs (group A: KLF1/2/4/17, group B: KLF3/8/12; group C: KLF5/5l; group D: KLF6/7; group E: KLF9/13/16; group F: KLF10/KLF11; group G: KLF15/15l) and 10 SPs (group A: SP1/2/3/4; group B: SP5/5l; group C: SP6/7/8/9), since they were found in both cartilaginous and boned fishes. Placental mammals have added KLF14 (group E) and KLF18 (group A), and lost KLF5l (KLF5-like) and KLF15l (KLF15-like). Multiple KLF members were found in basal metazoans (Ctenophora, Porifera, Placozoa, and Cnidaria). Ctenophora has the least number of KLFs and no SPs, which could be attributed to its proposed sister group relationship to other metazoans or gene loss. While SP, EGR and Klumpfuss were only detected in metazoans, KLF, WT1, and Huckebein are present in nonmetazoan holozoans. Of the seven metazoan KLF groups, only KLFG, represented by KLF15 in human, was found in nonmetazoans. In addition, two nonmetazoan groups of KLFs are present in Choanoflagellatea and Filasterea. WT1 could be evolutionarily the earliest among these GC/GT-box-binding families due to its sole presence in Ichthyosporea. PMID:26187067

  9. Isolation of a Novel Family of C2H2 Zinc Finger Proteins Implicated in Transcriptional Repression Mediated by Chicken Ovalbumin Upstream Promoter Transcription Factor (COUP-TF) Orphan Nuclear Receptors*

    OpenAIRE

    Avram, Dorina; Fields, Andrew; Top, Karen Pretty On; Nevrivy, Daniel J.; Ishmael, Jane E.; Leid, Mark

    2000-01-01

    Two novel and related C2H2 zinc finger proteins that are highly expressed in the brain, CTIP1 and CTIP2 (COUP TF-interacting proteins 1 and 2, respectively), were isolated and shown to interact with all members of the chicken ovalbumin upstream promoter transcription factor (COUP-TF) subfamily of orphan nuclear receptors. The interaction of CTIP1 with ARP1 was studied in detail, and CTIP1 was found to harbor two independent ARP1 interaction domains, ID1 and ID2, whereas the putative AF-2 of A...

  10. The transcriptional regulator c2h2 accelerates mushroom formation in Agaricus bisporus.

    Science.gov (United States)

    Pelkmans, Jordi F; Vos, Aurin M; Scholtmeijer, Karin; Hendrix, Ed; Baars, Johan J P; Gehrmann, Thies; Reinders, Marcel J T; Lugones, Luis G; Wösten, Han A B

    2016-08-01

    The Cys2His2 zinc finger protein gene c2h2 of Schizophyllum commune is involved in mushroom formation. Its inactivation results in a strain that is arrested at the stage of aggregate formation. In this study, the c2h2 orthologue of Agaricus bisporus was over-expressed in this white button mushroom forming basidiomycete using Agrobacterium-mediated transformation. Morphology, cap expansion rate, and total number and biomass of mushrooms were not affected by over-expression of c2h2. However, yield per day of the c2h2 over-expression strains peaked 1 day earlier. These data and expression analysis indicate that C2H2 impacts timing of mushroom formation at an early stage of development, making its encoding gene a target for breeding of commercial mushroom strains. PMID:27207144

  11. The interaction of DNA with multi-Cys2His2 zinc finger proteins

    Science.gov (United States)

    Liu, Lei; Heermann, Dieter W.

    2015-02-01

    The multi-Cys2His2 (mC2H2) zinc finger protein, like CTCF, plays a central role in the three-dimensional organization of chromatin and gene regulation. The interaction between DNA and mC2H2 zinc finger proteins becomes crucial to better understand how CTCF dynamically shapes the chromatin structure. Here, we study a coarse-grained model of the mC2H2 zinc finger proteins in complexes with DNA, and in particular, we study how a mC2H2 zinc finger protein binds to and searches for its target DNA loci. On the basis of coarse-grained molecular dynamics simulations, we present several interesting kinetic conformational properties of the proteins, such as the rotation-coupled sliding, the asymmetrical roles of different zinc fingers and the partial binding partial dangling mode. In addition, two kinds of studied mC2H2 zinc finger proteins, of CG-rich and AT-rich binding motif each, were able to recognize their target sites and slide away from their non-target sites, which shows a proper sequence specificity in our model and the derived force field for mC2H2-DNA interaction. A further application to CTCF shows that the protein binds to a specific DNA duplex only with its central zinc fingers. The zinc finger domains of CTCF asymmetrically bend the DNA, but do not form a DNA loop alone in our simulations.

  12. An expanded binding model for Cys2His2 zinc finger protein–DNA interfaces

    International Nuclear Information System (INIS)

    Cys2His2 zinc finger (C2H2-ZF) proteins comprise the largest class of eukaryotic transcription factors. The 'canonical model' for C2H2-ZF protein–DNA interaction consists of only four amino acid–nucleotide contacts per zinc finger domain, and this model has been the basis for several efforts for computationally predicting and experimentally designing protein–DNA interfaces. Here, we perform a systematic analysis of structural and experimental binding data and find that, in addition to the canonical contacts, several other amino acid and base pair combinations frequently play a role in C2H2-ZF protein–DNA binding. We suggest an expansion of the canonical C2H2-ZF model to include one to three additional contacts, and show that computational approaches including these additional contacts improve predictions of DNA targets of zinc finger proteins

  13. Zinc finger structure-function in Ikaros

    Institute of Scientific and Technical Information of China (English)

    Marvin; A; Payne

    2011-01-01

    The zinc finger motif was used as a vehicle for the initial discovery of Ikaros in the context of T-cell differentiation and has been central to all subsequent analyses of Ikaros function.The Ikaros gene is alternately spliced to produce several isoforms that confer diversity of function and consequently have complicated analysis of the function of Ikaros in vivo.Key features of Ikaros in vivo function are associated with six C2H2 zinc fingers;four of which are alternately incorporated in the production of the various Ikaros isoforms.Although no complete structures are available for the Ikaros protein or any of its family members,considerable evidence has accumulated about the structure of zinc fingers and the role that this structure plays in the functions of the Ikaros family of proteins.This review summarizes the structural aspects of Ikaros zinc fingers,individually,and in tandem to provide a structural context for Ikaros function and to provide a structural basis to inform the design of future experiments with Ikaros and its family members.

  14. Expression of putative zinc-finger protein lcn61 gene in lymphocystis disease virus China (LCDV-cn) genome

    Institute of Scientific and Technical Information of China (English)

    YAN Xiuying; SUN Xiuqin

    2009-01-01

    An open reading frame (lcn61) of iymphocystis disease virus China (LCDV-cn), probably responsible for encoding putative zinc-finger proteins was amplified and inserted into pET24a (+) vector.Then it expressed in E. coli BL21 (DE3), and His-tag fusion protein of high yield was obtained. It was found that the fusion protein existed in E. coli mainly as inclusion bodies. The bioinformatics analysis indicates that LCN61 is C2H2 type zinc-finger protein containing four C2H2 zinc-finger motifs. This work provides a theory for functional research of lcn61 gene.

  15. Zinc finger proteins in cancer progression

    OpenAIRE

    Jen, Jayu; Wang, Yi-Ching

    2016-01-01

    Zinc finger proteins are the largest transcription factor family in human genome. The diverse combinations and functions of zinc finger motifs make zinc finger proteins versatile in biological processes, including development, differentiation, metabolism and autophagy. Over the last few decades, increasing evidence reveals the potential roles of zinc finger proteins in cancer progression. However, the underlying mechanisms of zinc finger proteins in cancer progression vary in different cancer...

  16. Activation of transcriptional activity of HSE by a novel mouse zinc finger protein ZNFD specifically expressed in testis.

    Science.gov (United States)

    Xu, Fengqin; Wang, Weiping; Lei, Chen; Liu, Qingmei; Qiu, Hao; Muraleedharan, Vinaydhar; Zhou, Bin; Cheng, Hongxia; Huang, Zhongkai; Xu, Weian; Li, Bichun; Wang, Minghua

    2012-04-01

    Zinc finger proteins (ZFPs) that contain multiple cysteine and/or histidine residues perform important roles in various cellular functions, including transcriptional regulation, cell proliferation, differentiation, and apoptosis. The Cys-Cys-His-His (C(2)H(2)) type of ZFPs are the well-defined members of this super family and are the largest and most complex proteins in eukaryotic genomes. In this study, we identified a novel C(2)H(2) type of zinc finger gene ZNFD from mice which has a 1,002 bp open reading frame and encodes a protein with 333 amino acid residues. The predicted 37.4 kDa protein contains a C(2)H(2) zinc finger domain. ZNFD gene is located on chromosome 18qD1. RT-PCR analysis revealed that the ZNFD gene was specifically expressed in mouse testis but not in other tissues. Subcellular localization analysis demonstrated that ZNFD was localized in the nucleus. Reporter gene assays showed that overexpression of ZNFD in the COS7 cells activates the transcriptional activities of heat shock element (HSE). Overall, these results suggest that ZNFD is a member of the zinc finger transcription factor family and it participates in the transcriptional regulation of HSE. Many heat shock proteins regulated by HSE are involved in testicular development. Therefore, our results suggest that ZNFD may probably participate in the development of mouse testis and function as a transcription activator in HSE-mediated gene expression and signaling pathways.

  17. 15-zinc finger protein Bloody Fingers is required for zebrafish morphogenetic movements during neurulation.

    Science.gov (United States)

    Sumanas, Saulius; Zhang, Bo; Dai, Rujuan; Lin, Shuo

    2005-07-01

    A novel zebrafish gene bloody fingers (blf) encoding a 478 amino acid protein containing fifteen C(2)H(2) type zinc fingers was identified by expression screening. As determined by in situ hybridization, blf RNA displays strong ubiquitous early zygotic expression, while during late gastrulation and early somitogenesis, blf expression becomes transiently restricted to the posterior dorsal and lateral mesoderm. During later somitogenesis, blf expression appears only in hematopoietic cells. It is completely eliminated in cloche, moonshine but not in vlad tepes (gata1) mutant embryos. Morpholino (MO) knockdown of the Blf protein results in the defects of morphogenetic movements. Blf-MO-injected embryos (morphants) display shortened and widened axial tissues due to defective convergent extension. Unlike other convergent extension mutants, blf morphants display a split neural tube, resulting in a phenotype similar to the human open neural tube defect spina bifida. In addition, dorsal ectodermal cells delaminate in blf morphants during late somitogenesis. We propose a model explaining the role of blf in convergent extension and neurulation. We conclude that blf plays an important role in regulating morphogenetic movements during gastrulation and neurulation while its role in hematopoiesis may be redundant.

  18. Differential binding of monomethylarsonous acid compared to arsenite and arsenic trioxide with zinc finger peptides and proteins.

    Science.gov (United States)

    Zhou, Xixi; Sun, Xi; Mobarak, Charlotte; Gandolfi, A Jay; Burchiel, Scott W; Hudson, Laurie G; Liu, Ke Jian

    2014-04-21

    Arsenic is an environmental toxin that enhances the carcinogenic effect of DNA-damaging agents, such as ultraviolet radiation and benzo[a]pyrene. Interaction with zinc finger proteins has been shown to be an important molecular mechanism for arsenic toxicity and cocarcinogenesis. Arsenicals such as arsenite, arsenic trioxide (ATO), and monomethylarsonous acid (MMA(III)) have been reported to interact with cysteine residues of zinc finger domains, but little is known about potential differences in their selectivity of interaction. Herein we analyzed the interaction of arsenite, MMA(III), and ATO with C2H2, C3H1, and C4 configurations of zinc fingers using UV-vis, cobalt, fluorescence, and mass spectrometry. We observed that arsenite and ATO both selectively bound to C3H1 and C4 zinc fingers, while MMA(III) interacted with all three configurations of zinc finger peptides. Structurally and functionally, arsenite and ATO caused conformational changes and zinc loss on C3H1 and C4 zinc finger peptide and protein, respectively, whereas MMA(III) changed conformation and displaced zinc on all three types of zinc fingers. The differential selectivity was also demonstrated in zinc finger proteins isolated from cells treated with these arsenicals. Our results show that trivalent inorganic arsenic compounds, arsenite and ATO, have the same selectivity and behavior when interacting with zinc finger proteins, while methylation removes the selectivity. These findings provide insights on the molecular mechanisms underlying the differential effects of inorganic versus methylated arsenicals, as well as the role of in vivo arsenic methylation in arsenic toxicity and carcinogenesis.

  19. Characterization of 47 Cys2 -His2 zinc finger proteins required for the development and pathogenicity of the rice blast fungus Magnaporthe oryzae.

    Science.gov (United States)

    Cao, Huijuan; Huang, Pengyun; Zhang, Lilin; Shi, Yongkai; Sun, Dandan; Yan, Yuxin; Liu, Xiaohong; Dong, Bo; Chen, Guoqing; Snyder, John Hugh; Lin, Fucheng; Lu, Jianping

    2016-08-01

    The Cys2 -His2 (C2H2) zinc finger protein family is the second-largest family of transcription factors (TFs) in Magnaporthe oryzae, the causal fungus responsible for the destructive rice blast disease. However, little is known about the roles of most C2H2 TFs in the development and pathogenicity of M. oryzae. The roles of 47 C2H2 genes in development and pathogenicity were investigated by gene deletion in M. oryzae. The TF-dependent genes in mycelia or appressoria were analyzed with RNA sequencing and quantitative PCR (qPCR). Forty-four C2H2 genes are involved in growth (20 genes), conidiation (28 genes), appressorium formation (four genes) and pathogenicity (22 genes) in M. oryzae. Of these, MGG_14931, named as VRF1, is required for pathogenicity, specifically controlling appressorium maturation by affecting the expression of genes related to appressorial structure and function, including melanin biosynthesis, chitin catabolism, lipid metabolism, proteolysis, transmembrane transport, and response to oxidative stress; MGG_01776, named as VRF2, is required for plant penetration and invasive growth; conidiation-related gene CON7 is required for conidial differentiation; and MoCREA, encoding a carbon catabolite repression protein, is a novel repressor of lipid catabolism when glucose obtainable in M. oryzae. This study provides many insights into the regulation of growth, asexual development, appressorium formation, and pathogenicity by C2H2 TFs in M. oryzae. PMID:27041000

  20. Electron swarm parameters in pure C2H2 and in C2H2-Ar mixtures and electron collision cross sections for the C2H2 molecule

    Science.gov (United States)

    Nakamura, Yoshiharu

    2010-09-01

    Electron swarm parameters (the drift velocity and the longitudinal diffusion coefficient) were measured in pure C2H2 and also in C2H2-Ar mixtures containing 0.517% and 5.06% acetylene over wide E/N ranges. These swarm parameters were analysed using a Boltzmann equation analysis and a set of electron collision cross sections for the C2H2 molecule was derived so that it was consistent with the present swarm data and published ionization coefficient. The present result suggested the presence of a Ramsauer-Townsend minimum in the elastic momentum transfer cross section at 0.08 eV and prominent threshold and resonance peaks in the ν4/ν5 vibrational excitation cross section. The present cross section set was also confirmed to be consistent with the published experimental total cross section of C2H2.

  1. Prediction of DNA-binding specificity in zinc finger proteins

    Indian Academy of Sciences (India)

    Sumedha Roy; Shayoni Dutta; Kanika Khanna; Shruti Singla; Durai Sundar

    2012-07-01

    Zinc finger proteins interact via their individual fingers to three base pair subsites on the target DNA. The four key residue positions −1, 2, 3 and 6 on the alpha-helix of the zinc fingers have hydrogen bond interactions with the DNA. Mutating these key residues enables generation of a plethora of combinatorial possibilities that can bind to any DNA stretch of interest. Exploiting the binding specificity and affinity of the interaction between the zinc fingers and the respective DNA can help to generate engineered zinc fingers for therapeutic purposes involving genome targeting. Exploring the structure–function relationships of the existing zinc finger–DNA complexes can aid in predicting the probable zinc fingers that could bind to any target DNA. Computational tools ease the prediction of such engineered zinc fingers by effectively utilizing information from the available experimental data. A study of literature reveals many approaches for predicting DNA-binding specificity in zinc finger proteins. However, an alternative approach that looks into the physico-chemical properties of these complexes would do away with the difficulties of designing unbiased zinc fingers with the desired affinity and specificity. We present a physico-chemical approach that exploits the relative strengths of hydrogen bonding between the target DNA and all combinatorially possible zinc fingers to select the most optimum zinc finger protein candidate.

  2. Genome Engineering With Zinc-Finger Nucleases

    OpenAIRE

    Carroll, Dana

    2011-01-01

    Zinc-finger nucleases (ZFNs) are targetable DNA cleavage reagents that have been adopted as gene-targeting tools. ZFN-induced double-strand breaks are subject to cellular DNA repair processes that lead to both targeted mutagenesis and targeted gene replacement at remarkably high frequencies. This article briefly reviews the history of ZFN development and summarizes applications that have been made to genome editing in many different organisms and situations. Considerable progress has been mad...

  3. A Novel Zinc Finger Protein 219-like (ZNF219L) is Involved in the Regulation of Collagen Type 2 Alpha 1a (col2a1a) Gene Expression in Zebrafish Notochord

    OpenAIRE

    Lien, Huang-Wei; Yang, Chung-Hsiang; Cheng, Chia-Hsiung; Hung, Chin-Chun; Liao, Wei-Hao; Hwang, Pung-Pung; Han, Yu-San; Huang, Chang-Jen

    2013-01-01

    The notochord is required for body plan patterning in vertebrates, and defects in notochord development during embryogenesis can lead to diseases affecting the adult. It is therefore important to elucidate the gene regulatory mechanism underlying notochord formation. In this study, we cloned the zebrafish zinc finger 219-like (ZNF219L) based on mammalian ZNF219, which contains nine C2H2-type zinc finger domains. Through whole-mount in situ hybridization, we found that znf219L mRNA is mainly e...

  4. Zinc fingers, zinc clusters, and zinc twists in DNA-binding protein domains.

    OpenAIRE

    Vallee, B L; Coleman, J E; Auld, D S

    1991-01-01

    We now recognize three distinct motifs of DNA-binding zinc proteins: (i) zinc fingers, (ii) zinc clusters, and (iii) zinc twists. Until very recently, x-ray crystallographic or NMR three-dimensional structure analyses of DNA-binding zinc proteins have not been available to serve as standards of reference for the zinc binding sites of these families of proteins. Those of the DNA-binding domains of the fungal transcription factor GAL4 and the rat glucocorticoid receptor are the first to have be...

  5. Speed-stability paradox in DNA-scanning by zinc-finger proteins

    OpenAIRE

    Iwahara, Junji; Levy, Yaakov

    2013-01-01

    Extensive contact with DNA via multiple zinc fingers allows highly specific DNA-binding of zinc-finger-class transcription factors, but can also slow the target search process. Here we introduce recent insights into how zinc-finger proteins can rapidly scan DNA. Potential application of the new knowledge to the zinc-finger-based technology is also discussed.

  6. Terahertz Spectroscopy of the Bending Vibrations of Acetylene 12C2H2

    Science.gov (United States)

    Yu, Shanshan; Drouin, Brian J.; Pearson, John C.

    2009-11-01

    Twenty P-branch transitions of 12C2H2 have been measured in the 0.8-1.6 THz region of its bending vibrational difference band. The accuracy of these measurements is estimated to be 100 kHz. The 12C2H2 molecules were generated under room temperature by passing 150 mTorr H2O vapor through calcium carbide (CaC2) powder. The observed transitions were modeled together with prior far-infrared data involving the bending levels with ∑\

  7. Experimental and Kinetic Modeling Study of C2H2Oxidation at High Pressure

    DEFF Research Database (Denmark)

    Lopez, Jorge Gimenez; Rasmussen, Christian Tihic; Hashemi, Hamid;

    2016-01-01

    A detailed chemical kinetic model for oxidation of acetylene at intermediate temperatures and high pressure has been developed and evaluated experimentally. The rate coefficients for the reactions of C2H2 with HO2 and O2 were investigated, based on the recent analysis of the potential energy...... to be more than 50 kcal mol−1 and predictions of the initiation temperature were not sensitive to this reaction. Experiments were conducted with C2H2/O2 mixtures highly diluted in N2 in a high-pressure flow reactor at 600–900 K and 60 bar, varying the reaction stoichiometry from very lean to fuel...

  8. Investigation of Compton profiles of NO and C2H2%NO与C2H2的康普顿轮廓研究

    Institute of Scientific and Technical Information of China (English)

    马永朋; 赵小利; 刘亚伟; 徐龙泉; 康旭; 倪冬冬; 闫帅; 朱林繁; 杨科

    2015-01-01

    The Compton profiles of nitric oxide and acetylene molecules have been determined at an incident photon energy of 20 keV. Compton profile measurements are carried out with the beamline BL15U1 at the Shanghai Synchrotron Radiation Facility (SSRF). A dedicated gas cell is used, in which diffuse scattering is effectively suppressed. By considering that the statistical accuracy of 0.2%at pz ≈0 is achieved, the Compton profiles of NO and C2H2 determined in this paper can serve as the experimental benchmark data. Furthermore, the density functional theory (DFT) and HF calculation for different basis sets are used to calculate the Compton profiles of nitric oxide and acetylene. It is found that the DFT calculations with the diffuse basis sets are closer to the experimental results, indicating that the electronic density distribution of nitric oxide is more diffuse. For acetylene, the HF calculation is of better agreement with the experimental result. To better understand Compton profiles, we have compared them with distributions of electron density by theoretical calculation. There are clear correspondences between them: diffuse distribution is related to the localized profile and complex structure in electron density distribution, which also shows a subtle structure in profile. The present Compton profiles of nitric oxide and acetylene molecules achieved by synchrotron radiation are the most accurate up to now, as far as we know.%基于第三代同步辐射光源,在20 keV的入射X射线能量下测量了NO与C2H2分子的康普顿轮廓.考虑到本次实验结果在pz ≈0附近的统计精度达到了0.2%,本文报道的NO和C2H2的康普顿轮廓可以作为严格检验理论的实验基准.除此之外,还分别采用HF方法及密度泛函方法选用不同的基组计算了NO与C2 H2康普顿轮廓.通过对比实验结果与理论计算,发现对于NO分子,加入弥散函数基组理论计算结果与实验符合更好,说明NO分子基态的电子分布较为弥散.对于C

  9. Predicting success of oligomerized pool engineering (OPEN for zinc finger target site sequences

    Directory of Open Access Journals (Sweden)

    Goodwin Mathew J

    2010-11-01

    Full Text Available Abstract Background Precise and efficient methods for gene targeting are critical for detailed functional analysis of genomes and regulatory networks and for potentially improving the efficacy and safety of gene therapies. Oligomerized Pool ENgineering (OPEN is a recently developed method for engineering C2H2 zinc finger proteins (ZFPs designed to bind specific DNA sequences with high affinity and specificity in vivo. Because generation of ZFPs using OPEN requires considerable effort, a computational method for identifying the sites in any given gene that are most likely to be successfully targeted by this method is desirable. Results Analysis of the base composition of experimentally validated ZFP target sites identified important constraints on the DNA sequence space that can be effectively targeted using OPEN. Using alternate encodings to represent ZFP target sites, we implemented Naïve Bayes and Support Vector Machine classifiers capable of distinguishing "active" targets, i.e., ZFP binding sites that can be targeted with a high rate of success, from those that are "inactive" or poor targets for ZFPs generated using current OPEN technologies. When evaluated using leave-one-out cross-validation on a dataset of 135 experimentally validated ZFP target sites, the best Naïve Bayes classifier, designated ZiFOpT, achieved overall accuracy of 87% and specificity+ of 90%, with an ROC AUC of 0.89. When challenged with a completely independent test set of 140 newly validated ZFP target sites, ZiFOpT performance was comparable in terms of overall accuracy (88% and specificity+ (92%, but with reduced ROC AUC (0.77. Users can rank potentially active ZFP target sites using a confidence score derived from the posterior probability returned by ZiFOpT. Conclusion ZiFOpT, a machine learning classifier trained to identify DNA sequences amenable for targeting by OPEN-generated zinc finger arrays, can guide users to target sites that are most likely to function

  10. Gas-phase CO2, C2H2, and HCN toward Orion-KL

    Science.gov (United States)

    Boonman, A. M. S.; van Dishoeck, E. F.; Lahuis, F.; Doty, S. D.; Wright, C. M.; Rosenthal, D.

    2003-03-01

    The infrared spectra toward Orion-IRc2, Peak 1 and Peak 2 in the 13.5-15.5 mu m wavelength range are presented, obtained with the Short Wavelength Spectrometer on board the Infrared Space Observatory. The spectra show absorption and emission features of the vibration-rotation bands of gas-phase CO2, HCN, and C2H2, respectively. Toward the deeply embedded massive young stellar object IRc2 all three bands appear in absorption, while toward the shocked region Peak 2 CO2, HCN, and C2H2 are seen in emission. Toward Peak 1 only CO2 has been detected in emission. Analysis of these bands shows that the absorption features toward IRc2 are characterized by excitation temperatures of ~ 175-275 K, which can be explained by an origin in the shocked plateau gas. HCN and C2H2 are only seen in absorption in the direction of IRc2, whereas the CO2 absorption is probably more widespread. The CO2 emission toward Peak 1 and 2 is best explained with excitation by infrared radiation from dust mixed with the gas in the warm component of the shock. The similarity of the CO2 emission and absorption line shapes toward IRc2, Peak 1 and Peak 2 suggests that the CO2 is located in the warm component of the shock (T ~ 200 K) toward all three positions. The CO2 abundances of ~ 10-8 for Peak 1 and 2, and of a few times 10-7 toward IRc2 can be explained by grain mantle evaporation and/or reformation in the gas-phase after destruction by the shock. The HCN and C2H2 emission detected toward Peak 2 is narrower (T ~ 50-150 K) and originates either in the warm component of the shock or in the extended ridge. In the case of an origin in the warm component of the shock, the low HCN and C2H2 abundances of ~ 10-9 suggest that they are destroyed by the shock or have only been in the warm gas for a short time (t Germany, The Netherlands and the UK) and with the participation of ISAS and NASA.

  11. Genetic dissection of photoreceptor subtype specification by the Drosophila melanogaster zinc finger proteins elbow and no ocelli.

    Directory of Open Access Journals (Sweden)

    Mathias F Wernet

    2014-03-01

    Full Text Available The elbow/no ocelli (elb/noc complex of Drosophila melanogaster encodes two paralogs of the evolutionarily conserved NET family of zinc finger proteins. These transcriptional repressors share a conserved domain structure, including a single atypical C2H2 zinc finger. In flies, Elb and Noc are important for the development of legs, eyes and tracheae. Vertebrate NET proteins play an important role in the developing nervous system, and mutations in the homolog ZNF703 human promote luminal breast cancer. However, their interaction with transcriptional regulators is incompletely understood. Here we show that loss of both Elb and Noc causes mis-specification of polarization-sensitive photoreceptors in the 'dorsal rim area' (DRA of the fly retina. This phenotype is identical to the loss of the homeodomain transcription factor Homothorax (Hth/dMeis. Development of DRA ommatidia and expression of Hth are induced by the Wingless/Wnt pathway. Our data suggest that Elb/Noc genetically interact with Hth, and we identify two conserved domains crucial for this function. Furthermore, we show that Elb/Noc specifically interact with the transcription factor Orthodenticle (Otd/Otx, a crucial regulator of rhodopsin gene transcription. Interestingly, different Elb/Noc domains are required to antagonize Otd functions in transcriptional activation, versus transcriptional repression. We propose that similar interactions between vertebrate NET proteins and Meis and Otx factors might play a role in development and disease.

  12. The conserved Bud20 zinc finger protein is a new component of the ribosomal 60S subunit export machinery.

    Science.gov (United States)

    Bassler, Jochen; Klein, Isabella; Schmidt, Claudia; Kallas, Martina; Thomson, Emma; Wagner, Maria Anna; Bradatsch, Bettina; Rechberger, Gerald; Strohmaier, Heimo; Hurt, Ed; Bergler, Helmut

    2012-12-01

    The nuclear export of the preribosomal 60S (pre-60S) subunit is coordinated with late steps in ribosome assembly. Here, we show that Bud20, a conserved C(2)H(2)-type zinc finger protein, is an unrecognized shuttling factor required for the efficient export of pre-60S subunits. Bud20 associates with late pre-60S particles in the nucleoplasm and accompanies them into the cytoplasm, where it is released through the action of the Drg1 AAA-ATPase. Cytoplasmic Bud20 is then reimported via a Kap123-dependent pathway. The deletion of Bud20 induces a strong pre-60S export defect and causes synthetic lethality when combined with mutant alleles of known pre-60S subunit export factors. The function of Bud20 in ribosome export depends on a short conserved N-terminal sequence, as we observed that mutations or the deletion of this motif impaired 60S subunit export and generated the genetic link to other pre-60S export factors. We suggest that the shuttling Bud20 is recruited to the nascent 60S subunit via its central zinc finger rRNA binding domain to facilitate the subsequent nuclear export of the preribosome employing its N-terminal extension.

  13. Toward a Code for the Interactions of Zinc Fingers with DNA: Selection of Randomized Fingers Displayed on Phage

    Science.gov (United States)

    Choo, Yen; Klug, Aaron

    1994-11-01

    We have used two selection techniques to study sequence-specific DNA recognition by the zinc finger, a small, modular DNA-binding minidomain. We have chosen zinc fingers because they bind as independent modules and so can be linked together in a peptide designed to bind a predetermined DNA site. In this paper, we describe how a library of zinc fingers displayed on the surface of bacteriophage enables selection of fingers capable of binding to given DNA triplets. The amino acid sequences of selected fingers which bind the same triplet are compared to examine how sequence-specific DNA recognition occurs. Our results can be rationalized in terms of coded interactions between zinc fingers and DNA, involving base contacts from a few α-helical positions. In the paper following this one, we describe a complementary technique which confirms the identity of amino acids capable of DNA sequence discrimination from these positions.

  14. Dual-laser absorption spectroscopy of C2H2 at 1.4 μ m

    Science.gov (United States)

    Fasci, E.; Odintsova, T. A.; Castrillo, A.; De Vizia, M. D.; Merlone, A.; Bertiglia, F.; Moretti, L.; Gianfrani, L.

    2016-04-01

    Spectroscopic parameters (line intensity factor, pressure self-broadening, and shifting coefficients) of C2H2 at 1.4 μ m were accurately measured using a dual-laser approach, based upon the technique of optical phase locking. This generated an absolute frequency scale underneath the absorption spectra. A pair of extended-cavity diode lasers was used. One of them, the probe laser, is forced to maintain a precise frequency offset from a reference laser, which is an optical frequency standard based on noise-immune cavity-enhanced optical heterodyne molecular spectroscopy. Laser-gas interaction takes place inside an isothermal multipass cell that is stabilized at the temperature of the triple point of water. The fidelity in the observation of the shape associated to the Pe(14) line of the 2 ν3+ν5 band allowed us to measure the spectroscopic parameters, with a global uncertainty for the line strength of 0.22%.

  15. Direct growth of graphene on gallium nitride using C2H2 as carbon source

    Science.gov (United States)

    Wang, Bing; Zhao, Yun; Yi, Xiao-Yan; Wang, Guo-Hong; Liu, Zhi-Qiang; Duan, Rui-Rei; Huang, Peng; Wang, Jun-Xi; Li, Jin-Min

    2016-04-01

    Growing graphene on gallium nitride (GaN) at temperatures greater than 900°C is a challenge that must be overcome to obtain high quality of GaN epi-layers. We successfully met this challenge using C2H2 as the carbon source. We demonstrated that graphene can be grown both on copper and directly on GaN epi-layers. The Raman spectra indicated that the graphene films were about 4-5 layers thick. Meanwhile, the effects of the growth temperature on the growth of the graphene films were systematically studied, and 830°C was found to be the optimum growth temperature. We successfully grew high-quality graphene films directly on gallium nitride.

  16. Editing the Plasmodium vivax Genome, Using Zinc-Finger Nucleases

    OpenAIRE

    Moraes Barros, Roberto R.; Straimer, Judith; Sa, Juliana M; Salzman, Rebecca E.; Melendez-Muniz, Viviana A.; Mu, Jianbing; David A Fidock; Thomas E. Wellems

    2014-01-01

    Plasmodium vivax is a major cause of malaria morbidity worldwide yet has remained genetically intractable. To stably modify this organism, we used zinc-finger nucleases (ZFNs), which take advantage of homology-directed DNA repair mechanisms at the site of nuclease action. Using ZFNs specific to the gene encoding P. vivax dihydrofolate reductase (pvdhfr), we transfected blood specimens from Saimiri boliviensis monkeys infected with the pyrimethamine (Pyr)–susceptible Chesson strain with a ZFN ...

  17. Dopamine receptor regulating factor, DRRF: A zinc finger transcription factor

    OpenAIRE

    Hwang, Cheol Kyu; D'Souza, Ursula M.; Eisch, Amelia J.; Yajima, Shunsuke; Lammers, Claas-Hinrich; Yang, Young; Lee, Sang-Hyeon; Kim, Yong-Man; Nestler, Eric J.; Mouradian, M. Maral

    2001-01-01

    Dopamine receptor genes are under complex transcription control, determining their unique regional distribution in the brain. We describe here a zinc finger type transcription factor, designated dopamine receptor regulating factor (DRRF), which binds to GC and GT boxes in the D1A and D2 dopamine receptor promoters and effectively displaces Sp1 and Sp3 from these sequences. Consequently, DRRF can modulate the activity of these dopamine receptor promoters. Highest DRRF mRNA levels are found in ...

  18. Acetylene C2H 2 retrievals from MIPAS data and regions of enhanced upper tropospheric concentrations in August 2003

    Directory of Open Access Journals (Sweden)

    V. P. Kanawade

    2011-10-01

    Full Text Available Acetylene (C2H2 volume mixing ratios (VMRs have been successfully retrieved from the Michelson Interferometer for Passive Atmospheric Sounding (MIPAS Level 1B radiances during August 2003, providing the first global map of such data and ratios to CO in the literature. The data presented here contain most information between 300 hPa and 100 hPa with systematic errors less than 10% at the upper levels. Random errors per point are less than 15% at lower levels and are closer to 30% at 100 hPa. Global distributions of the C2H2 and C2H2/CO ratio confirm significant features associated with both the Asian monsoon anticyclone and biomass burning for this important hydrocarbon in a characteristic summer month (August 2003, showing tight correlations regionally, particularly at lower to medium values, but globally emphasising the differences between sources and lifetimes of CO and C2H2. The correlations are seen to be particularly disturbed in the regions of highest C2H2 concentrations, indicating variability in the surface emissions or fast processing. A strong isolation of C2H2 within the Asian monsoon anticyclone is observed, evidencing convective transport into the upper troposphere, horizontal advection within the anticyclone at 200 hPa, distinct gradients at the westward edge of the vortex and formation of a secondary dynamical feature from the eastward extension of the anticyclone outflow over the Asian Pacific. Ratios of C2H2/CO are consistent with the evidence from the cross-sections that the C2H2 is uplifted rapidly in convection. Observations are presented of enhanced C2H2 associated with the injection from biomass burning into the upper troposphere and the outflow from Africa at 200 hPa into both the Atlantic and Indian Oceans. In the biomass burning regions, C2H2 and CO are well correlated, but the uplift is less marked and peaks at lower altitudes compared to the strong effects observed in the Asian monsoon anticyclone. Ratios of C2H2/CO

  19. A FORMAÇÃO DE LIGAÇÕES DE HIDROGÊNIO π‧‧‧H, F‧‧‧H E C‧‧‧H NOS COMPLEXOS C2H2‧‧‧(HF, C2H2‧‧‧2(HF E C2H2‧‧‧3(HF

    Directory of Open Access Journals (Sweden)

    Boaz G. Oliveira

    2016-04-01

    Full Text Available In this work, a theoretical study on the basis of structural, vibrational, electronic and topological parameters of the C2H2‧‧‧(HF, C2H2‧‧‧2(HF and C2H2‧‧‧3(HF complexes concerning the formation of π‧‧‧H, F‧‧‧H and C‧‧‧H hydrogen bonds is presented. The main difference among these complexes is not properly the interaction strength, but the hydrogen bond type whose benchmark is ruled justly by the structure. Meanwhile, the occurrence of π‧‧‧H hydrogen bonds was unveiled in both C2H2‧‧‧(HF dimer and C2H2‧‧‧3(HF tetramer, although in latter, this interaction is stronger than C‧‧‧H of the C2H2‧‧‧2(HF trimer. However, the F‧‧‧H hydrogen bonds within the subunits of hydrofluoric acid are the strongest ones, reaching a partial covalent limit, and thereby contribute decisively to the stabilization of the tetramer structure. In line with this, the largest red-shifts were observed on the hydrofluoric acid trimer of the C2H2‧‧‧3(HF complex.

  20. CO2 and C2H2 in cold nanodroplets of oxygenated organic molecules and water

    Science.gov (United States)

    Devlin, J. Paul; Balcı, F. Mine; Maşlakcı, Zafer; Uras-Aytemiz, Nevin

    2014-11-01

    Recent demonstrations of subsecond and microsecond timescales for formation of clathrate hydrate nanocrystals hint at future methods of control of environmental and industrial gases such as CO2 and methane. Combined results from cold-chamber and supersonic-nozzle [A. S. Bhabhe, "Experimental study of condensation and freezing in a supersonic nozzle," Ph.D. thesis (Ohio State University, 2012), Chap. 7] experiments indicate extremely rapid encagement of components of all-vapor pre-mixtures. The extreme rates are derived from (a) the all-vapor premixing of the gas-hydrate components and (b) catalytic activity of certain oxygenated organic large-cage guests. Premixing presents no obvious barrier to large-scale conditions of formation. Further, from sequential efforts of the groups of Trout and Buch, a credible defect-based model of the catalysis mechanism exists for guidance. Since the catalyst-generated defects are both mobile and abundant, it is often unnecessary for a high percentage of the cages to be occupied by a molecular catalyst. Droplets represent the liquid phase that bridges the premixed vapor and clathrate hydrate phases but few data exist for the droplets themselves. Here we describe a focused computational and FTIR spectroscopic effort to characterize the aerosol droplets of the all-vapor cold-chamber methodology. Computational data for CO2 and C2H2, hetero-dimerized with each of the organic catalysts and water, closely match spectroscopic redshift patterns in both magnitude and direction. Though vibrational frequency shifts are an order of magnitude greater for the acetylene stretch mode, both CO2 and C2H2 experience redshift values that increase from that for an 80% water-methanol solvent through the solvent series to approximately doubled values for tetrahydrofuran and trimethylene oxide (TMO) droplets. The TMO solvent properties extend to a 50 mol.% solution of CO2, more than an order of magnitude greater than for the water-methanol solvent mixture

  1. Phylogenetic Analysis of the Plant-specific Zinc Finger-Homeobox and Mini Zinc Finger Gene Families

    Institute of Scientific and Technical Information of China (English)

    Wei Hu; Claude W.dePamphilis; Hong Ma

    2008-01-01

    Zinc finger-homaodomain proteins (ZHD) are present in many plants;however,the evolutionary history of the ZHD gene family remains largely unknown.We show here that ZHD genes are plant-specific,nearly all intronless,and related to MINI ZINC FINGER (MIF) genes that possess only the zinc finger.Phylogenetic analyses of ZHD genes from representative land plants suggest that non-seed plant ZHD genes occupy basal positions and angiosperm homologs form seven distinct clades.Several clades contain genes from two or more major angiosperm groups,including eudicots,monocots,magnoliids,and other basal angiosperms,indicating that several duplications occurred before the diversification of flowering plants.In addition,specific lineages have experienced more recent duplications.Unlike the ZHD genes,&fiFs are found only from seed plants,possibly derived from ZHDs by loss of the homeodomain before the divergence of seed plants.Moreover,the MIF genes have also undergone relatively recent gene duplications.Finally,genome duplication might have contributed substantially to the expansion of family size in angiosperms and caused a high level of functional redundancy/overlap in these genes.

  2. A Family of Zinc Finger Proteins Is Required forChromosome-specific Pairing and Synapsis during Meiosis in C.elegans

    Energy Technology Data Exchange (ETDEWEB)

    Phillips, Carolyn M.; Dernburg, Abby F.

    2006-06-07

    Homologous chromosome pairing and synapsis are prerequisitefor accurate chromosome segregation during meiosis. Here, we show that afamily of four related C2H2 zinc-finger proteins plays a central role inthese events in C. elegans. These proteins are encoded within a tandemgene cluster. In addition to the X-specific HIM-8 protein, threeadditional paralogs collectively mediate the behavior of the fiveautosomes. Each chromosome relies on a specific member of the family topair and synapse with its homolog. These "ZIM" proteins concentrate atspecial regions called meiotic pairing centers on the correspondingchromosomes. These sites are dispersed along the nuclear envelope duringearly meiotic prophase, suggesting a role analogous to thetelomere-mediated meiotic bouquet in other organisms. To gain insightinto the evolution of these components, wecharacterized homologs in C.briggsae and C. remanei, which revealed changes in copy number of thisgene family within the nematode lineage.

  3. Laser-induced nuclear motions in the Coulomb explosion of C2H2+ ions

    International Nuclear Information System (INIS)

    The laser-induced multifragmentation of C2H2 into protons and multicharged carbon ions is shown to be a direct instantaneous explosion of the molecule. The evolution of the overall nuclear structure is studied through ion-ion correlation peak shapes. The ratios of the maxima of the kinetic-energy release distributions to the Coulomb repulsion energies calculated at the equilibrium internuclear distances are measured to be 45% for the protons and 53% for the CZ+ ions for all the detected H++CZ'++CZ+ +H+ fragmentation channels. The time scale for electronic polarization and stripping compared with the intramolecular electronic and nuclear time evolutions does not allow using a frozen molecular ion structure for the description of the explosion, thus explaining in part the observed fragmentation pattern. During the laser-induced alignment and subsequent stabilization of the molecular frame around the laser polarization direction, the carbon-carbon axis undergoes small damped oscillations that remain larger than the corresponding oscillations of the hydrogen-hydrogen axis. This difference comes from the lower moment of inertia of the hydrogen atoms compared with that of the carbon atoms in the molecule. However, the subsequent deviation from the initial linear structure remains small and is observed when the molecular ion is not completely aligned along the laser electric field

  4. Infrared fingerprints and periodic formation of nanoparticles in Ar/C2H2 plasmas

    Science.gov (United States)

    Kovačević, Eva; Stefanović, Ilija; Berndt, Johannes; Winter, Jörg

    2003-03-01

    The formation of dust particles in argon diluted C2H2 plasmas was studied by means of Fourier transform infrared absorption spectroscopy and mass spectroscopy. The detection limit for infrared absorption was significantly improved by the use of a multipass technique. Measuring the intensity of the Rayleigh/Mie scattering of the infrared signal we found a periodicity of dust formation/vanishing (period of about 35 min in our experimental conditions). The fast disappearance of the dust from the plasma region at the end of every period is the evidence of a narrow particle size distribution, as confirmed by secondary electron micrographs of the collected powder. Characteristic infrared absorption features have their origin in absorption within the dust particles. Besides the strong presence of aliphatic hydrocarbons characteristic for amorphous hydrocarbon films, a significant amount of aromatic structures was detected. Heavy positive ions measured by ion-mass spectroscopy originate from polyacetilenic (C2nH2) and aromatic compounds. Time resolved mass spectra gave insight into the plasma response to the dust formation.

  5. Isomerization, Perturbations, Calculations and the S_{1} State of C_{2}H_{2}

    Science.gov (United States)

    Baraban, J. H.; Changala, P. B.; Berk, J. R. P.; Field, R. W.; Stanton, J. F.; Merer, A. J.

    2013-06-01

    Preliminary analysis of the energy region of the cis-trans isomerization transition state on the S_{1} surface of C_{2}H_{2} has revealed novel patterns and surprising perturbations, including unusually large (and high-order) anharmonicities, as well as K-staggerings of several vibrational levels. These effects complicate the analysis considerably, and require new models and calculations to account for and predict features of the observed spectra. The ˜{A}-˜{X} spectrum of acetylene has been studied both experimentally and theoretically for almost a century, and this cycle of unexpected phenomena eliciting innovative responses is found throughout its history. Especially in the last ten years, progress in understanding the S_{1} state rovibrational level structure and cis-trans isomerization has been accelerated by combining the information available from both ab initio computation and spectroscopic observations. The resulting dialogue has then frequently suggested fruitful avenues for further experiments and calculations. Current challenges and recent results in understanding the cis-trans isomerization transition state region will be discussed in this context.

  6. Preparation of hydrogenated diamond-like carbon films using high-density pulsed plasmas of Ar/C2H2 and Ne/C2H2 mixture

    Science.gov (United States)

    Kimura, Takashi; Kamata, Hikaru

    2016-07-01

    Hydrogenated diamond-like carbon films are prepared using reactive high-density pulsed plasmas of Ar/C2H2 and Ne/C2H2 mixture in the total pressure range from 0.5 to 2 Pa. The plasmas are produced using a reactive high-power impulse magnetron sputtering (HiPIMS) system. A negative pulse voltage of ‑500 V is applied to the substrate for a period of 15 µs in the afterglow mode. The growth rate does not strongly depend on the type of ambient gas but it markedly increases to about 2.7 µm/h at a C2H2 fraction of 10% and a total pressure of 2 Pa with increasing C2H2 fraction. The marked increase in the growth rate means that the HiPIMS system can be regarded as a plasma source for the chemical vapor deposition process. The hardness of the films prepared by Ne/C2H2 plasmas is somewhat higher than that of the films prepared by Ar/C2H2 plasmas under the same operating conditions, and the difference becomes larger as the pressure increases. The hardness of the films prepared by Ne/C2H2 plasmas ranges between 11 and 18 GPa. In the Raman spectra, two very broad overlapping bands are assigned as the G (graphite) and D (disorder) bands. The peak position of the G band is roughly independent of the total pressure, whereas the FWHM of the G peak decreases with increasing total pressure as a whole.

  7. C. elegans PAT-9 is a nuclear zinc finger protein critical for the assembly of muscle attachments

    Directory of Open Access Journals (Sweden)

    Liu Qian

    2012-05-01

    Full Text Available Abstract Background Caenorhabditis elegans sarcomeres have been studied extensively utilizing both forward and reverse genetic techniques to provide insight into muscle development and the mechanisms behind muscle contraction. A previous genetic screen investigating early muscle development produced 13 independent mutant genes exhibiting a Pat (paralyzed and arrested elongation at the two-fold length of embryonic development muscle phenotype. This study reports the identification and characterization of one of those genes, pat-9. Results Positional cloning, reverse genetics, and plasmid rescue experiments were used to identify the predicted C. elegans gene T27B1.2 (recently named ztf-19 as the pat-9 gene. Analysis of pat-9 showed it is expressed early in development and within body wall muscle lineages, consistent with a role in muscle development and producing a Pat phenotype. However, unlike most of the other known Pat gene family members, which encode structural components of muscle attachment sites, PAT-9 is an exclusively nuclear protein. Analysis of the predicted PAT-9 amino acid sequence identified one putative nuclear localization domain and three C2H2 zinc finger domains. Both immunocytochemistry and PAT-9::GFP fusion expression confirm that PAT-9 is primarily a nuclear protein and chromatin immunoprecipitation (ChIP experiments showed that PAT-9 is present on certain gene promoters. Conclusions We have shown that the T27B1.2 gene is pat-9. Considering the Pat-9 mutant phenotype shows severely disrupted muscle attachment sites despite PAT-9 being a nuclear zinc finger protein and not a structural component of muscle attachment sites, we propose that PAT-9 likely functions in the regulation of gene expression for some necessary structural or regulatory component(s of the muscle attachment sites.

  8. Metal Coupled Folding of Cys2His2 Zinc-Finger

    OpenAIRE

    Li, Wenfei; Zhang, Jian; Wang, Jun; Wang, Wei

    2008-01-01

    Zinc-fingers, which widely exist in eukaryotic cell and play crucial roles in life processes, depend on the binding of zinc ion for their proper folding. To computationally study the zinc coupled folding of the zinc-fingers, charge transfer and metal induced protonation/deprotonation effects have to be considered. Here, by attempting to implicitly account for such effects in classical molecular dynamics and performing intensive simulations with explicit solvent for the peptides with and witho...

  9. Chemically synthesized zinc finger molecules as nano-addressable probes for double-stranded DNAs

    Directory of Open Access Journals (Sweden)

    Christmann Alexander

    2005-06-01

    Full Text Available Abstract Our experiments describe an alternative method of dsDNA recognition using zinc finger (ZF molecules which bind DNA specifically and with high affinity. Our aim was to develop zinc finger probes which are able to bind to dsDNA molecules at predetermined sites. In our basic approach we used pairs of complementary oligonucleotides to form dsDNAs, containing one of the three SP1-transcription factor motifs as a zinc finger recognition site. Two zinc finger probes of the SP1 motif were chemically synthesized and modified with a Dy-633 fluorophore. The SP1 peptides were folded into functional zinc fingers using zinc chloride. The addressable dsDNAs were immobilized on optical fibres, and the kinetics and binding rates of the artificial zinc finger probes were measured by a fluorescence detecting device (photomultiplying tube. The two zinc fingers and their corresponding DNA recognition sites served as specific probes and controls for the matching site and vice versa. Our experiments showed that a variety of dsDNA-binding probes may be created by modification of the amino acid sequence of natural zinc finger proteins. Our findings offer an alternative approach of addressing dsDNA molecules, for example for use in a nanoarray device.

  10. ZifBASE: a database of zinc finger proteins and associated resources

    Directory of Open Access Journals (Sweden)

    Punetha Ankita

    2009-09-01

    Full Text Available Abstract Background Information on the occurrence of zinc finger protein motifs in genomes is crucial to the developing field of molecular genome engineering. The knowledge of their target DNA-binding sequences is vital to develop chimeric proteins for targeted genome engineering and site-specific gene correction. There is a need to develop a computational resource of zinc finger proteins (ZFP to identify the potential binding sites and its location, which reduce the time of in vivo task, and overcome the difficulties in selecting the specific type of zinc finger protein and the target site in the DNA sequence. Description ZifBASE provides an extensive collection of various natural and engineered ZFP. It uses standard names and a genetic and structural classification scheme to present data retrieved from UniProtKB, GenBank, Protein Data Bank, ModBase, Protein Model Portal and the literature. It also incorporates specialized features of ZFP including finger sequences and positions, number of fingers, physiochemical properties, classes, framework, PubMed citations with links to experimental structures (PDB, if available and modeled structures of natural zinc finger proteins. ZifBASE provides information on zinc finger proteins (both natural and engineered ones, the number of finger units in each of the zinc finger proteins (with multiple fingers, the synergy between the adjacent fingers and their positions. Additionally, it gives the individual finger sequence and their target DNA site to which it binds for better and clear understanding on the interactions of adjacent fingers. The current version of ZifBASE contains 139 entries of which 89 are engineered ZFPs, containing 3-7F totaling to 296 fingers. There are 50 natural zinc finger protein entries ranging from 2-13F, totaling to 307 fingers. It has sequences and structures from literature, Protein Data Bank, ModBase and Protein Model Portal. The interface is cross linked to other public

  11. Molecular Cloning and Expression Analysis of a Cys2/His2 Type Zinc Finger Protein Gene in Upland Cotton

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The zinc finger proteins belong to the largest family of transcription factors.But there is little research of Cys2/His2 type zinc finger proteins in cotton,and there is no submission of correlating ESTs

  12. Acetylene (C2H2 and hydrogen cyanide (HCN from IASI satellite observations: global distributions, validation, and comparison with model

    Directory of Open Access Journals (Sweden)

    V. Duflot

    2015-05-01

    Full Text Available We present global distributions of C2H2 and HCN total columns derived from the Infrared Atmospheric Sounding Interferometer (IASI. These distributions are obtained with a fast method allowing to retrieve C2H2 abundance globally with a 5% precision and HCN abundance in the tropical (subtropical belt with a 10% (30% precision. IASI data are compared for validation purposes with ground-based Fourier Transform Infrared (FTIR spectrometer measurements at four selected stations. We show that there is an overall agreement between the ground-based and space measurements. Global C2H2 and subtropical HCN abundances retrieved from IASI spectra show the expected seasonality linked to variations in the anthropogenic emissions and seasonal biomass burning activity, as well as exceptional events, and are in good agreement with previous spaceborne studies. IASI measurements are also compared to the distributions from the Model for Ozone and Related Chemical Tracers, version 4 (MOZART-4. Seasonal cycles observed from satellite data are reasonably well reproduced by the model. However, the model seems to overestimate (underestimate anthropogenic (biomass burning emissions and a negative global mean bias of 1% (16% of the model relative to the satellite observations was found for C2H2 (HCN.

  13. Experimental and computational study of C2H2 and CO in a laminar axisymmetric methane-air diffusion flame

    NARCIS (Netherlands)

    Mokhov, A. V.; Bennett, B. A. V.; Levinsky, H.B.; Smooke, M. D.

    2007-01-01

    Raman measurements of C2H2 and CO mole fractions in a laminar axisymmetric methane-air diffusion flame are compared with numerical predictions. A high-repetition-rate, high-average-power laser is used to increase signal-to-noise ratio to measure these minor flame species. Computationally, the system

  14. Identification and expression of C2H2 transcription factor genes in Carica papaya under abiotic and biotic stresses.

    Science.gov (United States)

    Jiang, Ling; Pan, Lin-jie

    2012-06-01

    C2H2 proteins belong to a group of transcription factors (TFs) existing as a superfamily that plays important roles in defense responses and various other physiological processes in plants. The present study aimed to screen for and identify C2H2 proteins associated with defense responses to abiotic and biotic stresses in Carica papaya L. Data were collected for 47,483 papaya-expressed sequence tags (ESTs). The full-length cDNA nucleotide sequences of 87 C2H2 proteins were predicated by BioEdit. All 91 C2H2 proteins were aligned, and a phylogenetic tree was constructed using DNAman. The expression levels of 42 C2H2 were analyzed under conditions of salt stress by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Methyl jasmonate treatment rapidly upregulated ZF(23.4) and ZF(30,912.1) by 18.6- and 21.7-fold, respectively. ZF(1.3), ZF(138.44), ZF(94.49), ZF(29.160), and ZF(20.206) were found to be downregulated after low temperature treatment at very significant levels (p papaya ringspot virus pathogen. ZF(30,912.1) was subcellularly localized in the nucleus by a transgenic fusion of pBS-ZF(30,912.1)-GFP into the protoplast of papaya. The results of the present study showed that ZF(30,912.1) could be an important TF that mediates responses to abiotic and biotic stresses in papaya.

  15. Editing the Plasmodium vivax genome, using zinc-finger nucleases.

    Science.gov (United States)

    Moraes Barros, Roberto R; Straimer, Judith; Sa, Juliana M; Salzman, Rebecca E; Melendez-Muniz, Viviana A; Mu, Jianbing; Fidock, David A; Wellems, Thomas E

    2015-01-01

    Plasmodium vivax is a major cause of malaria morbidity worldwide yet has remained genetically intractable. To stably modify this organism, we used zinc-finger nucleases (ZFNs), which take advantage of homology-directed DNA repair mechanisms at the site of nuclease action. Using ZFNs specific to the gene encoding P. vivax dihydrofolate reductase (pvdhfr), we transfected blood specimens from Saimiri boliviensis monkeys infected with the pyrimethamine (Pyr)-susceptible Chesson strain with a ZFN plasmid carrying a Pyr-resistant mutant pvdhfr sequence. We obtained Pyr-resistant parasites in vivo that carried mutant pvdhfr and additional silent mutations designed to confirm editing. These results herald the era of stable P. vivax genetic modifications.

  16. THE STARK EFFECT IN THE STIMULATED RAMAN SPECTRA (SRS) OF MOLECULE C2H2%C2H2分子受激Raman光谱中的光学Stark效应研究

    Institute of Scientific and Technical Information of China (English)

    高文斌; 杨永德; 徐娙梅

    2002-01-01

    本文研究了C2H2分子受激Raman光谱(SRS)中的光学斯塔克(Stark)效应,同时还研究了激光功率以及单一转动态量子数J与Stark效应的关系.实验表明:C2H2分子在高强度激光电场作用下,它的SRS由于光学Stark效应产生明显的非对称加宽,利用Stark线型加宽数值模拟,从理论上计算了Raman光谱线型,与实验的SRS线型比较,二者符合程度相当好.

  17. Can Co(II) or Cd(II) substitute for Zn(II) in zinc fingers?

    Indian Academy of Sciences (India)

    P Rabindra Reddy; M Radhika

    2001-02-01

    Zinc finger domains consist of sequences of amino acids containing cysteine and histidine residues tetrahedrally coordinated to a zinc ion. The role of zinc in a DNA binding finger was considered purely structural due to the absence of redox chemistry in zinc. However, whether other metals e.g. Co(II) or Cd(II) can substitute Zn(II) is not settled. For an answer the detailed interaction of Co(II) and Cd(II) with cysteine methylester and histidine methylester has been investigated as a model for the zinc core in zinc fingers. The study was extended to different temperatures to evaluate the thermodynamic parameters associated with these interactions. The results suggest that zinc has a unique role.

  18. Research Progress of PR Domain Zinc Finger Protein 14

    Directory of Open Access Journals (Sweden)

    Yudong HAN

    2016-02-01

    Full Text Available PR domain zinc finger protein 14 (PRDM14 is an important member of the PRDM family, PRDM14 plays a key role in the maintenance of cell integrity and differentiation, growth and apoptosis of the cell. It also plays an critical role in the formation of primordial germ cells, the maintenance of the totipotency of stem cells and the formation of tissues and organs. PRDM14 bears a single PR domain and six tandemly repeated zinc fingers, which is involved in the process of the deacetylation and methylation of the histone, and is involved in the formation of tumor trough the change level of methylation in the promoter region. The abnormal methylation of PRDM14 can change the chromatin structure, DNA conformation and the interaction mode of DNA and protein, it can suppress transcription and expression of the gene, which caused the occurrence, development and metastasis of tumor. The research progress of PRDM14 is reviewed based on the relevant literatures published in China and abroad.

  19. Zinc finger peptide based optic sensor for detection of zinc ions.

    Science.gov (United States)

    Verma, Neelam; Kaur, Gagandeep

    2016-12-15

    In the present work, polyacrylamide gel has been used as a matrix for the immobilization of zinc finger peptide and fluorescent dye acrydine orange on the micro well plate to fabricate the fluorescence based biosensor for the detection of zinc ions in milk samples. The fluorescent dye moves in the hydrophobic groove formed after folding of the peptide in the presence of zinc ions. Under optimized conditions, linear range was observed between 0.001µg/l to 10µg/l of Zinc ions, with a lowest detection limit of 0.001µg/l and response time of 5min. Presented biosensor has shown 20% decrease in fluorescent intensity values after 5 regenerations and stable for more than one month, stored at 4°C. Interference study with other metal ions like lead, cadmium and copper showed a negligible change in fluorescence intensity in comparison to zinc ions. Developed bio sensing system was found to be novel, quick, reliable, miniaturized, stable, reproducible and repeatable and specific for zinc ion, which has been applied to various milk samples. PMID:27424265

  20. Zinc finger peptide based optic sensor for detection of zinc ions.

    Science.gov (United States)

    Verma, Neelam; Kaur, Gagandeep

    2016-12-15

    In the present work, polyacrylamide gel has been used as a matrix for the immobilization of zinc finger peptide and fluorescent dye acrydine orange on the micro well plate to fabricate the fluorescence based biosensor for the detection of zinc ions in milk samples. The fluorescent dye moves in the hydrophobic groove formed after folding of the peptide in the presence of zinc ions. Under optimized conditions, linear range was observed between 0.001µg/l to 10µg/l of Zinc ions, with a lowest detection limit of 0.001µg/l and response time of 5min. Presented biosensor has shown 20% decrease in fluorescent intensity values after 5 regenerations and stable for more than one month, stored at 4°C. Interference study with other metal ions like lead, cadmium and copper showed a negligible change in fluorescence intensity in comparison to zinc ions. Developed bio sensing system was found to be novel, quick, reliable, miniaturized, stable, reproducible and repeatable and specific for zinc ion, which has been applied to various milk samples.

  1. Near-IR tunable laser based photoacoustic sensor for sub-ppb C2H2 detections

    Science.gov (United States)

    Wang, Jianwei; Wang, Huili

    2015-05-01

    A photoacoustic sensor for the detection of acetylene (C2H2) based on an EDFA-amplified tunable laser at 1530.37 nm is reported. This sensor consists of a near-IR tunable external cavity diode laser combined with a fiber amplifier and wavelength modulation mechanism. In order to improve detection sensitivity and accuracy, an optimized resonant photoacoustic cell based on an acoustic longitudinal mode and automatic tracking of the acoustic resonance frequency using a speaker and a FPGA-based servo controller is implemented. The minimum detection limit (signal-to-noise ratio = 1) of 0.8 ppb for C2H2 is demonstrated at room temperature and atmospheric pressure.

  2. Measurements of hydrogen cyanide (HCN and acetylene (C2H2 from the Infrared Atmospheric Sounding Interferometer (IASI

    Directory of Open Access Journals (Sweden)

    V. Duflot

    2013-04-01

    Full Text Available Hydrogen cyanide (HCN and acetylene (C2H2 are ubiquitous atmospheric trace gases with medium lifetime, which are frequently used as indicators of combustion sources and as tracers for atmospheric transport and chemistry. Because of their weak infrared absorption, overlapped by the CO2 Q branch near 720 cm−1, nadir sounders have up to now failed to measure these gases routinely. Taking into account CO2 line mixing, we provide for the first time extensive measurements of HCN and C2H2 total columns at Reunion Island (21° S, 55° E and Jungfraujoch (46° N, 8° E in 2009–2010 using observations from the Infrared Atmospheric Sounding Interferometer (IASI. A first order comparison with local ground-based Fourier transform infraRed (FTIR measurements has been carried out allowing tests of seasonal consistency which is reasonably captured, except for HCN at Jungfraujoch. The IASI data shows a greater tendency to high C2H2 values. We also examine a nonspecific biomass burning plume over austral Africa and show that the emission ratios with respect to CO agree with previously reported values.

  3. Comparative analysis of zinc finger proteins involved in plant disease resistance.

    Science.gov (United States)

    Gupta, Santosh Kumar; Rai, Amit Kumar; Kanwar, Shamsher Singh; Sharma, Tilak R

    2012-01-01

    A meta-analysis was performed to understand the role of zinc finger domains in proteins of resistance (R) genes cloned from different crops. We analyzed protein sequences of seventy R genes of various crops in which twenty six proteins were found to have zinc finger domains along with nucleotide binding sites - leucine rice repeats (NBS-LRR) domains. We identified thirty four zinc finger domains in the R proteins of nine crops and were grouped into 19 types of zinc fingers. The size of individual zinc finger domain within the R genes varied from 11 to 84 amino acids, whereas the size of proteins containing these domains varied from 263 to 1305 amino acids. The biophysical analysis revealed that molecular weight of Pi54 zinc finger was lowest whereas the highest one was found in rice Pib zinc finger named as Transposes Transcription Factor (TTF). The instability (R(2) =0.95) and the aliphatic (R(2) =0.94) indices profile of zinc finger domains follows the polynomial distribution pattern. The pairwise identity analysis showed that the Lin11, Isl-1 & Mec-3 (LIM) zinc finger domain of rice blast resistance protein pi21 have 12.3% similarity with the nuclear transcription factor, X-box binding-like 1 (NFX) type zinc finger domain of Pi54 protein. For the first time, we reported that Pi54 (Pi-k(h)-Tetep), a rice blast resistance (R) protein have a small zinc finger domain of NFX type located on the C-terminal in between NBS and LRR domains of the R-protein. Compositional analysis depicted by the helical wheel diagram revealed the presence of a hydrophobic region within this domain which might help in exposing the LRR region for a possible R-Avr interaction. This domain is unique among all other cloned plant disease resistance genes and might play an important role in broad-spectrum nature of rice blast resistance gene Pi54. PMID:22916136

  4. Dopamine receptor regulating factor, DRRF: a zinc finger transcription factor.

    Science.gov (United States)

    Hwang, C K; D'Souza, U M; Eisch, A J; Yajima, S; Lammers, C H; Yang, Y; Lee, S H; Kim, Y M; Nestler, E J; Mouradian, M M

    2001-06-19

    Dopamine receptor genes are under complex transcription control, determining their unique regional distribution in the brain. We describe here a zinc finger type transcription factor, designated dopamine receptor regulating factor (DRRF), which binds to GC and GT boxes in the D1A and D2 dopamine receptor promoters and effectively displaces Sp1 and Sp3 from these sequences. Consequently, DRRF can modulate the activity of these dopamine receptor promoters. Highest DRRF mRNA levels are found in brain with a specific regional distribution including olfactory bulb and tubercle, nucleus accumbens, striatum, hippocampus, amygdala, and frontal cortex. Many of these brain regions also express abundant levels of various dopamine receptors. In vivo, DRRF itself can be regulated by manipulations of dopaminergic transmission. Mice treated with drugs that increase extracellular striatal dopamine levels (cocaine), block dopamine receptors (haloperidol), or destroy dopamine terminals (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) show significant alterations in DRRF mRNA. The latter observations provide a basis for dopamine receptor regulation after these manipulations. We conclude that DRRF is important for modulating dopaminergic transmission in the brain. PMID:11390978

  5. Impact-induced chemisorption of C2H2 on diamond(0 0 1) surfaces: a molecular dynamics simulation

    International Nuclear Information System (INIS)

    This paper reports some new observations in studying the interaction between hyperthermal C2H2 and diamond (0 0 1)-2x1 surface at room temperature by means of molecular dynamics simulations with the many-body hydrocarbon potential. Six types of chemisorption configurations have been observed, some of these are reported for the first time. Also, we have observed threshold effect and hydrogen atom dissociation in the chemisorption processes. All these facts are expected to be useful in setting experimental condition in diamond synthesis

  6. Ion-neutral reactions of C2H2N+ with hydrocarbons : relevant to Titan’s ionosphere

    OpenAIRE

    Fathi, Pantea

    2015-01-01

    To provide some insights on the formation of long-chain carbon-nitrogen bearing ions in Titan’s atmosphere, laboratory measurements of ion-neutral reactions were conducted and results are presented in the following licentiate thesis. The product ions of the C2H2N+ reaction with saturated and unsaturated neutral hydrocarbons have been studied using guided ion beam mass spectrometer (GIB-MS). The data have been used to infer the product ion signal intensity dependences on pressure and collision...

  7. Measurements of hydrogen cyanide (HCN and acetylene (C2H2 from the Infrared Atmospheric Sounding Interferometer (IASI

    Directory of Open Access Journals (Sweden)

    C. Clerbaux

    2012-10-01

    Full Text Available Hydrogen cyanide (HCN and acetylene (C2H2 are ubiquitous atmospheric trace gases with medium lifetime, which are frequently used as indicators of combustion sources and as tracers for atmospheric transport and chemistry. Because of their weak infrared absorption, overlapped by the CO2 Q-branch near 720 cm−1, nadir sounders have up to now failed to measure these gases routinely. Taking into account CO2 line mixing we provide for the first time extensive measurements of HCN and C2H2 total columns at Reunion Island (21° S; 55° E and Jungfraujoch (46° N; 8° E in 2009–2010 using observations from the Infrared Atmospheric Sounding Interferometer (IASI. These are compared with local ground-based Fourier Transform InfraRed (FTIR measurements and we demonstrate that the seasonality is well captured, except for HCN at Jungfraujoch. We also examine a nonspecific biomass burning plume over austral Africa and show that the emission ratios with respect to CO agree with previously reported values.

  8. Molecular Cloning and Expression Analysis of a Cys2/His2 Type Zinc Finger Protein Gene in Upland Cotton

    Institute of Scientific and Technical Information of China (English)

    YANG Yu-wen; NI Wan-chao; ZHANG Bao-long; SHEN Xin-lian

    2008-01-01

    @@ The zinc finger proteins belong to the largest family of transcription factors.But there is little research of Cys2/His2 type zinc finger proteins in cotton,and there is no submission of correlating ESTs to GenBank.In this study,a full length of one Cys2/His2 type zinc finger protein (GZFP) and its 5 flanking sequence were obtained by RT-PCR and tail PCR.

  9. Metal Coupled Folding of Cys2His2 Zinc-Finger

    CERN Document Server

    Li, Wenfei; Wang, Jun; Wang, Wei

    2008-01-01

    Zinc-fingers, which widely exist in eukaryotic cell and play crucial roles in life processes, depend on the binding of zinc ion for their proper folding. To computationally study the zinc coupled folding of the zinc-fingers, charge transfer and metal induced protonation/deprotonation effects have to be considered. Here, by attempting to implicitly account for such effects in classical molecular dynamics and performing intensive simulations with explicit solvent for the peptides with and without zinc binding, we investigate the folding of the Cys2His2 type zinc-finger motif and the coupling between the peptide folding and zinc binding. We find that zinc ion not only stabilizes the native structure, but also participates in the whole folding process. It binds to the peptide at early stage of folding, and directs or modulates the folding and stabilizations of the component beta-hairpin and alpha-helix. Such a crucial role of zinc binding is mediated by the packing of the conserved hydrophobic residues. We also f...

  10. The artificial zinc finger coding gene 'Jazz' binds the utrophin promoter and activates transcription.

    Science.gov (United States)

    Corbi, N; Libri, V; Fanciulli, M; Tinsley, J M; Davies, K E; Passananti, C

    2000-06-01

    Up-regulation of utrophin gene expression is recognized as a plausible therapeutic approach in the treatment of Duchenne muscular dystrophy (DMD). We have designed and engineered new zinc finger-based transcription factors capable of binding and activating transcription from the promoter of the dystrophin-related gene, utrophin. Using the recognition 'code' that proposes specific rules between zinc finger primary structure and potential DNA binding sites, we engineered a new gene named 'Jazz' that encodes for a three-zinc finger peptide. Jazz belongs to the Cys2-His2 zinc finger type and was engineered to target the nine base pair DNA sequence: 5'-GCT-GCT-GCG-3', present in the promoter region of both the human and mouse utrophin gene. The entire zinc finger alpha-helix region, containing the amino acid positions that are crucial for DNA binding, was specifically chosen on the basis of the contacts more frequently represented in the available list of the 'code'. Here we demonstrate that Jazz protein binds specifically to the double-stranded DNA target, with a dissociation constant of about 32 nM. Band shift and super-shift experiments confirmed the high affinity and specificity of Jazz protein for its DNA target. Moreover, we show that chimeric proteins, named Gal4-Jazz and Sp1-Jazz, are able to drive the transcription of a test gene from the human utrophin promoter.

  11. Intense laser induced field ionization of C2H2, C2H4,and C2H6

    Institute of Scientific and Technical Information of China (English)

    GAO Lirong; JI Na; XONG Yijia; TANG Xiaoping; KONG Fan'ao

    2003-01-01

    Using HOMO Field Ionization Model, the tunneling probabilities and the theoretical threshold intensities of the field ionizations of acetylene, ethylene, and ethane in intense laser field are calculated. C2H2, C2H4, and C2H6 were irradiated by 800 nm, 100 fs laser pulses with the intensity range of 1013-1014 W/cm2. A TOF-mass spectrometer was coupled to the laser system and used to experimentally investigate the field ionization of these molecules. The experimental ionization threshold intensities are obtained. The calculating results of the three molecules agree well with the experimental results, indicating that HOMO Field Ionization Model is valid for the ionization of polyatomic molecules in intense laser field.

  12. Temperature Dependent Product Yields for the Spin Forbidden Singlet Channel of the C(3P) + C2H2 Reaction

    CERN Document Server

    Hickson, Kevin M; Wakelam, Valentine

    2016-01-01

    The atomic hydrogen formation channels of the C + C2H2 reaction have been investigated using a continuous supersonic flow reactor over the 52 K 296 K temperature range. H-atoms were detected directly at 121.567 nm by vacuum ultraviolet laser induced fluorescence. Absolute H-atom yields were determined by comparison with the H-atom signal generated by the C + C2H4 reaction. The product yields agree with earlier crossed beam experiments employing universal detection methods. Incorporating these branching ratios in a gas-grain model of dense interstellar clouds increases the cC3H abundance. This reaction is a minor source of C3 containing molecules in the present simulations.

  13. Simulating electron momentum spectra of iso-C2H2Cl2: A study of the core electronic structure

    International Nuclear Information System (INIS)

    Electron momentum spectroscopy (EMS) has been used for the first time to study core electronic structure of iso-C2H2Cl2. In the present work, the pronounced difference between ionization energies of two C1s core orbitals (2A1 and 3A1) is seen as a chemical shift of 3 eV, which is due to different chemical environments of the related carbon atoms. Both the calculated spherically averaged core electron momentum distributions (MDs) and three-dimensional electron momentum density maps show that these core molecular orbitals (MOs) 2A1 and 3A1 exhibit strong atomic orbital characteristics in real and momentum space. However, the core states 2B2 and 4A1, which are almost degenerate and related to two equivalent atoms, exhibit notable differences between the momentum and position depictions. In contrast to the position space, the momentum density maps of these two core MOs highlight the interference effects which are due to the nuclear positions. The 2B2 orbital of iso-C2H2Cl2 is the antisymmetric counterpart of the 4A1 core orbital in real space. However, it relates to the 4A1 orbital by an exchange of maxima and minima in momentum space. Due to interference effects between electrons scattered from different atomic centers, modulations with a periodicity of 1.12 a.u. can be seen in the computed momentum densities, which tend to decay with increasing electron momenta. Accordingly, the EMS can not only effectively image the electronic structure of compounds by studying valence orbitals, but also provides direct information on the nature of the nuclear geometry by investigating the core states. (atomic and molecular physics)

  14. Zinc-finger antiviral protein inhibits XMRV infection.

    Directory of Open Access Journals (Sweden)

    Xinlu Wang

    Full Text Available BACKGROUND: The zinc-finger antiviral protein (ZAP is a host factor that specifically inhibits the replication of certain viruses, including Moloney murine leukemia virus (MoMLV, HIV-1, and certain alphaviruses and filoviruses. ZAP binds to specific viral mRNAs and recruits cellular mRNA degradation machinery to degrade the target RNA. The common features of ZAP-responsive RNA sequences remain elusive and thus whether a virus is susceptible to ZAP can only be determined experimentally. Xenotropic murine leukemia virus-related virus (XMRV is a recently identified γ-retrovirus that was originally thought to be involved in prostate cancer and chronic fatigue syndrome but recently proved to be a laboratory artefact. Nonetheless, XMRV as a new retrovirus has been extensively studied. Since XMRV and MoMLV share only 67.9% sequence identity in the 3'UTRs, which is the target sequence of ZAP in MoMLV, whether XMRV is susceptible to ZAP remains to be determined. FINDINGS: We constructed an XMRV-luc vector, in which the coding sequences of Gag-Pol and part of Env were replaced with luciferase-coding sequence. Overexpression of ZAP potently inhibited the expression of XMRV-luc in a ZAP expression-level-dependent manner, while downregulation of endogenous ZAP rendered cells more sensitive to infection. Furthermore, ZAP inhibited the spreading of replication-competent XMRV. Consistent with the previously reported mechanisms by which ZAP inhibits viral infection, ZAP significantly inhibited the accumulation of XMRV-luc mRNA in the cytoplasm. The ZAP-responsive element in XMRV mRNA was mapped to the 3'UTR. CONCLUSIONS: ZAP inhibits XMRV replication by preventing the accumulation of viral mRNA in the cytoplasm. Documentation of ZAP inhibiting XMRV helps to broaden the spectrum of ZAP's antiviral activity. Comparison of the target sequences of ZAP in XMRV and MoMLV helps to better understand the features of ZAP-responsive elements.

  15. The (unusual) aspartic acid in the metal coordination sphere of the prokaryotic zinc finger domain.

    Science.gov (United States)

    D'Abrosca, Gianluca; Russo, Luigi; Palmieri, Maddalena; Baglivo, Ilaria; Netti, Fortuna; de Paola, Ivan; Zaccaro, Laura; Farina, Biancamaria; Iacovino, Rosa; Pedone, Paolo Vincenzo; Isernia, Carla; Fattorusso, Roberto; Malgieri, Gaetano

    2016-08-01

    The possibility of choices of protein ligands and coordination geometries leads to diverse Zn(II) binding sites in zinc-proteins, allowing a range of important biological roles. The prokaryotic Cys2His2 zinc finger domain (originally found in the Ros protein from Agrobacterium tumefaciens) tetrahedrally coordinates zinc through two cysteine and two histidine residues and it does not adopt a correct fold in the absence of the metal ion. Ros is the first structurally characterized member of a family of bacterial proteins that presents several amino acid changes in the positions occupied in Ros by the zinc coordinating residues. In particular, the second position is very often occupied by an aspartic acid although the coordination of structural zinc by an aspartate in eukaryotic zinc fingers is very unusual. Here, by appropriately mutating the protein Ros, we characterize the aspartate role within the coordination sphere of this family of proteins demonstrating how the presence of this residue only slightly perturbs the functional structure of the prokaryotic zinc finger domain while it greatly influences its thermodynamic properties. PMID:27238756

  16. Stress Responsive Zinc-finger Protein Gene of Populus euphratica in Tobacco Enhances Salt Tolerance

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The Populus euphratica stress responsive zinc-finger protein gene PSTZ, which encodes a protein including typical Cys2/His2 zinc finger structure, was isolated by reverse transcription-polymerase chain reaction from P. euphratica.Northern hybridization revealed that its expression was induced under drought and salt stress conditions. To examine its function, cDNA of the PSTZ gene, driven by the cauliflower mosaic virus 35S promoter, was cloned into a plant expression vector pBin438 and introduced into tobacco plants. Transgenic tobacco showed an enhanced salt tolerance, suggesting that PSTZ may play a role in plant responsiveness to salt stress.

  17. Metal binding and folding properties of a minimalist Cys2His2 zinc finger peptide.

    OpenAIRE

    Michael, S.F.; Kilfoil, V J; Schmidt, M.H.; Amann, B T; Berg, J M

    1992-01-01

    A minimalist Cys2His2 zinc finger peptide, Lys-Tyr-Ala-Cys-Ala-Ala-Cys-Ala-Ala-Ala-Phe-Ala-Ala-Lys-Ala-Ala-Leu-Ala- Ala-His-Ala-Ala-Ala-His-Ala-Lys, has been synthesized. Metal binding studies using Co2+ as a probe indicated that this peptide forms a 1:1 peptide/metal complex with a dissociation constant comparable to that observed for other zinc finger peptides. At high peptide concentrations, a 2:1 peptide/metal complex also forms, with four cysteinates coordinated to Co2+. Additional studi...

  18. Relationship between C2H2 reduction, H2 evolution and 15N2 fixation in root nodules of pea (Pisum sativum)

    DEFF Research Database (Denmark)

    Skøt, Leif

    1983-01-01

    after planting. After correction for H2 evolution in air, the factor (C2H2-H2)/N2 decreased to values near the theoretical value 3, or in one case to a value significantly (P electron flow through nitrogenase, which is not wasted in H2 production but used...... for N2 reduction, is often stated as the relative efficiency (1-H2/C2H2). This factor varied significantly (P electrons to H2 and N2, expressed as the H2/N2 ratio, was independent of plant age, however. This discrepancy and the observation...... that the (C2H2-H2)/N2 conversion factor tended to be lower than 3, suggests that the C2H2reduction assay underestimates the total electron flow through nitrogenase....

  19. 人工锌指蛋白随机库的构建及其在锌指核酸酶筛选中的应用%Construction of Artificial Zinc Finger Libraries and Application in Screening Zinc Finger Nucleases

    Institute of Scientific and Technical Information of China (English)

    李战伟; 王昕; 任刚; 王令; 杨涵江; 张智英

    2012-01-01

    利用开源式(Oligomerized pool engineering,OPEN)方法筛选具有较高特异性和亲和性的锌指蛋白(Zinc finger protein,ZFP).以已知三锌指蛋白为框架,使单个锌指关键位点氨基酸编码序列产生随机突变,构建3个人工锌指蛋白随机库,建立和完善应用人工锌指蛋白随机库筛选特异性锌指蛋白的技术平台.以人DYRK1A基因为例,测试和验证该技术平台的可行性和效率,分别获得对DYRK1A基因靶序列中左侧和右侧位点具有较高亲和性的50个三锌指蛋白;然后将得到的锌指蛋白与非限制性核酸内切酶FokⅠ连接,构建DYRK1A锌指核酸酶(Zinc finger nucleases,ZFN),应用酵母验证系统测试DYRK1A锌指核酸酶的活性,结果表明,90%的组装锌指核酸酶能够在靶位点进行切割.%The oligomerized pool engineering (OPEN) approach was applied to select specific zinc finger proteins (ZFP) with high affinity. We constructed three zinc finger random libraries, each based on a synthetic "framework" ZFP. In each library, recognition helix residues -1, 1, 2, 3, 4, 5, and 6 from a single finger were randomized by cassette mutagenesis. The three zinc finger random libraries were used to screen specific zinc finger proteins. Human DYRK1A gene was exemplified to examine the feasibility and efficiency of the technology platform for screening of zinc finger proteins. As a result 5o zinc finger proteins with higher affinity to the recognition sites of DYRK1A gene were obtained. The zinc finger proteins were fused to the non-specific endonuclease domain of Fok I to form zinc finger nucleases. Then the activity of zinc finger nucleases was examined in yeast validation system. The result showd that 90% zinc finger nucleases worked well in yeast cells. This research provided the technique support for the application of zinc finger nucleases to the editing of the genome.

  20. An aminopyrimidine-functionalized cage-based metal-organic framework exhibiting highly selective adsorption of C2H2 and CO2 over CH4.

    Science.gov (United States)

    Jiao, Jingjing; Dou, Li; Liu, Huimin; Chen, Fengli; Bai, Dongjie; Feng, Yunlong; Xiong, Shunshun; Chen, De-Li; He, Yabing

    2016-09-14

    There has been considerable interest in adsorptive separation of C2H2/CH4 and CO2/CH4 gas mixtures due to its industrial significance and scientific challenge. In this work, we have designed and synthesized a bent diisophthalate ligand functionalized with aminopyrimidine groups, and constructed via a solvothermal reaction, a porous copper-based framework. Single-crystal X-ray diffraction studies show that the framework is a three-dimensional network containing three different types of polyhedral nanocages, which are stacked together to form two distinct types of one-dimensional channels along the crystallographic c axis. The compound after activation shows exceptionally high C2H2 and CO2 uptakes of 211 and 120 cm(3) (STP) g(-1) at 295 K and 1 atm, as well as impressive adsorption selectivities towards C2H2 and CO2 over CH4. High C2H2 and CO2 uptake capacities as well as significant adsorption selectivities of C2H2 and CO2 over CH4 imply potential applications in the adsorptive separation and purification of C2H2/CH4 and CO2/CH4 gas mixtures, which have been verified by column breakthrough experiments. Several important binding sites for C2H2 and CO2 in ZJNU-54 were revealed by quantum chemical calculations, demonstrating that the organic linkers in ZJNU-54 form unique structures that facilitate the adsorption of C2H2, while the amine groups and the Lewis basic pyrimidine-ring nitrogen sites in the organic linker improve the adsorption energies for CO2, finally leading to the increase of adsorption capacities for these two gas molecules. This work provides an efficient strategy for incorporating specific functional groups into cage-based MOFs for generating new adsorbents for highly selective gas storage and separation. PMID:27483189

  1. Buckybowls as adsorbents for CO2, CH4, and C2H2: Binding and structural insights from computational study.

    Science.gov (United States)

    Hussain, M Althaf; Vijay, Dolly; Sastry, G Narahari

    2016-01-30

    Noncovalent functionalization of buckybowls sumanene (S), corannulene (R), and coronene (C) with greenhouse gases (GGs) such as CO2 , CH4 (M), and C2 H2 (A) has been studied using hybrid density functional theory. The propensity and preferences of these small molecules to interact with the concave and convex surfaces of the buckybowls has been quantitatively estimated. The results indicate that curvature plays a significant role in the adsorption of these small molecules on the π surface and it is observed that buckybowls have higher binding energies (BEs) compared with their planar counterpart coronene. The concave surface of the buckybowl is found to be more feasible for adsorption of small molecules. BEs of small molecules towards π systems is CO2  > A > M and the BEs of π systems toward small molecules is S > R > C. Obviously, the binding preference is dictated by the way in which various noncovalent interactions, such as π···π, lone pair···π, and CH···π manifest themselves on carbaneous surfaces. To delineate the intricate details of the interactions, we have employed Bader's quantum theory of atoms in molecule and localized molecular orbital energy decomposition analysis (LMO-EDA). LMO-EDA, which measures the contribution of various components and traces the physical origin of the interactions, indicates that the complexes are stabilized largely by dispersion interactions.

  2. High-frequency genome editing using ssDNA oligonucleotides with zinc-finger nucleases

    DEFF Research Database (Denmark)

    Chen, Fuqiang; Pruett-Miller, Shondra M; Huang, Yuping;

    2011-01-01

    Zinc-finger nucleases (ZFNs) have enabled highly efficient gene targeting in multiple cell types and organisms. Here we describe methods for using simple ssDNA oligonucleotides in tandem with ZFNs to efficiently produce human cell lines with three distinct genetic outcomes: (i) targeted point...

  3. DUF581 is plant specific FCS-like zinc finger involved in protein-protein interaction.

    Science.gov (United States)

    K, Muhammed Jamsheer; Laxmi, Ashverya

    2014-01-01

    Zinc fingers are a ubiquitous class of protein domain with considerable variation in structure and function. Zf-FCS is a highly diverged group of C2-C2 zinc finger which is present in animals, prokaryotes and viruses, but not in plants. In this study we identified that a plant specific domain of unknown function, DUF581 is a zf-FCS type zinc finger. Based on HMM-HMM comparison and signature motif similarity we named this domain as FCS-Like Zinc finger (FLZ) domain. A genome wide survey identified that FLZ domain containing genes are bryophytic in origin and this gene family is expanded in spermatophytes. Expression analysis of selected FLZ gene family members of A. thaliana identified an overlapping expression pattern suggesting a possible redundancy in their function. Unlike the zf-FCS domain, the FLZ domain found to be highly conserved in sequence and structure. Using a combination of bioinformatic and protein-protein interaction tools, we identified that FLZ domain is involved in protein-protein interaction.

  4. DUF581 is plant specific FCS-like zinc finger involved in protein-protein interaction.

    Directory of Open Access Journals (Sweden)

    Muhammed Jamsheer K

    Full Text Available Zinc fingers are a ubiquitous class of protein domain with considerable variation in structure and function. Zf-FCS is a highly diverged group of C2-C2 zinc finger which is present in animals, prokaryotes and viruses, but not in plants. In this study we identified that a plant specific domain of unknown function, DUF581 is a zf-FCS type zinc finger. Based on HMM-HMM comparison and signature motif similarity we named this domain as FCS-Like Zinc finger (FLZ domain. A genome wide survey identified that FLZ domain containing genes are bryophytic in origin and this gene family is expanded in spermatophytes. Expression analysis of selected FLZ gene family members of A. thaliana identified an overlapping expression pattern suggesting a possible redundancy in their function. Unlike the zf-FCS domain, the FLZ domain found to be highly conserved in sequence and structure. Using a combination of bioinformatic and protein-protein interaction tools, we identified that FLZ domain is involved in protein-protein interaction.

  5. Vertical spectrum of the C2H2+ system. An open shell (SC)2-CAS-SDCI study.

    Science.gov (United States)

    Pitarch-Ruiz, José; Sánchez-Marín, José; Maynau, Daniel

    2003-04-15

    The open shell (SC)(2)-CAS-SDCI method along with a basis set of atomic natural orbitals (ANO) has been applied for calculating the main ionization potentials of acetylene, as well as the manifold of excited states of the different symmetries up to 32 eV. In this method, the single and double excitations of a CAS space are generated and the corresponding CI matrix is corrected by means of the (SC)(2) procedure that cancels the size-extensivity error and adds some high order contributions. The mean absolute error for the outer-valence X (2)Pi(u)(1pi(u) (-1)), A (2)Sigma(g) (+)(3sigma(g) (-1)), and B (2)Sigma(u) (+)(2sigma(u) (-1)) states, and the inner-valence C (2)Sigma(g) (+)(2sigma(g) (-1)) state is 0.1 eV. The excited states of C(2)H(2) (+) corresponding to the Sigma(g) (+), Sigma(u) (+), Pi(g), Pi(u), Delta(g), and Delta(u) symmetries are reported and their composition is discussed. The results are thoroughly compared to the best available multireference CI calculations. Recent multichannel CI results by Wells and Lucchese [J Chem Phys 1999, 110, 6365] have been used also as a guide for the discussion of the results. Discrepancies in the description of many multiconfigurational states by means of the (SC)(2)-CAS-SDCI wave function as compared to previous large MR-CI calculations are significant and are, consequently, remarked on. A large mixing of the 3sigma(g) (-1) and 2sigma(g) (-1) processes is found in the A (2)Sigma(g) (+) and C (2)Sigma(g) (+) states, provided that the basis set is augmented with Rydberg functions. PMID:12632475

  6. Rapid mutation of endogenous zebrafish genes using zinc finger nucleases made by Oligomerized Pool ENgineering (OPEN.

    Directory of Open Access Journals (Sweden)

    Jonathan E Foley

    Full Text Available BACKGROUND: Customized zinc finger nucleases (ZFNs form the basis of a broadly applicable tool for highly efficient genome modification. ZFNs are artificial restriction endonucleases consisting of a non-specific nuclease domain fused to a zinc finger array which can be engineered to recognize specific DNA sequences of interest. Recent proof-of-principle experiments have shown that targeted knockout mutations can be efficiently generated in endogenous zebrafish genes via non-homologous end-joining-mediated repair of ZFN-induced DNA double-stranded breaks. The Zinc Finger Consortium, a group of academic laboratories committed to the development of engineered zinc finger technology, recently described the first rapid, highly effective, and publicly available method for engineering zinc finger arrays. The Consortium has previously used this new method (known as OPEN for Oligomerized Pool ENgineering to generate high quality ZFN pairs that function in human and plant cells. METHODOLOGY/PRINCIPAL FINDINGS: Here we show that OPEN can also be used to generate ZFNs that function efficiently in zebrafish. Using OPEN, we successfully engineered ZFN pairs for five endogenous zebrafish genes: tfr2, dopamine transporter, telomerase, hif1aa, and gridlock. Each of these ZFN pairs induces targeted insertions and deletions with high efficiency at its endogenous gene target in somatic zebrafish cells. In addition, these mutations are transmitted through the germline with sufficiently high frequency such that only a small number of fish need to be screened to identify founders. Finally, in silico analysis demonstrates that one or more potential OPEN ZFN sites can be found within the first three coding exons of more than 25,000 different endogenous zebrafish gene transcripts. CONCLUSIONS AND SIGNIFICANCE: In summary, our study nearly triples the total number of endogenous zebrafish genes successfully modified using ZFNs (from three to eight and suggests that OPEN

  7. Selective catalytic reduction of NO by C2H2 over Ce-Al2O3 catalyst with rate-determining step of NO oxidation

    Institute of Scientific and Technical Information of China (English)

    Suhua Yan; Xinping Wang; Wenchen Wang; Zequn Liu; Jiahao Niu

    2012-01-01

    Ce-Al2O3 catalysts prepared by co-precipitation are investigated both in NO oxidation by O2 and in selective catalytic reduction of NO by C2H2 (C2H2-SCR).It is found that C2H2-SCR is initiated and controlled by NO oxidation to NO2 over A12O3.Ce loading on Al2O3 is almost inactive for NO oxidation below 350 ℃,since NO2 strongly adsorbs on cerium oxide,leading to the active sites being blocked,which was characterized by temperature-programmed desorption of NO and NO2 and Fourier transform infrared spectroscopy after NO+O2 coadsorption over the samples.However,in the case of C2H2-SCR,Ce loading on Al2O3 significantly improves the reaction by accelerating the NO oxidation step in the temperature range of 250-450 ℃,since the nitrate species produced by NO2 adsorption is an active intermediate required by C2H2-SCR.

  8. Lead neurotoxicity: exploring the potential impact of lead substitution in zinc-finger proteins on mental health.

    Science.gov (United States)

    Ordemann, Jacqueline Michelle; Austin, Rachel Narehood

    2016-06-01

    Childhood lead poisoning is a costly and largely preventable public health problem that lowers IQs, decreases attention spans, and leads to the development of other childhood intellectual disabilities. Furthermore, recent evidence links developmental lead poisoning with the etiology of disorders that appear much later in life, such as Alzheimer's disease, Parkinson's disease, and schizophrenia. Little is known about how lead influences the onset of these disorders. This paper reviews the evidence that lead substitution for zinc in zinc-finger proteins contributes to the development of Alzheimer's disease, Parkinson's disease, and schizophrenia. The zinc-finger proteins potentially impacted by lead include DNA methyltransferase 1 (DNMT1) and Presenilin 1 and 2 (PSEN1/2) in Alzheimer's disease, the dopamine receptor in Parkinson's disease, and the NMDA receptor, zinc-finger protein 804A (ZNF804A), and disrupted-in-schizophrenia 1 (DISC1)-binding zinc-finger (DBZ) in schizophrenia. PMID:26745006

  9. Design, construction, and analysis of specific zinc finger nucleases for microphthalmia - associate transcription factor

    Directory of Open Access Journals (Sweden)

    Wenwen Wang

    2012-08-01

    Full Text Available This work studied the design, construction, and cleavage analysis of zinc finger nucleases (ZFNs that could cut the specific sequences within microphthalmia - associate transcription factor (mitfa of zebra fish. The target site and ZFPs were selected and designed with zinc finger tools, while the ZFPs were synthesized using DNAWorks and two-step PCR. The ZFNs were constructed, expressed, purified, and analyzed in vitro. As expected, the designed ZFNs could create a double-stand break (DSB at the target site in vitro. The DNAWorks, two-step PCR, and an optimized process of protein expression were firstly induced in the construction of ZFNs successfully, which was an effective and simplified protocol. These results could be useful for further application of ZFNs - mediated gene targeting.

  10. Site-Specific Editing of the Plasmodium falciparum Genome Using Engineered Zinc-Finger Nucleases

    OpenAIRE

    Straimer, Judith; Lee, Marcus CS; Lee, Andrew H.; Zeitler, Bryan; Williams, April E.; Pearl, Jocelynn R.; Zhang, Lei; Rebar, Edward J.; Gregory, Philip D.; Llinás, Manuel; Urnov, Fyodor D; David A Fidock

    2012-01-01

    Malaria afflicts over 200 million people worldwide and its most lethal etiologic agent, Plasmodium falciparum, is evolving to resist even the latest-generation therapeutics. Efficient tools for genome-directed investigations of P. falciparum pathogenesis, including drug resistance mechanisms, are clearly required. Here we report rapid and targeted genetic engineering of this parasite, using zinc-finger nucleases (ZFNs) that produce a double-strand break in a user-defined locus and trigger hom...

  11. Efficient targeted mutagenesis in the monarch butterfly using zinc-finger nucleases

    OpenAIRE

    Merlin, Christine; Beaver, Lauren E.; Taylor, Orley R.; Wolfe, Scot A.; Reppert, Steven M.

    2013-01-01

    The development of reverse-genetic tools in “nonmodel” insect species with distinct biology is critical to establish them as viable model systems. The eastern North American monarch butterfly (Danaus plexippus), whose genome is sequenced, has emerged as a model to study animal clocks, navigational mechanisms, and the genetic basis of long-distance migration. Here, we developed a highly efficient gene-targeting approach in the monarch using zinc-finger nucleases (ZFNs), engineered nucleases th...

  12. A study of the ground states of CaC2H+2,CaC2D+2 and CaC2H+4

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    The geometries, vibrational frequencies and bind energies are reported for the ground states of CaC2H+2, CaC2D+2 and CaC2H+4. CaC2H+2 and CaC2H+4 equilibrium geometries have C2v symmetry with the metal ion lying in the perpendicular bisector of the C-C bond. The ground state in both CaC2H+2 and CaC2H+4 molecules ia a 2A1 state and the binding in the ground state is mainly electrostatic. For both CaC2H+2 and CaC2H+4 the ligand is only slightly distorted from its free ligand structure, the C-C distance has hardly increased and there is only a very small bending of the H atom away from the Ca atom. This is consistent with the electrostatic nature of the bonding. Two different approaches-Hartree-Fock(HF) and density functional theory methods(DFT)-are used and basis sets here used is 6-311+G(3df,2p). The DFT results are in good agreement with experiments, namely, DFT methods provide the benefits that some more expensive ab initio methods can do, but at essentially HF cost. So it is important to include electron correlation for accurate results in this study.

  13. A rod-packing microporous hydrogen-bonded organic framework for highly selective separation of C2H2/CO2at room temperature

    KAUST Repository

    Li, Peng

    2014-11-13

    Self-assembly of a trigonal building subunit with diaminotriazines (DAT) functional groups leads to a unique rod-packing 3D microporous hydrogen-bonded organic framework (HOF-3). This material shows permanent porosity and demonstrates highly selective separation of C2H2/CO2 at ambient temperature and pressure.

  14. Positional cloning of zinc finger domain transcription factor Zfp69, a candidate gene for obesity-associated diabetes contributed by mouse locus Nidd/SJL.

    Directory of Open Access Journals (Sweden)

    Stephan Scherneck

    2009-07-01

    Full Text Available Polygenic type 2 diabetes in mouse models is associated with obesity and results from a combination of adipogenic and diabetogenic alleles. Here we report the identification of a candidate gene for the diabetogenic effect of a QTL (Nidd/SJL, Nidd1 contributed by the SJL, NON, and NZB strains in outcross populations with New Zealand Obese (NZO mice. A critical interval of distal chromosome 4 (2.1 Mbp conferring the diabetic phenotype was identified by interval-specific congenic introgression of SJL into diabetes-resistant C57BL/6J, and subsequent reporter cross with NZO. Analysis of the 10 genes in the critical interval by sequencing, qRT-PCR, and RACE-PCR revealed a striking allelic variance of Zfp69 encoding zinc finger domain transcription factor 69. In NZO and C57BL/6J, a retrotransposon (IAPLTR1a in intron 3 disrupted the gene by formation of a truncated mRNA that lacked the coding sequence for the KRAB (Krüppel-associated box and Znf-C2H2 domains of Zfp69, whereas the diabetogenic SJL, NON, and NZB alleles generated a normal mRNA. When combined with the B6.V-Lep(ob background, the diabetogenic Zfp69(SJL allele produced hyperglycaemia, reduced gonadal fat, and increased plasma and liver triglycerides. mRNA levels of the human orthologue of Zfp69, ZNF642, were significantly increased in adipose tissue from patients with type 2 diabetes. We conclude that Zfp69 is the most likely candidate for the diabetogenic effect of Nidd/SJL, and that retrotransposon IAPLTR1a contributes substantially to the genetic heterogeneity of mouse strains. Expression of the transcription factor in adipose tissue may play a role in the pathogenesis of type 2 diabetes.

  15. A unique sequence in the N-terminal regulatory region controls the nuclear localization of KLF8 by cooperating with the C-terminal zinc-fingers

    Institute of Scientific and Technical Information of China (English)

    Tina S Mehta; Heng Lu; Xianhui Wang; Alison M Urvalek; Kim-Hang H Nguyen; Farah Monzur; Jojo D Hammond; Jameson Q Ma; Jihe Zhao

    2009-01-01

    Kruppel-like factor 8 (KLF8) transcription factor plays a critical role in cell cycle progression, oncogenic trans-formation, epithelial to mesenchymal transition and invasion. However, its nuclear localization signal(s) (NLS) has not been identified. KLF8 shares with other KLFs monopartite NLSs (mNLS) and C2H2 zinc fingers (ZFs), both of which have been shown to be the NLSs for some other KLFs. In this report, using PCR-directed mutagenesis and immunofluorescent microscopy, we show that disruption of the mNLSs, deletion of any single ZF, or mutation of the Zn2+-binding or DNA-contacting motifs did not affect the nuclear localization of KLF8. Deletion of>1.5 ZFs from C-terminus, however, caused cytoplasmic accumulation of KLF8. Surprisingly, deletion of amino acid (aa) 151-200 re-gion almost eliminated KLF8 from the nucleus. S165A, K171E or K171R mutation, or treatment with PKC inhibitor led to partial cytoplasmic accumulation. Co-immunoprecipitation demonstrated that KLF8 interacted with importin-β and this interaction required the ZF motif. Deletion of aa 1-150 or 201-261 region alone did not alter the nuclear lo-calization. BrdU incorporation and cyclin D1 promoter luciferase assays showed that the KLF8 mutants defective in nuclear localization could not promote DNA synthesis or cyclin D1 promoter activation as the wild-type KLF8 did. Taken together, these results suggest that KLF8 has two NLSs, one surrounding S165 and K171 and the other being two tandem ZFs, which are critical for the regulation of KLF8 nuclear localization and its cellular functions.

  16. Transcripts from a novel human KRAB zinc finger gene contain spliced Alu and endogenous retroviral segments

    Energy Technology Data Exchange (ETDEWEB)

    Baban, S.; Freeman, J.D.; Mager, D.L. [Univ. of British Columbia, Vancouver, British Columbia (Canada)

    1996-05-01

    During the course of an investigation into the potential effects of endogenous retroviruses on adjacent gene expression, we isolated two cDNA clones containing a small sequence segment belonging to the human endogenous retrovirus family, HERV-H. Characterization of the clones revealed that they represent transcripts from a novel KRAB zinc finger gene termed ZNF177. The two cDNA clones differ at their 5{prime} termini and in the presence of a 559-bp internal exon. The clone containing this internal exon has six imperfect zinc finger motifs followed by seven perfect copies of the C{sub 2}H{sub 2} type but has a frame shift between the KRAB domain and the downstream zinc finger region. The smaller clone lacks the six imperfect motifs and has an intact ORF. The 5{prime} putative untranslated regions of both cDNAs contain an 86-bp HERV-H env segment and a segment of an Alu repeat, both in the antisense orientation, that have been incorporated by splicing. RT-PCR experiments show evidence of alternative splicing but the majority of transcripts appear to contain the Alu and env segments. Genomic PCR and hybridization experiments suggest that a partial HERV-H element is integrated within the ZNF177 locus, which Southern analysis has shown to be a single-copy gene. Northern and RT-PCR analyses suggest that ZNF177 is transcribed at a low level in a variety of cell types. 41 refs., 8 figs.

  17. Interaction of Sp1 zinc finger with transport factor in the nuclear localization of transcription factor Sp1

    Energy Technology Data Exchange (ETDEWEB)

    Ito, Tatsuo [Department of Medicinal Biotechnology, Institute for Medicinal Research, Graduate School of Pharmaceutical Sciences, The University of Tokushima, 1-78 Sho-machi, Tokushima 770-8505 (Japan); Kitamura, Haruka; Uwatoko, Chisana; Azumano, Makiko [Department of Molecular Biophysical Chemistry, Faculty of Pharmaceutical Sciences, Doshisha Women' s University, Kodo, Kyotanabe City, Kyoto 610-0395 (Japan); Itoh, Kohji, E-mail: kitoh@ph.tokushima-u.ac.jp [Department of Medicinal Biotechnology, Institute for Medicinal Research, Graduate School of Pharmaceutical Sciences, The University of Tokushima, 1-78 Sho-machi, Tokushima 770-8505 (Japan); Kuwahara, Jun, E-mail: jkuwahar@dwc.doshisha.ac.jp [Department of Molecular Biophysical Chemistry, Faculty of Pharmaceutical Sciences, Doshisha Women' s University, Kodo, Kyotanabe City, Kyoto 610-0395 (Japan)

    2010-12-10

    Research highlights: {yields} Sp1 zinc fingers themselves interact with importin {alpha}. {yields} Sp1 zinc finger domains play an essential role as a nuclear localization signal. {yields} Sp1 can be transported into the nucleus in an importin-dependent manner. -- Abstract: Transcription factor Sp1 is localized in the nucleus and regulates the expression of many cellular genes, but the nuclear transport mechanism of Sp1 is not well understood. In this study, we revealed that GST-fused Sp1 protein bound to endogenous importin {alpha} in HeLa cells via the Sp1 zinc finger domains, which comprise the DNA binding domain of Sp1. It was found that the Sp1 zinc finger domains directly interacted with a wide range of importin {alpha} including the armadillo (arm) repeat domain and the C-terminal acidic domain. Furthermore, it turned out that all three zinc fingers of Sp1 are essential for binding to importin {alpha}. Taken together, these results suggest that the Sp1 zinc finger domains play an essential role as a NLS and Sp1 can be transported into the nucleus in an importin-dependent manner even though it possesses no classical NLSs.

  18. Myocardial ischemic preconditioning upregulated protein 1(Mipu1):zinc finger protein 667 - a multifunctional KRAB/C{sub 2}H{sub 2} zinc finger protein

    Energy Technology Data Exchange (ETDEWEB)

    Han, D.; Zhang, C. [Institute of Cardiovascular Disease, Key Lab for Arteriosclerology of Hunan Province, Post-doctoral Mobile Stations for Basic Medicine, University of South China, Hengyang City, Hunan Province (China); Fan, W.J. [Institute of Cardiovascular Disease, Key Lab for Arteriosclerology of Hunan Province, Post-doctoral Mobile Stations for Basic Medicine, University of South China, Hengyang City, Hunan Province (China); The Second Affiliated Hospital, University of South China, Hengyang City, Hunan Province (China); Pan, W.J.; Feng, D.M.; Qu, S.L.; Jiang, Z.S. [Institute of Cardiovascular Disease, Key Lab for Arteriosclerology of Hunan Province, Post-doctoral Mobile Stations for Basic Medicine, University of South China, Hengyang City, Hunan Province (China)

    2014-10-31

    Myocardial ischemic preconditioning upregulated protein 1 (Mipu1) is a newly discovered upregulated gene produced in rats during the myocardial ischemic preconditioning process. Mipu1 cDNA contains a 1824-base pair open reading frame and encodes a 608 amino acid protein with an N-terminal Krüppel-associated box (KRAB) domain and classical zinc finger C{sub 2}H{sub 2} motifs in the C-terminus. Mipu1 protein is located in the cell nucleus. Recent studies found that Mipu1 has a protective effect on the ischemia-reperfusion injury of heart, brain, and other organs. As a nuclear factor, Mipu1 may perform its protective function through directly transcribing and repressing the expression of proapoptotic genes to repress cell apoptosis. In addition, Mipu1 also plays an important role in regulating the gene expression of downstream inflammatory mediators by inhibiting the activation of activator protein-1 and serum response element.

  19. Um estudo teórico relativo à não-linearidade da ligação de hidrogênio em sistemas heterocíclicos C2H4O-C2H2 e C2H4S-C2H2 A theoretical study about the non-linearity of the hydrogen bonding in the C2H4O-C2H2 and C2H4S-C2H2 heterocyclic systems

    Directory of Open Access Journals (Sweden)

    Boaz G. Oliveira

    2007-10-01

    Full Text Available B3LYP/6-31G(d,p calculations were used to determine the optimized geometries of the C2H4O-C2H2 and C2H4S-C2H2 heterocyclic hydrogen-bonded complexes. Results of structural, rotational, electronic and vibrational parameters indicate that the hydrogen bonding is non-linear due to the pi bond of the acetylene interacting with the hydrogen atoms of the methyl groups of the three-membered rings. Moreover, the theoretical investigation showed that the non-linearity is much more intriguing, since there is a structural disjunction on the acetylene within the heterocyclic system.

  20. Secondary Organic Aerosol Formation from Acetylene (C2H2: seed effect on SOA yields due to organic photochemistry in the aerosol aqueous phase

    Directory of Open Access Journals (Sweden)

    P. J. Ziemann

    2009-03-01

    Full Text Available The lightest Non Methane HydroCarbon (NMHC, i.e., acetylene (C2H2 is found to form secondary organic aerosol (SOA. Contrary to current belief, the number of carbon atoms, n, for a NMHC to act as SOA precursor is lowered to n=2 here. The OH-radical initiated oxidation of C2H2 forms glyoxal (CHOCHO as the highest yield product, and >99% of the SOA from C2H2 is attributed to CHOCHO. SOA formation from C2H2 and CHOCHO was studied in a photochemical and a dark simulation chamber. Further, the experimental conditions were varied with respect to the chemical composition of the seed aerosols, mild acidification with sulphuric acid (SA, 3

  1. Refined ab initio intermolecular ground-state potential energy surface for the He-C2H2 van der Waals complex

    DEFF Research Database (Denmark)

    Fernández, Berta; Henriksen, Christian; Farrelly, David

    2013-01-01

    A refined CCSD(T) intermolecular potential energy surface is developed for the He-C2H2 van der Waals complex. For this, 206 points on the intermolecular potential energy surface, evaluated using the CCSD(T) method and the aug-cc-pVQZ basis set extended with a set of 3s3p2d1f1g midbond functions...

  2. Electron correlation and hole localization in photon-stimulated ion desorption from solid N2, CO, and C2H2

    International Nuclear Information System (INIS)

    Results of photon-stimulated ion desorption measurements from solid N2, CO, and C2H2 are presented in the photon energy range 172, charge localization on the nitrogen atom nearest the surface causes the N+ spectral yield to differ from the N+ yield from gaseous N2 and to resemble the O+ yield from CO. The effect of hole migration on desorption from physisorbed systems is discussed

  3. Silicon Carbide Nanoparticles Produced by CO2 Laser Pyrolysis of SiH4/C2H2 Gas Mixtures in a Flow Reactor

    International Nuclear Information System (INIS)

    Pulsed CO2-laser-induced decomposition of different mixtures of SiH4 and C2H2 in a flow reactor has been employed to produce silicon carbide clusters and nanoparticles with varying content of carbon. The as-synthesized species were extracted from the reaction zone by a conical nozzle and expanded into the source chamber of a cluster beam apparatus where, after having traversed a differential chamber, they were analyzed with a time-of-flight mass spectrometer. Thin films of silicon carbide nanoclusters were produced by depositing the clusters at low energy on potassium bromide and sapphire windows mounted into the differential chamber. At the same time, Si and SiC nanoparticles were collected in a filter placed into the exhaust line of the flow reactor. Both beam and powder samples were characterized by FTIR spectroscopy. The close resemblance of the spectra suggests that the composition of the beam and powder particles obtained during the same run is nearly identical. XRD spectroscopy could only be employed for the investigation of the powders. It was found that CO2 laser pyrolysis is ideally suited to produce silicon carbide nanoparticles with a high degree of crystallinity. Nanopowders produced from the pyrolysis of a stoichiometric (2:1) mixture of SiH4/C2H2 were found to contain particles or domains of pure silicon. The characteristic silicon features in the FTIR and XRD spectra, however, disappeared when C2H2 was applied in excess

  4. Characterization of two novel nuclear BTB/POZ domain zinc finger isoforms. Association with differentiation of hippocampal neurons, cerebellar granule cells, and macroglia

    DEFF Research Database (Denmark)

    Mitchelmore, Cathy; Kjaerulff, Karen M; Pedersen, Hans C;

    2002-01-01

    BTB/POZ (broad complex tramtrack bric-a-brac/poxvirus and zinc finger) zinc finger factors are a class of nuclear DNA-binding proteins involved in development, chromatin remodeling, and cancer. However, BTB/POZ domain zinc finger factors linked to development of the mammalian cerebral cortex......, cerebellum, and macroglia have not been described previously. We report here the isolation and characterization of two novel nuclear BTB/POZ domain zinc finger isoforms, designated HOF(L) and HOF(S), that are specifically expressed in early hippocampal neurons, cerebellar granule cells, and gliogenic...

  5. The Electronic Behavior of Zinc-Finger Protein Binding Sites in the Context of the DNA Extended Ladder Model

    Science.gov (United States)

    Oiwa, Nestor; Cordeiro, Claudette; Heermann, Dieter

    2016-05-01

    Instead of ATCG letter alignments, typically used in bioinformatics, we propose a new alignment method using the probability distribution function of the bottom of the occupied molecular orbital (BOMO), highest occupied molecular orbital (HOMO) and lowest unoccupied orbital (LUMO). We apply the technique to transcription factors with Cys2His2 zinc fingers. These transcription factors search for binding sites, probing for the electronic patterns at the minor and major DNA groves. The eukaryotic Cys2His2 zinc finger proteins bind to DNA ubiquitously at highly conserved domains. They are responsible for gene regulation and the spatial organization of DNA. To study and understand these zinc finger DNA-protein interactions, we use the extended ladder in the DNA model proposed by Zhu, Rasmussen, Balatsky & Bishop (2007) te{Zhu-2007}. Considering one single spinless electron in each nucleotide π-orbital along a double DNA chain (dDNA), we find a typical pattern for the bottom of BOMO, HOMO and LUMO along the binding sites. We specifically looked at two members of zinc finger protein family: specificity protein 1 (SP1) and early grown response 1 transcription factors (EGR1). When the valence band is filled, we find electrons in the purines along the nucleotide sequence, compatible with the electric charges of the binding amino acids in SP1 and EGR1 zinc finger.

  6. ZFNGenome: A comprehensive resource for locating zinc finger nuclease target sites in model organisms

    Directory of Open Access Journals (Sweden)

    Voytas Daniel F

    2011-01-01

    Full Text Available Abstract Background Zinc Finger Nucleases (ZFNs have tremendous potential as tools to facilitate genomic modifications, such as precise gene knockouts or gene replacements by homologous recombination. ZFNs can be used to advance both basic research and clinical applications, including gene therapy. Recently, the ability to engineer ZFNs that target any desired genomic DNA sequence with high fidelity has improved significantly with the introduction of rapid, robust, and publicly available techniques for ZFN design such as the Oligomerized Pool ENgineering (OPEN method. The motivation for this study is to make resources for genome modifications using OPEN-generated ZFNs more accessible to researchers by creating a user-friendly interface that identifies and provides quality scores for all potential ZFN target sites in the complete genomes of several model organisms. Description ZFNGenome is a GBrowse-based tool for identifying and visualizing potential target sites for OPEN-generated ZFNs. ZFNGenome currently includes a total of more than 11.6 million potential ZFN target sites, mapped within the fully sequenced genomes of seven model organisms; S. cerevisiae, C. reinhardtii, A. thaliana, D. melanogaster, D. rerio, C. elegans, and H. sapiens and can be visualized within the flexible GBrowse environment. Additional model organisms will be included in future updates. ZFNGenome provides information about each potential ZFN target site, including its chromosomal location and position relative to transcription initiation site(s. Users can query ZFNGenome using several different criteria (e.g., gene ID, transcript ID, target site sequence. Tracks in ZFNGenome also provide "uniqueness" and ZiFOpT (Zinc Finger OPEN Targeter "confidence" scores that estimate the likelihood that a chosen ZFN target site will function in vivo. ZFNGenome is dynamically linked to ZiFDB, allowing users access to all available information about zinc finger reagents, such as the

  7. Zinc finger protein TTP interacts with CCL3 mRNA and regulates tissue inflammation*

    OpenAIRE

    Kang, Ju-Gyeong; Amar, Marcelo J.; Remaley, Alan T.; Kwon, Jaeyul; Blackshear, Perry J.; Wang, Ping-yuan; Hwang, Paul M.

    2011-01-01

    Zinc finger protein tristetraprolin (TTP) modulates macrophage inflammatory activity by destabilizing cytokine mRNAs. Here, through a screen of TTP-bound mRNAs in activated human macrophages, we have identified CC chemokine ligand 3 (CCL3) mRNA as the most abundantly bound TTP target mRNA and have characterized this interaction via conserved AU-rich elements. Compared to the wild-type cells, TTP−/− macrophages produced higher levels of LPS-induced CCL3. In addition, the plasma level of CCL3 i...

  8. Evaluation of a modular strategy for the construction of novel polydactyl zinc finger DNA-binding proteins.

    Science.gov (United States)

    Segal, David J; Beerli, Roger R; Blancafort, Pilar; Dreier, Birgit; Effertz, Karin; Huber, Adrian; Koksch, Beate; Lund, Caren V; Magnenat, Laurent; Valente, David; Barbas, Carlos F

    2003-02-25

    In previous studies, we have developed a technology for the rapid construction of novel DNA-binding proteins with the potential to recognize any unique site in a given genome. This technology relies on the modular assembly of modified zinc finger DNA-binding domains, each of which recognizes a three bp subsite of DNA. A complete set of 64 domains would provide comprehensive recognition of any desired DNA sequence, and new proteins could be assembled by any laboratory in a matter of hours. However, a critical parameter for this approach is the extent to which each domain functions as an independent, modular unit, without influence or dependence on its neighboring domains. We therefore examined the detailed binding behavior of several modularly assembled polydactyl zinc finger proteins. We first demonstrated that 80 modularly assembled 3-finger proteins can recognize their DNA target with very high specificity using a multitarget ELISA-based specificity assay. A more detailed analysis of DNA binding specificity for eight 3-finger proteins and two 6-finger proteins was performed using a target site selection assay. Results showed that the specificity of these proteins was as good or better than that of zinc finger proteins constructed using methods that allow for interdependency. In some cases, near perfect specificity was achieved. Complications due to target site overlap were found to be restricted to only one particular amino acid interaction (involving an aspartate in position 2 of the alpha-helix) that occurs in a minority of cases. As this is the first report of target site selection for designed, well characterized 6-finger proteins, unique insights are discussed concerning the relationship of protein length and specificity. These results have important implications for the design of proteins that can recognize extended DNA sequences, as well as provide insights into the general rules of recognition for naturally occurring zinc finger proteins.

  9. Zinc finger protein 278, a potential oncogene in human colorectal cancer

    Institute of Scientific and Technical Information of China (English)

    Xiaoqing Tian; Danfeng Sun; Yanjie Zhang; Shuliang Zhao; Hua Xiong; Jingyuan Fang

    2008-01-01

    Zinc finger protein 278 (ZNF278) is a novel Krueppel Cys2-His2-type zinc finger protein that is ubiquitously distributed in human tissues. Whether ZNF278 is related to the development of colorectal cancer is still unclear. The transcriptional level of ZNF278 was studied in colorectal cancer by real-time polymerase chain reaction. The results showed that ZNF278 expression was increased in 53% of colorectal cancer tissues compared to corresponding non-cancerous tissues. The transcriptional down-regulation of ZNF278 was detected in only three (6%) human colorectal cancer tissues compared to corresponding non-cancer tissues. No significant difference was detected in 19 (41%) pairs of samples.However, we failed to find a significant association between the up-regulation of ZNF278 transcription and age, sex, the degree of infiltration, or the tumor size of colorectal cancer.To study the function of ZNF278 in colorectal carcinogenesis,the colon cancer cell line SW1116 was stably transfected with a wild-type ZNF278 plasmid to construct an overexpression system, and was transiently transfected with the small interfering RNA of ZNF278 to construct a ZNF278 knockdown system. Cell proliferation was assessed with 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide dye and a cell counter. The results show that ZNF278 promotes cell growth, and its knockdown suppresses cell proliferation. ZNF278 could be a potential proto-oncogene in colorectal cancer.

  10. Zinc Finger Nuclease induced DNA double stranded breaks and rearrangements in MLL

    Energy Technology Data Exchange (ETDEWEB)

    Do, To Uyen [Graduate Group in Immunology, University of California Davis, Davis, CA 95616 (United States); Department of Radiation Oncology, University of California Davis, Sacramento CA 95817 (United States); Ho, Bay; Shih, Shyh-Jen [Department of Radiation Oncology, University of California Davis, Sacramento CA 95817 (United States); Vaughan, Andrew, E-mail: Andrew.vaughan@ucdmc.ucdavis.edu [Graduate Group in Immunology, University of California Davis, Davis, CA 95616 (United States); Department of Radiation Oncology, University of California Davis, Sacramento CA 95817 (United States)

    2012-12-15

    Highlights: ► A Zinc Finger Nuclease (ZFN) targeting a leukemogenic hot spot for rearrangement in MLL is created. ► The novel ZFN efficiently cleaves MLL exon 13. ► Despite MLL cleavage and evidence of mis-repair, no leukemogenic translocations were produced. ► MLL cleavage alone is insufficient to generate leukemogenic translocations. - Abstract: Radiation treatment or chemotherapy has been linked with a higher risk of secondary cancers such as therapy related Acute Myeloid Leukemia (tAML). Several of these cancers have been shown to be correlated to the introduction of double stranded breaks (DSB) and rearrangements within the Mixed Lineage Leukemia (MLL) gene. We used Zinc Finger Nucleases (ZFNs) to introduce precise cuts within MLL to examine how a single DNA DSB might lead to chromosomal rearrangements. A ZFN targeting exon 13 within the Breakpoint Cluster Region of MLL was transiently expressed in a human lymphoblast cell line originating from a CML patient. Although FISH analysis showed ZFN DSB at this region increased the rate of MLL fragmentation, we were unable to detect leukemogenic rearrangements or translocations via inverse PCR. Interestingly, gene fragmentation as well as small interstitial deletions, insertions and base substitutions increased with the inhibition of DNA-PK, suggesting repair of this particular DSB is linked to non-homologous end joining (NHEJ). Although mis-repair of DSBs may be necessary for the initiation of leukemogenic translocations, a MLL targeted DNA break alone is insufficient.

  11. Preparation of Carbon Nanotubes with High Yield and Narrow Diameter Distribution from C2H2 over LaCu0.2Ni0.8Ox

    Institute of Scientific and Technical Information of China (English)

    Chang Xing TANG; Mei Zhen QU; Bo Lan ZHANG; Zuo Long YU

    2004-01-01

    Carbon nanotubes (CNTs) were prepared by decomposition of C2H2 over newly developed LaCu0.2Ni0.8Ox in the temperature range from 600 to 850℃. The effect of the reaction temperature on the yield of CNTs was investigated in detail. At 680℃,the yield of CNTs reaches 17 g/g.catal. or so. The morphology of CNTs was examined by TEM. The diameter of CNTs rangs from 9 nm to 14 nm.

  12. New line intensity measurements for 12C2H2 around 7.7 µm and HITRAN format line list for applications

    OpenAIRE

    Gomez, Laura; Jacquemart, David; Lacome, Nelly; Mandin, Jean-Yves

    2010-01-01

    International audience Absolute intensities of 467 lines are measured in 9 bands of the 7.7 µm spectral region of the 12C2H2 molecule, with an average accuracy of 5%. For each band, the vibrational transition dipole moment squared and Herman-Wallis coefficients are obtained in order to model the rotational dependence of the transition dipole moment squared. These results are used to calculate a line list for atmospheric or astrophysical applications. Merged in the line list set up in a pre...

  13. Classification of the treble clef zinc finger: noteworthy lessons for structure and function evolution.

    Science.gov (United States)

    Kaur, Gurmeet; Subramanian, Srikrishna

    2016-01-01

    Treble clef (TC) zinc fingers constitute a large fold-group of structural zinc-binding protein domains that mediate numerous cellular functions. We have analysed the sequence, structure, and function relationships among all TCs in the Protein Data Bank. This led to the identification of novel TCs, such as lsr2, YggX and TFIIIC τ 60 kDa subunit, and prediction of a nuclease-like function for the DUF1364 family. The structural malleability of TCs is evident from the many examples with variations to the core structural elements of the fold. We observe domains wherein the structural core of the TC fold is circularly permuted, and also some examples where the overall fold resembles both the TC motif and another unrelated fold. All extant TC families do not share a monophyletic origin, as several TC proteins are known to have been present in the last universal common ancestor and the last eukaryotic common ancestor. We identify several TCs where the zinc-chelating site and residues are not merely responsible for structure stabilization but also perform other functions, such as being redox active in C1B domain of protein kinase C, a nucleophilic acceptor in Ada and catalytic in organomercurial lyase, MerB. PMID:27562564

  14. Classification of the treble clef zinc finger: noteworthy lessons for structure and function evolution

    Science.gov (United States)

    Kaur, Gurmeet; Subramanian, Srikrishna

    2016-01-01

    Treble clef (TC) zinc fingers constitute a large fold-group of structural zinc-binding protein domains that mediate numerous cellular functions. We have analysed the sequence, structure, and function relationships among all TCs in the Protein Data Bank. This led to the identification of novel TCs, such as lsr2, YggX and TFIIIC τ 60 kDa subunit, and prediction of a nuclease-like function for the DUF1364 family. The structural malleability of TCs is evident from the many examples with variations to the core structural elements of the fold. We observe domains wherein the structural core of the TC fold is circularly permuted, and also some examples where the overall fold resembles both the TC motif and another unrelated fold. All extant TC families do not share a monophyletic origin, as several TC proteins are known to have been present in the last universal common ancestor and the last eukaryotic common ancestor. We identify several TCs where the zinc-chelating site and residues are not merely responsible for structure stabilization but also perform other functions, such as being redox active in C1B domain of protein kinase C, a nucleophilic acceptor in Ada and catalytic in organomercurial lyase, MerB. PMID:27562564

  15. C2H2流量对SiCN 薄膜结构及阻挡性能的影响%Influence of C2H2 Flow Rate on Microstructures and Electronic Properties of SiCN Diffusion Barriers

    Institute of Scientific and Technical Information of China (English)

    张治超; 周继承; 彭银桥

    2011-01-01

    The SiCN films were grown by RF reactive magnetrmn sputtering with acetylene and nitrogen as the reactive gases,and polycrystalline silicon as the target material. The impacts of the film growth conditions,including the C2H2 flow rate, annealing temperature on the properties of the SiCN film were studied. Its microstructures and electronic properties were characterized with Fourier transform infrared spectroscopy, X-ray diffraction, and conventional surface probes.The results show that the C2H2 flow rate and annealing temperature strongly affect its microstructures, dielectric constant and diffusion barrier efficiency. For instance,as C2H2 flow rate increases,the grain size on surfaces of the highly stable,amorphous SiCN film increases. The C-N bonds dominate the surface matrix possibly because of substitution of Si for C atom. Its dielectric constant was estimated to be 4.2 ~ 5.8, depending on contents C and N, and on defect-density of the films. We found that high temperature inter-diffusion and reaction of Cu and Si are responsible for the breakdown of the SiCN diffusion barrier%采用C2H2和N2作为反应气体、多晶Si作为靶材,利用射频磁控溅射系统沉积了SiCN薄膜.利用傅里叶红外光谱仪、X射线衍射仪、四探针测试仪等研究了C2H2流量对薄膜结构、介电常数以及阻挡性能的影响.结果表明,薄膜为非晶结构,1000℃退火下未出现结晶,稳定性很好;随着C2H2流量的增大,薄膜表面颗粒呈现增大趋势;C原子取代Si原子占据薄膜中的网络位置,薄膜形成了以C-N键为主的网络结构;制得的SiCN薄膜介电常数在4.2~5.8之间,C,N含量以及薄膜结构是影响介电性能的关键因素,高温使得Cu穿过薄膜中的缺陷与Si发生互扩散是薄膜阻挡性能失效的主要原因.

  16. Modular system for the construction of zinc-finger libraries and proteins.

    Science.gov (United States)

    Gonzalez, Beatriz; Schwimmer, Lauren J; Fuller, Roberta P; Ye, Yongjun; Asawapornmongkol, Lily; Barbas, Carlos F

    2010-04-01

    Engineered zinc-finger transcription factors (ZF-TF) are powerful tools to modulate the expression of specific genes. Complex libraries of ZF-TF can be delivered into cells to scan the genome for genes responsible for a particular phenotype or to select the most effective ZF-TF to regulate an individual gene. In both cases, the construction of highly representative and unbiased libraries is critical. In this protocol, we describe a user-friendly ZF technology suitable for the creation of complex libraries and the construction of customized ZF-TFs. The new technology described here simplifies the building of ZF libraries, avoids PCR-introduced bias and ensures equal representation of every module. We also describe the construction of a customized ZF-TF that can be transferred to a number of expression vectors. This protocol can be completed in 9-11 d.

  17. Site-specific genome editing in Plasmodium falciparum using engineered zinc-finger nucleases.

    Science.gov (United States)

    Straimer, Judith; Lee, Marcus C S; Lee, Andrew H; Zeitler, Bryan; Williams, April E; Pearl, Jocelynn R; Zhang, Lei; Rebar, Edward J; Gregory, Philip D; Llinás, Manuel; Urnov, Fyodor D; Fidock, David A

    2012-10-01

    Malaria afflicts over 200 million people worldwide, and its most lethal etiologic agent, Plasmodium falciparum, is evolving to resist even the latest-generation therapeutics. Efficient tools for genome-directed investigations of P. falciparum-induced pathogenesis, including drug-resistance mechanisms, are clearly required. Here we report rapid and targeted genetic engineering of this parasite using zinc-finger nucleases (ZFNs) that produce a double-strand break in a user-defined locus and trigger homology-directed repair. Targeting an integrated egfp locus, we obtained gene-deletion parasites with unprecedented speed (2 weeks), both with and without direct selection. ZFNs engineered against the parasite gene pfcrt, responsible for escape under chloroquine treatment, rapidly produced parasites that carried either an allelic replacement or a panel of specified point mutations. This method will enable a diverse array of genome-editing approaches to interrogate this human pathogen.

  18. Knockout of Myostatin by Zinc-finger Nuclease in Sheep Fibroblasts and Embryos.

    Science.gov (United States)

    Zhang, Xuemei; Wang, Liqin; Wu, Yangsheng; Li, Wenrong; An, Jing; Zhang, Fuchun; Liu, Mingjun

    2016-10-01

    Myostatin (MSTN) can negatively regulate the growth and development of skeletal muscle, and natural mutations can cause "double-muscling" trait in animals. In order to block the inhibiting effect of MSTN on muscle growth, we transferred zinc-finger nucleases (ZFN) which targeted sheep MSTN gene into cultured fibroblasts. Gene targeted colonies were isolated from transfected fibroblasts by serial dilution culture and screened by sequencing. Two colonies were identified with mono-allele mutation and one colony with bi-allelic deletion. Further, we introduced the MSTN-ZFN mRNA into sheep embryos by microinjection. Thirteen of thirty-seven parthenogenetic embryos were targeted by ZFN, with the efficiency of 35%. Our work established the technical foundation for generation of MSTN gene editing sheep by somatic cloning and microinjection ZFN into embryos. PMID:27189642

  19. Functional promoter variant in zinc finger protein 202 predicts severe atherosclerosis and ischemic heart disease

    DEFF Research Database (Denmark)

    Frikke-Schmidt, R.; Nordestgaard, Børge; Grande, Peer;

    2008-01-01

    Objectives This study was designed to test the hypotheses that single nucleotide polymorphisms ( SNPs), in zinc finger protein 202 ( ZNF202), predict severe atherosclerosis and ischemic heart disease ( IHD). Background ZNF202 is a transcriptional repressor controlling promoter elements in genes...... involved in vascular maintenance and lipid metabolism. Methods We first determined genotype association for 9 ZNF202 SNPs with severe atherosclerosis ( ankle brachial index >0.7 vs. ...,998 controls. Finally, we determined whether g. -660A>G altered transcriptional activity of the ZNF202 promoter in vitro. Results Cross-sectionally, ZNF202 g. -660 GG versus AA homozygosity predicted an odds ratio for severe atherosclerosis of 2.01 ( 95% confidence interval [CI]: 1.34 to 3.01). Prospectively...

  20. The functional significance of common polymorphisms in zinc finger transcription factors.

    Science.gov (United States)

    Lockwood, Sarah H; Guan, Anna; Yu, Abigail S; Zhang, Chi; Zykovich, Artem; Korf, Ian; Rannala, Bruce; Segal, David J

    2014-09-01

    Variants that alter the DNA-binding specificity of transcription factors could affect the specificity for and expression of potentially many target genes, as has been observed in several tumor-derived mutations. Here we examined if such trans expression quantitative trait loci (trans-eQTLs) could similarly result from common genetic variants. We chose to focus on the Cys2-His2 class of zinc finger transcription factors because they are the most abundant superfamily of transcription factors in human and have well-characterized DNA binding interactions. We identified 430 SNPs that cause missense substitutions in the DNA-contacting residues. Fewer common missense SNPs were found at DNA-contacting residues compared with non-DNA-contacting residues (P = 0.00006), consistent with possible functional selection against SNPs at DNA-contacting positions. Functional predictions based on zinc finger transcription factor (ZNF) DNA binding preferences also suggested that many common substitutions could potentially alter binding specificity. However, Hardy-Weinberg Equilibrium analysis and examination of seven orthologs within the primate lineage failed to find evidence of trans-eQTLs associated with the DNA-contacting positions or evidence of a different selection pressure on a contemporary and evolutionary timescales. The overall conclusion was that common SNPs that alter the DNA-contacting residues of these factors are unlikely to produce strong trans-eQTLs, consistent with the observations by others that trans-eQTLs in humans tend to be few and weak. Some rare SNPs might alter specificity and remained rare due to purifying selection. The study also underscores the need for large-scale eQTLs mapping efforts that might provide experimental evidence for SNPs that alter the choice of transcription factor binding sites. PMID:24970883

  1. Automatic polymerase chain reaction product detection system for food safety monitoring using zinc finger protein fused to luciferase

    Energy Technology Data Exchange (ETDEWEB)

    Yoshida, Wataru; Kezuka, Aki; Murakami, Yoshiyuki; Lee, Jinhee; Abe, Koichi [Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 (Japan); Motoki, Hiroaki; Matsuo, Takafumi; Shimura, Nobuaki [System Instruments Co., Ltd., 776-2 Komiya-cho, Hachioji, Tokyo 192-0031 (Japan); Noda, Mamoru; Igimi, Shizunobu [Division of Biomedical Food Research, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501 (Japan); Ikebukuro, Kazunori, E-mail: ikebu@cc.tuat.ac.jp [Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 (Japan)

    2013-11-01

    Graphical abstract: -- Highlights: •Zif268 fused to luciferase was used for E. coli O157, Salmonella and coliform detection. •Artificial zinc finger protein fused to luciferase was constructed for Norovirus detection. •An analyzer that automatically detects PCR products by zinc finger protein fused to luciferase was developed. •Target pathogens were specifically detected by the automatic analyzer with zinc finger protein fused to luciferase. -- Abstract: An automatic polymerase chain reaction (PCR) product detection system for food safety monitoring using zinc finger (ZF) protein fused to luciferase was developed. ZF protein fused to luciferase specifically binds to target double stranded DNA sequence and has luciferase enzymatic activity. Therefore, PCR products that comprise ZF protein recognition sequence can be detected by measuring the luciferase activity of the fusion protein. We previously reported that PCR products from Legionella pneumophila and Escherichia coli (E. coli) O157 genomic DNA were detected by Zif268, a natural ZF protein, fused to luciferase. In this study, Zif268–luciferase was applied to detect the presence of Salmonella and coliforms. Moreover, an artificial zinc finger protein (B2) fused to luciferase was constructed for a Norovirus detection system. In the luciferase activity detection assay, several bound/free separation process is required. Therefore, an analyzer that automatically performed the bound/free separation process was developed to detect PCR products using the ZF–luciferase fusion protein. By means of the automatic analyzer with ZF–luciferase fusion protein, target pathogenic genomes were specifically detected in the presence of other pathogenic genomes. Moreover, we succeeded in the detection of 10 copies of E. coli BL21 without extraction of genomic DNA by the automatic analyzer and E. coli was detected with a logarithmic dependency in the range of 1.0 × 10 to 1.0 × 10{sup 6} copies.

  2. Automatic polymerase chain reaction product detection system for food safety monitoring using zinc finger protein fused to luciferase

    International Nuclear Information System (INIS)

    Graphical abstract: -- Highlights: •Zif268 fused to luciferase was used for E. coli O157, Salmonella and coliform detection. •Artificial zinc finger protein fused to luciferase was constructed for Norovirus detection. •An analyzer that automatically detects PCR products by zinc finger protein fused to luciferase was developed. •Target pathogens were specifically detected by the automatic analyzer with zinc finger protein fused to luciferase. -- Abstract: An automatic polymerase chain reaction (PCR) product detection system for food safety monitoring using zinc finger (ZF) protein fused to luciferase was developed. ZF protein fused to luciferase specifically binds to target double stranded DNA sequence and has luciferase enzymatic activity. Therefore, PCR products that comprise ZF protein recognition sequence can be detected by measuring the luciferase activity of the fusion protein. We previously reported that PCR products from Legionella pneumophila and Escherichia coli (E. coli) O157 genomic DNA were detected by Zif268, a natural ZF protein, fused to luciferase. In this study, Zif268–luciferase was applied to detect the presence of Salmonella and coliforms. Moreover, an artificial zinc finger protein (B2) fused to luciferase was constructed for a Norovirus detection system. In the luciferase activity detection assay, several bound/free separation process is required. Therefore, an analyzer that automatically performed the bound/free separation process was developed to detect PCR products using the ZF–luciferase fusion protein. By means of the automatic analyzer with ZF–luciferase fusion protein, target pathogenic genomes were specifically detected in the presence of other pathogenic genomes. Moreover, we succeeded in the detection of 10 copies of E. coli BL21 without extraction of genomic DNA by the automatic analyzer and E. coli was detected with a logarithmic dependency in the range of 1.0 × 10 to 1.0 × 106 copies

  3. Measurement of drift velocity and amplification coefficient in C2H2F4-Isobutane mixtures for avalanche operated Resistive Plate Counters

    CERN Document Server

    Colucci, A; Grancagnolo, F; Primavera, M

    1998-01-01

    The knowledge of the transport and amplification parameters (drift velocity, first Townsend and attachment coefficients) of the gas mixtures used for avalanche operated Resistive Plate Counters gives the possibility ofbetter understanding the underlying physical processes of these detectors and of developping Montecarlo simulations of their behaviour.We present here a measurement of the drift velocity $v_{d}$and of the amplification coefficient $\\eta$ in $C_{2}H_{2}F_{4}$-Isobutanemixtures. The ionization in the gas, contained in a 6$\\times$22~cm$^2$, 2~mm gap RPC, is obtained by means of a pulsed $N_{2}$ laser,via double photon ionization. Results are presented for $v_{d}$ and $\\eta$ as a function of the applied electric field.

  4. Analysis of the High-Resolution Fourier Spectrum of the ν6 Band of the cis-C2h2d2 Molecule

    Science.gov (United States)

    Konov, I. A.; Chertavskikh, Yu. V.; Fomchenko, A. L.; Aslapovskaya, Yu. S.; Zhdanovich, S. A.; Sydow, C.

    2016-03-01

    The spectrum of the ν6 band of the cis-ethylene-d2 molecule (cis-C2H2D2) is recorded with a Bruker IFS 125 HR Fourier spectrometer in the range 580-1210 cm-1 with resolution of 0.0021 cm-1. An analysis of the experimental spectrum allows more than 1500 transitions belonging to this band to be assigned that by more than 2.5 times greater than it has been known in the literature so far. The obtained experimental data are then used to determine the model parameters of the molecule (the effective Hamiltonian in the A-reduction and I'- representation). Strong resonance interaction with the band ν4 forbidden in absorption by the symmetry of a molecule is taken into account. 10 parameters of the Hamiltonian obtained by solving inverse spectroscopic problem reproduce 427 initial experimental energies (more than 1500 transitions) with accuracy close to the experimental uncertainty.

  5. Feedback Neural Network Power Transformer Fault Diagnosis Based on Classification of Fault Feature Gases and C2H2/C2H4 Ratio%基于故障特征气体与C2H2/C2H4比值分类的反馈型神经网络电力变压器故障诊断

    Institute of Scientific and Technical Information of China (English)

    黄祥柠; 马腾; 苏骏

    2014-01-01

    分析了五种特征气体及C2H2/C2H4的比值与变压器故障类型之间的关系,将变压器故障进行分类.依据C2H2/C2H4的比值将故障分为两类,第一类包含低温过热、中温过热、高温过热和局部放电,第二类包含低能放电、高能放电两类,并将C2H2/C2H4的比值过高的诊断为高能放电.再依据氢气对五种特征气体的百分比,从第一类中区分出局部放电.最后用反馈型神经网络中的Elman网络确定具体故障类型.

  6. Embryonic neural inducing factor churchill is not a DNA-binding zinc finger protein: solution structure reveals a solvent-exposed beta-sheet and zinc binuclear cluster.

    Science.gov (United States)

    Lee, Brian M; Buck-Koehntop, Bethany A; Martinez-Yamout, Maria A; Dyson, H Jane; Wright, Peter E

    2007-08-31

    Churchill is a zinc-containing protein that is involved in neural induction during embryogenesis. At the time of its discovery, it was thought on the basis of sequence alignment to contain two zinc fingers of the C4 type. Further, binding of an N-terminal GST-Churchill fusion protein to a particular DNA sequence was demonstrated by immunoprecipitation selection assay, suggesting that Churchill may function as a transcriptional regulator by sequence-specific DNA binding. We show by NMR solution structure determination that, far from containing canonical C4 zinc fingers, the protein contains three bound zinc ions in novel coordination sites, including an unusual binuclear zinc cluster. The secondary structure of Churchill is also unusual, consisting of a highly solvent-exposed single-layer beta-sheet. Hydrogen-deuterium exchange and backbone relaxation measurements reveal that Churchill is unusually dynamic on a number of time scales, with the exception of regions surrounding the zinc coordinating sites, which serve to stabilize the otherwise unstructured N terminus and the single-layer beta-sheet. No binding of Churchill to the previously identified DNA sequence could be detected, and extensive searches using DNA sequence selection techniques could find no other DNA sequence that was bound by Churchill. Since the N-terminal amino acids of Churchill form part of the zinc-binding motif, the addition of a fusion protein at the N terminus causes loss of zinc and unfolding of Churchill. This observation most likely explains the published DNA-binding results, which would arise due to non-specific interaction of the unfolded protein in the immunoprecipitation selection assay. Since Churchill does not appear to bind DNA, we suggest that it may function in embryogenesis as a protein-interaction factor.

  7. Deducing the Energetic Cost of Protein Folding in Zinc Finger Proteins Using Designed Metallopeptides

    International Nuclear Information System (INIS)

    Zinc finger transcription factors represent the largest single class of metalloproteins in the human genome. Binding of Zn(II) to their canonical Cys4, Cys3His1, or Cys2His2 sites results in metal-induced protein folding events required to achieve their proper structure for biological activity. The thermodynamic contribution of Zn(II) in each of these coordination spheres toward protein folding is poorly understood because of the coupled nature of the metal-ligand and protein-protein interactions. Using an unstructured peptide scaffold, GGG, we have employed fluorimetry, potentiometry, and calorimetry to determine the thermodynamics of Zn(II) binding to the Cys4, Cys3His1, and Cys2His2 ligand sets with minimal interference from protein folding effects. The data show that Zn(II) complexation is entropy driven and modulated by proton release. The formation constants for Zn(II)-GGG with a Cys4, Cys3His1, or Cys2His2 site are 5.6 x 1016, 1.5 x 1015, or 2.5 x 1013 M-1, respectively. Thus, the Zn(II)-Cys4, Zn(II)-Cys3His1, and Zn(II)-Cys2His2 interactions can provide up to 22.8, 20.7, and 18.3 kcal/mol, respectively, in driving force for protein stabilization, folding, and/or assembly at pH values above the ligand pKa values. While the contributions from the three coordination motifs differ by 4.5 kcal/mol in Zn(II) affinity at pH 9.0, they are equivalent at physiological pH, ?G = -16.8 kcal/mol or a Ka = 2.0 x 1012 M-1. Calorimetric data show that this is due to proton-based enthalpy-entropy compensation between the favorable entropic term from proton release and the unfavorable enthalpic term due to thiol deprotonation. Since protein folding effects have been minimized in the GGG scaffold, these peptides possess nearly the tightest Zn(II) affinities possible for their coordination motifs. The Zn(II) affinities in each coordination motif are compared between the GGG scaffold and natural zinc finger proteins to determine the free energy required to fold the latter

  8. A poxvirus protein with a RING finger motif binds zinc and localizes in virus factories.

    Science.gov (United States)

    Upton, C; Schiff, L; Rice, S A; Dowdeswell, T; Yang, X; McFadden, G

    1994-07-01

    Shope fibroma virus (SFV) is a Leporipoxvirus closely related to the highly virulent myxoma virus. The DNA sequence of the BamHI N fragment of the SFV DNA genome was determined, and the single complete open reading frame (N1R) was characterized. The protein encoded by the N1R gene was found to contain a C3HC4 RING finger motif at the C terminus. This C3HC4 motif is the hallmark of a growing family of proteins, many of which are involved in regulation of gene expression, DNA repair, or DNA recombination. Complete homologs of the SFV N1R gene were also detected in variola virus, myxoma virus, and vaccinia virus strain IHD-W. In contrast, the gene is completely absent from vaccinia virus strain Copenhagen, and in vaccinia virus strain WR, the open reading frame is truncated prior to the zinc binding domain because of an 11-bp deletion, thus producing a frameshift and premature stop codon. Recombinant N1R protein from SFV was expressed in Escherichia coli and shown to bind zinc in a specific manner. Using fluorescence microscopy to visualize a peptide epitope tag (derived from ICP27 of herpes simplex virus) fused to the N terminus of the poxvirus proteins, we observed that the N1R protein of SFV and its homologs in myxoma virus and vaccinia virus IHD-W were localized primarily to the virus factories in the cytoplasm of infected cells and, to a lesser degree, the host cell nucleus. The truncated protein of vaccinia virus strain WR failed to localize in this manner but instead was observed throughout the cytoplasm.

  9. Total Cross Sections for Electron Scattering from the Isoelectronic (Z = 14) Molecules (C2H2, CO, HCN and N2) at 100-5000 eV

    Institute of Scientific and Technical Information of China (English)

    SHI De-Heng; ZHU Zun-Lue; SUN Jin-Feng; YANG Xiang-Dong; LIU Yu-Fang; ZHAO Ye

    2004-01-01

    @@ A complex optical model potential correlated by the conception of bonded atom, which considers the overlapping effect of electron clouds between the two atoms in a molecule, is firstly employed to calculate the total cross sections for electron scattering from the isoelectronic (Z = 14) molecules (C2H2, CO, HCN, and N2) at 100-5000eV using the additivity rule at the Hartree-Fock level The difference between the bonded atom and the free one is that the overlapping effect of electron clouds of bonded atoms in molecules is considered. The quantitative molecular total cross section results are compared with the experimental data and with the other calculations wherever available and good agreement is obtained above 100eV. It is shown that the additivity rule along with the complex optical model potential considering the overlapping effect of electron clouds can give the results better than that uncorrelated by it. The correlating calculations are much closer to the experiments than the spherical-complex-optical-potential results in the lower energy region [Phys. Rev. A 45 (1992) 202]. Therefore,considering the overlapping effect of electron clouds in the complex optical potential could be helpful for the better accuracy of the total cross section calculations of electron scattering from molecules.

  10. Characterization of mid-infrared emissions from C2H2, CO, CO2, and HCN-filled hollow fiber lasers

    Science.gov (United States)

    Jones, A. M.; Fourcade-Dutin, C.; Mao, C.; Baumgart, B.; Nampoothiri, A. V. V.; Campbell, N.; Wang, Y.; Benabid, F.; Rudolph, W.; Washburn, B. R.; Corwin, K. L.

    2012-02-01

    We have now demonstrated and characterized gas-filled hollow-core fiber lasers based on population inversion from acetylene (12C2H2) and HCN gas contained within the core of a kagome-structured hollow-core photonic crystal fiber. The gases are optically pumped via first order rotational-vibrational overtones near 1.5 μm using 1-ns pulses from an optical parametric amplifier. Transitions from the pumped overtone modes to fundamental C-H stretching modes in both molecules create narrow-band laser emissions near 3 μm. High gain resulting from tight confinement of the pump and laser light together with the active gas permits us to operate these lasers in a single pass configuration, without the use of any external resonator structure. A delay between the emitted laser pulse and the incident pump pulse has been observed and is shown to vary with pump pulse energy and gas pressure. Furthermore, we have demonstrated lasing beyond 4 μm from CO and CO2 using silver-coated glass capillaries, since fused silica based fibers do not transmit in this spectral region and chalcogenide fibers are not yet readily available. Studies of the laser pulse energy as functions of the pump pulse energy and gas pressure were performed. Efficiencies reaching ~ 20% are observed for both acetylene and CO2.

  11. 10 kHz accuracy of an optical frequency reference based on (12)C2H2-filled large-core kagome photonic crystal fibers.

    Science.gov (United States)

    Knabe, Kevin; Wu, Shun; Lim, Jinkang; Tillman, Karl A; Light, Philip S; Couny, Francois; Wheeler, Natalie; Thapa, Rajesh; Jones, Andrew M; Nicholson, Jeffrey W; Washburn, Brian R; Benabid, Fetah; Corwin, Kristan L

    2009-08-31

    Saturated absorption spectroscopy reveals the narrowest features so far in molecular gas-filled hollow-core photonic crystal fiber. The 48-68 mum core diameter of the kagome-structured fiber used here allows for 8 MHz full-width half-maximum sub-Doppler features, and its wavelength-insensitive transmission is suitable for high-accuracy frequency measurements. A fiber laser is locked to the (12)C2H2 nu(1); + nu(3) P(13) transition inside kagome fiber, and compared with frequency combs based on both a carbon nanotube fiber laser and a Cr:forsterite laser, each of which are referenced to a GPS-disciplined Rb oscillator. The absolute frequency of the measured line center agrees with those measured in power build-up cavities to within 9.3 kHz (1 sigma error), and the fractional frequency instability is less than 1.2 x 10(-11) at 1 s averaging time.

  12. Absolute Intensities Measurements in the nu(4) + nu(5) Band of (12)C(2)H(2): Analysis of Herman-Wallis Effects and Forbidden Transitions.

    Science.gov (United States)

    Vander Auwera J

    2000-05-01

    We measured absolute line intensities in two bands of (12)C(2)H(2) near 7.5 µm, namely the nu(4) + nu(5)(Sigma(+)(u))-0(Sigma(+)(g)) and nu(4) + nu(5)(Delta(u))-0(Sigma(+)(g)) bands, using Fourier transform spectroscopy with an accuracy estimated to be better than 2%. Using theoretical predictions from Watson [J. K. G. Watson, J. Mol. Spectrosc. 188, 78 (1998)], the observation of the forbidden nu(4) + nu(5)(Delta(u))-0(Sigma(+)(g)) band and the Herman-Wallis behavior exhibited by its rotational lines were studied quantitatively in terms of two types of interactions affecting the levels involved by the band: l-type resonance and Coriolis interaction. In the case of the nu(4) + nu(5)(Sigma(+)(u))-0(Sigma(+)(g)) band, the influence of l-type resonance is also confirmed. We also attributed the intensity asymmetry observed between the R and P branches of that latter band to a Coriolis interaction with l = 1 levels. We did not observe the nu(4) + nu(5)(Sigma(-)(u))-0(Sigma(+)(g)) band, consisting only of a Q branch, in agreement with Watson's prediction. Copyright 2000 Academic Press.

  13. Zinc finger nucleases and their application%锌指核酶技术的原理及应用

    Institute of Scientific and Technical Information of China (English)

    邓珊珊; 王颖芝; 马端

    2010-01-01

    锌指核酶(zinc finger nucleases,ZFNs)是将锌指蛋白的DNA识别域和非特异性核酸内切酶FokⅠ人工连接而构成的一种核酶.1对ZFNs能在DNA上产生双链断裂(double-strand breaks,DSB),诱导细胞发生同源重组(homology recombination,HR)或非同源末端连接(nonhomologous end joining,NHEJ).最近锌指核酶技术在基因功能的研究中受到重视.作者就ZFNs的作用原理、关键技术及其应用领域进行介绍.%Zinc finger nuclease (ZFN), which is a chimeric fusion structure between a Cys2-His2 zinc-finger protein (ZFP) and the cleavage domain of Fok Ⅰ endonuclease,can be used to introduce targeted double-stranded breaks (DSBs). ZFN-mediated cleavage leads to mutations when double-stranded breaks are repaired by homologous recombination (HR) or nonhomologous end joining (NHEJ). In recent years, ZFNs are widely used in the fields of genetic research. In this review, the methodology and technical advantages of ZFNs were briefly discussed.

  14. Zinc finger protein 521 overexpression increased transcript levels of Fndc5 in mouse embryonic stem cells

    Indian Academy of Sciences (India)

    Motahere-Sadat Hashemi; Abbas Kiani Esfahani; Maryam Peymani; Alireza Shoaraye Nejati; Kamran Ghaedi; Mohammad Hossein Nasr-Esfahani; Hossein Baharvand

    2016-03-01

    Zinc finger protein 521 is highly expressed in brain, neural stem cells and early progenitors of the human hematopoietic cells. Zfp521 triggers the cascade of neurogenesis inmouse embryonic stemcells through inducing expression of the early neuroectodermal genes Sox1, Sox3 and Pax6. Fndc5, a precursor of Irisin has inducing effects on the expression level of brain derived neurotrophic factor in hippocampus. Therefore, it is most likely that Fndc5 may play an important role in neural differentiation. To exhibit whether the expression of this protein is under regulation with Zfp521, we overexpressed Zfp521 in a stable transformants of mESCs expressing EGFP under control of Fndc5 promoter. Increased expression of Zfp521 enhanced transcription levels of both EGFP and endogenous Fndc5. This result was confirmed by overexpression the aforementioned vectors in HEK cells and indicated that Zfp521 functions upstream of Fndc5 expression. It is most likely that Zfp521 may act through the binding to its response element on Fndc5 core promoter. Therefore it is concluding that an enhanced expression of Fndc5 in neural progenitor cells is stimulated by Zfp521 overexpression in these cells.

  15. Identification of SCAN domain zinc-finger gene ZNF449 as a novel factor of chondrogenesis.

    Directory of Open Access Journals (Sweden)

    Keita Okada

    Full Text Available Transcription factors SOX9, SOX5 and SOX6 are indispensable for generation and differentiation of chondrocytes. However, molecular mechanisms to induce the SOX genes are poorly understood. To address this issue, we previously determined the human embryonic enhancer of SOX6 by 5'RACE analysis, and identified the 46-bp core enhancer region (CES6. We initially performed yeast one-hybrid assay for screening other chondrogenic factors using CES6 as bait, and identified a zinc finger protein ZNF449. ZNF449 and Zfp449, a counterpart in mouse, transactivated enhancers or promoters of SOX6, SOX9 and COL2A1. Zfp449 was expressed in mesenchyme-derived tissues including cartilage, calvaria, muscle and tendon, as well as in other tissues including brain, lung and kidney. In limb cartilage of mouse embryo, Zfp449 protein was abundantly located in periarticular chondrocytes, and decreased in accordance with the differentiation. Zfp449 protein was also detected in articular cartilage of an adult mouse. During chondrogenic differentiation of human mesenchymal stem cells, ZNF449 was increased at an early stage, and its overexpression enhanced SOX9 and SOX6 only at the initial stage of the differentiation. We further generated Zfp449 knockout mice to examine the in vivo roles; however, no obvious abnormality was observed in skeletal development or articular cartilage homeostasis. ZNF449 may regulate chondrogenic differentiation from mesenchymal progenitor cells, although the underlying mechanisms are still unknown.

  16. Origin and evolution of new exons in the rodent zinc finger protein 39 gene

    Institute of Scientific and Technical Information of China (English)

    PENG Lixin; ZHENG Hongkun; LI Xin; YANG Shuang; CHEN Hong; WANG Wen

    2005-01-01

    The origin of new structures and functions is an important process in evolution. In the past decades, we have obtained some preliminary knowledge of the origin and evolution of new genes. However, as the basic unit of genes, the origin and evolution of exons remain unclear. Because young exons retain the footprints of origination, they can be good materials for studying origin and evolution of new exons. In this paper, we report two young exons in a zinc finger protein gene of rodents. Since they are unique sequences in mouse and rat genome and no homologous sequences were found in the orthologous genes of human and pig, the young exons might originate after the divergence of primates and rodents through exonization of intronic sequences. Strong positive selection was detected in the new exons between mouse and rat, suggesting that these exons have undergone significant functional divergence after the separation of the two species. On the other hand, population genetics data of mouse demonstrate that the new exons have been subject to functional constraint, indicating an important function of the new exons in mouse. Functional analyses suggest that these new exons encode a nuclear localization signal peptide, which may mediate new ways of nuclear protein transport. To our knowledge, this is the first example of the origin and evolution of young exons.

  17. Zinc finger protein 148 is dispensable for primitive and definitive hematopoiesis in mice.

    Directory of Open Access Journals (Sweden)

    Anna Nilton

    Full Text Available Hematopoiesis is regulated by transcription factors that induce cell fate and differentiation in hematopoietic stem cells into fully differentiated hematopoietic cell types. The transcription factor zinc finger protein 148 (Zfp148 interacts with the hematopoietic transcription factor Gata1 and has been implicated to play an important role in primitive and definitive hematopoiesis in zebra fish and mouse chimeras. We have recently created a gene-trap knockout mouse model deficient for Zfp148, opening up for analyses of hematopoiesis in a conventional loss-of-function model in vivo. Here, we show that Zfp148-deficient neonatal and adult mice have normal or slightly increased levels of hemoglobin, hematocrit, platelets and white blood cells, compared to wild type controls. Hematopoietic lineages in bone marrow, thymus and spleen from Zfp148 (gt/gt mice were further investigated by flow cytometry. There were no differences in T-cells (CD4 and CD8 single positive cells, CD4 and CD8 double negative/positive cells in either organ. However, the fraction of CD69- and B220-positive cells among lymphocytes in spleen was slightly lower at postnatal day 14 in Zfp148 (gt/gt mice compared to wild type mice. Our results demonstrate that Zfp148-deficient mice generate normal mature hematopoietic populations thus challenging earlier studies indicating that Zfp148 plays a critical role during hematopoietic development.

  18. Efficient targeted mutagenesis in the monarch butterfly using zinc-finger nucleases.

    Science.gov (United States)

    Merlin, Christine; Beaver, Lauren E; Taylor, Orley R; Wolfe, Scot A; Reppert, Steven M

    2013-01-01

    The development of reverse-genetic tools in "nonmodel" insect species with distinct biology is critical to establish them as viable model systems. The eastern North American monarch butterfly (Danaus plexippus), whose genome is sequenced, has emerged as a model to study animal clocks, navigational mechanisms, and the genetic basis of long-distance migration. Here, we developed a highly efficient gene-targeting approach in the monarch using zinc-finger nucleases (ZFNs), engineered nucleases that generate mutations at targeted genomic sequences. We focused our ZFN approach on targeting the type 2 vertebrate-like cryptochrome gene of the monarch (designated cry2), which encodes a putative transcriptional repressor of the monarch circadian clockwork. Co-injections of mRNAs encoding ZFNs targeting the second exon of monarch cry2 into "one nucleus" stage embryos led to high-frequency nonhomologous end-joining-mediated, mutagenic lesions in the germline (up to 50%). Heritable ZFN-induced lesions in two independent lines produced truncated, nonfunctional CRY2 proteins, resulting in the in vivo disruption of circadian behavior and the molecular clock mechanism. Our work genetically defines CRY2 as an essential transcriptional repressor of the monarch circadian clock and provides a proof of concept for the use of ZFNs for manipulating genes in the monarch butterfly genome. Importantly, this approach could be used in other lepidopterans and "nonmodel" insects, thus opening new avenues to decipher the molecular underpinnings of a variety of biological processes. PMID:23009861

  19. Editing T cell specificity towards leukemia by zinc-finger nucleases and lentiviral gene transfer

    Science.gov (United States)

    Lombardo, Angelo; Magnani, Zulma; Liu, Pei-Qi; Reik, Andreas; Chu, Victoria; Paschon, David E.; Zhang, Lei; Kuball, Jurgen; Camisa, Barbara; Bondanza, Attilio; Casorati, Giulia; Ponzoni, Maurilio; Ciceri, Fabio; Bordignon, Claudio; Greenberg, Philip D.; Holmes, Michael C.; Gregory, Philip D.; Naldini, Luigi; Bonini, Chiara

    2016-01-01

    The transfer of high-avidity T-cell receptor (TCR) genes isolated from rare tumor-specific lymphocytes into polyclonal T cells is an attractive cancer immunotherapy strategy. However, TCR gene transfer results in competition for surface expression and inappropriate pairing between the exogenous and endogenous TCR chains, resulting in suboptimal activity and potentially harmful unpredicted specificities. We designed zinc-finger nucleases (ZFNs) promoting the disruption of endogenous TCR β and α chain genes. ZFN-treated lymphocytes lacked CD3/TCR surface expression and expanded with IL-7 and IL-15. Upon lentiviral transfer of a TCR for the WT1 tumor antigen, these TCR-edited cells expressed the new TCR at high levels, were easily expanded to near-purity, and proved superior in specific antigen recognition to matched TCR-transferred cells. In contrast to TCR-transferred cells, TCR edited lymphocytes did not mediate off-target reactivity while maintaining anti-tumor activity in vivo, thus demonstrating that complete editing of T-cell specificity generate tumor-specific lymphocytes with improved biosafety profile. PMID:22466705

  20. Mutagenesis of Genes for Starch Debranching Enzyme Isoforms in Pea by Zinc-Finger Endonucleases

    International Nuclear Information System (INIS)

    Starch debranching enzymes in plants are divided into two groups based on their ability to hydrolyze different substrates. The first group, pullulanases, hydrolyze α-1,6-glucosidic linkages in substrates such as pullulan, amylopectin and glycogen. The second group of debranching enzymes, isoamylases, hydrolyze glycogen and amylopectin and are not active on pullulan. Three isoforms of isoamylase and a pullulanase have been isolated from a cDNA library of Pisum sativum. These isoamylases have been characterized following their heterologous expression in E. coli. Based on the DNA sequence that encodes these debranching enzymes, a specific mutagenesis targeting these enzymes will be attempted. The technique involves the homologous recombination of DNA mediated by zinc-finger endonucleases. Vectors will be constructed to include a fragment that will modify these genes. Using this technique, it is hoped that null mutants for each enzyme will be created and the exact role of these enzymes for the synthesis and degradation of starch in plants will be elucidated. (author)

  1. An elastic-network based local molecular field analysis of zinc-finger proteins

    CERN Document Server

    Dixit, Purushottam D

    2011-01-01

    We study two designed and one natural zinc-finger peptide each with the Cys2His2 (CCHH) type of metal binding motif. In the approach we have developed, we describe the role of the protein and solvent outside the Zn(II)-CCHH metal-residue cluster by a molecular field represented by generalized harmonic restraints. The strength of the field is adjusted to reproduce the binding energy distribution of the metal with the cluster obtained in a reference all-atom simulation with empirical potentials. The quadratic field allows us to investigate analytically the protein restraints on the binding site in terms of its eigenmodes. Examining these eigenmodes suggests, consistent with experimental observations, the importance of the first histidine (H) in the CCHH cluster in metal binding. Further, the eigenvalues corresponding to these modes also indicate that the designed proteins form a tighter complex with the metal. We find that the bulk protein and solvent response tends to destabilize metal-binding, emphasizing tha...

  2. Different binding properties and function of CXXC zinc finger domains in Dnmt1 and Tet1.

    Directory of Open Access Journals (Sweden)

    Carina Frauer

    Full Text Available Several mammalian proteins involved in chromatin and DNA modification contain CXXC zinc finger domains. We compared the structure and function of the CXXC domains in the DNA methyltransferase Dnmt1 and the methylcytosine dioxygenase Tet1. Sequence alignment showed that both CXXC domains have a very similar framework but differ in the central tip region. Based on the known structure of a similar MLL1 domain we developed homology models and designed expression constructs for the isolated CXXC domains of Dnmt1 and Tet1 accordingly. We show that the CXXC domain of Tet1 has no DNA binding activity and is dispensable for catalytic activity in vivo. In contrast, the CXXC domain of Dnmt1 selectively binds DNA substrates containing unmethylated CpG sites. Surprisingly, a Dnmt1 mutant construct lacking the CXXC domain formed covalent complexes with cytosine bases both in vitro and in vivo and rescued DNA methylation patterns in dnmt1⁻/⁻ embryonic stem cells (ESCs just as efficiently as wild type Dnmt1. Interestingly, neither wild type nor ΔCXXC Dnmt1 re-methylated imprinted CpG sites of the H19a promoter in dnmt1⁻/⁻ ESCs, arguing against a role of the CXXC domain in restraining Dnmt1 methyltransferase activity on unmethylated CpG sites.

  3. A dual role for zinc fingers in both DNA binding and zinc sensing by the Zap1 transcriptional activator

    OpenAIRE

    Bird, Amanda J.; Zhao, Hui; Luo, Huan; Jensen, Laran T.; Srinivasan, Chandra; Evans-Galea, Marguerite; Winge, Dennis R.; Eide, David J.

    2000-01-01

    The Zap1 transcriptional activator of Saccharomyces cerevisiae controls zinc homeostasis. Zap1 induces target gene expression in zinc-limited cells and is repressed by high zinc. One such target gene is ZAP1 itself. In this report, we examine how zinc regulates Zap1 function. First, we show that transcriptional autoregulation of Zap1 is a minor component of zinc responsiveness; most regulation of Zap1 activity occurs post-translationally. Secondly, nuclear localization of Zap1 does not change...

  4. C6H6/C2H2…(HX)2(X=F,Cl,Br,I)分子间相互作用研究%Research on the interaction between molecules of C6H6/C2H2···(HX)2

    Institute of Scientific and Technical Information of China (English)

    阳杰; 叶青; 肖扬; 赵培培; 张霞; 李少波; 叶朗; 席小倩

    2015-01-01

    采用从头算MP2方法,对乙炔分子中的π键与(HX)2(X=F,Cl,Br,I)形成的典型X-H···π键复合物进行了系统的研究,并对构建的复合物结构、相互作用能和BSSE进行了优化计算,对计算结果在结构、分子间相互作用能等参数进行分析,最后对C6H6中的π键与(HCl)2形成的典型X-H…π键复合物进行计算研究,并比较炔烃和芳香烃中的两种π键与卤化氢形成的复合物在氢键性质差异。研究结果表明,C2H2···(HX)2复合物体系随着卤素原子序数的递增,结构参数都出现了周期性增加,复合物的相互作用能在整体上呈现减小的总趋势;C6H6···(HCl)2复合物相互作用明显强于C2H2···(HCl)2复合物。%Applyingab initio theoretical MP2 method studies on the complex of C2H2 and(HX)2(X=F,Cl,Br,I),which formed by X-H···π bond.They optimized the complexes,then,calculated the BSSE and the binding energy of different systems, analysis the calculation results in the geometry, intermolecular interaction energy parameters, and finally the typicalπ bond ofC6H6molecule and (HCl)2 form ofX-H···π bond complexes have been studied, composites and compare alkyne to aromatic hydrocarbon twoπ bond with a hydrogen halide formed by the difference in the nature of hydrogen bonding.The results demonstrated that C2H2···(HX)2,with increasing halogen atomic number,from HF to HI,the bond length have all increasing,however,the binding energy of complex and X-H···π interaction present in the overall decreasing trend, correspondingX-H···πinteraction weakened;C6H6···(HCl)2 complex interaction was stronger than C2H2···(HCl)2 complex.

  5. Vibration-rotation alchemy in acetylene (12C2H2), ? at low vibrational excitation: from high resolution spectroscopy to fast intramolecular dynamics

    Science.gov (United States)

    Perry, David S.; Miller, Anthony; Amyay, Badr; Fayt, André; Herman, Michel

    2010-04-01

    The link between energy-resolved spectra and time-resolved dynamics is explored quantitatively for acetylene (12C2H2), ? with up to 8600 cm-1 of vibrational energy. This comparison is based on the extensive and reliable knowledge of the vibration-rotation energy levels and on the model Hamiltonian used to fit them to high precision [B. Amyay, S. Robert, M. Herman, A. Fayt, B. Raghavendra, A. Moudens, J. Thiévin, B. Rowe, and R. Georges, J. Chem. Phys. 131, 114301 (2009)]. Simulated intensity borrowing features in high resolution absorption spectra and predicted survival probabilities in intramolecular vibrational redistribution (IVR) are first investigated for the v 4 + v 5 and v 3 bright states, for J = 2, 30 and 100. The dependence of the results on the rotational quantum number and on the choice of vibrational bright state reflects the interplay of three kinds of off-diagonal resonances: anharmonic, rotational l-type, and Coriolis. The dynamical quantities used to characterize the calculated time-dependent dynamics are the dilution factor φ d, the IVR lifetime τ IVR , and the recurrence time τ rec. For the two bright states v 3 + 2v 4 and 7v 4, the collisionless dynamics for thermally averaged rotational distributions at T = 27, 270 and 500 K were calculated from the available spectroscopic data. For the 7v 4 bright state, an apparent irreversible decay of is found. In all cases, the model Hamiltonian allows a detailed calculation of the energy flow among all of the coupled zeroth-order vibration-rotation states.

  6. Characterization of the target DNA sequence for the DNA-binding domain of zinc finger protein 191

    Institute of Scientific and Technical Information of China (English)

    Haoyue Wang; Ruilin Sun; Guoxiang Liu; Minghui Yao; Jian Fei; Hebai Shen

    2008-01-01

    Studies on the DNA-binding properties of transcription factors are important in searching for the downstream genes regulated by these factors. In the present study, we report on the DNA-binding property of a Cys2His2-type transcription factor, zinc finger protein 191 (Zfp191), which has been newly found to play a significant role in mice.By constructing a fusion protein containing the DNA-binding domain of Zfp191,we characterized target DNA by determining the protein's binding specificity and dependence on zinc.The data showed that the DNA-binding domain of Zfp191can specifically bind to the TCAT repeat motif and that there is a cooperative effect among the target DNA's multiple binding sites.Furthermore,the binding reaction is dependent on zinc.This work provides a foundation for further studies on the role of Zfp191 in gene regulation and development.

  7. Efficient immunoglobulin gene disruption and targeted replacement in rabbit using zinc finger nucleases.

    Directory of Open Access Journals (Sweden)

    Tatiana Flisikowska

    Full Text Available Rabbits are widely used in biomedical research, yet techniques for their precise genetic modification are lacking. We demonstrate that zinc finger nucleases (ZFNs introduced into fertilized oocytes can inactivate a chosen gene by mutagenesis and also mediate precise homologous recombination with a DNA gene-targeting vector to achieve the first gene knockout and targeted sequence replacement in rabbits. Two ZFN pairs were designed that target the rabbit immunoglobulin M (IgM locus within exons 1 and 2. ZFN mRNAs were microinjected into pronuclear stage fertilized oocytes. Founder animals carrying distinct mutated IgM alleles were identified and bred to produce offspring. Functional knockout of the immunoglobulin heavy chain locus was confirmed by serum IgM and IgG deficiency and lack of IgM(+ and IgG(+ B lymphocytes. We then tested whether ZFN expression would enable efficient targeted sequence replacement in rabbit oocytes. ZFN mRNA was co-injected with a linear DNA vector designed to replace exon 1 of the IgM locus with ∼1.9 kb of novel sequence. Double strand break induced targeted replacement occurred in up to 17% of embryos and in 18% of fetuses analyzed. Two major goals have been achieved. First, inactivation of the endogenous IgM locus, which is an essential step for the production of therapeutic human polyclonal antibodies in the rabbit. Second, establishing efficient targeted gene manipulation and homologous recombination in a refractory animal species. ZFN mediated genetic engineering in the rabbit and other mammals opens new avenues of experimentation in immunology and many other research fields.

  8. AAV-mediated delivery of zinc finger nucleases targeting hepatitis B virus inhibits active replication.

    Directory of Open Access Journals (Sweden)

    Nicholas D Weber

    Full Text Available Despite an existing effective vaccine, hepatitis B virus (HBV remains a major public health concern. There are effective suppressive therapies for HBV, but they remain expensive and inaccessible to many, and not all patients respond well. Furthermore, HBV can persist as genomic covalently closed circular DNA (cccDNA that remains in hepatocytes even during otherwise effective therapy and facilitates rebound in patients after treatment has stopped. Therefore, the need for an effective treatment that targets active and persistent HBV infections remains. As a novel approach to treat HBV, we have targeted the HBV genome for disruption to prevent viral reactivation and replication. We generated 3 zinc finger nucleases (ZFNs that target sequences within the HBV polymerase, core and X genes. Upon the formation of ZFN-induced DNA double strand breaks (DSB, imprecise repair by non-homologous end joining leads to mutations that inactivate HBV genes. We delivered HBV-specific ZFNs using self-complementary adeno-associated virus (scAAV vectors and tested their anti-HBV activity in HepAD38 cells. HBV-ZFNs efficiently disrupted HBV target sites by inducing site-specific mutations. Cytotoxicity was seen with one of the ZFNs. scAAV-mediated delivery of a ZFN targeting HBV polymerase resulted in complete inhibition of HBV DNA replication and production of infectious HBV virions in HepAD38 cells. This effect was sustained for at least 2 weeks following only a single treatment. Furthermore, high specificity was observed for all ZFNs, as negligible off-target cleavage was seen via high-throughput sequencing of 7 closely matched potential off-target sites. These results show that HBV-targeted ZFNs can efficiently inhibit active HBV replication and suppress the cellular template for HBV persistence, making them promising candidates for eradication therapy.

  9. AAV-mediated delivery of zinc finger nucleases targeting hepatitis B virus inhibits active replication.

    Science.gov (United States)

    Weber, Nicholas D; Stone, Daniel; Sedlak, Ruth Hall; De Silva Feelixge, Harshana S; Roychoudhury, Pavitra; Schiffer, Joshua T; Aubert, Martine; Jerome, Keith R

    2014-01-01

    Despite an existing effective vaccine, hepatitis B virus (HBV) remains a major public health concern. There are effective suppressive therapies for HBV, but they remain expensive and inaccessible to many, and not all patients respond well. Furthermore, HBV can persist as genomic covalently closed circular DNA (cccDNA) that remains in hepatocytes even during otherwise effective therapy and facilitates rebound in patients after treatment has stopped. Therefore, the need for an effective treatment that targets active and persistent HBV infections remains. As a novel approach to treat HBV, we have targeted the HBV genome for disruption to prevent viral reactivation and replication. We generated 3 zinc finger nucleases (ZFNs) that target sequences within the HBV polymerase, core and X genes. Upon the formation of ZFN-induced DNA double strand breaks (DSB), imprecise repair by non-homologous end joining leads to mutations that inactivate HBV genes. We delivered HBV-specific ZFNs using self-complementary adeno-associated virus (scAAV) vectors and tested their anti-HBV activity in HepAD38 cells. HBV-ZFNs efficiently disrupted HBV target sites by inducing site-specific mutations. Cytotoxicity was seen with one of the ZFNs. scAAV-mediated delivery of a ZFN targeting HBV polymerase resulted in complete inhibition of HBV DNA replication and production of infectious HBV virions in HepAD38 cells. This effect was sustained for at least 2 weeks following only a single treatment. Furthermore, high specificity was observed for all ZFNs, as negligible off-target cleavage was seen via high-throughput sequencing of 7 closely matched potential off-target sites. These results show that HBV-targeted ZFNs can efficiently inhibit active HBV replication and suppress the cellular template for HBV persistence, making them promising candidates for eradication therapy. PMID:24827459

  10. Knockout of exogenous EGFP gene in porcine somatic cells using zinc-finger nucleases

    International Nuclear Information System (INIS)

    Research highlights: → EGFP gene integrated in porcine somatic cells could be knocked out using the ZFN-KO system. → ZFNs induced targeted mutations in porcine primary cultured cells. → Complete absence of EGFP fluorescence was confirmed in ZFN-treated cells. -- Abstract: Zinc-finger nucleases (ZFNs) are expected as a powerful tool for generating gene knockouts in laboratory and domestic animals. Currently, it is unclear whether this technology can be utilized for knocking-out genes in pigs. Here, we investigated whether knockout (KO) events in which ZFNs recognize and cleave a target sequence occur in porcine primary cultured somatic cells that harbor the exogenous enhanced green fluorescent protein (EGFP) gene. ZFN-encoding mRNA designed to target the EGFP gene was introduced by electroporation into the cell. Using the Surveyor nuclease assay and flow cytometric analysis, we confirmed ZFN-induced cleavage of the target sequence and the disappearance of EGFP fluorescence expression in ZFN-treated cells. In addition, sequence analysis revealed that ZFN-induced mutations such as base substitution, deletion, or insertion were generated in the ZFN cleavage site of EGFP-expression negative cells that were cloned from ZFN-treated cells, thereby showing it was possible to disrupt (i.e., knock out) the function of the EGFP gene in porcine somatic cells. To our knowledge, this study provides the first evidence that the ZFN-KO system can be applied to pigs. These findings may open a new avenue to the creation of gene KO pigs using ZFN-treated cells and somatic cell nuclear transfer.

  11. Establishment of transgenic mice carrying gene encoding human zinc finger protein 191

    Institute of Scientific and Technical Information of China (English)

    Jian-Zhong Li; Ji-Liang Fu; Xia Chen; Hua Yang; Shui-Liang Wang; Xue-Lian Gong; Hao Feng; Bao-Yu Guo; Long Yu; Zhu-Gang Wang

    2004-01-01

    AIM: Human zinc finger protein 191 (ZNF191) was cloned and characterized as a Kruppel-like transcription factor, which might be relevant to many diseases such as liver cancer,neuropsychiatric and cardiovascular diseases. Although progress has been made recently, the biological function of ZNF191 remains largely unidentified. The aim of this study was to establish a ZNF191 transgenic mouse model, which would promote the functional study of ZNF191.METHODS: Transgene fragments were microinjected into fertilized eggs of mice. The manipulated embryos were transferred into the oviducts of pseudo-pregnant female mice. The offsprings were identified by PCR and Southern blot analysis. ZNF 191 gene expression was analyzed by RT-PCR. Transgenic founder mice were used to establish transgenic mouse lineages. The first generation (F1) and the second generation (F2) mice were identified by PCR analysis. Ten-week transgenic mice were used for pathological examination.RESULTS: Four mice were identified as carrying copies of ZNF191 gene. The results of RT-PCR showed that ZNF191gene was expressed in the liver, testis and brain in one of the transgenic mouse lineages. Genetic analysis of transgenic mice demonstrated that ZNF191 gene was integrated into the chromosome at a single site and could be transmitted stably. Pathological analysis showed that the expression of ZNF 191 did not cause obvious pathological changes in multiple tissues of transgenic mice.CONCLUSION: ZNF 191 transgenic mouse model would facilitate the investigation of biological functions of ZNF191 in vivo.

  12. Zinc finger protein A20 protects rats against chronic liver allograft dysfunction

    Institute of Scientific and Technical Information of China (English)

    Jie Yang; Ming-Qing Xu; Lu-Nan Yan; Xiao-Bo Chen; Jiao Liu

    2012-01-01

    AIM:To investigate the effect of zinc finger protein A20 on chronic liver allograft dysfunction in rats.METHODS:Allogeneic liver transplantation from DA rats to Lewis rats was performed.Chronic liver allograft dysfunction was induced in the rats by administering low-dose tacrolimus at postoperative day (POD) 5.Hepatic overexpression of A20 was achieved by recombinant adenovirus (rAd.)-mediated gene transfer administered intravenously every 10 d starting from POD 10.The recipient rats were injected with physiological saline,rAdEasy-A20 (1 x 109 pfu/30 g weight) or rAdEasy (1 x 109 pfu/30 g weight) every 10 d through the tail vein for 3 mo starting from POD 10.Liver tissue samples were harvested on POD 30 and POD 60.RESULTS:Liver-transplanted rats treated with only tacrolimus showed chronic allograft dysfunction with severe hepatic fibrosis.A20 overexpression ameliorated the effects on liver function,attenuated liver allograft fibrosis and prolonged the survival of the recipient rats.Treatment with A20 suppressed hepatic protein production of tumor growth factor (TGF)-β1,interleukin1β,caspase-8,CD40,CD40L,intercellular adhesion molecule-1,vascular cell adhesion molecule-1 and E-selectin.A20 treatment suppressed liver cell apoptosis and inhibited nuclear factor-κB activation of Kupffer cells (KCs),liver sinusoidal endothelial cells (LSECs)and hepatic stellate cells (HSCs),and it subsequently decreased cytokine mRNA expression in KCs and LSECs and reduced the production of TGF-β1 in HSCs.CONCLUSION:A20 might prevent chronic liver allograft dysfunction by re-establishing functional homeostasis of KCs,LSECs and HSCs.

  13. Zinc finger transcription factors displaced SREBP proteins as the major Sterol regulators during Saccharomycotina evolution.

    Directory of Open Access Journals (Sweden)

    Sarah L Maguire

    2014-01-01

    Full Text Available In most eukaryotes, including the majority of fungi, expression of sterol biosynthesis genes is regulated by Sterol-Regulatory Element Binding Proteins (SREBPs, which are basic helix-loop-helix transcription activators. However, in yeasts such as Saccharomyces cerevisiae and Candida albicans sterol synthesis is instead regulated by Upc2, an unrelated transcription factor with a Gal4-type zinc finger. The SREBPs in S. cerevisiae (Hms1 and C. albicans (Cph2 have lost a domain, are not major regulators of sterol synthesis, and instead regulate filamentous growth. We report here that rewiring of the sterol regulon, with Upc2 taking over from SREBP, likely occurred in the common ancestor of all Saccharomycotina. Yarrowia lipolytica, a deep-branching species, is the only genome known to contain intact and full-length orthologs of both SREBP (Sre1 and Upc2. Deleting YlUPC2, but not YlSRE1, confers susceptibility to azole drugs. Sterol levels are significantly reduced in the YlUPC2 deletion. RNA-seq analysis shows that hypoxic regulation of sterol synthesis genes in Y. lipolytica is predominantly mediated by Upc2. However, YlSre1 still retains a role in hypoxic regulation; growth of Y. lipolytica in hypoxic conditions is reduced in a Ylupc2 deletion and is abolished in a Ylsre1/Ylupc2 double deletion, and YlSre1 regulates sterol gene expression during hypoxia adaptation. We show that YlSRE1, and to a lesser extent YlUPC2, are required for switching from yeast to filamentous growth in hypoxia. Sre1 appears to have an ancestral role in the regulation of filamentation, which became decoupled from its role in sterol gene regulation by the arrival of Upc2 in the Saccharomycotina.

  14. ZFN-Site searches genomes for zinc finger nuclease target sites and off-target sites

    Directory of Open Access Journals (Sweden)

    Iseli Christian

    2011-05-01

    Full Text Available Abstract Background Zinc Finger Nucleases (ZFNs are man-made restriction enzymes useful for manipulating genomes by cleaving target DNA sequences. ZFNs allow therapeutic gene correction or creation of genetically modified model organisms. ZFN specificity is not absolute; therefore, it is essential to select ZFN target sites without similar genomic off-target sites. It is important to assay for off-target cleavage events at sites similar to the target sequence. Results ZFN-Site is a web interface that searches multiple genomes for ZFN off-target sites. Queries can be based on the target sequence or can be expanded using degenerate specificity to account for known ZFN binding preferences. ZFN off-target sites are outputted with links to genome browsers, facilitating off-target cleavage site screening. We verified ZFN-Site using previously published ZFN half-sites and located their target sites and their previously described off-target sites. While we have tailored this tool to ZFNs, ZFN-Site can also be used to find potential off-target sites for other nucleases, such as TALE nucleases. Conclusions ZFN-Site facilitates genome searches for possible ZFN cleavage sites based on user-defined stringency limits. ZFN-Site is an improvement over other methods because the FetchGWI search engine uses an indexed search of genome sequences for all ZFN target sites and possible off-target sites matching the half-sites and stringency limits. Therefore, ZFN-Site does not miss potential off-target sites.

  15. Zebrafish foxP2 zinc finger nuclease mutant has normal axon pathfinding.

    Directory of Open Access Journals (Sweden)

    Lingyan Xing

    Full Text Available foxP2, a forkhead-domain transcription factor, is critical for speech and language development in humans, but its role in the establishment of CNS connectivity is unclear. While in vitro studies have identified axon guidance molecules as targets of foxP2 regulation, and cell culture assays suggest a role for foxP2 in neurite outgrowth, in vivo studies have been lacking regarding a role for foxP2 in axon pathfinding. We used a modified zinc finger nuclease methodology to generate mutations in the zebrafish foxP2 gene. Using PCR-based high resolution melt curve analysis (HRMA of G0 founder animals, we screened and identified three mutants carrying nonsense mutations in the 2(nd coding exon: a 17 base-pair (bp deletion, an 8bp deletion, and a 4bp insertion. Sequence analysis of cDNA confirmed that these were frameshift mutations with predicted early protein truncations. Homozygous mutant fish were viable and fertile, with unchanged body morphology, and no apparent differences in CNS apoptosis, proliferation, or patterning at embryonic stages. There was a reduction in expression of the known foxP2 target gene cntnap2 that was rescued by injection of wild-type foxP2 transcript. When we examined axon pathfinding using a pan-axonal marker or transgenic lines, including a foxP2-neuron-specific enhancer, we did not observe any axon guidance errors. Our findings suggest that foxP2 is not necessary for axon pathfinding during development.

  16. Hydrogen sulfide represses androgen receptor transactivation by targeting at the second zinc finger module.

    Science.gov (United States)

    Zhao, Kexin; Li, Shuangshuang; Wu, Lingyun; Lai, Christopher; Yang, Guangdong

    2014-07-25

    Androgen receptor (AR) signaling is indispensable for the development of prostate cancer from the initial androgen-dependent state to a later aggressive androgen-resistant state. This study examined the role of hydrogen sulfide (H(2)S), a novel gasotransmitter, in the regulation of AR signaling as well as its mediation in androgen-independent cell growth in prostate cancer cells. Here we found that H(2)S inhibits cell proliferation of both androgen-dependent (LNCaP) and antiandrogen-resistant prostate cancer cells (LNCaP-B), with more significance on the latter, which was established by long term treatment of parental LNCaP cells with bicalutamide. The expression of cystathionine γ-lyase (CSE), a major H(2)S producing enzyme in prostate tissue, was reduced in both human prostate cancer tissues and LNCaP-B cells. LNCaP-B cells were resistant to bicalutamide-induced cell growth inhibition, and CSE overexpression could rebuild the sensitivity of LNCaP-B cells to bicalutamide. H(2)S significantly repressed the expression of prostate-specific antigen (PSA) and TMPRSS2, two AR-targeted genes. In addition, H(2)S inhibited AR binding with PSA promoter and androgen-responsive element (ARE) luciferase activity. We further found that AR is post-translationally modified by H(2)S through S-sulfhydration. Mutation of cysteine 611 and cysteine 614 in the second zinc finger module of AR-DNA binding domain diminished the effects of H(2)S on AR S-sulfhydration and AR dimerization. These data suggest that reduced CSE/H2S signaling contributes to antiandrogen-resistant status, and sufficient level of H(2)S is able to inhibit AR transactivation and treat castration-resistant prostate cancer.

  17. Knockout of exogenous EGFP gene in porcine somatic cells using zinc-finger nucleases

    Energy Technology Data Exchange (ETDEWEB)

    Watanabe, Masahito [Japan Science and Technology Agency (JST), ERATO, Nakauchi Stem Cell and Organ Regeneration Project, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Department of Life Sciences, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571 (Japan); Umeyama, Kazuhiro [Department of Life Sciences, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571 (Japan); International Cluster for Bio-Resource Research, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571 (Japan); Matsunari, Hitomi [Department of Life Sciences, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571 (Japan); Takayanagi, Shuko [Japan Science and Technology Agency (JST), ERATO, Nakauchi Stem Cell and Organ Regeneration Project, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Department of Life Sciences, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571 (Japan); Haruyama, Erika; Nakano, Kazuaki; Fujiwara, Tsukasa; Ikezawa, Yuka [Department of Life Sciences, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571 (Japan); Nakauchi, Hiromitsu [Japan Science and Technology Agency (JST), ERATO, Nakauchi Stem Cell and Organ Regeneration Project, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, Tokyo University, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); and others

    2010-11-05

    Research highlights: {yields} EGFP gene integrated in porcine somatic cells could be knocked out using the ZFN-KO system. {yields} ZFNs induced targeted mutations in porcine primary cultured cells. {yields} Complete absence of EGFP fluorescence was confirmed in ZFN-treated cells. -- Abstract: Zinc-finger nucleases (ZFNs) are expected as a powerful tool for generating gene knockouts in laboratory and domestic animals. Currently, it is unclear whether this technology can be utilized for knocking-out genes in pigs. Here, we investigated whether knockout (KO) events in which ZFNs recognize and cleave a target sequence occur in porcine primary cultured somatic cells that harbor the exogenous enhanced green fluorescent protein (EGFP) gene. ZFN-encoding mRNA designed to target the EGFP gene was introduced by electroporation into the cell. Using the Surveyor nuclease assay and flow cytometric analysis, we confirmed ZFN-induced cleavage of the target sequence and the disappearance of EGFP fluorescence expression in ZFN-treated cells. In addition, sequence analysis revealed that ZFN-induced mutations such as base substitution, deletion, or insertion were generated in the ZFN cleavage site of EGFP-expression negative cells that were cloned from ZFN-treated cells, thereby showing it was possible to disrupt (i.e., knock out) the function of the EGFP gene in porcine somatic cells. To our knowledge, this study provides the first evidence that the ZFN-KO system can be applied to pigs. These findings may open a new avenue to the creation of gene KO pigs using ZFN-treated cells and somatic cell nuclear transfer.

  18. Molecular cloning and characterization of a gene encoding RING zinc finger ankyrin protein from drought-tolerant Artemisia desertorum

    Indian Academy of Sciences (India)

    Xiuhong Yang; Chao Sun; Yuanlei Hun; Zhongping Lin

    2008-03-01

    A RING zinc finger ankyrin protein gene, designated AdZFP1, was isolated from drought-tolerant Artemisia desertorum Spreng by mRNA differential display and RACE. Its cDNA was 1723 bp and encoded a putative protein of 445 amino acids with a predicted molecular mass of 47.9 kDa and an isoelectric point (pI) of 7.49. A typical C3HC4-type RING finger domain was found at the C-terminal region of the AdZFP1 protein, and several groups of ankyrin repeats were found at the N-terminal region. Alignments of amino acid sequence showed that AdZFP1 was 66% identical to the Arabidopsis thaliana putative RING zinc finger ankyrin protein AAN31869. Transcriptional analysis showed that AdZFP1 was inducible under drought stress in root, stem and leaf of the plant. Semi-quantitative reverse-transcriptase-polymerase chain reaction (RT-PCR) analysis showed that the transcript of AdZFP1 was strongly induced by exogenous abscisic acid (ABA) and also by salinity, cold and heat to some extent. Overexpression of the AdZFP1 gene in transgenic tobacco enhanced their tolerance to drought stress.

  19. N2O-C2H2火焰原子吸收光谱法测定膨胀合金中钴%Measurement of Cobalt in the Expanded Alloy by N2O-C2H2 Flame Atomic Absorption Spectrometer

    Institute of Scientific and Technical Information of China (English)

    薛光荣

    2005-01-01

    采用了N2O-C2H2火焰原子吸收光谱法测定膨胀合金中钴.介绍了钴的最佳测定条件,并在样品测定中对干扰因素进行了综合考虑.该方法灵敏度高、干扰小,操作步骤简单、分析周期快等特点.其方法的相对标准偏差均小于1.0%.加标回收率均为97.0%~102.0%范围内,测定结果令人满意.

  20. Chanabayaite, Cu2(N3C2H2)Cl(NH3,Cl,H2O,□)4, a new mineral containing triazolate anion

    Science.gov (United States)

    Chukanov, N. V.; Zubkova, N. V.; Möhn, G.; Pekov, I. V.; Pushcharovsky, D. Yu.; Zadov, A. E.

    2015-12-01

    A new mineral, chanabayaite, has been discovered at a guano deposit located at Mt. Pabellón de Pica near the village of Chanabaya, Iquique Province, Tarapacá region, Chile. It is associated with salammoniac, halite, joanneumite, nitratine and earlier chalcopyrite. Chanabayaite occurs as blue translucent imperfect prismatic crystals, up to 0.05 × 0.1 × 0.5 mm in size, and their radial aggregates. Chanabayaite is brittle, with a Mohs' hardness of 2. The cleavage is perfect on (001) and imperfect on (100) and (010). D meas = 1.48(2) g/cm3, D calc = 1.464 g/cm3. The mineral is optically biaxial (-), α = 1.561(2), β = 1.615(3), γ = 1.620(2), 2 V meas = 25(10)°, 2 V calc = 33°. Pleochroism is strong, Z ≈ Y (deep blue) ≫ X (pale blue with gray tint). IR spectrum is given. The chemical composition (electron microprobe data for Cu, Fe and Cl; gas chromatography data for H, N, C and O) is as follows (wt %): 32.23 Cu, 1.14 Fe, 16.13 Cl, 3.1 H, 29.9 N, 12.2 C, 3.4 O, total is 98.1. The empirical formula is ( Z = 4): Cu1.92Fe0.08Cl1.72N8.09C3.85H11.66O0.81. The structural model was based on the single-crystal X-ray diffraction data ( R = 0.1627). Chanabayaite is orthorhombic, space group Imma, a = 19.484(3), b = 7.2136(10), c = 11.999(4) Å, V = 1686.5(7) Å3, Z = 2. In chanabayaite, chains of the corner-sharing Cu(l)-centered octahedra and single Cu(2)-centered octahedra are linked via 1,2,4-triazolate anions C2N3H2 -. NH3 and Cl- are additional ligands coordinating Cu2+. Chanabayaite is a transformational mineral species formed by leaching of Na and one third of Cl and partial dehydration of the protophase Na2Cu2Cl3(N3C2H2)2(NH3)2 • 4H2O. The strongest reflections in the powder X-ray diffraction pattern [ d, Å ( I, %) ( hkl)] are detected: 10.19 (100) (101), 6.189 (40) (011), 5.729 (23) (301), 5.216 (75) (211, 202), 4.964 (20) (400), 2.830 (20) (602, 413, 503), 2.611 (24) (123, 422, 404).

  1. Darling-Dennison resonance and Coriolis coupling in the bending overtones of the A 1A(u) state of acetylene, C2H2.

    Science.gov (United States)

    Merer, Anthony J; Yamakita, Nami; Tsuchiya, Soji; Steeves, Adam H; Bechtel, Hans A; Field, Robert W

    2008-08-01

    Rotational analyses have been carried out for the overtones of the nu(4) (torsion) and nu(6) (in-plane cis-bend) vibrations of the A (1)A(u) state of C(2)H(2). The v(4)+v(6)=2 vibrational polyad was observed in high-sensitivity one-photon laser-induced fluorescence spectra and the v(4)+v(6)=3 polyad was observed in IR-UV double resonance spectra via the ground state nu(3) (Sigma(+) (u)) and nu(3)+nu(4) (Pi(u)) vibrational levels. The structures of these polyads are dominated by the effects of vibrational angular momentum: Vibrational levels of different symmetry interact via strong a-and b-axis Coriolis coupling, while levels of the same symmetry interact via Darling-Dennison resonance, where the interaction parameter has the exceptionally large value K(4466)=-51.68 cm(-1). The K-structures of the polyads bear almost no resemblance to the normal asymmetric top patterns, and many local avoided crossings occur between close-lying levels with nominal K-values differing by one or more units. Least squares analysis shows that the coupling parameters change only slightly with vibrational excitation, which has allowed successful predictions of the structures of the higher polyads: A number of weak bands from the v(4)+v(6)=4 and 5 polyads have been identified unambiguously. The state discovered by Scherer et al. [J. Chem. Phys. 85, 6315 (1986)], which appears to interact with the K=1 levels of the 3(3) vibrational state at low J, is identified as the second highest of the five K=1 members of the v(4)+v(6)=4 polyad. After allowing for the Darling-Dennison resonance, the zero-order bending structure can be represented by omega(4)=764.71, omega(6)=772.50, x(44)=0.19, x(66)=-4.23, and x(46)=11.39 cm(-1). The parameters x(46) and K(4466) are both sums of contributions from the vibrational angular momentum and from the anharmonic force field. For x(46) these contributions are 14.12 and -2.73 cm(-1), respectively, while the corresponding values for K(4466) are -28.24 and -23

  2. Expression and purification of a novel ZNF191 zinc finger protein——ZNF191 protein and its truncated zinc finger region ZNF191 (243—368)

    Institute of Scientific and Technical Information of China (English)

    刘玉奇; 余龙; 余文浩; 施少林; 孙炳耘; 吴国俊; 黄仲贤

    1999-01-01

    ZNF191 is a new zine finger gene whieh has a 1107 bp open reading frame (ORF) and eneodes a 368 amino avid protein including a putative DNA-binding domain of four Cys2 His2 zine finger motifs at its C-terminal region. The ZNF191 cDNA is loeated in the chromosome 18q12.1 region where it is known that a variety of hereditary diseases are related to. Probably, this protein has potential function of stimulating the gene franscription. In order to examine the function and structure of ZNF191 protein, the ORF of ZNF191 and its zine finger region ZNF191(243--368) genes were inserted into PTSA-18 expression vector by PCR amplification, then the constructed genes were expressed in the PTSA-18/B121 (DE3) system. The two proteins were purified by DEAE-52, CM-23 and Heparin-Sepharose 4B columns. The pooled proteins showed a single band as assayed by Coomasie Brillian Blue Staining of an SDS/polyacryamide gel.

  3. The LSD1-type zinc finger motifs of Pisum sativa LSD1 are a novel nuclear localization signal and interact with importin alpha.

    Directory of Open Access Journals (Sweden)

    Shanping He

    Full Text Available BACKGROUND: Genetic studies of the Arabidopsis mutant lsd1 highlight the important role of LSD1 in the negative regulation of plant programmed cell death (PCD. Arabidopsis thaliana LSD1 (AtLSD1 contains three LSD1-type zinc finger motifs, which are involved in the protein-protein interaction. METHODOLOGY/PRINCIPAL FINDINGS: To further understand the function of LSD1, we have analyzed cellular localization and functional localization domains of Pisum sativa LSD1 (PsLSD1, which is a homolog of AtLSD1. Subcellular localization analysis of green fluorescent protein (GFP-tagged PsLSD1 indicates that PsLSD1 is localized in the nucleus. Using a series of GFP-tagged PsLSD1 deletion mutants, we found that the three LSD1-type zinc finger motifs of PsLSD1 alone can target GFP to the nucleus, whereas deletion of the three zinc finger motifs or any individual zinc finger motif causes PsLSD1 to lose its nuclear localization, indicating that the three zinc finger motifs are necessary and sufficient for its nuclear localization. Moreover, site-directed mutagenesis analysis of GFP-tagged PsLSD1 indicates that tertiary structure and basic amino acids of each zinc finger motif are necessary for PsLSD1 nuclear localization. In addition, yeast two-hybrid, pull-down, and BiFC assays demonstrate that the three zinc finger motifs of PsLSD1 directly bind to importin α in vitro and in vivo. CONCLUSIONS/SIGNIFICANCE: Our data demonstrate that the LSD1-type zinc finger motifs of PsLSD1 are a novel nuclear localization signal and directly bind to importin α, and suggest that the nuclear import of LSD1 may rely on the interaction between its zinc finger motifs and importin α. Moreover, the nuclear localization of PsLSD1 suggests that LSD1 may function as a transcription regulator involved in negatively regulating PCD.

  4. Theoretical Study of the Structure and π-H Bond Properties for Dimers of Halogenated Ethene with Hydrogen Fluoride g-C2H2X2-HF (X=F, Cl)%卤代乙烯与氟化氢二聚体g-C2H2X2-HF(X=F,Cl)的结构和氢键性质的理论研究

    Institute of Scientific and Technical Information of China (English)

    李瑞延; 陈建林; 吴迪

    2006-01-01

    本文作者使用了Counterpoise(CP)校正方法,在MP2/aug-cc-pVDZ水平,计算得到了实频的C2H4-HF和g-C2H2X2-HF(X=F,Cl)的优化结构.由于X原子的作用,二聚体g-C2H2X2-HF(X=F,Cl)的π-H键被延长,并且在结构中发生了倾斜和弯曲.X原子的作用降低了π-H键相互作用能,而电子相关效应增加了π-H键相互作用能.g-C2H2X2-HF(X=F,Cl)结构中存在着π-type H-bond.

  5. Direct binding of specific AUF1 isoforms to tandem zinc finger domains of tristetraprolin (TTP) family proteins.

    Science.gov (United States)

    Kedar, Vishram P; Zucconi, Beth E; Wilson, Gerald M; Blackshear, Perry J

    2012-02-17

    Tristetraprolin (TTP) is the prototype of a family of CCCH tandem zinc finger proteins that can bind to AU-rich elements in mRNAs and promote their decay. TTP binds to mRNA through its central tandem zinc finger domain; it then promotes mRNA deadenylation, considered to be the rate-limiting step in eukaryotic mRNA decay. We found that TTP and its related family members could bind to certain isoforms of another AU-rich element-binding protein, HNRNPD/AUF1, as well as a related protein, laAUF1. The interaction domain within AUF1p45 appeared to be a C-terminal "GY" region, and the interaction domain within TTP was the tandem zinc finger domain. Surprisingly, binding of AUF1p45 to TTP occurred even with TTP mutants that lacked RNA binding activity. In cell extracts, binding of AUF1p45 to TTP potentiated TTP binding to ARE-containing RNA probes, as determined by RNA gel shift assays; AUF1p45 did not bind to the RNA probes under these conditions. Using purified, recombinant proteins and a synthetic RNA target in FRET assays, we demonstrated that AUF1p45, but not AUF1p37, increased TTP binding affinity for RNA ∼5-fold. These data suggest that certain isoforms of AUF1 can serve as "co-activators" of TTP family protein binding to RNA. The results raise interesting questions about the ability of AUF1 isoforms to regulate the mRNA binding and decay-promoting activities of TTP and its family members as well as the ability of AUF1 proteins to serve as possible physical links between TTP and other mRNA decay proteins and structures.

  6. Molecular cloning and characterization of a novel RING zinc-finger protein gene up-regulated under in vitro salt stress in cassava.

    Science.gov (United States)

    dos Reis, Sávio Pinho; Tavares, Liliane de Souza Conceição; Costa, Carinne de Nazaré Monteiro; Brígida, Aílton Borges Santa; de Souza, Cláudia Regina Batista

    2012-06-01

    Cassava (Manihot esculenta Crantz) is one of the world's most important food crops. It is cultivated mainly in developing countries of tropics, since its root is a major source of calories for low-income people due to its high productivity and resistance to many abiotic and biotic factors. A previous study has identified a partial cDNA sequence coding for a putative RING zinc finger in cassava storage root. The RING zinc finger protein is a specialized type of zinc finger protein found in many organisms. Here, we isolated the full-length cDNA sequence coding for M. esculenta RZF (MeRZF) protein by a combination of 5' and 3' RACE assays. BLAST analysis showed that its deduced amino acid sequence has a high level of similarity to plant proteins of RZF family. MeRZF protein contains a signature sequence motif for a RING zinc finger at its C-terminal region. In addition, this protein showed a histidine residue at the fifth coordination site, likely belonging to the RING-H2 subgroup, as confirmed by our phylogenetic analysis. There is also a transmembrane domain in its N-terminal region. Finally, semi-quantitative RT-PCR assays showed that MeRZF expression is increased in detached leaves treated with sodium chloride. Here, we report the first evidence of a RING zinc finger gene of cassava showing potential role in response to salt stress.

  7. The maize INDETERMINATE1 flowering time regulator defines a highly conserved zinc finger protein family in higher plants

    Directory of Open Access Journals (Sweden)

    Colasanti Joseph

    2006-06-01

    Full Text Available Abstract Background The maize INDETERMINATE1 gene, ID1, is a key regulator of the transition to flowering and the founding member of a transcription factor gene family that encodes a protein with a distinct arrangement of zinc finger motifs. The zinc fingers and surrounding sequence make up the signature ID domain (IDD, which appears to be found in all higher plant genomes. The presence of zinc finger domains and previous biochemical studies showing that ID1 binds to DNA suggests that members of this gene family are involved in transcriptional regulation. Results Comparison of IDD genes identified in Arabidopsis and rice genomes, and all IDD genes discovered in maize EST and genomic databases, suggest that ID1 is a unique member of this gene family. High levels of sequence similarity amongst all IDD genes from maize, rice and Arabidopsis suggest that they are derived from a common ancestor. Several unique features of ID1 suggest that it is a divergent member of the maize IDD family. Although no clear ID1 ortholog was identified in the Arabidopsis genome, highly similar genes that encode proteins with identity extending beyond the ID domain were isolated from rice and sorghum. Phylogenetic comparisons show that these putative orthologs, along with maize ID1, form a group separate from other IDD genes. In contrast to ID1 mRNA, which is detected exclusively in immature leaves, several maize IDD genes showed a broad range of expression in various tissues. Further, Western analysis with an antibody that cross-reacts with ID1 protein and potential orthologs from rice and sorghum shows that all three proteins are detected in immature leaves only. Conclusion Comparative genomic analysis shows that the IDD zinc finger family is highly conserved among both monocots and dicots. The leaf-specific ID1 expression pattern distinguishes it from other maize IDD genes examined. A similar leaf-specific localization pattern was observed for the putative ID1 protein

  8. Characterization of the contradictory chromatin signatures at the 3' exons of zinc finger genes.

    Directory of Open Access Journals (Sweden)

    Kimberly R Blahnik

    Full Text Available The H3K9me3 histone modification is often found at promoter regions, where it functions to repress transcription. However, we have previously shown that 3' exons of zinc finger genes (ZNFs are marked by high levels of H3K9me3. We have now further investigated this unusual location for H3K9me3 in ZNF genes. Neither bioinformatic nor experimental approaches support the hypothesis that the 3' exons of ZNFs are promoters. We further characterized the histone modifications at the 3' ZNF exons and found that these regions also contain H3K36me3, a mark of transcriptional elongation. A genome-wide analysis of ChIP-seq data revealed that ZNFs constitute the majority of genes that have high levels of both H3K9me3 and H3K36me3. These results suggested the possibility that the ZNF genes may be imprinted, with one allele transcribed and one allele repressed. To test the hypothesis that the contradictory modifications are due to imprinting, we used a SNP analysis of RNA-seq data to demonstrate that both alleles of certain ZNF genes having H3K9me3 and H3K36me3 are transcribed. We next analyzed isolated ZNF 3' exons using stably integrated episomes. We found that although the H3K36me3 mark was lost when the 3' ZNF exon was removed from its natural genomic location, the isolated ZNF 3' exons retained the H3K9me3 mark. Thus, the H3K9me3 mark at ZNF 3' exons does not impede transcription and it is regulated independently of the H3K36me3 mark. Finally, we demonstrate a strong relationship between the number of tandemly repeated domains in the 3' exons and the H3K9me3 mark. We suggest that the H3K9me3 at ZNF 3' exons may function to protect the genome from inappropriate recombination rather than to regulate transcription.

  9. Single Photon K-2 and K-1K-1 Double Core Ionization in C2H2n (n=1-3), CO, and N2 as a Potential New Tool for Chemical Analysis

    Science.gov (United States)

    Nakano, M.; Penent, F.; Tashiro, M.; Grozdanov, T. P.; Žitnik, M.; Carniato, S.; Selles, P.; Andric, L.; Lablanquie, P.; Palaudoux, J.; Shigemasa, E.; Iwayama, H.; Hikosaka, Y.; Soejima, K.; Suzuki, I. H.; Kouchi, N.; Ito, K.

    2013-04-01

    We have observed single photon double K-shell photoionization in the C2H2n (n=1-3) hydrocarbon sequence and in N2 and CO, using synchrotron radiation and electron coincidence spectroscopy. Our previous observations of the K-2 process in these molecules are extended by the observations of a single photon double photoionization with one core hole created at each of the two neighboring atoms in the molecule (K-1K-1 process). In the C2H2n sequence, the spectroscopy of K-1K-1 states is much more sensitive to the bond length than conventional electron spectroscopy for chemical analysis spectroscopy based on single K-shell ionization. The cross section variation for single photon K-1K-1 double core ionization in the C2H2n sequence and in the isoelectronic C2H2, N2 and CO molecules validates a knock-out mechanism in which a primary ionized 1s photoelectron ejects another 1s electron of the neighbor atom. The specific Auger decay from such states is clearly observed in the CO case.

  10. A novel genetic system based on zinc finger nucleases for the identification of interactions between proteins in vivo.

    Directory of Open Access Journals (Sweden)

    Ling Wang

    Full Text Available Yeast two-hybrid (Y2H methods are powerful tools for detecting protein-protein interactions. The traditional Y2H method has been widely applied to screen novel protein interactions since it was established two decades ago. The high false-positive rate of the traditional method drove the development of modified Y2H systems. Here, we describe a novel Y2H system using zinc-finger nucleases (ZFNs. ZFNs contain two functional domains, a zinc-finger DNA-binding domain (ZFP and a non-specific nuclease domain (FokI. In this system, the bait is expressed as a fusion protein with a specific ZFP, and the prey is fused to the FokI. A reporter vector is designed such that the ZFN target site disrupts the Gal4 open reading frame. By transforming the three plasmids into a yeast strain (AH109, the interaction between the bait and prey proteins reconstitutes ZFN function and generates the double-strand break (DSB on its target site. The DNA DSB repair restores Gal4 function, which activates the expression of the four reporter genes. We used p53-SV40LT interacting proteins to prove the concept. In addition, 80% positive rate was observed in a cDNA screening test against WDSV orfA protein. Our results strongly suggested that this Y2H system could increase screening reliability and reproducibility, and provide a novel approach for interactomics research.

  11. The DnaJ-like zinc finger domain protein PSA2 affects light acclimation and chloroplast development in Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Yan-Wen eWang

    2016-03-01

    Full Text Available The biosynthesis of chlorophylls and carotenoids and the assembly of thylakoid membranes are critical for the photoautotrophic growth of plants. Different factors are involved in these two processes. In recent years, members of the DnaJ-like zinc finger domain proteins have been found to take part in the biogenesis and/or the maintenance of plastids. One member of this family of proteins, PSA2, was recently found to localize to the thylakoid lumen and regulate the accumulation of photosystem I. In this study, we report that the silencing of PSA2 in Arabidopsis thaliana resulted in variegated leaves and retarded growth. Although both chlorophylls and total carotenoids decreased in the psa2 mutant, violaxanthin and zeaxanthin accumulated in the mutant seedlings grown under growth condition. Lower levels of non-photochemical quenching and electron transport rate were also found in the psa2 mutant seedlings under growth condition compared with those of the wild-type plants, indicating an impaired capability to acclimate to normal light irradiance when PSA2 was silenced. Moreover, we also observed an abnormal assembly of grana thylakoids and poorly developed stroma thylakoids in psa2 chloroplasts. Taken together, our results demonstrate that PSA2 is a member of the DnaJ-like zinc finger domain protein family that affects light acclimation and chloroplast development.

  12. Automatic polymerase chain reaction product detection system for food safety monitoring using zinc finger protein fused to luciferase.

    Science.gov (United States)

    Yoshida, Wataru; Kezuka, Aki; Murakami, Yoshiyuki; Lee, Jinhee; Abe, Koichi; Motoki, Hiroaki; Matsuo, Takafumi; Shimura, Nobuaki; Noda, Mamoru; Igimi, Shizunobu; Ikebukuro, Kazunori

    2013-11-01

    An automatic polymerase chain reaction (PCR) product detection system for food safety monitoring using zinc finger (ZF) protein fused to luciferase was developed. ZF protein fused to luciferase specifically binds to target double stranded DNA sequence and has luciferase enzymatic activity. Therefore, PCR products that comprise ZF protein recognition sequence can be detected by measuring the luciferase activity of the fusion protein. We previously reported that PCR products from Legionella pneumophila and Escherichia coli (E. coli) O157 genomic DNA were detected by Zif268, a natural ZF protein, fused to luciferase. In this study, Zif268-luciferase was applied to detect the presence of Salmonella and coliforms. Moreover, an artificial zinc finger protein (B2) fused to luciferase was constructed for a Norovirus detection system. In the luciferase activity detection assay, several bound/free separation process is required. Therefore, an analyzer that automatically performed the bound/free separation process was developed to detect PCR products using the ZF-luciferase fusion protein. By means of the automatic analyzer with ZF-luciferase fusion protein, target pathogenic genomes were specifically detected in the presence of other pathogenic genomes. Moreover, we succeeded in the detection of 10 copies of E. coli BL21 without extraction of genomic DNA by the automatic analyzer and E. coli was detected with a logarithmic dependency in the range of 1.0×10 to 1.0×10(6) copies.

  13. Engineering HIV-Resistant Human CD4+ T Cells with CXCR4-Specific Zinc-Finger Nucleases

    Science.gov (United States)

    Wilen, Craig B.; Wang, Jianbin; Tilton, John C.; Miller, Jeffrey C.; Kim, Kenneth A.; Rebar, Edward J.; Sherrill-Mix, Scott A.; Patro, Sean C.; Secreto, Anthony J.; Jordan, Andrea P. O.; Lee, Gary; Kahn, Joshua; Aye, Pyone P.; Bunnell, Bruce A.; Lackner, Andrew A.; Hoxie, James A.; Danet-Desnoyers, Gwenn A.; Bushman, Frederic D.; Riley, James L.; Gregory, Philip D.; June, Carl H.; Holmes, Michael C.; Doms, Robert W.

    2011-01-01

    HIV-1 entry requires the cell surface expression of CD4 and either the CCR5 or CXCR4 coreceptors on host cells. Individuals homozygous for the ccr5Δ32 polymorphism do not express CCR5 and are protected from infection by CCR5-tropic (R5) virus strains. As an approach to inactivating CCR5, we introduced CCR5-specific zinc-finger nucleases into human CD4+ T cells prior to adoptive transfer, but the need to protect cells from virus strains that use CXCR4 (X4) in place of or in addition to CCR5 (R5X4) remains. Here we describe engineering a pair of zinc finger nucleases that, when introduced into human T cells, efficiently disrupt cxcr4 by cleavage and error-prone non-homologous DNA end-joining. The resulting cells proliferated normally and were resistant to infection by X4-tropic HIV-1 strains. CXCR4 could also be inactivated in ccr5Δ32 CD4+ T cells, and we show that such cells were resistant to all strains of HIV-1 tested. Loss of CXCR4 also provided protection from X4 HIV-1 in a humanized mouse model, though this protection was lost over time due to the emergence of R5-tropic viral mutants. These data suggest that CXCR4-specific ZFNs may prove useful in establishing resistance to CXCR4-tropic HIV for autologous transplant in HIV-infected individuals. PMID:21533216

  14. Engineering HIV-resistant human CD4+ T cells with CXCR4-specific zinc-finger nucleases.

    Directory of Open Access Journals (Sweden)

    Craig B Wilen

    2011-04-01

    Full Text Available HIV-1 entry requires the cell surface expression of CD4 and either the CCR5 or CXCR4 coreceptors on host cells. Individuals homozygous for the ccr5Δ32 polymorphism do not express CCR5 and are protected from infection by CCR5-tropic (R5 virus strains. As an approach to inactivating CCR5, we introduced CCR5-specific zinc-finger nucleases into human CD4+ T cells prior to adoptive transfer, but the need to protect cells from virus strains that use CXCR4 (X4 in place of or in addition to CCR5 (R5X4 remains. Here we describe engineering a pair of zinc finger nucleases that, when introduced into human T cells, efficiently disrupt cxcr4 by cleavage and error-prone non-homologous DNA end-joining. The resulting cells proliferated normally and were resistant to infection by X4-tropic HIV-1 strains. CXCR4 could also be inactivated in ccr5Δ32 CD4+ T cells, and we show that such cells were resistant to all strains of HIV-1 tested. Loss of CXCR4 also provided protection from X4 HIV-1 in a humanized mouse model, though this protection was lost over time due to the emergence of R5-tropic viral mutants. These data suggest that CXCR4-specific ZFNs may prove useful in establishing resistance to CXCR4-tropic HIV for autologous transplant in HIV-infected individuals.

  15. The RNA Binding Zinc Finger Protein Tristetraprolin Regulates AU-Rich mRNAs Involved in Breast Cancer-Related Processes

    OpenAIRE

    Al-Souhibani, Norah; Al-Ahmadi, Wijdan; Hesketh, John E.; Blackshear, Perry J.; Khabar, Khalid S.A.

    2010-01-01

    Tristetraprolin (TTP or ZFP36) is a tandem CCCH zinc finger RNA binding protein that regulates the stability of certain AU-rich mRNAs. Recent work suggests that TTP is deficient in cancer cells when compared to normal cell types. Here we found that TTP expression was lower in invasive breast cancer cells (MDA-MB-231) compared to normal breast cell lines, MCF12A and MCF-10. TTP targets were probed using a novel approach by expressing the C124R zinc finger TTP mutant that act as dominant negati...

  16. Structural and dynamical characterization of the Miz-1 zinc fingers 5-8 by solution-state NMR

    Energy Technology Data Exchange (ETDEWEB)

    Bernard, David; Bedard, Mikaeel; Bilodeau, Josee; Lavigne, Pierre, E-mail: pierre.lavigne@usherbrooke.ca [Universite de Sherbrooke, Departement de Biochimie, Faculte de Medecine et des Sciences de la Sante, Institut de Pharmacologie de Sherbrooke (Canada)

    2013-10-15

    Myc-interacting zinc finger protein-1 (Miz-1) is a BTB/POZ transcription factor that activates the transcription of cytostatic genes, such as p15{sup INK4B} or p21{sup CIP1}. The C-terminus of Miz-1 contains 13 consensus C{sub 2}H{sub 2} zinc finger domains (ZF). ZFs 1-4 have been shown to interact with SMAD3/4, while the remaining ZFs are expected to bind the promoters of target genes. We have noted unusual features in ZF 5 and the linker between ZFs 5 and 6. Indeed, a glutamate is found instead of the conserved basic residue two positions before the second zinc-coordinating histidine on the ZF 5 helix, and the linker sequence is DTDKE in place of the classical TGEKP sequence. In a canonical {beta}{beta}{alpha} fold, such unusual primary structure elements should cause severe electrostatic repulsions. In this context, we have characterized the structure and the dynamics of a Miz-1 construct comprising ZFs 5-8 (Miz 5-8) by solution-state NMR. Whilst ZFs 5, 7 and 8 were shown to adopt the classical {beta}{beta}{alpha} fold for C{sub 2}H{sub 2} ZFs, the number of long-range NOEs was insufficient to define a classical fold for ZF 6. We show by using {sup 15}N-relaxation dispersion experiments that this lack of NOEs is due to the presence of extensive motions on the {mu}s-ms timescale. Since this negatively charged region would have to be located near the phosphodiester backbone in a DNA complex, we propose that in addition to promoting conformational searches, it could serve as a hinge region to keep ZFs 1-4 away from DNA.

  17. Air-C2H2-O2 FAAS determination of tin in lead-calcium-tin Alloy%富氧火焰原子吸收光谱法测定铅钙锡合金中的锡

    Institute of Scientific and Technical Information of China (English)

    王嘉盛

    2003-01-01

    采用富氧火焰(Air-C2H2-O2)原子吸收光谱法直接测定铅钙锡合金中0.1%以上的锡.对仪器工作条件、测定体系酸度、共存离子干扰等作了研究.方法准确度高,结果重现性好.

  18. Arabidopsis VARIEGATED 3 encodes a chloroplast-targeted, zinc-finger protein required for chloroplast and palisade cell development

    DEFF Research Database (Denmark)

    Næsted, Henrik; Holm, Agnethe; Jenkins, Tom;

    2004-01-01

    The stable, recessive Arabidopsis variegated 3 (var3) mutant exhibits a variegated phenotype due to somatic areas lacking or containing developmentally retarded chloroplasts and greatly reduced numbers of palisade cells. The VAR3 gene, isolated by transposon tagging, encodes the 85.9 kDa VAR3...... protein containing novel repeats and zinc fingers described as protein interaction domains. VAR3 interacts specifically in yeast and in vitro with NCED4, a putative polyene chain or carotenoid dioxygenase, and both VAR3 and NCED4 accumulate in the chloroplast stroma. Metabolic profiling demonstrates...... that pigment profiles are qualitatively similar in wild type and var3, although var3 accumulates lower levels of chlorophylls and carotenoids. These results indicate that VAR3 is a part of a protein complex required for normal chloroplast and palisade cell development....

  19. Arabidopsis VARIEGATED 3 encodes a chloroplasttargeted, zinc-finger protein required for chloroplast and palisade cell development

    DEFF Research Database (Denmark)

    Næsted, Henrik; Holm, A.; Jenkins, T.;

    2004-01-01

    The stable, recessive Arabidopsis variegated 3 (var3) mutant exhibits a variegated phenotype due to somatic areas lacking or containing developmentally retarded chloroplasts and greatly reduced numbers of palisade cells. The VAR3 gene, isolated by transposon tagging, encodes the 85.9 kDa VAR3...... protein containing novel repeats and zinc fingers described as protein interaction domains. VAR3 interacts specifically in yeast and in vitro with NCED4, a putative polyene chain or carotenoid dioxygenase, and both VAR3 and NCED4 accumulate in the chloroplast stroma. Metabolic profiling demonstrates...... that pigment profiles are qualitatively similar in wild type and var3, although var3 accumulates lower levels of chlorophylls and carotenoids. These results indicate that VAR3 is a part of a protein complex required for normal chloroplast and palisade cell development....

  20. The Role of Cdkn1A-Interacting Zinc Finger Protein 1 (CIZ1 in DNA Replication and Pathophysiology

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    Qiang Liu

    2016-02-01

    Full Text Available Cdkn1A-interacting zinc finger protein 1 (CIZ1 was first identified in a yeast-2-hybrid system searching for interacting proteins of CDK2 inhibitor p21Cip1/Waf1. Ciz1 also binds to CDK2, cyclin A, cyclin E, CDC6, PCNA, TCF4 and estrogen receptor-α. Recent studies reveal numerous biological functions of CIZ1 in DNA replication, cell proliferation, and differentiation. In addition, splicing variants of CIZ1 mRNA is associated with a variety of cancers and Alzheimer’s disease, and mutations of the CIZ1 gene lead to cervical dystonia. CIZ1 expression is increased in cancers and rheumatoid arthritis. In this review, we will summarize the biological functions and molecular mechanisms of CIZ1 in these physiological and pathological processes.

  1. Sensitive and direct electrochemical detection of double-stranded DNA utilizing alkaline phosphatase-labelled zinc finger proteins.

    Science.gov (United States)

    Noh, Soodong; Ha, Dat Thinh; Yang, Haesik; Kim, Moon-Soo

    2015-06-21

    Direct detection of double-stranded DNA (dsDNA) using zinc finger proteins (ZFPs) is of great importance in biomedical applications such as identifying pathogens and circulating DNAs. However, its sensitivity is still not sufficiently high because limited signalling labels can be conjugated or fused. Herein, we report sensitive and direct detection of dsDNA using (i) alkaline phosphatase (ALP) as a fast catalytic label conjugated to ZFPs along with (ii) electrochemical measurement of an ALP product (l-ascorbic acid) at the indium-tin oxide electrode with a high signal-to-background ratio. ALP is simply conjugated to a ZFP through lysine residues in a ZFP purification tag, a maltose binding protein (MBP). Sandwich-type electrochemical detection of dsDNA allows a detection limit of ca. 100 fM without using DNA amplification. PMID:25969923

  2. Evolutionary expansion and divergence in a large family of primate-specific zinc finger transcription factor genes

    Energy Technology Data Exchange (ETDEWEB)

    Hamilton, A T; Huntley, S; Tran-Gyamfi, M; Baggott, D; Gordon, L; Stubbs, L

    2005-09-28

    Although most genes are conserved as one-to-one orthologs in different mammalian orders, certain gene families have evolved to comprise different numbers and types of protein-coding genes through independent series of gene duplications, divergence and gene loss in each evolutionary lineage. One such family encodes KRAB-zinc finger (KRAB-ZNF) genes, which are likely to function as transcriptional repressors. One KRAB-ZNF subfamily, the ZNF91 clade, has expanded specifically in primates to comprise more than 110 loci in the human genome, yielding large gene clusters in human chromosomes 19 and 7 and smaller clusters or isolated copies at other chromosomal locations. Although phylogenetic analysis indicates that many of these genes arose before the split between old world monkeys and new world monkeys, the ZNF91 subfamily has continued to expand and diversify throughout the evolution of apes and humans. The paralogous loci are distinguished by sequence divergence within their zinc finger arrays indicating a selection for proteins with different DNA binding specificities. RT-PCR and in situ hybridization data show that some of these ZNF genes can have tissue-specific expression patterns, however many KRAB-ZNFs that are near-ubiquitous could also be playing very specific roles in halting target pathways in all tissues except for a few, where the target is released by the absence of its repressor. The number of variant KRAB-ZNF proteins is increased not only because of the large number of loci, but also because many loci can produce multiple splice variants, which because of the modular structure of these genes may have separate and perhaps even conflicting regulatory roles. The lineage-specific duplication and rapid divergence of this family of transcription factor genes suggests a role in determining species-specific biological differences and the evolution of novel primate traits.

  3. TIF1alpha: a possible link between KRAB zinc finger proteins and nuclear receptors

    DEFF Research Database (Denmark)

    Le Douarin, B; You, J; Nielsen, Anders Lade;

    1998-01-01

    Ligand-induced gene activation by nuclear receptors (NRs) is thought to be mediated by transcriptional intermediary factors (TIFs), that interact with their ligand-dependent AF-2 activating domain. Included in the group of the putative AF-2 TIFs identified so far is TIF1alpha, a member of a new...... family of proteins which contains an N-terminal RBCC (RING finger-B boxes-coiled coil) motif and a C-terminal bromodomain preceded by a PHD finger. In addition to these conserved domains present in a number of transcriptional regulatory proteins, TIF1alpha was found to contain several protein......-protein interaction sites. Of these, one specifically interacts with NRs bound to their agonistic ligand and not with NR mutants that are defective in the AF-2 activity. Immediately adjacent to this 'NR box', TIF1alpha contains an interaction site for members of the chromatin organization modifier (chromo) family, HP...

  4. Chorioallantoic Fusion Defects and Embryonic Lethality Resulting from Disruption of Zfp36L1, a Gene Encoding a CCCH Tandem Zinc Finger Protein of the Tristetraprolin Family

    OpenAIRE

    Stumpo, Deborah J.; Byrd, Noah A.; Phillips, Ruth S.; Ghosh, Sanjukta; Maronpot, Robert R.; Castranio, Trisha; Meyers, Erik N.; Mishina, Yuji; Blackshear, Perry J.

    2004-01-01

    The mouse gene Zfp36L1 encodes zinc finger protein 36-like 1 (Zfp36L1), a member of the tristetraprolin (TTP) family of tandem CCCH finger proteins. TTP can bind to AU-rich elements within the 3′-untranslated regions of the mRNAs encoding tumor necrosis factor (TNF) and granulocyte-macrophage colony-stimulating factor (GM-CSF), leading to accelerated mRNA degradation. TTP knockout mice exhibit an inflammatory phenotype that is largely due to increased TNF secretion. Zfp36L1 has activities sim...

  5. Heat of Mixing and Solution of 1,1,2,2-Tetrachloroethane C2H2Cl4 + C6H10O Cyclohexanone (HMSD1111, LB3666_H)

    Science.gov (United States)

    Cibulka, I.; Fontaine, J.-C.; Sosnkowska-Kehiaian, K.; Kehiaian, H. V.

    This document is part of Subvolume C 'Binary Liquid Systems of Nonelectrolytes III' of Volume 26 'Heats of Mixing, Vapor-Liquid Equilibrium, and Volumetric Properties of Mixtures and Solutions' of Landolt-Börnstein Group IV 'Physical Chemistry'. It contains the Chapter 'heat of Mixing and Solution of 1,1,2,2-Tetrachloroethane C2H2Cl4 + C6H10O Cyclohexanone (HMSD1111, LB3666_H)' providing data from direct low-pressure calorimetric measurement of molar excess enthalpy at variable mole fraction and constant temperature.

  6. Proviral HIV-genome-wide and pol-gene specific Zinc Finger Nucleases: Usability for targeted HIV gene therapy

    Directory of Open Access Journals (Sweden)

    Wayengera Misaki

    2011-07-01

    Full Text Available Abstract Background Infection with HIV, which culminates in the establishment of a latent proviral reservoir, presents formidable challenges for ultimate cure. Building on the hypothesis that ex-vivo or even in-vivo abolition or disruption of HIV-gene/genome-action by target mutagenesis or excision can irreversibly abrogate HIV's innate fitness to replicate and survive, we previously identified the isoschizomeric bacteria restriction enzymes (REases AcsI and ApoI as potent cleavers of the HIV-pol gene (11 and 9 times in HIV-1 and 2, respectively. However, both enzymes, along with others found to cleave across the entire HIV-1 genome, slice (SX at palindromic sequences that are prevalent within the human genome and thereby pose the risk of host genome toxicity. A long-term goal in the field of R-M enzymatic therapeutics has thus been to generate synthetic restriction endonucleases with longer recognition sites limited in specificity to HIV. We aimed (i to assemble and construct zinc finger arrays and nucleases (ZFN with either proviral-HIV-pol gene or proviral-HIV-1 whole-genome specificity respectively, and (ii to advance a model for pre-clinically testing lentiviral vectors (LV that deliver and transduce either ZFN genotype. Methods and Results First, we computationally generated the consensus sequences of (a 114 dsDNA-binding zinc finger (Zif arrays (ZFAs or ZifHIV-pol and (b two zinc-finger nucleases (ZFNs which, unlike the AcsI and ApoI homeodomains, possess specificity to >18 base-pair sequences uniquely present within the HIV-pol gene (ZifHIV-polFN. Another 15 ZFNs targeting >18 bp sequences within the complete HIV-1 proviral genome were constructed (ZifHIV-1FN. Second, a model for constructing lentiviral vectors (LVs that deliver and transduce a diploid copy of either ZifHIV-polFN or ZifHIV-1FN chimeric genes (termed LV- 2xZifHIV-polFN and LV- 2xZifHIV-1FN, respectively is proposed. Third, two preclinical models for controlled testing of

  7. Molecular Characterization of the Schistosoma mansoni Zinc Finger Protein SmZF1 as a Transcription Factor

    Science.gov (United States)

    D'Astolfo, Diego S.; Cardoso, Fernanda C.; Rajão, Matheus A.; Mourão, Marina M.; Gava, Elisandra; Oliveira, Sérgio C.; Macedo, Andréa M.; Machado, Carlos R.; Pena, Sérgio D. J.; Kitten, Gregory T.; Franco, Glória R.

    2009-01-01

    Background During its development, the parasite Schistosoma mansoni is exposed to different environments and undergoes many morphological and physiological transformations as a result of profound changes in gene expression. Characterization of proteins involved in the regulation of these processes is of importance for the understanding of schistosome biology. Proteins containing zinc finger motifs usually participate in regulatory processes and are considered the major class of transcription factors in eukaryotes. It has already been shown, by EMSA (Eletrophoretic Mobility Shift Assay), that SmZF1, a S. mansoni zinc finger (ZF) protein, specifically binds both DNA and RNA oligonucleotides. This suggests that this protein might act as a transcription factor in the parasite. Methodology/Principal Findings In this study we extended the characterization of SmZF1 by determining its subcellular localization and by verifying its ability to regulate gene transcription. We performed immunohistochemistry assays using adult male and female worms, cercariae and schistosomula to analyze the distribution pattern of SmZF1 and verified that the protein is mainly detected in the cells nuclei of all tested life cycle stages except for adult female worms. Also, SmZF1 was heterologously expressed in mammalian COS-7 cells to produce the recombinant protein YFP-SmZF1, which was mainly detected in the nucleus of the cells by confocal microscopy and Western blot assays. To evaluate the ability of this protein to regulate gene transcription, cells expressing YFP-SmZF1 were tested in a luciferase reporter system. In this system, the luciferase gene is downstream of a minimal promoter, upstream of which a DNA region containing four copies of the SmZF1 putative best binding site (D1-3DNA) was inserted. SmZF1 increased the reporter gene transcription by two fold (p≤0.003) only when its specific binding site was present. Conclusion Taken together, these results strongly support the hypothesis

  8. Structural and functional organization of the HF.10 human zinc finger gene (ZNF35) located on chromosome 3p21-p22

    DEFF Research Database (Denmark)

    Lanfrancone, L; Pengue, G; Pandolfi, P P;

    1992-01-01

    We report the structural and functional characterization of the HF.10 zinc finger gene (ZNF35) in normal human cells, as well as a processed pseudogene. The HF.10 gene spans about 13 kb and it is interrupted by three introns. All 11 zinc finger DNA-binding domains are contiguously encoded within...... and partial nucleotide sequencing of the HF.10 pseudogene indicated that it has arisen by retroposition of spliced HF.10 mRNA. In situ hybridization experiments revealed that both the functional locus and the pseudogene map to chromosome 3p21p22, a region that is frequently deleted in small cell lung...... and renal carcinomas. Hybridization of the HF.10 gene and the HF.10 pseudogene DNA probes to metaphases from a small cell lung carcinoma cell line with the 3p deletion revealed that both loci are part of the deleted chromosome region....

  9. 乙炔与H2O分子之间的氢键结构与性质%The structures and properties of the hydrogen bond interactions between C2H2 and H2O

    Institute of Scientific and Technical Information of China (English)

    刘艳芝; 袁煜; 吕玲玲; 李志锋; 朱元成

    2011-01-01

    The full geometry optimizations for H2O… C2H2 complexes were carried out using the method of DFT B3LYP at the 6 - 311 ++ G** level. And the σ-n and H-π type hydrogen bond complexes were obtained. Frequency analysis showed that the IR spectrum of the related convent bond occurs red shift. The charge transfer has been discussed via natural bond orbit theory (NBO). The results of the interaction energies showed that the σ-n type is more stable than the H-π type hydrogen bond complex.%采用B3LYP方法,在6-311++G水平上优化得到了H2O…C2H2氢键复合物的σ-n和H-π型两种稳定构型,并进行频率分析,讨论了相关自然键红外振动光谱的红移现象.采用NBO理论对σ-n和H-π氢键复合物形成过程中的电荷转移的类型进行了分析讨论.分子间的氢键相互作用能结果表明,σ-n型比H-π型氢键复合物更稳定.

  10. Overexpression of a zinc-finger protein gene from rice confers tolerance to cold, dehydration, and salt stress in transgenic tobacco

    OpenAIRE

    Mukhopadhyay, Arnab; Vij, Shubha; Tyagi, Akhilesh K

    2004-01-01

    Stress perception and signal transduction leading to tolerance involve a complex interplay of different gene products. We describe here the isolation and characterization of an intronless gene (OSISAP1) from rice encoding a zinc-finger protein that is induced after different types of stresses, namely cold, desiccation, salt, submergence, and heavy metals as well as injury. The gene is also induced by stress hormone abscisic acid. Overexpression of the gene in transgenic tobacco conferred tole...

  11. Localization and Differential Expression of the Krüppel-Associated Box Zinc Finger Proteins 1 and 54 in Early Mouse Development

    DEFF Research Database (Denmark)

    Albertsen, Maria; Teperek, Marta; Elholm, Grethe;

    2010-01-01

    -fused reporter gene into zygotes demonstrated the intracellular distribution of ZFP1-green fluorescent protein (GFP) and ZFP54-GFP colocalized with a DNA marker in the two-cell embryo. The KRAB domain was essential to colocalize with DNA, and deletion of the KRAB domain in ZFP1-GFP and ZFP54-GFP localized...... transcriptional repressors, zinc finger protein (ZFP1) and ZFP54, belonging to the Krüppel-associated box (KRAB) zinc finger family, were isolated. ZFP1 and ZFP54 contain an N-terminally located KRAB repressor domain followed by 8 and 12 repeats of Krüppel zinc-finger motifs, respectively. Reverse transcription...... (RT) and quantitative (q) PCR show that maternally contributed Zfp1 and Zfp54 mRNA are detected throughout preimplantation development. α-Amanitin-treated zygotes revealed that maternal Zfp1 and Zfp54 are fully degraded at the two-cell stage. Microinjections of in vitro-transcribed mRNA encoding a gfp...

  12. Zinc Finger Domain of the PRDM9 Gene on Chromosome 1 Exhibits High Diversity in Ruminants but Its Paralog PRDM7 Contains Multiple Disruptive Mutations

    Science.gov (United States)

    Ahlawat, Sonika; Sharma, Priyanka; Sharma, Rekha; Arora, Reena; De, Sachinandan

    2016-01-01

    PRDM9 is the sole hybrid sterility gene identified so far in vertebrates. PRDM9 gene encodes a protein with an immensely variable zinc-finger (ZF) domain that determines the site of meiotic recombination hotspots genome-wide. In this study, the terminal ZF domain of PRDM9 on bovine chromosome 1 and its paralog on chromosome 22 were characterized in 225 samples from five ruminant species (cattle, yak, mithun, sheep and goat). We found extraordinary variation in the number of PRDM9 zinc fingers (6 to 12). We sequenced PRDM9 ZF encoding region from 15 individuals (carrying the same ZF number in both copies) and found 43 different ZF domain sequences. Ruminant zinc fingers of PRDM9 were found to be diversifying under positive selection and concerted evolution, specifically at positions involved in defining their DNA-binding specificity, consistent with the reports from other vertebrates such as mice, humans, equids and chimpanzees. ZF-encoding regions of the PRDM7, a paralog of PRDM9 on bovine chromosome 22 and on unknown chromosomes in other studied species were found to contain 84 base repeat units as in PRDM9, but there were multiple disruptive mutations after the first repeat unit. The diversity of the ZFs suggests that PRDM9 may activate recombination hotspots that are largely unique to each ruminant species. PMID:27203728

  13. Zinc Finger Domain of the PRDM9 Gene on Chromosome 1 Exhibits High Diversity in Ruminants but Its Paralog PRDM7 Contains Multiple Disruptive Mutations.

    Science.gov (United States)

    Ahlawat, Sonika; Sharma, Priyanka; Sharma, Rekha; Arora, Reena; De, Sachinandan

    2016-01-01

    PRDM9 is the sole hybrid sterility gene identified so far in vertebrates. PRDM9 gene encodes a protein with an immensely variable zinc-finger (ZF) domain that determines the site of meiotic recombination hotspots genome-wide. In this study, the terminal ZF domain of PRDM9 on bovine chromosome 1 and its paralog on chromosome 22 were characterized in 225 samples from five ruminant species (cattle, yak, mithun, sheep and goat). We found extraordinary variation in the number of PRDM9 zinc fingers (6 to 12). We sequenced PRDM9 ZF encoding region from 15 individuals (carrying the same ZF number in both copies) and found 43 different ZF domain sequences. Ruminant zinc fingers of PRDM9 were found to be diversifying under positive selection and concerted evolution, specifically at positions involved in defining their DNA-binding specificity, consistent with the reports from other vertebrates such as mice, humans, equids and chimpanzees. ZF-encoding regions of the PRDM7, a paralog of PRDM9 on bovine chromosome 22 and on unknown chromosomes in other studied species were found to contain 84 base repeat units as in PRDM9, but there were multiple disruptive mutations after the first repeat unit. The diversity of the ZFs suggests that PRDM9 may activate recombination hotspots that are largely unique to each ruminant species. PMID:27203728

  14. Zinc Finger Domain of the PRDM9 Gene on Chromosome 1 Exhibits High Diversity in Ruminants but Its Paralog PRDM7 Contains Multiple Disruptive Mutations.

    Directory of Open Access Journals (Sweden)

    Sonika Ahlawat

    Full Text Available PRDM9 is the sole hybrid sterility gene identified so far in vertebrates. PRDM9 gene encodes a protein with an immensely variable zinc-finger (ZF domain that determines the site of meiotic recombination hotspots genome-wide. In this study, the terminal ZF domain of PRDM9 on bovine chromosome 1 and its paralog on chromosome 22 were characterized in 225 samples from five ruminant species (cattle, yak, mithun, sheep and goat. We found extraordinary variation in the number of PRDM9 zinc fingers (6 to 12. We sequenced PRDM9 ZF encoding region from 15 individuals (carrying the same ZF number in both copies and found 43 different ZF domain sequences. Ruminant zinc fingers of PRDM9 were found to be diversifying under positive selection and concerted evolution, specifically at positions involved in defining their DNA-binding specificity, consistent with the reports from other vertebrates such as mice, humans, equids and chimpanzees. ZF-encoding regions of the PRDM7, a paralog of PRDM9 on bovine chromosome 22 and on unknown chromosomes in other studied species were found to contain 84 base repeat units as in PRDM9, but there were multiple disruptive mutations after the first repeat unit. The diversity of the ZFs suggests that PRDM9 may activate recombination hotspots that are largely unique to each ruminant species.

  15. Multiple interferon stimulated genes synergize with the zinc finger antiviral protein to mediate anti-alphavirus activity.

    Directory of Open Access Journals (Sweden)

    Sophiya Karki

    Full Text Available The zinc finger antiviral protein (ZAP is a host factor that mediates inhibition of viruses in the Filoviridae, Retroviridae and Togaviridae families. We previously demonstrated that ZAP blocks replication of Sindbis virus (SINV, the prototype Alphavirus in the Togaviridae family at an early step prior to translation of the incoming genome and that synergy between ZAP and one or more interferon stimulated genes (ISGs resulted in maximal inhibitory activity. The present study aimed to identify those ISGs that synergize with ZAP to mediate Alphavirus inhibition. Using a library of lentiviruses individually expressing more than 350 ISGs, we screened for inhibitory activity in interferon defective cells with or without ZAP overexpression. Confirmatory tests of the 23 ISGs demonstrating the largest infection reduction in combination with ZAP revealed that 16 were synergistic. Confirmatory tests of all potentially synergistic ISGs revealed 15 additional ISGs with a statistically significant synergistic effect in combination with ZAP. These 31 ISGs are candidates for further mechanistic studies. The number and diversity of the identified ZAP-synergistic ISGs lead us to speculate that ZAP may play an important role in priming the cell for optimal ISG function.

  16. Conversion of Human Fibroblasts to Stably Self-Renewing Neural Stem Cells with a Single Zinc-Finger Transcription Factor

    Directory of Open Access Journals (Sweden)

    Ebrahim Shahbazi

    2016-04-01

    Full Text Available Direct conversion of somatic cells into neural stem cells (NSCs by defined factors holds great promise for mechanistic studies, drug screening, and potential cell therapies for different neurodegenerative diseases. Here, we report that a single zinc-finger transcription factor, Zfp521, is sufficient for direct conversion of human fibroblasts into long-term self-renewable and multipotent NSCs. In vitro, Zfp521-induced NSCs maintained their characteristics in the absence of exogenous factor expression and exhibited morphological, molecular, developmental, and functional properties that were similar to control NSCs. In addition, the single-seeded induced NSCs were able to form NSC colonies with efficiency comparable with control NSCs and expressed NSC markers. The converted cells were capable of surviving, migrating, and attaining neural phenotypes after transplantation into neonatal mouse and adult rat brains, without forming tumors. Moreover, the Zfp521-induced NSCs predominantly expressed rostral genes. Our results suggest a facilitated approach for establishing human NSCs through Zfp521-driven conversion of fibroblasts.

  17. Mutagenesis of genes for starch debranching enzyme isoforms in pea by means of zinc-finger endonucleases

    International Nuclear Information System (INIS)

    Starch debranching enzymes in plants are divided into two groups based on their ability to hydrolyse different substrates. The first group, pullulanases, hydrolyses α-1,6-glucosidic linkages in substrates such as pullulan, amylopectin and glycogen. The second group of debranching enzymes, isoamylases, hydrolyse glycogen and amylopectin and are not active on pullulan. Three isoforms of isoamylase and a pullulanase have been isolated from cDNA library of Pisum sativum. These isoamylases have been characterised based on the their heterologous expression in E coli. Based on the DNA sequence that encodes these debranching enzyme, a specific mutagenesis targeting at these DNA will be attempted. The method that will be employed are based on the techniques developed by Wright et al. (2005). This technique involves the homologous recombination of DNA that is mediated by zinc-finger endonucleases. Vectors will be constructed to include a fragment that will modify these genes. Microinjection technique will be used to insert these vectors into pollen which then will be fertilized. Using this technique, it is hoped that null mutant for each enzyme will be created and the exact role of these enzymes for the synthesis and degradation of starch in plants will be elucidate. (author)

  18. Suppression of zinc finger protein 467 alleviates osteoporosis through promoting differentiation of adipose derived stem cells to osteoblasts

    Directory of Open Access Journals (Sweden)

    You Li

    2012-01-01

    Full Text Available Abstract Osteoblast and adipocyte are derived from common mesenchymal progenitor cells. The bone loss of osteoporosis is associated with altered progenitor differentiation from an osteoblastic to an adipocytic lineage. In this study, a comparative analysis of gene expression profiling using cDNA microarray and realtime-PCR indicated that Zinc finger protein 467 (Zfp467 involved in adipocyte and osteoblast differentiation of cultured adipose derived stem cells (ADSCs. Our results showed that RNA interference for Zfp467 in ADSCs inhibited adipocyte formation and stimulated osteoblast commitment. The mRNA levels of osteogenic and adipogenic markers in ADSCs were regulated by si-Zfp467. Zfp467 RNAi in ADSCs could restore bone function and structure in an ovariectomized (OVX-induced osteoporotic mouse model. Thus Zfp467 play an important role in ADSCs differentiation to adipocyte and osteoblast. This has relevance to therapeutic interventions in osteoporosis, including si-Zfp467-based therapies currently available, and may be of relevance for the use of adipose-derived stem cells for tissue engineering.

  19. The Transcriptional Repressive Activity of KRAB Zinc Finger Proteins Does Not Correlate with Their Ability to Recruit TRIM28

    Science.gov (United States)

    Murphy, Kristin E.; Shylo, Natalia A.; Alexander, Katherine A.; Churchill, Angela J.; Copperman, Cecilia; García-García, María J.

    2016-01-01

    KRAB domain Zinc finger proteins are one of the most abundant families of transcriptional regulators in higher vertebrates. The prevailing view is that KRAB domain proteins function as potent transcriptional repressors by recruiting TRIM28 and promoting heterochromatin spreading. However, the extent to which all KRAB domain proteins are TRIM28-dependent transcriptional repressors is currently unclear. Our studies on mouse ZFP568 revealed that TRIM28 recruitment by KRAB domain proteins is not sufficient to warrant transcriptional repressive activity. By using luciferase reporter assays and yeast two-hybrid experiments, we tested the ability of ZFP568 and other mouse KRAB domain proteins to repress transcription and bind TRIM28. We found that some mouse KRAB domain proteins are poor transcriptional repressors despite their ability to recruit TRIM28, while others showed strong KRAB-dependent transcriptional repression, but no TRIM28 binding. Together, our results show that the transcriptional repressive activity of KRAB-ZNF proteins does not correlate with their ability to recruit TRIM28, and provide evidence that KRAB domains can regulate transcription in a TRIM28-independent fashion. Our findings challenge the current understanding of the molecular mechanisms used by KRAB domain proteins to control gene expression and highlight that a high percentage of KRAB domain proteins in the mouse genome differ from the consensus KRAB sequence at amino acid residues that are critical for TRIM28 binding and/or repressive activity. PMID:27658112

  20. Zinc Finger Homeodomain Factor Zfhx3 Is Essential for Mammary Lactogenic Differentiation by Maintaining Prolactin Signaling Activity.

    Science.gov (United States)

    Zhao, Dan; Ma, Gui; Zhang, Xiaolin; He, Yuan; Li, Mei; Han, Xueying; Fu, Liya; Dong, Xue-Yuan; Nagy, Tamas; Zhao, Qiang; Fu, Li; Dong, Jin-Tang

    2016-06-10

    The zinc finger homeobox 3 (ZFHX3, also named ATBF1 for AT motif binding factor 1) is a transcription factor that suppresses prostatic carcinogenesis and induces neuronal differentiation. It also interacts with estrogen receptor α to inhibit cell proliferation and regulate pubertal mammary gland development in mice. In the present study, we examined whether and how Zfhx3 regulates lactogenic differentiation in mouse mammary glands. At different stages of mammary gland development, Zfhx3 protein was expressed at varying levels, with the highest level at lactation. In the HC11 mouse mammary epithelial cell line, an in vitro model of lactogenesis, knockdown of Zfhx3 attenuated prolactin-induced β-casein expression and morphological changes, indicators of lactogenic differentiation. In mouse mammary tissue, knock-out of Zfhx3 interrupted lactogenesis, resulting in underdeveloped glands with much smaller and fewer alveoli, reduced β-casein expression, accumulation of large cytoplasmic lipid droplets in luminal cells after parturition, and failure in lactation. Mechanistically, Zfhx3 maintained the expression of Prlr (prolactin receptor) and Prlr-Jak2-Stat5 signaling activity, whereas knockdown and knock-out of Zfhx3 in HC11 cells and mammary tissues, respectively, decreased Prlr expression, Stat5 phosphorylation, and the expression of Prlr-Jak2-Stat5 target genes. These findings indicate that Zfhx3 plays an essential role in proper lactogenic development in mammary glands, at least in part by maintaining Prlr expression and Prlr-Jak2-Stat5 signaling activity. PMID:27129249

  1. Generation of PPARγ mono-allelic knockout pigs via zinc-finger nucleases and nuclear transfer cloning

    Institute of Scientific and Technical Information of China (English)

    Dongshan Yang; Jiangtian Tian; Feng Li; Jifeng Zhang; Lin Chang; Duanqing Pei; Y Eugene Chen; Liangxue Lai; Huaqiang Yang; Wei Li; Bentian Zhao; Zhen Ouyang; Zhaoming Liu; Yu Zhao; Nana Fan; Jun Song

    2011-01-01

    @@ Dear Editor, Gene targeting in mouse embryonic stem (ES) cells has revolutionized the field of mouse genetics and allowed for the analysis of diverse aspects of gene function in vivo.For more than two decades,researchers have been actively searching for ES cells from large animals such as pigs and cattle.Unfortunately,to date,no ES cell lines from large animals have passed the crucial test of germ line contribution.The sole method of gene targeting to date in these species remains somatic cell nuclear transfer (SCNT) combined with DNA homologous recombination (HR).Due to the limited proliferation competency and extremely low frequency of HR in somatic cells (less than 10-6),this process is highly inefficient and only a few successful examples have been achieved,even though enrichment strategies such as positivenegative marker selection,promoter-trap and adenoassociated viral delivery were previously used [1-3].The low efficiency of gene targeting in cultured somatic cells is the main barrier for gene targeting in large animals.Recently,zinc-finger nuclease (ZFN) technology has emerged as a powerful tool for genome editing.The success of ZFN technology for gene targeting in fruit flies,zebra fish,rodents as well as human cell lines encouraged us to establish a high-efficiency gene-targeting platform in large animals such as pigs [4-8].

  2. Covariance mapping of two-photon double core hole states in C 2 H 2 and C 2 H 6 produced by an x-ray free electron laser

    International Nuclear Information System (INIS)

    Few-photon ionization and relaxation processes in acetylene (C2H2) and ethane (C2H6) were investigated at the linac coherent light source x-ray free electron laser (FEL) at SLAC, Stanford using a highly efficient multi-particle correlation spectroscopy technique based on a magnetic bottle. The analysis method of covariance mapping has been applied and enhanced, allowing us to identify electron pairs associated with double core hole (DCH) production and competing multiple ionization processes including Auger decay sequences. The experimental technique and the analysis procedure are discussed in the light of earlier investigations of DCH studies carried out at the same FEL and at third generation synchrotron radiation sources. In particular, we demonstrate the capability of the covariance mapping technique to disentangle the formation of molecular DCH states which is barely feasible with conventional electron spectroscopy methods

  3. H2, He, and CO2 line-broadening coefficients, pressure shifts and temperature-dependence exponents for the HITRAN database. Part 1: SO2, NH3, HF, HCl, OCS and C2H2

    Science.gov (United States)

    Wilzewski, Jonas S.; Gordon, Iouli E.; Kochanov, Roman V.; Hill, Christian; Rothman, Laurence S.

    2016-01-01

    To increase the potential for use of the HITRAN database in astronomy, experimental and theoretical line-broadening coefficients, line shifts and temperature-dependence exponents of molecules of planetary interest broadened by H2, He, and CO2 have been assembled from available peer-reviewed sources. The collected data were used to create semi-empirical models so that every HITRAN line of the studied molecules has corresponding parameters. Since H2 and He are major constituents in the atmospheres of gas giants, and CO2 predominates in atmospheres of some rocky planets with volcanic activity, these spectroscopic data are important for remote sensing studies of planetary atmospheres. In this paper we make the first step in assembling complete sets of these parameters, thereby creating datasets for SO2, NH3, HF, HCl, OCS and C2H2.

  4. Role of Bmznf-2, a Bombyx mori CCCH zinc finger gene, in masculinisation and differential splicing of Bmtra-2.

    Science.gov (United States)

    Gopinath, Gajula; Arunkumar, Kallare P; Mita, Kazuei; Nagaraju, Javaregowda

    2016-08-01

    Deciphering the regulatory factors involved in Bombyx mori sex determination has been a puzzle, challenging researchers for nearly a century now. The pre-mRNA of B. mori doublesex (Bmdsx), a master regulator gene of sexual differentiation, is differentially spliced, producing Bmdsxm and Bmdsxf transcripts in males and females respectively. The putative proteins encoded by these differential transcripts orchestrate antagonistic functions, which lead to sexual differentiation. A recent study in B. mori illustrated the role of a W-derived fem piRNA in conferring femaleness. In females, the fem piRNA was shown to suppress the activity of a Z-linked CCCH type zinc finger (znf) gene, Masculiniser (masc), which indirectly promotes the Bmdsxm type of splicing. In this study, we report a novel autosomal (Chr 25) CCCH type znf motif encoding gene Bmznf-2 as one of the potential factors in the Bmdsx sex specific differential splicing, and we also provide insights into its role in the alternative splicing of Bmtra2 by using ovary derived BmN cells. Over-expression of Bmznf-2 induced Bmdsxm type of splicing (masculinisation) with a correspondingly reduced expression of Bmdsxf type isoform in BmN cells. Further, the site-directed mutational studies targeting the tandem CCCH znf motifs revealed their indispensability in the observed phenotype of masculinisation. Additionally, the dual luciferase assays in BmN cells using 5' UTR region of the Bmznf-2 strongly implied the existence of a translational repression over this gene. From these findings, we propose Bmznf-2 to be one of the potential factors of masculinisation similar to Masc. From the growing number of Bmdsx splicing regulators, we assume that the sex determination cascade of B. mori is quite intricate in nature; hence, it has to be further investigated for its comprehensive understanding. PMID:27260399

  5. Promyelocytic leukemia zinc finger protein activates GATA4 transcription and mediates cardiac hypertrophic signaling from angiotensin II receptor 2.

    Directory of Open Access Journals (Sweden)

    Ning Wang

    Full Text Available BACKGROUND: Pressure overload and prolonged angiotensin II (Ang II infusion elicit cardiac hypertrophy in Ang II receptor 1 (AT(1 null mouse, whereas Ang II receptor 2 (AT(2 gene deletion abolishes the hypertrophic response. The roles and signals of the cardiac AT(2 receptor still remain unsettled. Promyelocytic leukemia zinc finger protein (PLZF was shown to bind to the AT(2 receptor and transmit the hypertrophic signal. Using PLZF knockout mice we directed our studies on the function of PLZF concerning the cardiac specific transcription factor GATA4, and GATA4 targets. METHODOLOGY AND PRINCIPAL FINDINGS: PLZF knockout and age-matched wild-type (WT mice were treated with Ang II, infused at a rate of 4.2 ng·kg(-1·min(-1 for 3 weeks. Ang II elevated systolic blood pressure to comparable levels in PLZF knockout and WT mice (140 mmHg. WT mice developed prominent cardiac hypertrophy and fibrosis after Ang II infusion. In contrast, there was no obvious cardiac hypertrophy or fibrosis in PLZF knockout mice. An AT(2 receptor blocker given to Ang II-infused wild type mice prevented hypertrophy, verifying the role of AT(2 receptor for cardiac hypertrophy. Chromatin immunoprecipitation and electrophoretic mobility shift assay showed that PLZF bound to the GATA4 gene regulatory region. A Luciferase assay verified that PLZF up-regulated GATA4 gene expression and the absence of PLZF expression in vivo produced a corresponding repression of GATA4 protein. CONCLUSIONS: PLZF is an important AT(2 receptor binding protein in mediating Ang II induced cardiac hypertrophy through an AT(2 receptor-dependent signal pathway. The angiotensin II-AT(2-PLZF-GATA4 signal may further augment Ang II induced pathological effects on cardiomyocytes.

  6. The zinc finger transcription factor SlZFP2 negatively regulates abscisic acid biosynthesis and fruit ripening in tomato.

    Science.gov (United States)

    Weng, Lin; Zhao, Fangfang; Li, Rong; Xu, Changjie; Chen, Kunsong; Xiao, Han

    2015-03-01

    Abscisic acid (ABA) regulates plant development and adaptation to environmental conditions. Although the ABA biosynthesis pathway in plants has been thoroughly elucidated, how ABA biosynthetic genes are regulated at the molecular level during plant development is less well understood. Here, we show that the tomato (Solanum lycopersicum) zinc finger transcription factor SlZFP2 is involved in the regulation of ABA biosynthesis during fruit development. Overexpression of SlZFP2 resulted in multiple phenotypic changes, including more branches, early flowering, delayed fruit ripening, lighter seeds, and faster seed germination, whereas down-regulation of its expression caused problematic fruit set, accelerated ripening, and inhibited seed germination. SlZFP2 represses ABA biosynthesis during fruit development through direct suppression of the ABA biosynthetic genes NOTABILIS, SITIENS, and FLACCA and the aldehyde oxidase SlAO1. We also show that SlZFP2 regulates fruit ripening through transcriptional suppression of the ripening regulator COLORLESS NON-RIPENING. Using bacterial one-hybrid screening and a selected amplification and binding assay, we identified the (A/T)(G/C)TT motif as the core binding sequence of SlZFP2. Furthermore, by RNA sequencing profiling, we found that 193 genes containing the SlZFP2-binding motifs in their promoters were differentially expressed in 2 d post anthesis fruits between the SlZFP2 RNA interference line and its nontransgenic sibling. We propose that SlZFP2 functions as a repressor to fine-tune ABA biosynthesis during fruit development and provides a potentially valuable tool for dissecting the role of ABA in fruit ripening.

  7. Probing isoform-specific functions of polypeptide GalNAc-transferases using zinc finger nuclease glycoengineered SimpleCells.

    Science.gov (United States)

    Schjoldager, Katrine T-B G; Vakhrushev, Sergey Y; Kong, Yun; Steentoft, Catharina; Nudelman, Aaron S; Pedersen, Nis B; Wandall, Hans H; Mandel, Ulla; Bennett, Eric P; Levery, Steven B; Clausen, Henrik

    2012-06-19

    Our knowledge of the O-glycoproteome [N-acetylgalactosamine (GalNAc) type] is highly limited. The O-glycoproteome is differentially regulated in cells by dynamic expression of a subset of 20 polypeptide GalNAc-transferases (GalNAc-Ts), and methods to identify important functions of individual GalNAc-Ts are largely unavailable. We recently introduced SimpleCells, i.e., human cell lines made deficient in O-glycan extension by zinc finger nuclease targeting of a key gene in O-glycan elongation (Cosmc), which allows for proteome-wide discovery of O-glycoproteins. Here we have extended the SimpleCell concept to include proteome-wide discovery of unique functions of individual GalNAc-Ts. We used the GalNAc-T2 isoform implicated in dyslipidemia and the human HepG2 liver cell line to demonstrate unique functions of this isoform. We confirm that GalNAc-T2-directed site-specific O-glycosylation inhibits proprotein activation of the lipase inhibitor ANGPTL3 in HepG2 cells and further identify eight O-glycoproteins exclusively glycosylated by T2 of which one, ApoC-III, is implicated in dyslipidemia. Our study supports an essential role for GalNAc-T2 in lipid metabolism, provides serum biomarkers for GalNAc-T2 enzyme function, and validates the use of GALNT gene targeting with SimpleCells for broad discovery of disease-causing deficiencies in O-glycosylation. The presented glycoengineering strategy opens the way for proteome-wide discovery of functions of GalNAc-T isoforms and their role in congenital diseases and disorders.

  8. The rice B-box zinc finger gene family: genomic identification, characterization, expression profiling and diurnal analysis.

    Directory of Open Access Journals (Sweden)

    Jianyan Huang

    Full Text Available BACKGROUND: The B-box (BBX -containing proteins are a class of zinc finger proteins that contain one or two B-box domains and play important roles in plant growth and development. The Arabidopsis BBX gene family has recently been re-identified and renamed. However, there has not been a genome-wide survey of the rice BBX (OsBBX gene family until now. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we identified 30 rice BBX genes through a comprehensive bioinformatics analysis. Each gene was assigned a uniform nomenclature. We described the chromosome localizations, gene structures, protein domains, phylogenetic relationship, whole life-cycle expression profile and diurnal expression patterns of the OsBBX family members. Based on the phylogeny and domain constitution, the OsBBX gene family was classified into five subfamilies. The gene duplication analysis revealed that only chromosomal segmental duplication contributed to the expansion of the OsBBX gene family. The expression profile of the OsBBX genes was analyzed by Affymetrix GeneChip microarrays throughout the entire life-cycle of rice cultivar Zhenshan 97 (ZS97. In addition, microarray analysis was performed to obtain the expression patterns of these genes under light/dark conditions and after three phytohormone treatments. This analysis revealed that the expression patterns of the OsBBX genes could be classified into eight groups. Eight genes were regulated under the light/dark treatments, and eleven genes showed differential expression under at least one phytohormone treatment. Moreover, we verified the diurnal expression of the OsBBX genes using the data obtained from the Diurnal Project and qPCR analysis, and the results indicated that many of these genes had a diurnal expression pattern. CONCLUSIONS/SIGNIFICANCE: The combination of the genome-wide identification and the expression and diurnal analysis of the OsBBX gene family should facilitate additional functional studies of the Os

  9. The role of Zic family zinc finger transcription factors in the proliferation and differentiation of retinal progenitor cells

    International Nuclear Information System (INIS)

    Highlights: ► Zic transcription factors expressed early retinal progenitor cells. ► Zics sustain proliferation activity of retinal progenitor cells. ► Overexpression of Zic in retinal progenitors perturbed rod differentiation. ► Fate determination to rod photoreceptor was not affected. -- Abstract: Members of the Zic family of zinc finger transcription factors play critical roles in a variety of developmental processes. Using DNA microarray analysis, we found that Zics are strongly expressed in SSEA-1-positive early retinal progenitors in the peripheral region of the mouse retina. Reverse-transcription polymerase chain reaction using mRNA from the retina at various developmental stages showed that Zic1 and Zic2 are expressed in the embryonic retina and then gradually disappear during retinal development. Zic3 is also expressed in the embryonic retina; its expression level slightly decreases but it is expressed until adulthood. We overexpressed Zic1, Zic2, or Zic3 in retinal progenitors at embryonic day 17.5 and cultured the retina as explants for 2 weeks. The number of rod photoreceptors was fewer than in the control, but no other cell types showed significant differences between control and Zic overexpressing cells. The proliferation activity of normal retinal progenitors decreased after 5 days in culture, as observed in normal in vivo developmental processes. However, Zic expressing retinal cells continued to proliferate at days 5 and 7, suggesting that Zics sustain the proliferation activities of retinal progenitor cells. Since the effects of Zic1, 2, and 3 are indistinguishable in terms of differentiation and proliferation of retinal progenitors, the redundant function of Zics in retinal development is suggested.

  10. Canine uterine bacterial infection induces upregulation of proteolysis-related genes and downregulation of homeobox and zinc finger factors.

    Directory of Open Access Journals (Sweden)

    Ragnvi Hagman

    Full Text Available BACKGROUND: Bacterial infection with the severe complication of sepsis is a frequent and serious condition, being a major cause of death worldwide. To cope with the plethora of occurring bacterial infections there is therefore an urgent need to identify molecular mechanisms operating during the host response, in order both to identify potential targets for therapeutic intervention and to identify biomarkers for disease. Here we addressed this issue by studying global gene expression in uteri from female dogs suffering from spontaneously occurring uterine bacterial infection. PRINCIPAL FINDINGS: The analysis showed that almost 800 genes were significantly (p2-fold in the uteri of diseased animals. Among these were numerous chemokine and cytokine genes, as well as genes associated with inflammatory cell extravasation, anti-bacterial action, the complement system and innate immune responses, as well as proteoglycan-associated genes. There was also a striking representation of genes associated with proteolysis. Robust upregulation of immunoglobulin components and genes involved in antigen presentation was also evident, indicating elaboration of a strong adaptive immune response. The bacterial infection was also associated with a significant downregulation of almost 700 genes, of which various homeobox and zinc finger transcription factors were highly represented. CONCLUSIONS/SIGNIFICANCE: Together, these finding outline the molecular patterns involved in bacterial infection of the uterus. The study identified altered expression of numerous genes not previously implicated in bacterial disease, and several of these may be evaluated for potential as biomarkers of disease or as therapeutic targets. Importantly, since humans and dogs show genetic similarity and develop diseases that share many characteristics, the molecular events identified here are likely to reflect the corresponding situation in humans afflicted by similar disease.

  11. The role of Zic family zinc finger transcription factors in the proliferation and differentiation of retinal progenitor cells

    Energy Technology Data Exchange (ETDEWEB)

    Watabe, Yui [Division of Molecular and Developmental Biology, Institute of Medical Science, University of Tokyo (Japan); Department of Ophthalmology, Teikyo University School of Medicine, Tokyo (Japan); Division of Orthoptics, Teikyo University School of Medical Care and Technology, Tokyo (Japan); Baba, Yukihiro [Division of Molecular and Developmental Biology, Institute of Medical Science, University of Tokyo (Japan); Nakauchi, Hiromitsu [Division of Stem Cell Therapy, Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, University of Tokyo (Japan); Mizota, Atsushi [Department of Ophthalmology, Teikyo University School of Medicine, Tokyo (Japan); Watanabe, Sumiko, E-mail: sumiko@ims.u-tokyo.ac.jp [Division of Molecular and Developmental Biology, Institute of Medical Science, University of Tokyo (Japan)

    2011-11-11

    Highlights: Black-Right-Pointing-Pointer Zic transcription factors expressed early retinal progenitor cells. Black-Right-Pointing-Pointer Zics sustain proliferation activity of retinal progenitor cells. Black-Right-Pointing-Pointer Overexpression of Zic in retinal progenitors perturbed rod differentiation. Black-Right-Pointing-Pointer Fate determination to rod photoreceptor was not affected. -- Abstract: Members of the Zic family of zinc finger transcription factors play critical roles in a variety of developmental processes. Using DNA microarray analysis, we found that Zics are strongly expressed in SSEA-1-positive early retinal progenitors in the peripheral region of the mouse retina. Reverse-transcription polymerase chain reaction using mRNA from the retina at various developmental stages showed that Zic1 and Zic2 are expressed in the embryonic retina and then gradually disappear during retinal development. Zic3 is also expressed in the embryonic retina; its expression level slightly decreases but it is expressed until adulthood. We overexpressed Zic1, Zic2, or Zic3 in retinal progenitors at embryonic day 17.5 and cultured the retina as explants for 2 weeks. The number of rod photoreceptors was fewer than in the control, but no other cell types showed significant differences between control and Zic overexpressing cells. The proliferation activity of normal retinal progenitors decreased after 5 days in culture, as observed in normal in vivo developmental processes. However, Zic expressing retinal cells continued to proliferate at days 5 and 7, suggesting that Zics sustain the proliferation activities of retinal progenitor cells. Since the effects of Zic1, 2, and 3 are indistinguishable in terms of differentiation and proliferation of retinal progenitors, the redundant function of Zics in retinal development is suggested.

  12. KRAB-zinc finger proteins and KAP1 can mediate long-range transcriptional repression through heterochromatin spreading.

    Directory of Open Access Journals (Sweden)

    Anna C Groner

    2010-03-01

    Full Text Available Krüppel-associated box domain-zinc finger proteins (KRAB-ZFPs are tetrapod-specific transcriptional repressors encoded in the hundreds by the human genome. In order to explore their as yet ill-defined impact on gene expression, we developed an ectopic repressor assay, allowing the study of KRAB-mediated transcriptional regulation at hundreds of different transcriptional units. By targeting a drug-controllable KRAB-containing repressor to gene-trapping lentiviral vectors, we demonstrate that KRAB and its corepressor KAP1 can silence promoters located several tens of kilobases (kb away from their DNA binding sites, with an efficiency which is generally higher for promoters located within 15 kb or less. Silenced promoters exhibit a loss of histone H3-acetylation, an increase in H3 lysine 9 trimethylation (H3K9me3, and a drop in RNA Pol II recruitment, consistent with a block of transcriptional initiation following the establishment of silencing marks. Furthermore, we reveal that KRAB-mediated repression is established by the long-range spreading of H3K9me3 and heterochromatin protein 1 beta (HP1beta between the repressor binding site and the promoter. We confirm the biological relevance of this phenomenon by documenting KAP1-dependent transcriptional repression at an endogenous KRAB-ZFP gene cluster, where KAP1 binds to the 3' end of genes and mediates propagation of H3K9me3 and HP1beta towards their 5' end. Together, our data support a model in which KRAB/KAP1 recruitment induces long-range repression through the spread of heterochromatin. This finding not only suggests auto-regulatory mechanisms in the control of KRAB-ZFP gene clusters, but also provides important cues for interpreting future genome-wide DNA binding data of KRAB-ZFPs and KAP1.

  13. The ZNF75 zinc finger gene subfamily: Isolation and mapping of the four members in humans and great apes

    Energy Technology Data Exchange (ETDEWEB)

    Villa, A.; Strina, D.; Frattini, A. [Consiglio Nazionale delle Ricerche, Milan (Italy)] [and others

    1996-07-15

    We have previously reported the characterization of the human ZNF75 gene located on Xq26, which has only limited homology (less than 65%) to other ZF genes in the databases. Here, we describe three human zinc finger genes with 86 to 95% homology to ZNF75 at the nucleotide level, which represent all the members of the human ZNF75 subfamily. One of these, ZNF75B, is a pseudogene mapped to chromosome 12q13. The other two, ZNF75A and ZNF75C, maintain on ORF in the sequenced region, and at least the latter is expressed in the U937 cell line. They were mapped to chromosomes 16 and 11, respectively. All these genes are conserved in chimpanzees, gorillas, and orangutans. The ZNF75B homologue is a pseudogene in all three great apes, and in chimpanzee it is located on chromosome 10 (phylogenetic XII), at p13 (corresponding to the human 12q13). The chimpanzee homologue of ZNF75 is also located on the Xq26 chromosome, in the same region, as detected by in situ hybridization. As expected, nucleotide changes were clearly more abundant between human and organutan than between human and chimpanzee or gorilla homologues. Members of the same class were more similar to each other than to the other homologues within the same species. This suggests that the duplication and/or retrotranscription events occurred in a common ancestor long before great ape speciation. This, together with the existance of at least two genes in cows and horses, suggests a relatively high conservation of this gene family. 20 refs., 5 figs., 1 tab.

  14. Comprehensive analysis of CCCH-type zinc finger gene family in citrus (Clementine mandarin) by genome-wide characterization.

    Science.gov (United States)

    Liu, Shengrui; Khan, Muhammad Rehman Gul; Li, Yongping; Zhang, Jinzhi; Hu, Chungen

    2014-10-01

    The CCCH-type zinc finger proteins comprise a large gene family of regulatory proteins and are widely distributed in eukaryotic organisms. The CCCH proteins have been implicated in multiple biological processes and environmental responses in plants. Little information is available, however, about CCCH genes in plants, especially in woody plants such as citrus. The release of the whole-genome sequence of citrus allowed us to perform a genome-wide analysis of CCCH genes and to compare the identified proteins with their orthologs in model plants. In this study, 62 CCCH genes and a total of 132 CCCH motifs were identified, and a comprehensive analysis including the chromosomal locations, phylogenetic relationships, functional annotations, gene structures and conserved motifs was performed. Distribution mapping revealed that 54 of the 62 CCCH genes are unevenly dispersed on the nine citrus chromosomes. Based on phylogenetic analysis and gene structural features, we constructed 5 subfamilies of 62 CCCH members and integrative subfamilies from citrus, Arabidopsis, and rice, respectively. Importantly, large numbers of SNPs and InDels in 26 CCCH genes were identified from Poncirus trifoliata and Fortunella japonica using whole-genome deep re-sequencing. Furthermore, citrus CCCH genes showed distinct temporal and spatial expression patterns in different developmental processes and in response to various stress conditions. Our comprehensive analysis of CleC3Hs is a valuable resource that further elucidates the roles of CCCH family members in plant growth and development. In addition, variants and comparative genomics analyses deepen our understanding of the evolution of the CCCH gene family and will contribute to further genetics and genomics studies of citrus and other plant species.

  15. 补锌对轮状病毒肠炎患儿末梢血锌水平的影响%Influence of zinc supplementation on blood levels of zinc in finger tips of children with rotavirus enteritis

    Institute of Scientific and Technical Information of China (English)

    周雄飞; 郑君; 孙艳萍; 欧阳静萍

    2011-01-01

    目的 探讨口服补锌对轮状病毒肠炎患儿末梢血锌水平的变化.方法 收集52例6个月~2岁健康儿童(健康组)手指末梢血,测定血锌水平,将其作为本地区该年龄段儿童血锌正常值.随机将82例轮状病毒肠炎患儿分为治疗组42例,对照组40例.对照组给予常规治疗,治疗组在常规治疗的基础上加服葡萄糖酸锌片20 mg/d,连续治疗14 d.检测治疗前、治疗后手指末梢血血锌水平.结果 轮状病毒肠炎患儿末梢血锌水平明显低于本地区同龄健康儿童,有显著性差异(P0.05).治疗组总有效率显著高于对照组(95% vs 78%,P<0.01).结论 轮状病毒肠炎患儿末梢血锌水平明显降低,加用锌剂可提高轮状病毒肠炎患儿的血锌水平.%Objective It is to approach the changes of zinc supplementation on blood levels of zinc in finger tips of children with rotavirus enteritis. Methods Fifty-two finger tips blood specimens were collected from healthy children during 6 months to 2 years ( health group ) to measure the levels of zinc, and the obtained value was deemed as normal zinc value of children in this region. 82 children with rotavirus enteritis were randomly divided into treatment group ( n =42 ) and control group ( n = 40 ). The control group was given conventional therapy. The treatment group was given oral Zinc Gluconate tahlets 20 mg/d for 14 days based on conventional therapy. The finger tips hlood levels of zinc were detected before and after treatment. Results The finger tips blood levels of zinc in the children with rotavirus enteritis were obviously lower than that in the healthy children at the same age in this region, and the difference was significant ( P < 0.01 ). There was no significant difference between the treatment group after treatment and the healthy group ( P > 0.05 ). The total effective rate in the treatment group was significantly higher than that in the control group ( 95% vs 78% , P <0.01 ). Conclusion

  16. UtroUp is a novel six zinc finger artificial transcription factor that recognises 18 base pairs of the utrophin promoter and efficiently drives utrophin upregulation

    Directory of Open Access Journals (Sweden)

    Onori Annalisa

    2013-01-01

    Full Text Available Abstract Background Duchenne muscular dystrophy (DMD is the most common X-linked muscle degenerative disease and it is due to the absence of the cytoskeletal protein dystrophin. Currently there is no effective treatment for DMD. Among the different strategies for achieving a functional recovery of the dystrophic muscle, the upregulation of the dystrophin-related gene utrophin is becoming more and more feasible. Results We have previously shown that the zinc finger-based artificial transcriptional factor “Jazz” corrects the dystrophic pathology in mdx mice by upregulating utrophin gene expression. Here we describe a novel artificial transcription factor, named “UtroUp”, engineered to further improve the DNA-binding specificity. UtroUp has been designed to recognise an extended DNA target sequence on both the human and mouse utrophin gene promoters. The UtroUp DNA-binding domain contains six zinc finger motifs in tandem, which is able to recognise an 18-base-pair DNA target sequence that statistically is present only once in the human genome. To achieve a higher transcriptional activation, we coupled the UtroUp DNA-binding domain with the innovative transcriptional activation domain, which was derived from the multivalent adaptor protein Che-1/AATF. We show that the artificial transcription factor UtroUp, due to its six zinc finger tandem motif, possesses a low dissociation constant that is consistent with a strong affinity/specificity toward its DNA-binding site. When expressed in mammalian cell lines, UtroUp promotes utrophin transcription and efficiently accesses active chromatin promoting accumulation of the acetylated form of histone H3 in the utrophin promoter locus. Conclusions This novel artificial molecule may represent an improved platform for the development of future applications in DMD treatment.

  17. In Vivo Zinc Finger Nuclease-mediated Targeted Integration of a Glucose-6-phosphatase Transgene Promotes Survival in Mice With Glycogen Storage Disease Type IA.

    Science.gov (United States)

    Landau, Dustin J; Brooks, Elizabeth Drake; Perez-Pinera, Pablo; Amarasekara, Hiruni; Mefferd, Adam; Li, Songtao; Bird, Andrew; Gersbach, Charles A; Koeberl, Dwight D

    2016-04-01

    Glycogen storage disease type Ia (GSD Ia) is caused by glucose-6-phosphatase (G6Pase) deficiency in association with severe, life-threatening hypoglycemia that necessitates lifelong dietary therapy. Here we show that use of a zinc-finger nuclease (ZFN) targeted to the ROSA26 safe harbor locus and a ROSA26-targeting vector containing a G6PC donor transgene, both delivered with adeno-associated virus (AAV) vectors, markedly improved survival of G6Pase knockout (G6Pase-KO) mice compared with mice receiving the donor vector alone (P Ia, as compared with normal littermates, at 8 months following vector administration (P Ia.

  18. Scientific opinion addressing the safety assessment of plants developed using Zinc Finger Nuclease 3 and other Site-Directed Nucleases with similar function

    OpenAIRE

    EFSA Panel on Genetically Modified Organisms (GMO)

    2012-01-01

    The European Commission requested that the EFSA Panel on Genetically Modified Organisms deliver a scientific opinion related to risk assessment of plants developed using the zinc finger nuclease 3 technique (ZFN-3) which allows the integration of gene(s) in a predefined insertion site in the genome of the recipient species. Since other nucleases with a similar function to ZFN are considered in this opinion the term site-directed nuclease 3 (SDN-3) is used to describe the technique ra...

  19. NF-κB activation and zinc finger protein A20 expression in mature dendritic cells derived from liver allografts undergoing acute rejection

    Institute of Scientific and Technical Information of China (English)

    Ming-Qing Xu; Wei Wang; Lan Xue; Lv-Nan Yan

    2003-01-01

    AIM: To investigate the role of NF-κB activation and zinc finger protein A20 expression in the regulation of maturation of dendritic cells (DCs) derived from liver allografts undergoing acute rejection. METHODS: Sixty donor male SD rats and sixty recipient male LEW rats weighing 220-300 g were randomly divided into whole liver transplantation group and partial liver transplantation group. Allogeneic (SD rat to LEW rat) whole and 50 % partial liver transplantation were performed. DCs from liver grafts 0 hour and 4 days after transplantation were isolated and propagated in the presence of GM-CSF in vitro. Morphological characteristics and phenotypical features of DCs propagated for 10 days were analyzed by electron microscopy and flow cytometry, respectively. NF-κB binding activity, IL-12p70 protein and zinc finger protein A20expression in these DCs were measured by EMSA and Western blotting, respectively. Histological grading of rejection was determined. RESULTS: Allogeneic whole liver grafts showed no signs of rejection on day 4 after the transplantation. In contrast,allogeneic partial liver grafts demonstrated moderate to severe rejection on day 4 after the transplantation. After propagation for 10 days in the presence of GM-CSF in vitro,DCs from allogeneic whole liver grafts exhibited features of immature DC with absence of CD40 surface expression,these DCs were found to exhibit detectable but very low level of NF-κB activity, IL-12 p70 protein and zinc finger protein A20 expression. Whereas, DCs from allogeneic partial liver graft 4 days after transplantation displayed features of mature DC, with high level of CD40 surface expression, and as a consequence, higher expression of IL-12p70 protein, higher activities of NF-κB and higher expression of zinc finger protein A20 compared with those of DCs from whole liver grafts (P<0.001). CONCLUSION: These results suggest that A20expression is up-regulated in response to NF-κB activation in mature DCs derived from

  20. The LSD1-Type Zinc Finger Motifs of Pisum sativa LSD1 Are a Novel Nuclear Localization Signal and Interact with Importin Alpha

    OpenAIRE

    Shanping He; Kuowei Huang; Xu Zhang; Xiangchun Yu; Ping Huang; Chengcai An

    2011-01-01

    BACKGROUND: Genetic studies of the Arabidopsis mutant lsd1 highlight the important role of LSD1 in the negative regulation of plant programmed cell death (PCD). Arabidopsis thaliana LSD1 (AtLSD1) contains three LSD1-type zinc finger motifs, which are involved in the protein-protein interaction. METHODOLOGY/PRINCIPAL FINDINGS: To further understand the function of LSD1, we have analyzed cellular localization and functional localization domains of Pisum sativa LSD1 (PsLSD1), which is a homolog ...

  1. Gene targeting technologies in rats: zinc finger nucleases, transcription activator-like effector nucleases, and clustered regularly interspaced short palindromic repeats.

    Science.gov (United States)

    Mashimo, Tomoji

    2014-01-01

    The laboratory rat has been widely used as an animal model in biomedical science for more than 150 years. Applying zinc-finger nucleases or transcription activator-like effector nucleases to rat embryos via microinjection is an efficient genome editing tool for generating targeted knockout rats. Recently, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated endonucleases have been used as an effective tool for precise and multiplex genome editing in mice and rats. In this review, the advantages and disadvantages of these site-specific nuclease technologies for genetic analysis and manipulation in rats are discussed.

  2. High-efficiency genome editing via 2A-coupled co-expression of fluorescent proteins and zinc finger nucleases or CRISPR/Cas9 nickase pairs

    DEFF Research Database (Denmark)

    Duda, Katarzyna; Lonowski, Lindsey A; Kofoed-Nielsen, Michael;

    2014-01-01

    Targeted endonucleases including zinc finger nucleases (ZFNs) and clustered regularly interspaced short palindromic repeats (CRISPRs)/Cas9 are increasingly being used for genome editing in higher species. We therefore devised a broadly applicable and versatile method for increasing editing...... were minimal, and when occurring, our data suggest that they may be counteracted by selecting intermediate nuclease levels where off-target mutagenesis is low, but on-target mutagenesis remains relatively high. The method was also applicable to the CRISPR/Cas9 system, including CRISPR/Cas9 mutant...

  3. Promiscuous and specific phospholipid binding by domains in ZAC, a membrane-associated Arabidopsis protein with an ARF GAP zinc finger and a C2 domain

    DEFF Research Database (Denmark)

    Jensen, R B; Lykke-Andersen, K; Frandsen, G I;

    2000-01-01

    Arabidopsis proteins were predicted which share an 80 residue zinc finger domain known from ADP-ribosylation factor GTPase-activating proteins (ARF GAPs). One of these is a 37 kDa protein, designated ZAC, which has a novel domain structure in which the N-terminal ARF GAP domain and a C-terminal C2...... and plasma membrane marker proteins. ZAC membrane association was confirmed in assays by a fusion between ZAC and the green fluorescence protein and prompted an analysis of the in vitro phospholipid-binding ability of ZAC. Phospholipid dot-blot and liposome-binding assays indicated that fusion proteins...

  4. Synthesis and Crystal Structure of α-(-(1,2,4-Triazol-1H-yl)-ρ-chloro Acetophenone, [ClC6H4COCH2(C2H2N3)

    Institute of Scientific and Technical Information of China (English)

    建方方; 李艳; 肖海玲; 孙萍萍

    2003-01-01

    The title compound [ClC6H4COCH2(C2H2N3)] has been prepared and characterized by elemental analysis, IR spectrum and X-ray studies. It crystallizes in the monoclinic system, space group P21/c with Mr = 221.64 (C10H8ClN3O), a = 13.420(3), b = 9.720(2), c = 7.900(2)(A°),β= 92.00(3)o, V = 1029.9(4)(A°)3, Z = 4, Dc = 1.429 g/cm3, F(000) = 456,μ = 0.345 mm-1, R = 0.0435 and wR=0.0894. The total reflections were 1949 and the independent ones were 1805 (Rint=0.0340), of which 800 were observed with I > 2σ(I). The crystal structure consists of α-(1,2,4-triazol-1H- yl)-(-chloro-acetophenone. The existence of π conjugated systems in the molecule affects partly the bond lengths. The triazole and phenyl rings form the dihedral angle of 77.34o. The molecules of the title compound connect to each other through extensive hydrogen bonds to form a two-dimensional structure. Elemental analysis and IR spectra are in good agreement with the structure data.

  5. Optical emission spectroscopy study of premixed C2H4/O2 and C2H4/C2H2/O2 flames for diamond growth with and without CO2 laser excitation

    Science.gov (United States)

    He, X. N.; Gebre, T.; Shen, X. K.; Xie, Z. Q.; Zhou, Y. S.; Lu, Y. F.

    2010-02-01

    Optical emission spectroscopy (OES) measurements were carried out to study premixed C2H4/O2 and C2H4/C2H2/O2 combustion flame for diamond deposition with and without a CO2 laser excitation. Strong emissions from radicals C2 and CH were observed in the visible range in all the OES spectra acquired. By adding a continuous-wave CO2 laser to irradiate the flame at a wavelength of 10.591 μm, the common CO2 laser wavelength, it was discovered that the emission intensities of the C2 and CH radicals were increased due to the laser beam induced excitation. OES measurements of the C2 and CH radicals were performed using different gas combinations and laser powers. The rotational temperatures in the flame were determined by analyzing the spectra of the R-branch of the A2Δ-->X2Π (0, 0) electronic transition near 430 nm (CH band head). Information obtained from the OES spectra, including the emission intensities of the C2 and CH radicals, the intensity ratios, and the rotational temperatures, was integrated into the study of diamond deposition on tungsten carbide substrates for mechanism analysis of the laser induced vibrational excitation and laser-assisted diamond deposition.

  6. The role of isovalency in the reactions of the cyano (CN), boron monoxide (BO), silicon nitride (SiN), and ethynyl (C2H) radicals with unsaturated hydrocarbons acetylene (C2H2) and ethylene (C2H4).

    Science.gov (United States)

    Parker, D S N; Mebel, A M; Kaiser, R I

    2014-04-21

    The classification of chemical reactions based on shared characteristics is at the heart of the chemical sciences, and is well exemplified by Langmuir's concept of isovalency, in which 'two molecular entities with the same number of valence electrons have similar chemistries'. Within this account we further investigate the ramifications of the isovalency of four radicals with the same X(2)Σ(+) electronic structure - cyano (CN), boron monoxide (BO), silicon nitride (SiN), and ethynyl (C2H), and their reactions with simple prototype hydrocarbons acetylene (C2H2) and ethylene (C2H4). The fact that these four reactants own the same X(2)Σ(+) electronic ground state should dictate the outcome of their reactions with prototypical hydrocarbons holding a carbon-carbon triple and double bond. However, we find that other factors come into play, namely, atomic radii, bonding orbital overlaps, and preferential location of the radical site. These doublet radical reactions with simple hydrocarbons play significant roles in extreme environments such as the interstellar medium and planetary atmospheres (CN, SiN and C2H), and combustion flames (C2H, BO). PMID:24418936

  7. The C2H2-type transcription factor, FlbC, is involved in the transcriptional regulation of Aspergillus oryzae glucoamylase and protease genes specifically expressed in solid-state culture.

    Science.gov (United States)

    Tanaka, Mizuki; Yoshimura, Midori; Ogawa, Masahiro; Koyama, Yasuji; Shintani, Takahiro; Gomi, Katsuya

    2016-07-01

    Aspergillus oryzae produces a large amount of secreted proteins in solid-state culture, and some proteins such as glucoamylase (GlaB) and acid protease (PepA) are specifically produced in solid-state culture, but rarely in submerged culture. From the disruption mutant library of A. oryzae transcriptional regulators, we successfully identified a disruption mutant showing an extremely low production level of GlaB but a normal level of α-amylase production. This strain was a disruption mutant of the C2H2-type transcription factor, FlbC, which is reported to be involved in the regulation of conidiospore development. Disruption mutants of other upstream regulators comprising a conidiation regulatory network had no apparent effect on GlaB production in solid-state culture. In addition to GlaB, the production of acid protease in solid-state culture was also markedly decreased by flbC disruption. Northern blot analyses revealed that transcripts of glaB and pepA were significantly decreased in the flbC disruption strain. These results suggested that FlbC is involved in the transcriptional regulation of genes specifically expressed under solid-state cultivation conditions, possibly independent of the conidiation regulatory network.

  8. Probing cis-trans isomerization in the S1 state of C2H2 via H-atom action and hot band-pumped IR-UV double resonance spectroscopies.

    Science.gov (United States)

    Changala, P Bryan; Baraban, Joshua H; Merer, Anthony J; Field, Robert W

    2015-08-28

    We report novel experimental strategies that should prove instrumental in extending the vibrational and rotational assignments of the S1 state of acetylene, C2H2, in the region of the cis-trans isomerization barrier. At present, the assignments are essentially complete up to ∼500 cm(-1) below the barrier. Two difficulties arise when the assignments are continued to higher energies. One is that predissociation into C2H + H sets in roughly 1100 cm(-1) below the barrier; the resulting quenching of laser-induced fluorescence (LIF) reduces its value for recording spectra in this region. The other difficulty is that tunneling through the barrier causes a staggering in the K-rotational structure of isomerizing vibrational levels. The assignment of these levels requires data for K values up to at least 3. Given the rotational selection rule K' - ℓ('') = ± 1, such data must be obtained via excited vibrational levels of the ground state with ℓ('') > 0. In this paper, high resolution H-atom resonance-enhanced multiphoton ionization spectra are demonstrated to contain predissociated bands which are almost invisible in LIF spectra, while preliminary data using a hyperthermal pulsed nozzle show that ℓ('') = 2 states can be selectively populated in a jet, giving access to K' = 3 states in IR-UV double resonance. PMID:26328846

  9. Crossed beam reaction of the cyanogen radical, CN(X 2Σ+), with acetylene, C2H2(X 1Σg+): Observation of cyanoacetylene, HCCCN(X 1Σ+)

    Science.gov (United States)

    Huang, L. C. L.; Lee, Y. T.; Kaiser, R. I.

    1999-04-01

    The chemical dynamics to cyanoacetylene, HCCCN(X 1Σ+), formation via the neutral-neutral reaction of cyanogen, CN(X 2Σ+), with acetylene, C2H2(X 1Σg+), is investigated in a crossed molecular beams experiment at a collision energy of 21.1 kJ mol-1. The laboratory angular distribution and time-of-flight spectra of the HCCCN product are recorded at m/e=51 and 50. Forward-convolution fitting of our data reveals that the reaction dynamics are governed by an initial attack of the CN radical to the π electron density of the acetylene molecule to form a HCCHCN collision complex on the 2A' surface. The four heavy atoms are rotating in plane almost perpendicular to the total angular momentum vector J around the C axis of the complex which undergoes C-H bond rupture through a tight transition state to HCCCN and H. The H atom is emitted almost perpendicular to the HCCCN axis to yield a nearly "sideways" peaking of T(θ). The explicit identification of the cyanoacetylene reaction product represents a solid background for the title reaction to be included with more confidence in reaction networks modeling the chemistry in dark, molecular clouds, outflow of dying carbon stars, hot molecular cores, as well as the atmosphere of hydrocarbon rich planets and satellites such as the Saturnian moon Titan.

  10. Zinc

    Science.gov (United States)

    ... Consumer Datos en español Health Professional Other Resources Zinc Fact Sheet for Consumers What is zinc and what does it do? Zinc is a ... find out more about zinc? Disclaimer How much zinc do I need? The amount of zinc you ...

  11. Overexpression of myeloid zinc finger 1 suppresses matrix metalloproteinase-2 expression and reduces invasiveness of SiHa human cervical cancer cells.

    Science.gov (United States)

    Tsai, Su-Ju; Hwang, Jin-Ming; Hsieh, Shu-Ching; Ying, Tsung-Ho; Hsieh, Yi-Hsien

    2012-08-24

    Myeloid zinc finger 1 (MZF1) gene belongs to the Kruppel family of zinc finger transcription factors. MZF1 has been suggested to play an important role in the tumorigenesis, invasion, and apoptosis of various tumor cells. However, the role of MZF1 in human cervical cancer remains unclear. To investigate the molecular mechanisms of MZF1 and its functional role in human cervical cancer cell migration and invasion, we experimented on stable SiHa cells overexpressing MZF1. We found that MZF1 overexpression inhibits the migratory and invasive abilities of SiHa cervical cancer cells. In addition, the overexpression of MZF1 significantly reduces MMP-2 protein and mRNA levels. Luciferase and ChIP assays suggested that MZF1 directly binds to MMP-2 gene regulatory sequences in vivo and suppresses MMP-2 promoter activity in vitro. This study shows that MZF-1 represses MMP-2 transcription and suggests that this repression may be linked to inhibition of human cervical cancer cell migration and metastasis.

  12. An A20/AN1-type zinc finger protein modulates gibberellins and abscisic acid contents and increases sensitivity to abiotic stress in rice (Oryza sativa).

    Science.gov (United States)

    Zhang, Ye; Lan, Hongxia; Shao, Qiaolin; Wang, Ruqin; Chen, Hui; Tang, Haijuan; Zhang, Hongsheng; Huang, Ji

    2016-01-01

    The plant hormones gibberellins (GA) and abscisic acid (ABA) play important roles in plant development and stress responses. Here we report a novel A20/AN1-type zinc finger protein ZFP185 involved in GA and ABA signaling in the regulation of growth and stress response. ZFP185 was constitutively expressed in various rice tissues. Overexpression of ZFP185 in rice results in a semi-dwarfism phenotype, reduced cell size, and the decrease of endogenous GA3 content. By contrast, higher GA3 content was observed in RNAi plants. The application of exogenous GA3 can fully rescue the semi-dwarfism phenotype of ZFP185 overexpressing plants, suggesting the negative role of ZFP185 in GA biosynthesis. Besides GA, overexpression of ZFP185 decreased ABA content and expression of several ABA biosynthesis-related genes. Moreover, it was found that ZFP185, unlike previously known A20/AN1-type zinc finger genes, increases sensitivity to drought, cold, and salt stresses, implying the negative role of ZFP185 in stress tolerance. ZFP185 was localized in the cytoplasm and lacked transcriptional activation potential. Our study suggests that ZFP185 regulates plant growth and stress responses by affecting GA and ABA biosynthesis in rice.

  13. ZNF198 stabilizes the LSD1-CoREST-HDAC1 complex on chromatin through its MYM-type zinc fingers.

    Directory of Open Access Journals (Sweden)

    Christian B Gocke

    Full Text Available Histone modifications in chromatin regulate gene expression. A transcriptional co-repressor complex containing LSD1-CoREST-HDAC1 (termed LCH hereafter for simplicity represses transcription by coordinately removing histone modifications associated with transcriptional activation. RE1-silencing transcription factor (REST recruits LCH to the promoters of neuron-specific genes, thereby silencing their transcription in non-neuronal tissues. ZNF198 is a member of a family of MYM-type zinc finger proteins that associate with LCH. Here, we show that ZNF198-like proteins are required for the repression of E-cadherin (a gene known to be repressed by LSD1, but not REST-responsive genes. ZNF198 binds preferentially to the intact LCH ternary complex, but not its individual subunits. ZNF198- and REST-binding to the LCH complex are mutually exclusive. ZNF198 associates with chromatin independently of LCH. Furthermore, modification of HDAC1 by small ubiquitin-like modifier (SUMO in vitro weakens its interaction with CoREST whereas sumoylation of HDAC1 stimulates its binding to ZNF198. Finally, we mapped the LCH- and HDAC1-SUMO-binding domains of ZNF198 to tandem repeats of MYM-type zinc fingers. Therefore, our results suggest that ZNF198, through its multiple protein-protein interaction interfaces, helps to maintain the intact LCH complex on specific, non-REST-responsive promoters and may also prevent SUMO-dependent dissociation of HDAC1.

  14. 青花菜C3H型锌指蛋白基因 BoCCCH2的克隆与表达%Cloning and expression of a C3H-type zinc finger protein gene BoCCCH2 from Brassica oleracea var . italica

    Institute of Scientific and Technical Information of China (English)

    蒋明; 刘青娥; 章燕如; 祝琦; 龚秀; 俞可可; 周秀倩

    2016-01-01

    , translation , repair , metabolism and signaling . According to the number and order of cysteine and histidine residues , zinc finger proteins were classified into several different types , such as C2H2 , C2C2 , C2C2C2 , C2HC and C3H . For example , C3H‐type ones contain one to six typical motifs with three cysteine residues and one histidine residue . However , their functions are little known , and no gene has been reported in broccoli . In this study , a C3H‐type zinc finger protein gene BoCCCH2 was isolated from broccoli , and later the expression patterns in different organs as well as leaves infected by H . parasitica and B . cinerea were studied . Results indicated that BoCCCH2 contained no intron , and the full length of coding sequence was 1 740 bp encoding 579 amino acids . The deduced protein sequence contained two ANK domains and two CCCH zinc finger structures , respectively , and the CCCH zinc finger types were C —X8 —C—X5 —C—X3 —H and C—X5 —C—X4 —C—X3 — H . Reverse transcription‐polymerase chain reaction results showed that the BoCCCH2 was expressed in roots , leaves , stalks , young siliques , flower buds and flowers , with highest level in roots . Expression levels increased when challenged by both H . parasitica and B . cinerea . When infected by H . parasitica , expression levels increased after 24 h , and decreased after 72 h , while infected by B . cinerea , the highest level was detected after 6 h , and slowed down in 12 h . Homologous sequences were downloaded from NCBI ( National Center for Biotechnology Information) website , including Citrus sinensis , Gossypium raimondii , Populus trichocarpa , Ricinus communis , Prunus persica , P . mume , Malus domestica , Fragaria vesca , Phaseolus vulgaris , Glycine soja , B . rapa, Camelinasativa , Capsella rubella , Arabidopsis thaliana and Eutrema salsugineum . Phylogenetic analysis results revealed that BoCCCH2 was grouped with homogeneous sequences from other Cruciferae plants

  15. Crossed beam reaction of cyano radicals with hydrocarbon molecules. IV. Chemical dynamics of cyanoacetylene (HCCCN; X 1Σ+) formation from reaction of CN(X 2Σ+) with acetylene, C2H2(X 1Σg+)

    Science.gov (United States)

    Huang, L. C. L.; Asvany, O.; Chang, A. H. H.; Balucani, N.; Lin, S. H.; Lee, Y. T.; Kaiser, R. I.; Osamura, Y.

    2000-11-01

    The chemical reaction dynamics to form cyanoacetylene, HCCCN (X 1Σ+), via the radical-neutral reaction of cyano radicals, CN(X 2Σ+;ν=0), with acetylene, C2H2(X 1Σg+), are unraveled in crossed molecular beam experiments at two collision energies of 21.1 and 27.0 kJ mol-1. Laboratory angular distributions and time-of-flight spectra of the HCCCN product are recorded at m/e=51 and 50. Experiments were supplemented by electronic structure calculations on the doublet C3H2N potential energy surface and RRKM investigations. Forward-convolution fitting of the crossed beam data combined with our theoretical investigations shows that the reaction has no entrance barrier and is initiated by an attack of the CN radical to the π electron density of the acetylene molecule to form a doublet cis/trans HCCHCN collision complex on the 2A' surface via indirect reactive scattering dynamics. Here 85% of the collision complexes undergo C-H bond rupture through a tight transition state located 22 kJ mol-1 above the cyanoacetylene, HCCCN (X 1Σ+) and H(2S1/2) products (microchannel 1). To a minor amount (15%) trans HCCHCN shows a 1,2-H shift via a 177 kJ mol-1 barrier to form a doublet H2CCCN radical, which is 46 kJ mol-1 more stable than the initial reaction intermediate (microchannel 2). The H2CCCN complex decomposes via a rather loose exit transition state situated only 7 kJ mol-1 above the reaction products HCCCN (X 1Σ+) and H(2S1/2). In both cases the geometry of the exit transition states is reflected in the observed center-of-mass angular distributions showing a mild forward/sideways peaking. The explicit identification of the cyanoacetylene as the only reaction product represents a solid background for the title reaction to be included in reaction networks modeling the chemistry in dark, molecular clouds, outflow of dying carbon stars, hot molecular cores, as well as the atmosphere of hydrocarbon rich planets and satellites such as the Saturnian moon Titan.

  16. Genetic analysis of Kruppel-like zinc finger 11 variants in 5864 Danish individuals: potential effect on insulin resistance and modified signal transducer and activator of transcription-3 binding by promoter variant -1659G>C

    DEFF Research Database (Denmark)

    Gutiérrez-Aguilar, Ruth; Froguel, Philippe; Hamid, Yasmin H;

    2008-01-01

    CONTEXT: The transcription factor Krüppel-like zinc finger 11 (KLF11) has been suggested to contribute to genetic risk of type 2 diabetes (T2D). Our previous results showed that four KLF11 variants, in strong linkage disequilibrium (LD block including +185 A>G/Gln62Arg and -1659 G>C) were associa...

  17. Structure and expression of major histocompatibility complex-binding protein 2, a 275-kDa zinc finger protein that binds to an enhancer of major histocompatibility complex class I genes

    NARCIS (Netherlands)

    Veer, L.J. van 't; Lutz, P.M.; Isselbacher, K.J.; Bernards, R.A.

    1992-01-01

    We have isolated a cDNA encoding a transcription factor that binds to the enhancer of major histocompatibility complex (MHC) class I genes. MHC-binding protein 2 (MBP-2) is a 275-kDa protein, containing two sets of widely separated zinc fingers and a stretch of highly acidic amino acids, a putative

  18. The human zinc-finger protein-7 gene is located 90 kb 3' of MYC and is not expressed in Burkitt lymphoma cell lines.

    Science.gov (United States)

    Feduchi, E; Gallego, M I; Lazo, P A

    1994-09-15

    The zinc-finger gene-7 (ZNF7) was located 90 kb 3' of MYC on human chromosome 8 band q24 by pulsed-field gel electrophoresis (PFGE). This position lies between the MLV14 and BVR1 loci, 2 variant translocation breakpoints in Burkitt lymphomas. The structure of the ZNF7 gene was not altered by translocations in Burkitt-lymphoma cell lines as shown by its germline-restriction map configuration. The chromosomal region surrounding the ZNF7 gene was extensively methylated. The ZNF7 gene was not expressed in 19 BL cell lines. Expression was detected only in the BL41 and BL47 cell lines and in the SW756 cervical-carcinoma cell line. The RNA in each was of a different size. We postulate that the lack of ZNF7 expression in Burkitt lymphomas might contribute to the tumor phenotype.

  19. ErbB2-Driven Breast Cancer Cell Invasion Depends on a Complex Signaling Network Activating Myeloid Zinc Finger-1-Dependent Cathepsin B Expression

    DEFF Research Database (Denmark)

    Rafn, Bo; Nielsen, Christian Thomas Friberg; Andersen, Sofie Hagel;

    2012-01-01

    signaling network activates the transcription of cathepsin B gene (CTSB) via myeloid zinc finger-1 transcription factor that binds to an ErbB2-responsive enhancer element in the first intron of CTSB. This work provides a model system for ErbB2-induced breast cancer cell invasiveness, reveals a signaling......Aberrant ErbB2 receptor tyrosine kinase activation in breast cancer is strongly linked to an invasive disease. The molecular basis of ErbB2-driven invasion is largely unknown. We show that cysteine cathepsins B and L are elevated in ErbB2 positive primary human breast cancer and function...... as effectors of ErbB2-induced invasion in vitro. We identify Cdc42-binding protein kinase beta, extracellular regulated kinase 2, p21-activated protein kinase 4, and protein kinase C alpha as essential mediators of ErbB2-induced cysteine cathepsin expression and breast cancer cell invasiveness. The identified...

  20. A Comprehensive Catalog of Human KRAB-associated Zinc Finger Genes: Insights into the Evolutionary History of a Large Family of Transcriptional Repressors

    Energy Technology Data Exchange (ETDEWEB)

    Huntley, S; Baggott, D M; Hamilton, A T; Tran-Gyamfi, M; Yang, S; Kim, J; Gordon, L; Branscomb, E; Stubbs, L

    2005-09-30

    Krueppel-type zinc finger (ZNF) motifs are prevalent components of transcription factor proteins in all eukaryotic species. In mammals, most ZNF proteins comprise a single class of transcriptional repressors in which a chromatin interaction domain, called the Krueppel-associated box (KRAB) is attached to a tandem array of DNA-binding zinc-finger motifs. KRAB-ZNF loci are specific to tetrapod vertebrates, but have expanded dramatically in numbers through repeated rounds of segmental duplication to create a gene family with hundreds of members in mammals. To define the full repertoire of human KRAB-ZNF proteins, we searched the human genome for key motifs and used them to construct and manually curate gene models. The resulting KRAB-ZNF gene catalog includes 326 known genes, 243 of which were structurally corrected by manual annotation, and 97 novel KRAB-ZNF genes; this single family therefore comprises 20% of all predicted human transcription factor genes. Many of the genes are alternatively spliced, yielding a total of 743 distinct predicted proteins. Although many human KRAB-ZNF genes are conserved in mammals, at least 136 and potentially more than 200 genes of this type are primate-specific including many recent segmental duplicates. KRAB-ZNF genes are active in a wide variety of human tissues suggesting roles in many key biological processes, but most member genes remain completely uncharacterized. Because of their sheer numbers, wide-ranging tissue-specific expression patterns, and remarkable evolutionary divergence we predict that KRAB-ZNF transcription factors have played critical roles in crafting many aspects of human biology, including both deeply conserved and primate-specific traits.

  1. Gain, loss and divergence in primate zinc-finger genes: a rich resource for evolution of gene regulatory differences between species.

    Directory of Open Access Journals (Sweden)

    Katja Nowick

    Full Text Available The molecular changes underlying major phenotypic differences between humans and other primates are not well understood, but alterations in gene regulation are likely to play a major role. Here we performed a thorough evolutionary analysis of the largest family of primate transcription factors, the Krüppel-type zinc finger (KZNF gene family. We identified and curated gene and pseudogene models for KZNFs in three primate species, chimpanzee, orangutan and rhesus macaque, to allow for a comparison with the curated set of human KZNFs. We show that the recent evolutionary history of primate KZNFs has been complex, including many lineage-specific duplications and deletions. We found 213 species-specific KZNFs, among them 7 human-specific and 23 chimpanzee-specific genes. Two human-specific genes were validated experimentally. Ten genes have been lost in humans and 13 in chimpanzees, either through deletion or pseudogenization. We also identified 30 KZNF orthologs with human-specific and 42 with chimpanzee-specific sequence changes that are predicted to affect DNA binding properties of the proteins. Eleven of these genes show signatures of accelerated evolution, suggesting positive selection between humans and chimpanzees. During primate evolution the most extensive re-shaping of the KZNF repertoire, including most gene additions, pseudogenizations, and structural changes occurred within the subfamily homininae. Using zinc finger (ZNF binding predictions, we suggest potential impact these changes have had on human gene regulatory networks. The large species differences in this family of TFs stands in stark contrast to the overall high conservation of primate genomes and potentially represents a potent driver of primate evolution.

  2. Um estudo teórico de propriedades moleculares em complexos de hidrogênio trimoleculares C2H4···2HF, C2H2···2HF e C3h6···2HF A theoretical study of molecular properties of C2H4···2HF, C2H2···2HF AND C3H6···2HF trimolecular hydrogen-bonded complexes

    Directory of Open Access Journals (Sweden)

    Boaz G. Oliveira

    2008-01-01

    Full Text Available We present a theoretical study of molecular properties in C2H4···2HF, C2H2···2HF and C3H6···2HF trimolecular hydrogen-bonded complexes. From B3LYP/6-311++G(d,p calculations, the most important structural deformations are related to the C=C (C2H4, C≡C (C2H2, C-C (C3H6 and HF bond lengths. According to the Bader's atoms in molecules and CHELPG calculations, it was identified a tertiary interaction between the fluorine atom of the second hydrofluoric acid molecule and hydrogen atoms of the ethylene and acetylene within the C2H4···2HF and C2H2···2HF complexes, respectively. Additionally, the evaluation of the infrared spectrum characterized the new vibrational modes and bathochromic effect of the HF molecules.

  3. Aromatic Character Studies on Divalent 3, 5 and 7-membered Rings C2H2M,C4H4M and C6H6M(M=C,Si,Ge,Sn and Pb)via Nucleus-independent Chemical Shifts (NICS) Calculation%通过核独立化学位移(NICS)计算研究二价三、五、七元环C2H2M,C4H4M and C6H6M(M=C,Si,Ge,Sn and Pb)的芳香族特性

    Institute of Scientific and Technical Information of China (English)

    Vessally E; Nikoorazm M; Ramazani A

    2008-01-01

    The aromatic character of divalent three, five and seven-membered rings C2H2M,C4H4 and C6H6M(M=C,Si, Ge, Sn and Pb) is investigated through magnetic and geometric criteria by Density Functional Theory (DFT) method using 6-311++G (3df,2p) basis set of the GAUSSIAN 98 program. The result of Nucleus-independent Chemical Shifts (NICS) (0.5) calculations show an aromatic character for singlet state of C2H2M,(M=C, Si, Ge, Sn and Sn) and nonaromatic character for triplet states of C2H2M (except M =Ge and Pb). NICS (0.5) calculations show nonaromatic character for the singlet state of C4H4C and antiaromatic character for C4H4M (M=Si, Ge, Sn and Pb). In contrast, NICS (0.5) calculations indicate antiaromatic character for the triplet state of C4H4C and nonaromatic character to C4H4M (M=Si, Ge, Sn and Pb). NICS (0.5) calculations show a slightly homoaromatic character for the singlet state of C6H6M and anti-aromatic character for triplet state of C6H6M.

  4. Zinc

    Science.gov (United States)

    ... women when used in the recommended daily amounts (RDA). However, zinc is POSSIBLY UNSAFE when used in ... older infants, children, and adults, Recommended Dietary Allowance (RDA) quantities of zinc have been established: infants and ...

  5. Structures of three members of Pfam PF02663 (FmdE) implicated in microbial methanogenesis reveal a conserved α+β core domain and an auxiliary C-terminal treble-clef zinc finger

    International Nuclear Information System (INIS)

    The first structures from the FmdE Pfam family (PF02663) reveal that some members of this family form tightly intertwined dimers consisting of two domains (N-terminal α+β core and C-terminal zinc-finger domains), whereas others contain only the core domain. The presence of the zinc-finger domain suggests that some members of this family may perform functions associated with transcriptional regulation, protein–protein interaction, RNA binding or metal-ion sensing. Examination of the genomic context for members of the FmdE Pfam family (PF02663), such as the protein encoded by the fmdE gene from the methanogenic archaeon Methanobacterium thermoautotrophicum, indicates that 13 of them are co-transcribed with genes encoding subunits of molybdenum formylmethanofuran dehydrogenase (EC 1.2.99.5), an enzyme that is involved in microbial methane production. Here, the first crystal structures from PF02663 are described, representing two bacterial and one archaeal species: B8FYU2-DESHY from the anaerobic dehalogenating bacterium Desulfitobacterium hafniense DCB-2, Q2LQ23-SYNAS from the syntrophic bacterium Syntrophus aciditrophicus SB and Q9HJ63-THEAC from the thermoacidophilic archaeon Thermoplasma acidophilum. Two of these proteins, Q9HJ63-THEAC and Q2LQ23-SYNAS, contain two domains: an N-terminal thioredoxin-like α+β core domain (NTD) consisting of a five-stranded, mixed β-sheet flanked by several α-helices and a C-terminal zinc-finger domain (CTD). B8FYU2-DESHY, on the other hand, is composed solely of the NTD. The CTD of Q9HJ63-THEAC and Q2LQ23-SYNAS is best characterized as a treble-clef zinc finger. Two significant structural differences between Q9HJ63-THEAC and Q2LQ23-SYNAS involve their metal binding. First, zinc is bound to the putative active site on the NTD of Q9HJ63-THEAC, but is absent from the NTD of Q2LQ23-SYNAS. Second, whereas the structure of the CTD of Q2LQ23-SYNAS shows four Cys side chains within coordination distance of the Zn atom, the structure

  6. Finger pain

    Science.gov (United States)

    ... numbness and pain in the hand and fingers Raynaud's phenomenon , a condition that results in blocked blood flow to the fingers when it is cold Trigger finger , when a swollen finger ... the thumb side of the wrist from overuse. Infections Tumors

  7. Mycobacterial and HIV infections up-regulated human zinc finger protein 134, a novel positive regulator of HIV-1 LTR activity and viral propagation.

    Directory of Open Access Journals (Sweden)

    Ronald Benjamin

    Full Text Available BACKGROUND: Concurrent occurrence of HIV and Tuberculosis (TB infections influence the cellular environment of the host for synergistic existence. An elementary approach to understand such coalition at the molecular level is to understand the interactions of the host and the viral factors that subsequently effect viral replication. Long terminal repeats (LTR of HIV genome serve as a template for binding trans-acting viral and cellular factors that regulate its transcriptional activity, thereby, deciding the fate of HIV pathogenesis, making it an ideal system to explore the interplay between HIV and the host. METHODOLOGY/PRINCIPAL FINDINGS: In this study, using biotinylated full length HIV-1 LTR sequence as bait followed by MALDI analyses, we identified and further characterized human-Zinc-finger-protein-134 (hZNF-134 as a novel positive regulator of HIV-1 that promoted LTR-driven transcription and viral production. Over-expression of hZNF-134 promoted LTR driven luciferase activity and viral transcripts, resulting in increased virus production while siRNA mediated knockdown reduced both the viral transcripts and the viral titers, establishing hZNF-134 as a positive effector of HIV-1. HIV, Mycobacteria and HIV-TB co-infections increased hZNF-134 expressions in PBMCs, the impact being highest by mycobacteria. Corroborating these observations, primary TB patients (n = 22 recorded extraordinarily high transcript levels of hZNF-134 as compared to healthy controls (n = 16. CONCLUSIONS/SIGNIFICANCE: With these observations, it was concluded that hZNF-134, which promoted HIV-1 LTR activity acted as a positive regulator of HIV propagation in human host. High titers of hZNF-134 transcripts in TB patients suggest that up-regulation of such positive effectors of HIV-1 upon mycobacterial infection can be yet another mechanism by which mycobacteria assists HIV-1 propagation during HIV-TB co-infections. hZNF-134, an uncharacterized host protein, thus

  8. Transcriptional activation is a conserved feature of the early embryonic factor Zelda that requires a cluster of four zinc fingers for DNA binding and a low-complexity activation domain.

    Science.gov (United States)

    Hamm, Danielle C; Bondra, Eliana R; Harrison, Melissa M

    2015-02-01

    Delayed transcriptional activation of the zygotic genome is a nearly universal phenomenon in metazoans. Immediately following fertilization, development is controlled by maternally deposited products, and it is not until later stages that widespread activation of the zygotic genome occurs. Although the mechanisms driving this genome activation are currently unknown, the transcriptional activator Zelda (ZLD) has been shown to be instrumental in driving this process in Drosophila melanogaster. Here we define functional domains of ZLD required for both DNA binding and transcriptional activation. We show that the C-terminal cluster of four zinc fingers mediates binding to TAGteam DNA elements in the promoters of early expressed genes. All four zinc fingers are required for this activity, and splice isoforms lacking three of the four zinc fingers fail to activate transcription. These truncated splice isoforms dominantly suppress activation by the full-length, embryonically expressed isoform. We map the transcriptional activation domain of ZLD to a central region characterized by low complexity. Despite relatively little sequence conservation within this domain, ZLD orthologs from Drosophila virilis, Anopheles gambiae, and Nasonia vitripennis activate transcription in D. melanogaster cells. Transcriptional activation by these ZLD orthologs suggests that ZLD functions through conserved interactions with a protein cofactor(s). We have identified distinct DNA-binding and activation domains within the critical transcription factor ZLD that controls the initial activation of the zygotic genome.

  9. Generation of Interleukin-2 Receptor Gamma Gene Knockout Pigs from Somatic Cells Genetically Modified by Zinc Finger Nuclease-Encoding mRNA

    Science.gov (United States)

    Watanabe, Masahito; Nakano, Kazuaki; Matsunari, Hitomi; Matsuda, Taisuke; Maehara, Miki; Kanai, Takahiro; Kobayashi, Mirina; Matsumura, Yukina; Sakai, Rieko; Kuramoto, Momoko; Hayashida, Gota; Asano, Yoshinori; Takayanagi, Shuko; Arai, Yoshikazu; Umeyama, Kazuhiro; Nagaya, Masaki; Hanazono, Yutaka; Nagashima, Hiroshi

    2013-01-01

    Zinc finger nuclease (ZFN) is a powerful tool for genome editing. ZFN-encoding plasmid DNA expression systems have been recently employed for the generation of gene knockout (KO) pigs, although one major limitation of this technology is the use of potentially harmful genome-integrating plasmid DNAs. Here we describe a simple, non-integrating strategy for generating KO pigs using ZFN-encoding mRNA. The interleukin-2 receptor gamma (IL2RG) gene was knocked out in porcine fetal fibroblasts using ZFN-encoding mRNAs, and IL2RG KO pigs were subsequently generated using these KO cells through somatic cell nuclear transfer (SCNT). The resulting IL2RG KO pigs completely lacked a thymus and were deficient in T and NK cells, similar to human X-linked SCID patients. Our findings demonstrate that the combination of ZFN-encoding mRNAs and SCNT provides a simple robust method for producing KO pigs without genomic integration. PMID:24130776

  10. Chemical shift as a probe of molecular interfaces: NMR studies of DNA binding by the three amino-terminal zinc finger domains from transcription factor IIIA

    Energy Technology Data Exchange (ETDEWEB)

    Foster, Mark P.; Wuttke, Deborah S.; Clemens, Karen R.; Jahnke, Wolfgang; Radhakrishnan, Ishwar; Tennant, Linda; Reymond, Martine; Chung, John; Wright, Peter E. [Scripps Research Institute, Department of Molecular Biology and Skaggs Institute for Chemical Biology (United States)

    1998-07-15

    We report the NMR resonance assignments for a macromolecular protein/DNA complex containing the three amino-terminal zinc fingers (92 amino acid residues) of Xenopus laevis TFIIIA (termed zf1-3) bound to the physiological DNA target (15 base pairs), and for the free DNA. Comparisons are made of the chemical shifts of protein backbone{sup 1} H{sup N}, {sup 15}N,{sup 13} C{sup {alpha}} and{sup 13} C{sup {beta}} and DNA base and sugar protons of the free and bound species. Chemical shift changes are analyzed in the context of the structures of the zf1-3/DNA complex to assess the utility of chemical shift change as a probe of molecular interfaces. Chemical shift perturbations that occur upon binding in the zf1-3/DNA complex do not correspond directly to the structural interface, but rather arise from a number of direct and indirect structural and dynamic effects.

  11. Engineered zinc-finger proteins can compensate genetic haploinsufficiency by transcriptional activation of the wild-type allele: application to Willams-Beuren syndrome and supravalvular aortic stenosis.

    Science.gov (United States)

    Zhang, Pei; Huang, Angela; Morales-Ruiz, Manuel; Starcher, Barry C; Huang, Yan; Sessa, William C; Niklason, Laura E; Giordano, Frank J

    2012-11-01

    Williams-Beuren syndrome (WBS) and supravalvular aortic stenosis (SVAS) are genetic syndromes marked by the propensity to develop severe vascular stenoses. Vascular lesions in both syndromes are caused by haploinsufficiency of the elastin gene. We used these distinct genetic syndromes as models to evaluate the feasibility of using engineered zinc-finger protein transcription factors (ZFPs) to achieve compensatory expression of haploinsufficient genes by inducing augmented expression from the remaining wild-type allele. For complex genes with multiple splice variants, this approach could have distinct advantages over cDNA-based gene replacement strategies. Targeting the elastin gene, we show that transcriptional activation by engineered ZFPs can induce compensatory expression from the wild-type allele in the setting of classic WBS and SVAS genetic mutations, increase elastin expression in wild-type cells, induce expression of the major elastin splice variants, and recapitulate their natural stoichiometry. Further, we establish that transcriptional activation of the mutant allele in SVAS does not overcome nonsense-mediated decay, and thus ZFP-mediated transcriptional activation is not likely to induce production of a mutant protein, a crucial consideration. Finally, we show in bioengineered blood vessels that ZFP-mediated induction of elastin expression is capable of stimulating functional elastogenesis. Haploinsufficiency is a common mechanism of genetic disease. These findings have significant implications for WBS and SVAS, and establish that haploinsufficiency can be overcome by targeted transcriptional activation without inducing protein expression from the mutant allele.

  12. Highly efficient homology-driven genome editing in human T cells by combining zinc-finger nuclease mRNA and AAV6 donor delivery.

    Science.gov (United States)

    Wang, Jianbin; DeClercq, Joshua J; Hayward, Samuel B; Li, Patrick Wai-Lun; Shivak, David A; Gregory, Philip D; Lee, Gary; Holmes, Michael C

    2016-02-18

    The adoptive transfer of engineered T cells for the treatment of cancer, autoimmunity, and infectious disease is a rapidly growing field that has shown great promise in recent clinical trials. Nuclease-driven genome editing provides a method in which to precisely target genetic changes to further enhance T cell function in vivo. We describe the development of a highly efficient method to genome edit both primary human CD8 and CD4 T cells by homology-directed repair at a pre-defined site of the genome. Two different homology donor templates were evaluated, representing both minor gene editing events (restriction site insertion) to mimic gene correction, or the more significant insertion of a larger gene cassette. By combining zinc finger nuclease mRNA delivery with AAV6 delivery of a homologous donor we could gene correct 41% of CCR5 or 55% of PPP1R12C (AAVS1) alleles in CD8(+) T cells and gene targeting of a GFP transgene cassette in >40% of CD8(+) and CD4(+) T cells at both the CCR5 and AAVS1 safe harbor locus, potentially providing a robust genome editing tool for T cell-based immunotherapy. PMID:26527725

  13. Sda1, a Cys2-His2 zinc finger transcription factor, is involved in polyol metabolism and fumonisin B1 production in Fusarium verticillioides.

    Directory of Open Access Journals (Sweden)

    Martha Malapi-Wight

    Full Text Available The ubiquitous ascomycete Fusarium verticillioides causes ear rot and stalk rot of maize, both of which reduce grain quality and yield. Additionally, F. verticillioides produces the mycotoxin fumonisin B1 (FB1 during infection of maize kernels, and thus potentially compromises human and animal health. The current knowledge is fragmentary regarding the regulation of FB1 biosynthesis, particularly when considering interplay with environmental factors such as nutrient availability. In this study, SDA1 of F. verticillioides, predicted to encode a Cys-2 His-2 zinc finger transcription factor, was shown to play a key role in catabolizing select carbon sources. Growth of the SDA1 knock-out mutant (Δsda1 was completely inhibited when sorbitol was the sole carbon source and was severely impaired when exclusively provided mannitol or glycerol. Deletion of SDA1 unexpectedly increased FB1 biosynthesis, but reduced arabitol and mannitol biosynthesis, as compared to the wild-type progenitor. Trichoderma reesei ACE1, a regulator of cellulase and xylanase expression, complemented the F. verticillioides Δsda1 mutant, which indicates that Ace1 and Sda1 are functional orthologs. Taken together, the data indicate that Sda1 is a transcriptional regulator of carbon metabolism and toxin production in F. verticillioides.

  14. A Novel Role for the Zinc-Finger Transcription Factor EGL-46 in the Differentiation of Gas-Sensing Neurons in Caenorhabditis elegans

    DEFF Research Database (Denmark)

    Rojo Romanos, Teresa; Gramstrup Petersen, Jakob; Redo Riveiro, Alba;

    2014-01-01

    Oxygen (O2) and carbon dioxide (CO2) provoke distinct olfactory behaviors via specialized sensory neurons across metazoa. In the nematode Caenorhabditis elegans, the BAG sensory neurons are specialized to sense changes in both O2 and CO2 levels in the environment. The precise functionality of the......-13. Thereby, three conserved transcription factors collaborate to ensure neuron type-specific identity features of the BAG gas-sensing neurons.......Oxygen (O2) and carbon dioxide (CO2) provoke distinct olfactory behaviors via specialized sensory neurons across metazoa. In the nematode Caenorhabditis elegans, the BAG sensory neurons are specialized to sense changes in both O2 and CO2 levels in the environment. The precise functionality...... is partially, though not completely, controlled by ETS-5, an ETS-domain-containing transcription factor, and EGL-13, a Sox transcription factor. We report here, the identification of EGL-46, a zinc-finger transcription factor, which regulates BAG gas-sensing fate in partially parallel pathways to ETS-5 and EGL...

  15. A Missense Mutation in the Zinc Finger Domain of OsCESA7 Deleteriously Affects Cellulose Biosynthesis and Plant Growth in Rice.

    Science.gov (United States)

    Wang, Daofeng; Qin, Yanling; Fang, Jingjing; Yuan, Shoujiang; Peng, Lixiang; Zhao, Jinfeng; Li, Xueyong

    2016-01-01

    Rice is a model plant species for the study of cellulose biosynthesis. We isolated a mutant, S1-24, from ethyl methanesulfonate (EMS)-treated plants of the japonica rice cultivar, Nipponbare. The mutant exhibited brittle culms and other pleiotropic phenotypes such as dwarfism and partial sterility. The brittle culms resulted from reduced mechanical strength due to a defect in thickening of the sclerenchyma cell wall and reduced cellulose content in the culms of the S1-24 mutant. Map-based gene cloning and a complementation assay showed that phenotypes of the S1-24 mutant were caused by a recessive point mutation in the OsCESA7 gene, which encodes cellulose synthase A subunit 7. The missense mutation changed the highly conserved C40 to Y in the zinc finger domain. The OsCESA7 gene is expressed predominantly in the culm at the mature stage, particularly in mechanical tissues such as vascular bundles and sclerenchyma cells, consistent with the brittle phenotype in the culm. These results indicate that OsCESA7 plays an important role in cellulose biosynthesis and plant growth. PMID:27092937

  16. The Arabidopsis a zinc finger domain protein ARS1 is essential for seed germination and ROS homeostasis in response to ABA and oxidative stress

    Directory of Open Access Journals (Sweden)

    Dongwon eBaek

    2015-11-01

    Full Text Available The phytohormone abscisic acid (ABA induces accumulation of reactive oxygen species (ROS, which can disrupt seed dormancy and plant development. Here, we report the isolation and characterization of an Arabidopsis thaliana mutant called ars1 (aba and ros sensitive 1 that showed hypersensitivity to ABA during seed germination and to methyl viologen (MV at the seedling stage. ARS1 encodes a nuclear protein with one zinc finger domain, two nuclear localization signal (NLS domains, and one nuclear export signal (NES. The ars1 mutants showed reduced expression of a gene for superoxide dismutase (CSD3 and enhanced accumulation of ROS after ABA treatment. Transient expression of ARS1 in Arabidopsis protoplasts strongly suppressed ABA-mediated ROS production. Interestingly, nuclear-localized ARS1 translocated to the cytoplasm in response to treatment with ABA, H2O2, or MV. Taken together, these results suggest that ARS1 modulates seed germination and ROS homeostasis in response to ABA and oxidative stress in plants.

  17. Identification of a putative DEAD-box RNA helicase and a zinc-finger protein in Candida albicans by functional complementation of the S. cerevisiae rok1 mutation.

    Science.gov (United States)

    Kim, W I; Lee, W B; Song, K; Kim, J

    2000-03-30

    We identified two novel genes, CHR1 and CSR1, of the fungal pathogen Candida albicans, by functional complementation of the Saccharomyces cerevisiae rok1 mutation. The Rok1 protein is a member of the DEAD protein family of ATP-dependent RNA helicases. ROK1 is required for cell cycle progression and also for rRNA processing. The CHR1 gene product of 578 amino acids is highly homologous to the Rok1 protein (54% identity) and is considered to be a putative DEAD-box RNA helicase. We predict that the CSR1 gene encodes a 73 kDa protein of 612 amino acids with five zinc-finger motifs at the C-terminal region. CHR1 or CSR1 on a high-copy number plasmid showed a slow-growth phenotype in a condition where the ROK1 expression is turned on from the GAL1 promoter. This result is consistent with the lethality caused by the ROK1 overexpression. We conclude that CHR1 encodes a functional homologue of Rok1 protein and CSR1 is a heterologous suppressor of the rok1 mutation. PMID:10705369

  18. The zinc fingers of the SR-like protein ZRANB2 are single-stranded RNA-binding domains that recognize 5′ splice site-like sequences

    Energy Technology Data Exchange (ETDEWEB)

    Loughlin, Fionna E.; Mansfield, Robyn E.; Vaz, Paula M.; McGrath, Aaron P.; Setiyaputra, Surya; Gamsjaeger, Roland; Chen, Eva S.; Morris, Brian J.; Guss, J. Mitchell; Mackay, Joel P.; (Sydney)

    2009-09-02

    The alternative splicing of mRNA is a critical process in higher eukaryotes that generates substantial proteomic diversity. Many of the proteins that are essential to this process contain arginine/serine-rich (RS) domains. ZRANB2 is a widely-expressed and highly-conserved RS-domain protein that can regulate alternative splicing but lacks canonical RNA-binding domains. Instead, it contains 2 RanBP2-type zinc finger (ZnF) domains. We demonstrate that these ZnFs recognize ssRNA with high affinity and specificity. Each ZnF binds to a single AGGUAA motif and the 2 domains combine to recognize AGGUAA(N{sub x})AGGUAA double sites, suggesting that ZRANB2 regulates alternative splicing via a direct interaction with pre-mRNA at sites that resemble the consensus 5{prime} splice site. We show using X-ray crystallography that recognition of an AGGUAA motif by a single ZnF is dominated by side-chain hydrogen bonds to the bases and formation of a guanine-tryptophan-guanine 'ladder.' A number of other human proteins that function in RNA processing also contain RanBP2 ZnFs in which the RNA-binding residues of ZRANB2 are conserved. The ZnFs of ZRANB2 therefore define another class of RNA-binding domain, advancing our understanding of RNA recognition and emphasizing the versatility of ZnF domains in molecular recognition.

  19. Sda1, a Cys2-His2 zinc finger transcription factor, is involved in polyol metabolism and fumonisin B1 production in Fusarium verticillioides.

    Science.gov (United States)

    Malapi-Wight, Martha; Smith, Jonathon; Campbell, Jacquelyn; Bluhm, Burton H; Shim, Won-Bo

    2013-01-01

    The ubiquitous ascomycete Fusarium verticillioides causes ear rot and stalk rot of maize, both of which reduce grain quality and yield. Additionally, F. verticillioides produces the mycotoxin fumonisin B1 (FB1) during infection of maize kernels, and thus potentially compromises human and animal health. The current knowledge is fragmentary regarding the regulation of FB1 biosynthesis, particularly when considering interplay with environmental factors such as nutrient availability. In this study, SDA1 of F. verticillioides, predicted to encode a Cys-2 His-2 zinc finger transcription factor, was shown to play a key role in catabolizing select carbon sources. Growth of the SDA1 knock-out mutant (Δsda1) was completely inhibited when sorbitol was the sole carbon source and was severely impaired when exclusively provided mannitol or glycerol. Deletion of SDA1 unexpectedly increased FB1 biosynthesis, but reduced arabitol and mannitol biosynthesis, as compared to the wild-type progenitor. Trichoderma reesei ACE1, a regulator of cellulase and xylanase expression, complemented the F. verticillioides Δsda1 mutant, which indicates that Ace1 and Sda1 are functional orthologs. Taken together, the data indicate that Sda1 is a transcriptional regulator of carbon metabolism and toxin production in F. verticillioides.

  20. A Zinc-Finger-Family Transcription Factor, AbVf19, Is Required for the Induction of a Gene Subset Important for Virulence in Alternaria brassicicola

    Energy Technology Data Exchange (ETDEWEB)

    Srivastava, Akhil [Univ. of Hawaii, Manoa, HI (United States); Ohm, Robin A. [USDOE Joint Genome Inst., Walnut Creek, CA (United States); Oxiles, Lindsay [Univ. of Hawaii, Manoa, HI (United States); Brooks, Fred [Univ. of Hawaii, Manoa, HI (United States); Lawrence, Christopher B. [Virginia Polytechnic Inst. and State Univ. (Virginia Tech), Blacksburg, VA (United States); Grigoriev, Igor V. [USDOE Joint Genome Inst., Walnut Creek, CA (United States); Cho, Yangrae [Univ. of Hawaii, Manoa, HI (United States)

    2011-10-26

    Alternaria brassicicola is a successful saprophyte and necrotrophic plant pathogen with a broad host range within the family Brassicaceae. It produces secondary metabolites that marginally affect virulence. Cell wall degrading enzymes (CDWE) have been considered important for pathogenesis but none of them individually have been identified as significant virulence factors in A. brassicicola. In this study, knockout mutants of a gene, AbVf19, were created and produced considerably smaller lesions than the wild type on inoculated host plants. The presence of tandem zinc-finger domains in the predicted amino acid sequence and nuclear localization of AbVf19- reporter protein suggested that it was a transcription factor. Gene expression comparisons using RNA-seq identified 74 genes being downregulated in the mutant during a late stage of infection. Among the 74 downregulated genes, 28 were putative CWDE genes. These were hydrolytic enzyme genes that composed a small fraction of genes within each family of cellulases, pectinases, cutinases, and proteinases. The mutants grew slower than the wild type on an axenic medium with pectin as a major carbon source. This study demonstrated the existence and the importance of a transcription factor that regulates a suite of genes that are important for decomposing and utilizing plant material during the late stage of plant infection.

  1. Loss of c-Kit and bone marrow failure upon conditional removal of the GATA-2 C-terminal zinc finger domain in adult mice.

    Science.gov (United States)

    Li, Haiyan S; Jin, Jin; Liang, Xiaoxuan; Matatall, Katie A; Ma, Ying; Zhang, Huiyuan; Ullrich, Stephen E; King, Katherine Y; Sun, Shao-Cong; Watowich, Stephanie S

    2016-09-01

    Heterozygous mutations in the transcriptional regulator GATA-2 associate with multilineage immunodeficiency, myelodysplastic syndrome (MDS), and acute myeloid leukemia (AML). The majority of these mutations localize in the zinc finger (ZnF) domains, which mediate GATA-2 DNA binding. Deregulated hematopoiesis with GATA-2 mutation frequently develops in adulthood, yet GATA-2 function in the bone marrow remains unresolved. To investigate this, we conditionally deleted the GATA-2 C-terminal ZnF (C-ZnF) coding sequences in adult mice. Upon Gata2 C-ZnF deletion, we observed rapid peripheral cytopenia, bone marrow failure, and decreased c-Kit expression on hematopoietic progenitors. Transplant studies indicated GATA-2 has a cell-autonomous role in bone marrow hematopoiesis. Moreover, myeloid lineage populations were particularly sensitive to Gata2 hemizygosity, while molecular assays indicated GATA-2 regulates c-Kit expression in multilineage progenitor cells. Enforced c-Kit expression in Gata2 C-ZnF-deficient hematopoietic progenitors enhanced myeloid colony activity, suggesting GATA-2 sustains myelopoiesis via a cell intrinsic role involving maintenance of c-Kit expression. Our results provide insight into mechanisms regulating hematopoiesis in bone marrow and may contribute to a better understanding of immunodeficiency and bone marrow failure associated with GATA-2 mutation.

  2. Expression Profiling and Functional Implications of a Set of Zinc Finger Proteins, ZNF423, ZNF470, ZNF521, and ZNF780B, in Primary Osteoarthritic Articular Chondrocytes

    Directory of Open Access Journals (Sweden)

    Maria Mesuraca

    2014-01-01

    Full Text Available Articular chondrocytes are responsible for the maintenance of healthy articulations; indeed, dysregulation of their functions, including the production of matrix proteins and matrix-remodeling proteases, may result in fraying of the tissue and development of osteoarthritis (OA. To explore transcriptional mechanisms that contribute to the regulation of chondrocyte homeostasis and may be implicated in OA development, we compared the gene expression profile of a set of zinc finger proteins potentially linked to the control of chondrocyte differentiation and/or functions (ZNF423, ZNF470, ZNF521, and ZNF780B in chondrocytes from patients affected by OA and from subjects not affected by OA. This analysis highlighted a significantly lower expression of the transcript encoding ZNF423 in chondrocytes from OA, particularly in elderly patients. Interestingly, this decrease was mirrored by the similarly reduced expression of PPARγ, a known target of ZNF423 with anti-inflammatory and chondroprotective properties. The ZNF521 mRNA instead was abundant in all primary chondrocytes studied; the RNAi-mediated silencing of this gene significantly altered the COL2A/COL1 expression ratio, associated with the maintenance of the differentiated phenotype, in chondrocytes cultivated in alginate beads. These results suggest a role for ZNF423 and ZNF521 in the regulation of chondrocyte homeostasis and warrant further investigations to elucidate their mechanism of action.

  3. Identification of ‘safe harbor’ loci in indica rice genome by harnessing the property of zinc-finger nucleases to induce DNA damage and repair.

    Directory of Open Access Journals (Sweden)

    Christian eCantos

    2014-06-01

    Full Text Available Zinc-finger nucleases (ZFNs have proved to be successful tools for targeted genome manipulation in several organisms. Their main property is the induction of double-strand breaks (DSBs at specific sites, which are further repaired through homologous recombination (HR or non-homologous end joining (NHEJ. However, for the appropriate integration of genes at specific chromosomal locations, proper sites for gene integration need to be identified. These regions, hereby named safe harbor loci, must be localized in non-coding regions and possess high gene expression. In the present study, three different ZFN constructs (pZFN1, pZFN2, pZFN3, harboring β-glucuronidase (GUS as a reporter gene, were used to identify safe harbor loci regions on rice chromosomes. The constructs were delivered into IR64 rice by using an improved Agrobacterium-mediated transformation protocol, based on the use of immature embryos. Gene expression was measured by histochemical GUS activity and the flanking regions were determined through thermal-asymmetric interlaced polymerase chain reaction (TAIL PCR. Following sequencing, 28 regions were identified as putative sites for safe integration, but only one was localized in a non-coding region and it also possessed high GUS expression. These findings have significant applicability to create crops with new and valuable traits, since the site can be subsequently used to stably introduce one or more genes in a targeted manner.

  4. Clinical Scale Zinc Finger Nuclease-mediated Gene Editing of PD-1 in Tumor Infiltrating Lymphocytes for the Treatment of Metastatic Melanoma

    Science.gov (United States)

    Beane, Joal D; Lee, Gary; Zheng, Zhili; Mendel, Matthew; Abate-Daga, Daniel; Bharathan, Mini; Black, Mary; Gandhi, Nimisha; Yu, Zhiya; Chandran, Smita; Giedlin, Martin; Ando, Dale; Miller, Jeff; Paschon, David; Guschin, Dmitry; Rebar, Edward J; Reik, Andreas; Holmes, Michael C; Gregory, Philip D; Restifo, Nicholas P; Rosenberg, Steven A; Morgan, Richard A; Feldman, Steven A

    2015-01-01

    Programmed cell death-1 (PD-1) is expressed on activated T cells and represents an attractive target for gene-editing of tumor targeted T cells prior to adoptive cell transfer (ACT). We used zinc finger nucleases (ZFNs) directed against the gene encoding human PD-1 (PDCD-1) to gene-edit melanoma tumor infiltrating lymphocytes (TIL). We show that our clinical scale TIL production process yielded efficient modification of the PD-1 gene locus, with an average modification frequency of 74.8% (n = 3, range 69.9–84.1%) of the alleles in a bulk TIL population, which resulted in a 76% reduction in PD-1 surface-expression. Forty to 48% of PD-1 gene-edited cells had biallelic PD-1 modification. Importantly, the PD-1 gene-edited TIL product showed improved in vitro effector function and a significantly increased polyfunctional cytokine profile (TNFα, GM-CSF, and IFNγ) compared to unmodified TIL in two of the three donors tested. In addition, all donor cells displayed an effector memory phenotype and expanded approximately 500–2,000-fold in vitro. Thus, further study to determine the efficiency and safety of adoptive cell transfer using PD-1 gene-edited TIL for the treatment of metastatic melanoma is warranted. PMID:25939491

  5. Screening and validation of specific zinc finger nucleases targeted pig CFTR gene%猪CFTR基因特异性锌指核酸酶的筛选与鉴定

    Institute of Scientific and Technical Information of China (English)

    王瑞; 王令; 林娟; 张存芳; 张智英

    2012-01-01

    【目的】获得靶向猪囊性纤维化跨膜传导调节因子(Cystic fibrosis transmembrane conductance regulator,CFTR)基因的特异锌指核酸酶(Zinc finger nuclease,ZFN),为建立CFTR基因敲除猪细胞系提供技术支持。【方法】通过开源式(Oligomerized pool engineering,OPEN)方法筛选,首先以已知三锌指蛋白为框架,随机突变单个锌指关键位点氨基酸编码序列,建立人工三锌指蛋白随机库;然后应用细菌双杂交技术,从库中筛选出能够结合CFTR基因靶位点的三锌指蛋白;最后将获得的锌指蛋白与非限制性核酸内切酶FokⅠ组装成特异ZFN,通过酵母验证体系,检测ZFN靶向切割其识别序列的效率。【结果】获得3个人工三锌指蛋白随机库,每个库约含有2×106个单克隆,通过2轮细菌双杂交筛选,分别获得48个针对CFTR基因左右两侧靶位点的三锌指蛋白。ZFN活性验证结果表明,约90%的ZFN能够在酵母细胞内实现靶向切割,获得了高效特异的ZFN。【结论】获得了猪CFTR基因高效特异的ZFN。%【Objective】Specific zinc finger nucleases(ZFN)were screened to target pig cystic fibrosis transmembrane conductance regulator(CFTR)gene for the preparation of generating CFTR gene knockout pig cell line.【Method】Upon the oligomerized pool engineering(OPEN)strategy,firstly,three libraries of 3-zinc finger proteins were constructed.The DNA sequences of recognition amino acid residues of a finger motif were randomized by cassette mutagenesis.Secondly,bacterial twohybrid(B2H)system was used to screen for ZFPs from the three zinc finger protein libraries.Finally,selected ZFPs were fused to FokⅠdomain to generate specific ZFN for subsequent validation of ZFN cleavage activity in yeast.【Result】Three artificial randomly libraries of 3-zinc finger proteins were obtained,and each contained about 2×106 colonies.The three zinc finger random libraries were used

  6. ZIP4 Regulates Pancreatic Cancer Cell Growth by Activating IL-6/STAT3 Pathway via Zinc Finger Transcription Factor CREB

    Science.gov (United States)

    Zhang, Yuqing; Bharadwaj, Uddalak; Logsdon, Craig D.; Chen, Changyi; Yao, Qizhi; Li, Min

    2010-01-01

    Purpose Recent studies indicate a strong correlation of zinc transporter ZIP4 and pancreatic cancer progression; however, the underlying mechanisms are unclear. We have recently found that ZIP4 is overexpressed in pancreatic cancer. In this study, we investigated the signaling pathway through which ZIP4 regulates pancreatic cancer growth. Experimental Design The expression of cyclin D1, IL-6, and STAT3 in pancreatic cancer xenografts and cells were examined by real time PCR, Bio-Plex cytokine assay, and Western blot, respectively. The activity of CREB is examined by a promoter activity assay. Results Cyclin D1 was significantly increased in the ZIP4 overexpressing MIA PaCa-2 cells (MIA-ZIP4)-injected orthotopic xenografts and was downregulated in the ZIP4 silenced ASPC-1 (ASPC-shZIP4) group. The phosphorylation of signal transducer and activator of transcription 3 (STAT3), an upstream activator of cyclin D1, was increased in MIA-ZIP4 cells, and decreased in ASPC-shZIP4 cells. IL-6, a known upstream activator for STAT3, was also found to be significantly increased in the MIA-ZIP4 cells and xenografts, and decreased in the ASPC-shZIP4 group. Overexpression of ZIP4 led to a 75% increase of IL-6 promoter activity, and caused increased phosphorylation of cAMP response element binding protein (CREB). Conclusions Our study suggest that ZIP4 overexpression causes increased IL-6 transcription via CREB, which in turn activates STAT3, and leads to increased cyclin D1 expression, resulting in increased cell proliferation and tumor progression in pancreatic cancer. These results elucidated a novel pathway in ZIP4-mediated pancreatic cancer growth, and suggest new therapeutic targets including ZIP4, IL-6, and STAT3 in pancreatic cancer treatment. PMID:20160059

  7. Identity of zinc finger nucleases with specificity to herpes simplex virus type II genomic DNA: novel HSV-2 vaccine/therapy precursors

    Directory of Open Access Journals (Sweden)

    Wayengera Misaki

    2011-06-01

    Full Text Available Abstract Background Herpes simplex type II (HSV-2 is a member of the family herpesviridae. Human infection with this double stranded linear DNA virus causes genital ulcerative disease and existing treatment options only serve to resolve the symptomatology (ulcers associated with active HSV-2 infection but do not eliminate latent virus. As a result, infection with HSV-2 follows a life-long relapsing (active versus latent course. On the basis of a primitive bacterium anti-phage DNA defense, the restriction modification (R-M system, we previously identified the Escherichia coli restriction enzyme (REase EcoRII as a novel peptide to excise or irreversibly disrupt latent HSV-2 DNA from infected cells. However, sequences of the site specificity palindrome of EcoRII 5'-CCWGG-3' (W = A or T are equally present within the human genome and are a potential source of host-genome toxicity. This feature has limited previous HSV-2 EcoRII based therapeutic models to microbicides only, and highlights the need to engineer artificial REases (zinc finger nucleases-ZFNs with specificity to HSV-2 genomic-DNA only. Herein, the therapeutic-potential of zinc finger arrays (ZFAs and ZFNs is identified and modeled, with unique specificity to the HSV-2 genome. Methods and results Using the whole genome of HSV-2 strain HG52 (Dolan A et al.,, and with the ZFN-consortium's CoDA-ZiFiT software pre-set at default, more than 28,000 ZFAs with specificity to HSV-2 DNA were identified. Using computational assembly (through in-silico linkage to the Flavobacterium okeanokoites endonuclease Fok I of the type IIS class, 684 ZFNs with specificity to the HSV-2 genome, were constructed. Graphic-analysis of the HSV-2 genome-cleavage pattern using the afore-identified ZFNs revealed that the highest cleavage-incidence occurred within the 30,950 base-pairs (~between the genomic context coordinates 0.80 and 1.00 at the 3' end of the HSV-2 genome. At approximately 3,095 bp before and after the

  8. Editing of the Luteinizing Hormone Gene to Sterilize Channel Catfish, Ictalurus punctatus, Using a Modified Zinc Finger Nuclease Technology with Electroporation.

    Science.gov (United States)

    Qin, Zhenkui; Li, Yun; Su, Baofeng; Cheng, Qi; Ye, Zhi; Perera, Dayan A; Fobes, Michael; Shang, Mei; Dunham, Rex A

    2016-04-01

    Channel catfish (Ictalurus punctatus) is the most important freshwater aquaculture species in the USA. Genetically enhanced fish that are sterile could both profit the catfish industry and reduce potential environmental and ecological risks. As the first step to generate sterile channel catfish, three sets of zinc finger nuclease (ZFN) plasmids targeting the luteinizing hormone (LH) gene were designed and electroporated into one-cell embryos, different concentrations were introduced, and the Cel-I assay was conducted to detect mutations. Channel catfish carrying the mutated LH gene were sterile, as confirmed by DNA sequencing and mating experiments. The overall mutation rate was 19.7 % for 66 channel catfish, and the best treatment was ZFN set 1 at the concentration 25 μg/ml. To our knowledge, this is the first instance of gene editing of fish via plasmid introduction instead of mRNA microinjection. The introduction of the ZFN plasmids may have reduced mosaicism, as mutated individuals were gene edited in every tissue evaluated. Apparently, the plasmids were eventually degraded without integration, as they were not detectable in mutated individuals using PCR. Carp pituitary extract failed to induce spawning and restoration of fertility, indicating the need for developing other hormone therapies to achieve reversal of sterility upon demand. This is the first sterilization achieved using ZFN technology in an aquaculture species and the first successful gene editing of channel catfish. Our results will help understand the roles of the LH gene, purposeful sterilization of teleost fishes, and is a step towards control of domestic, hybrid, exotic, invasive, and transgenic fishes. PMID:26846523

  9. ZINC FINGER OF ARABIDOPSIS THALIANA12 (ZAT12) Interacts with FER-LIKE IRON DEFICIENCY-INDUCED TRANSCRIPTION FACTOR (FIT) Linking Iron Deficiency and Oxidative Stress Responses.

    Science.gov (United States)

    Le, Cham Thi Tuyet; Brumbarova, Tzvetina; Ivanov, Rumen; Stoof, Claudia; Weber, Eva; Mohrbacher, Julia; Fink-Straube, Claudia; Bauer, Petra

    2016-01-01

    Plants grown under iron (Fe)-deficient conditions induce a set of genes that enhance the efficiency of Fe uptake by the roots. In Arabidopsis (Arabidopsis thaliana), the central regulator of this response is the basic helix-loop-helix transcription factor FER-LIKE IRON DEFICIENCY-INDUCED TRANSCRIPTION FACTOR (FIT). FIT activity is regulated by protein-protein interactions, which also serve to integrate external signals that stimulate and possibly inhibit Fe uptake. In the search of signaling components regulating FIT function, we identified ZINC FINGER OF ARABIDOPSIS THALIANA12 (ZAT12), an abiotic stress-induced transcription factor. ZAT12 interacted with FIT, dependent on the presence of the ethylene-responsive element-binding factor-associated amphiphilic repression motif. ZAT12 protein was found expressed in the root early differentiation zone, where its abundance was modulated in a root layer-specific manner. In the absence of ZAT12, FIT expression was upregulated, suggesting a negative effect of ZAT12 on Fe uptake. Consistently, zat12 loss-of-function mutants had higher Fe content than the wild type at sufficient Fe. We found that under Fe deficiency, hydrogen peroxide (H2O2) levels were enhanced in a FIT-dependent manner. FIT protein, in turn, was stabilized by H2O2 but only in the presence of ZAT12, showing that H2O2 serves as a signal for Fe deficiency responses. We propose that oxidative stress-induced ZAT12 functions as a negative regulator of Fe acquisition. A model where H2O2 mediates the negative regulation of plant responses to prolonged stress might be applicable to a variety of stress conditions.

  10. The brain-specific neural zinc finger transcription factor 2b (NZF-2b/7ZFMyt1 suppresses cocaine self-administration in rats

    Directory of Open Access Journals (Sweden)

    Vijay Chandrasekar

    2010-04-01

    Full Text Available Brain-specific neural-zinc-finger transcription factor-2b (NZF2b/7ZFMyt1 is induced in the mesolimbic dopaminergic region after chronic cocaine exposure and lentiviral-mediated expression of NZF2b/7ZFMyt1 in the nucleus accumbens results in decreased locomotor activity (Chandrasekar and Dreyer, 2009. In this study the role of NZF2b/7ZFMyt1 in active cocaine seeking and of its interaction with histone deacetylase on the altered behavior has been observed. Localized expression of NZF2b/7ZFMyt1 in the nucleus accumbens resulted in attenuated cocaine self-administration, whereas silencing this transcription factor with lentiviruses expressing siRNAs increased the animal′s motivation to self-infuse cocaine. Low doses of sodium butyrate, a potent inhibitor of histone deacetylase, were sufficient to reverse the NZF2b/7ZFMyt1-mediated decrease in cocaine self-administration. NZF2b/7ZFMyt1 expression resulted in strong induction of transcription factors REST1 and NAC1 and of the dopamine D2 receptor, with concomitant inhibition of BDNF and its receptor TrkB. We show that NZF2b/7ZFMyt1 colocalizes with histone deacetylase-2 (HDAC2, probably overcoming the suppression of transcriptional activity caused by Lingo1. These findings show that molecular adaptations mediated by NZF2b/7ZFMyt1 expression possibly lead to decreased responsiveness to the reinforcing properties of cocaine and play a prominent role in affecting the behavioral changes induced by the drug.

  11. ZINC FINGER OF ARABIDOPSIS THALIANA12 (ZAT12) Interacts with FER-LIKE IRON DEFICIENCY-INDUCED TRANSCRIPTION FACTOR (FIT) Linking Iron Deficiency and Oxidative Stress Responses.

    Science.gov (United States)

    Le, Cham Thi Tuyet; Brumbarova, Tzvetina; Ivanov, Rumen; Stoof, Claudia; Weber, Eva; Mohrbacher, Julia; Fink-Straube, Claudia; Bauer, Petra

    2016-01-01

    Plants grown under iron (Fe)-deficient conditions induce a set of genes that enhance the efficiency of Fe uptake by the roots. In Arabidopsis (Arabidopsis thaliana), the central regulator of this response is the basic helix-loop-helix transcription factor FER-LIKE IRON DEFICIENCY-INDUCED TRANSCRIPTION FACTOR (FIT). FIT activity is regulated by protein-protein interactions, which also serve to integrate external signals that stimulate and possibly inhibit Fe uptake. In the search of signaling components regulating FIT function, we identified ZINC FINGER OF ARABIDOPSIS THALIANA12 (ZAT12), an abiotic stress-induced transcription factor. ZAT12 interacted with FIT, dependent on the presence of the ethylene-responsive element-binding factor-associated amphiphilic repression motif. ZAT12 protein was found expressed in the root early differentiation zone, where its abundance was modulated in a root layer-specific manner. In the absence of ZAT12, FIT expression was upregulated, suggesting a negative effect of ZAT12 on Fe uptake. Consistently, zat12 loss-of-function mutants had higher Fe content than the wild type at sufficient Fe. We found that under Fe deficiency, hydrogen peroxide (H2O2) levels were enhanced in a FIT-dependent manner. FIT protein, in turn, was stabilized by H2O2 but only in the presence of ZAT12, showing that H2O2 serves as a signal for Fe deficiency responses. We propose that oxidative stress-induced ZAT12 functions as a negative regulator of Fe acquisition. A model where H2O2 mediates the negative regulation of plant responses to prolonged stress might be applicable to a variety of stress conditions. PMID:26556796

  12. Zinc-Finger Transcription Factor ZAT6 Positively Regulates Cadmium Tolerance through the Glutathione-Dependent Pathway in Arabidopsis1[OPEN

    Science.gov (United States)

    Chen, Jian; Yan, Xingxing; Liu, Yunlei; Wang, Ren; Fan, Tingting; Ren, Yongbing; Tang, Xiaofeng; Xiao, Fangming

    2016-01-01

    Cadmium (Cd) is an environmental pollutant with high toxicity to animals and plants. It has been established that the glutathione (GSH)-dependent phytochelatin (PC) synthesis pathway is one of the most important mechanisms contributing to Cd accumulation and tolerance in plants. However, the transcription factors involved in regulating GSH-dependent PC synthesis pathway remain largely unknown. Here, we identified an Arabidopsis (Arabidopsis thaliana) Cd-resistant mutant xcd2-D (XVE system-induced cadmium-tolerance2) using a forward genetics approach. The mutant gene underlying xcd2-D mutation was revealed to encode a known zinc-finger transcription factor, ZAT6. Transgenic plants overexpressing ZAT6 showed significant increase of Cd tolerance, whereas loss of function of ZAT6 led to decreased Cd tolerance. Increased Cd accumulation and tolerance in ZAT6-overexpressing lines was GSH dependent and associated with Cd-activated synthesis of PC, which was correlated with coordinated activation of PC-synthesis related gene expression. By contrast, loss of function of ZAT6 reduced Cd accumulation and tolerance, which was accompanied by abolished PC synthesis and gene expression. Further analysis revealed that ZAT6 positively regulates the transcription of GSH1, GSH2, PCS1, and PCS2, but ZAT6 is capable of specifically binding to GSH1 promoter in vivo. Consistently, overexpression of GSH1 has been shown to restore Cd sensitivity in the zat6-1 mutant, suggesting that GSH1 is a key target of ZAT6. Taken together, our data provide evidence that ZAT6 coordinately activates PC synthesis-related gene expression and directly targets GSH1 to positively regulate Cd accumulation and tolerance in Arabidopsis. PMID:26983992

  13. Cytotoxic Effects during Knock Out of Multiple Porcine Endogenous Retrovirus (PERV Sequences in the Pig Genome by Zinc Finger Nucleases (ZFN.

    Directory of Open Access Journals (Sweden)

    Marwan Semaan

    Full Text Available Xenotransplantation has been proposed as a solution to the shortage of suitable human donors for transplantation and pigs are currently favoured as donor animals. However, xenotransplantation may be associated with the transmission of zoonotic microorganisms. Whereas most porcine microorganisms representing a risk for the human recipient may be eliminated by designated pathogen free breeding, multiple copies of porcine endogenous retroviruses (PERVs are integrated in the genome of all pigs and cannot be eliminated this way. PERVs are released as infectious particles and infect human cells. The zinc finger nuclease (ZFN technology allows knocking out specifically cellular genes, however it was not yet used to eliminate multiple integrated proviral sequences with a strong conservation in the target sequence. To reduce the risk of horizontal PERV transmission and to knock out as many as possible proviruses, for the first time the powerful tool of the ZFN technology was used. ZFN were designed to bind specifically to sequences conserved in all known replication-competent proviruses. Expression and transport of the ZFN into the nucleus was shown by Western blot analysis, co-localisation analysis, PLA and FRET. Survival of transfected cells was analysed using fluorescent ZFN and cell counting. After transfection a strong expression of the ZFN proteins and a co-localisation of the expressed ZFN proteins were shown. However, expression of the ZFN was found to be extremely toxic for the transfected cells. The induced cytotoxicity was likely due to the specific cutting of the high copy number of the PERV proviruses, which is also commonly observed when ZFN with low specificity cleave numerous off-target sites in a genome. This is the first attempt to knock out multiple, nearly identical, genes in a cellular genome using ZFN. The attempt failed, and other strategies should be used to prevent PERV transmission.

  14. Wheat zinc finger protein TaLSD1, a negative regulator of programmed cell death, is involved in wheat resistance against stripe rust fungus.

    Science.gov (United States)

    Guo, Jun; Bai, Pengfei; Yang, Qian; Liu, Furong; Wang, Xiaodong; Huang, Lili; Kang, Zhensheng

    2013-10-01

    Genetic characterization of the Arabidopsis lesion simulating disease 1 (lsd1) mutant, a lesion mimic mutant (LMM), has revealed the essential role of AtLSD1 in the negative regulation of cell death and disease resistance. The three zinc-finger motifs found in AtLSD1 revealed a novel plant-specific gene family, whose members are significantly related to programmed cell death (PCD). In this study, we characterized a functional homologue to AtLSD1, TaLSD1, in the wheat-stripe rust fungus pathosystem. The expression of TaLSD1 was differentially induced during incompatible and compatible interactions between wheat and Puccinia striiformis f. sp. tritici (Pst) and was up-regulated by oxidative stress generated by methyl viologen (MV). TaLSD1 was found to be predominately localized in the nucleus of onion epidermal cell. Transient overexpression assays in Nicotiana benthamiana demonstrated that TaLSD1 partially inhibited programmed cell death triggered by a mouse Bax protein, whereas expression of TaLSD1 alone had no influence on the phenotype of tobacco. Knocking down the expression of TaLSD1 through virus-induced gene silencing (VIGS) increased wheat resistance against Pst accompanied by an enhanced hypersensitive response (HR), an increase in PR1 gene expression and a reduction in Pst hyphal growth. Our results suggest that TaLSD1 functions negatively in regulating the plant hypersensitive cell death and is involved in disease resistance of wheat against the stripe rust pathogen.

  15. Scientific opinion addressing the safety assessment of plants developed using Zinc Finger Nuclease 3 and other Site-Directed Nucleases with similar function

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Genetically Modified Organisms (GMO

    2012-10-01

    Full Text Available

    The European Commission requested that the EFSA Panel on Genetically Modified Organisms deliver a scientific opinion related to risk assessment of plants developed using the zinc finger nuclease 3 technique (ZFN-3 which allows the integration of gene(s in a predefined insertion site in the genome of the recipient species. Since other nucleases with a similar function to ZFN are considered in this opinion the term site-directed nuclease 3 (SDN-3 is used to describe the technique rather than ZFN-3 specifically. The EFSA GMO Panel considers that its guidance documents are applicable for the evaluation of food and feed products derived from plants developed using the SDN-3 technique and for performing an environmental risk assessment. However, on a case-by-case basis lesser amounts of event specific data may be needed for the risk assessment of plants developed using the SDN-3 technique. The EFSA GMO Panel compared the hazards associated with plants produced by the SDN-3 technique with those obtained by conventional plant breeding techniques and by currently used transgenesis. With respect to the genes introduced, the SDN-3 technique does not differ from transgenesis or from the other genetic modification techniques currently used, and can be used to introduce transgenes, intragenes or cisgenes. The main difference between the SDN-3 technique and transgenesis is that the insertion of DNA is targeted to a predefined region of the genome. Therefore, the SDN-3 technique can minimise hazards associated with the disruption of genes and/or regulatory elements in the recipient genome. Whilst the SDN-3 technique can induce off-target changes in the genome of the recipient plant these would be fewer than those occurring with most mutagenesis techniques. Furthermore, where such changes occur they would be of the same types as those produced by conventional breeding techniques.

  16. Diversity of Prdm9 zinc finger array in wild mice unravels new facets of the evolutionary turnover of this coding minisatellite.

    Directory of Open Access Journals (Sweden)

    Jérôme Buard

    Full Text Available In humans and mice, meiotic recombination events cluster into narrow hotspots whose genomic positions are defined by the PRDM9 protein via its DNA binding domain constituted of an array of zinc fingers (ZnFs. High polymorphism and rapid divergence of the Prdm9 gene ZnF domain appear to involve positive selection at DNA-recognition amino-acid positions, but the nature of the underlying evolutionary pressures remains a puzzle. Here we explore the variability of the Prdm9 ZnF array in wild mice, and uncovered a high allelic diversity of both ZnF copy number and identity with the caracterization of 113 alleles. We analyze features of the diversity of ZnF identity which is mostly due to non-synonymous changes at codons -1, 3 and 6 of each ZnF, corresponding to amino-acids involved in DNA binding. Using methods adapted to the minisatellite structure of the ZnF array, we infer a phylogenetic tree of these alleles. We find the sister species Mus spicilegus and M. macedonicus as well as the three house mouse (Mus musculus subspecies to be polyphyletic. However some sublineages have expanded independently in Mus musculus musculus and M. m. domesticus, the latter further showing phylogeographic substructure. Compared to random genomic regions and non-coding minisatellites, none of these patterns appears exceptional. In silico prediction of DNA binding sites for each allele, overlap of their alignments to the genome and relative coverage of the different families of interspersed repeated elements suggest a large diversity between PRDM9 variants with a potential for highly divergent distributions of recombination events in the genome with little correlation to evolutionary distance. By compiling PRDM9 ZnF protein sequences in Primates, Muridae and Equids, we find different diversity patterns among the three amino-acids most critical for the DNA-recognition function, suggesting different diversification timescales.

  17. Improvement of acetic acid tolerance of Saccharomyces cerevisiae using a zinc-finger-based artificial transcription factor and identification of novel genes involved in acetic acid tolerance.

    Science.gov (United States)

    Ma, Cui; Wei, Xiaowen; Sun, Cuihuan; Zhang, Fei; Xu, Jianren; Zhao, Xinqing; Bai, Fengwu

    2015-03-01

    Acetic acid is present in cellulosic hydrolysate as a potent inhibitor, and the superior acetic acid tolerance of Saccharomyces cerevisiae ensures good cell viability and efficient ethanol production when cellulosic raw materials are used as substrates. In this study, a mutant strain of S. cerevisiae ATCC4126 (Sc4126-M01) with improved acetic acid tolerance was obtained through screening strains transformed with an artificial zinc finger protein transcription factor (ZFP-TF) library. Further analysis indicated that improved acetic acid tolerance was associated with improved catalase (CAT) activity. The ZFP coding sequence associated with the improved phenotype was identified, and real-time RT-PCR analysis revealed that three of the possible genes involved in the enhanced acetic acid tolerance regulated by this ZFP-TF, namely YFL040W, QDR3, and IKS1, showed decreased transcription levels in Sc4126-M01 in the presence of acetic acid, compared to those in the control strain. Sc4126-M01 mutants having QDR3 and IKS1 deletion (ΔQDR3 and ΔIKS1) exhibited higher acetic acid tolerance than the wild-type strain under acetic acid treatment. Glucose consumption rate and ethanol productivity in the presence of 5 g/L acetic acid were improved in the ΔQDR3 mutant compared to the wild-type strain. Our studies demonstrated that the synthetic ZFP-TF library can be used to improve acetic acid tolerance of S. cerevisiae and that the employment of an artificial transcription factor can facilitate the exploration of novel functional genes involved in stress tolerance of S. cerevisiae. PMID:25698512

  18. The BTB-zinc finger transcription factor abrupt acts as an epithelial oncogene in Drosophila melanogaster through maintaining a progenitor-like cell state.

    Directory of Open Access Journals (Sweden)

    Nezaket Turkel

    Full Text Available The capacity of tumour cells to maintain continual overgrowth potential has been linked to the commandeering of normal self-renewal pathways. Using an epithelial cancer model in Drosophila melanogaster, we carried out an overexpression screen for oncogenes capable of cooperating with the loss of the epithelial apico-basal cell polarity regulator, scribbled (scrib, and identified the cell fate regulator, Abrupt, a BTB-zinc finger protein. Abrupt overexpression alone is insufficient to transform cells, but in cooperation with scrib loss of function, Abrupt promotes the formation of massive tumours in the eye/antennal disc. The steroid hormone receptor coactivator, Taiman (a homologue of SRC3/AIB1, is known to associate with Abrupt, and Taiman overexpression also drives tumour formation in cooperation with the loss of Scrib. Expression arrays and ChIP-Seq indicates that Abrupt overexpression represses a large number of genes, including steroid hormone-response genes and multiple cell fate regulators, thereby maintaining cells within an epithelial progenitor-like state. The progenitor-like state is characterised by the failure to express the conserved Eyes absent/Dachshund regulatory complex in the eye disc, and in the antennal disc by the failure to express cell fate regulators that define the temporal elaboration of the appendage along the proximo-distal axis downstream of Distalless. Loss of scrib promotes cooperation with Abrupt through impaired Hippo signalling, which is required and sufficient for cooperative overgrowth with Abrupt, and JNK (Jun kinase signalling, which is required for tumour cell migration/invasion but not overgrowth. These results thus identify a novel cooperating oncogene, identify mammalian family members of which are also known oncogenes, and demonstrate that epithelial tumours in Drosophila can be characterised by the maintenance of a progenitor-like state.

  19. The BTB-zinc Finger Transcription Factor Abrupt Acts as an Epithelial Oncogene in Drosophila melanogaster through Maintaining a Progenitor-like Cell State

    Science.gov (United States)

    Turkel, Nezaket; Sahota, Virender K.; Bolden, Jessica E.; Goulding, Karen R.; Doggett, Karen; Willoughby, Lee F.; Blanco, Enrique; Martin-Blanco, Enrique; Corominas, Montserrat; Ellul, Jason; Aigaki, Toshiro; Richardson, Helena E.; Brumby, Anthony M.

    2013-01-01

    The capacity of tumour cells to maintain continual overgrowth potential has been linked to the commandeering of normal self-renewal pathways. Using an epithelial cancer model in Drosophila melanogaster, we carried out an overexpression screen for oncogenes capable of cooperating with the loss of the epithelial apico-basal cell polarity regulator, scribbled (scrib), and identified the cell fate regulator, Abrupt, a BTB-zinc finger protein. Abrupt overexpression alone is insufficient to transform cells, but in cooperation with scrib loss of function, Abrupt promotes the formation of massive tumours in the eye/antennal disc. The steroid hormone receptor coactivator, Taiman (a homologue of SRC3/AIB1), is known to associate with Abrupt, and Taiman overexpression also drives tumour formation in cooperation with the loss of Scrib. Expression arrays and ChIP-Seq indicates that Abrupt overexpression represses a large number of genes, including steroid hormone-response genes and multiple cell fate regulators, thereby maintaining cells within an epithelial progenitor-like state. The progenitor-like state is characterised by the failure to express the conserved Eyes absent/Dachshund regulatory complex in the eye disc, and in the antennal disc by the failure to express cell fate regulators that define the temporal elaboration of the appendage along the proximo-distal axis downstream of Distalless. Loss of scrib promotes cooperation with Abrupt through impaired Hippo signalling, which is required and sufficient for cooperative overgrowth with Abrupt, and JNK (Jun kinase) signalling, which is required for tumour cell migration/invasion but not overgrowth. These results thus identify a novel cooperating oncogene, identify mammalian family members of which are also known oncogenes, and demonstrate that epithelial tumours in Drosophila can be characterised by the maintenance of a progenitor-like state. PMID:23874226

  20. Absolute Integral Cross Sections for the State-selected Ion–Molecule Reaction N2+(X2Σg+ v+ = 0–2) + C2H2 in the Collision Energy Range of 0.03–10.00 eV

    Science.gov (United States)

    Xu, Yuntao; Xiong, Bo; Chung Chang, Yih; Ng, C. Y.

    2016-08-01

    Using the vacuum ultraviolet laser pulsed field ionization-photoion source, together with the double-quadrupole–double-octopole mass spectrometer developed in our laboratory, we have investigated the state-selected ion–molecule reaction {{{{N}}}2}+({X}2{{{{Σ }}}{{g}}}+; v + = 0–2, N+ = 0–9) + C2H2, achieving high internal-state selectivity and high kinetic energy resolution for reactant {{{{N}}}2}+ ions. The charge transfer (CT) and hydrogen-atom transfer (HT) channels, which lead to the respective formation of product {{{C}}}2{{{{H}}}2}+ and N2H+ ions, are observed. The vibrationally selected absolute integral cross sections for the CT [σ CT(v +)] and HT [[σ HT(v +)] channels obtained in the center-of-mass collision energy (E cm) range of 0.03–10.00 eV reveal opposite E cm dependences. The σ CT(v +) is found to increase as E cm is decreased, and is consistent with the long-range exothermic CT mechanism, whereas the E cm enhancement observed for the σ HT(v +) suggests effective coupling of kinetic energy to internal energy, enhancing the formation of N2H+. The σ HT(v +) curve exhibits a step at E cm = 0.70–1.00 eV, suggesting the involvement of the excited {{{C}}}2{{{{H}}}2}+({A}2{{{{Σ }}}{{g}}}+) state in the HT reaction. Contrary to the strong E cm dependences for σ CT(v +) and σ HT(v +), the effect of vibrational excitation of {{{{N}}}2}+ on both the CT and HT channels is marginal. The branching ratios and cross sections for the CT and HT channels determined in the present study are useful for modeling the atmospheric compositions of Saturn's largest moon, Titan. These cross sections and branching ratios are also valuable for benchmarking theoretical calculations on chemical dynamics of the titled reaction.

  1. Expression, Purification and Characterization of the Zinc Finger Domain of PLZF Protein%重组PLZF蛋白锌指结构域的表达、提纯和活性分析

    Institute of Scientific and Technical Information of China (English)

    李伟

    2011-01-01

    PLZF is an important transcription repressor. It consists of an N-terminal BTB domain and a C-terminal zinc finger domain. To date, the three dimensional structure of the zinc finger domain is still not clear. Hence, expression and purification studies were performed. To produce the zinc finger domain of PLZF protein, the sequence encoding zinc finger domain fragment with a start codon added to the 5' was inserted into PET-lla expression vector. The expression plasmid was transformed into E. Coli BL21 ( DE3) strain and protein expressionwas induced by IPTG. The recombinant protein was found to be expressed as inclusion bodies. The inclusion body was solubilized using buffers containing SDS detergent and proteins were purified to more than 96 % homogeneity through gel filtration. The purified proteins were denatured and then refolded by reverse dialysis. The refolded proteins were characterized by DNA electrophoretic mobility shift assay ( EMS A) and found to be active. An important foundation has been laid for further three-dimensional structural study on the zinc finger domain of PLZF.%PLZF(promyelocytic leukaemia zinc finger protein)是一种重要的转录抑制因子,它由位于N端的BTB结构域和C端的锌指结构域构成.鉴于目前对于锌指结构域的立体结构还不是十分清楚,对其进行了高效表达和提纯.为了表达PLZF蛋白的锌指结构域,在其编码序列的5'端加上起始密码ATG后插入到表达载体PET-11a的多克隆位点.构建好的表达质粒转化到BI21( DE3)大肠杆菌内并用IPTG诱导表达,发现重组蛋白主要以不溶性的包涵体形式在胞内表达.用含有SDS变性剂的缓冲液溶解包涵体后,采用凝胶过滤方法将重组蛋白纯化到纯度达96%以上.对纯化后的蛋白质用反透析的方法进行复性,然后用DNA结合实验进行活性分析,发现复性后的蛋白质具有特异的DNA结合活性,这为进一步研究PLZF蛋白锌指结构域的立体结构打下了重要基础.

  2. Arabidopsis CDS blastp result: AK287566 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287566 J065027L04 At1g34790.1 68414.m04337 transparent testa 1 protein (TT1) / zi...nc finger (C2H2 type) protein TT1 identical to transparent testa 1 GI:18253279 from [Arabidopsis thaliana]; contains Pfam profile PF00096: Zinc finger, C2H2 type 2e-77 ...

  3. Arabidopsis CDS blastp result: AK289209 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK289209 J100058I16 At1g34790.1 68414.m04337 transparent testa 1 protein (TT1) / zi...nc finger (C2H2 type) protein TT1 identical to transparent testa 1 GI:18253279 from [Arabidopsis thaliana]; contains Pfam profile PF00096: Zinc finger, C2H2 type 1e-12 ...

  4. Arabidopsis CDS blastp result: AK243285 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243285 J100051N01 At1g34790.1 68414.m04337 transparent testa 1 protein (TT1) / zi...nc finger (C2H2 type) protein TT1 identical to transparent testa 1 GI:18253279 from [Arabidopsis thaliana]; contains Pfam profile PF00096: Zinc finger, C2H2 type 1e-24 ...

  5. Trigger finger

    Science.gov (United States)

    Digital stenosing tenosynovitis; Trigger digit; Trigger finger release ... JH. Carpal tunnel syndrome, ulnar tunnel syndrome, and stenosing tenosynovitis. In: Canale ST, Beaty JH, eds. Campbell's Operative ...

  6. A Zinc Finger Transcription Factor, αA-Crystallin Binding Protein 1, Is a Negative Regulator of the Chondrocyte-Specific Enhancer of the α1(II) Collagen Gene

    OpenAIRE

    Tanaka, Kazuhiro; Matsumoto, Yoshihiro; Nakatani, Fumihiko; Iwamoto, Yukihide; Yamada, Yoshihiko

    2000-01-01

    Transcription of the type II collagen gene (Col2a1) is regulated by multiple cis-acting sites. The enhancer element, which is located in the first intron, is necessary for high-level and cartilage-specific expression of Col2a1. A mouse limb bud cDNA expression library was screened by the Saccharomyces cerevisiae one-hybrid screening method to identify protein factors bound to the enhancer. A zinc finger protein, αA-crystallin binding protein 1 (CRYBP1), which had been reported to bind to the ...

  7. U7 snRNP-specific Lsm11 protein: dual binding contacts with the 100 kDa zinc finger processing factor (ZFP100) and a ZFP100-independent function in histone RNA 3′ end processing

    OpenAIRE

    Azzouz, Teldja N.; Gruber, Andreas; Schümperli, Daniel

    2005-01-01

    The 3' cleavage generating non-polyadenylated animal histone mRNAs depends on the base pairing between U7 snRNA and a conserved histone pre-mRNA downstream element. This interaction is enhanced by a 100 kDa zinc finger protein (ZFP100) that forms a bridge between an RNA hairpin element upstream of the processing site and the U7 small nuclear ribonucleoprotein (snRNP). The N-terminus of Lsm11, a U7-specific Sm-like protein, was shown to be crucial for histone RNA processing and to bind ZFP100....

  8. Study on a cDNA sequence of cold inducible zinc finger protein in albinism tea cultivar "Xiaoxueya"%茶树品种“小雪芽”冷诱导锌指蛋白基因cDNA研究

    Institute of Scientific and Technical Information of China (English)

    王开荣; 李娜娜; 陆建良; 郑新强; 梁月荣; 吴颖; 李明

    2012-01-01

    A cDNA sequence of cold inducible zinc finger protein in leaf of albinism tea cultivar " Xiaoxueya" was investigated. The results shows that the cDNA sequence had 698 bp in length, with 83% and 82% identity to zinc finger protein mRNA of Glycine max and Ricinus communis respectively. It had an opening reading frame encoding 230 amino acids. Compared to that of common tea cuhivar " Fudingdabai" , there were three loci of nucleotide deletion and one locus of nucleotide substitution. Its deduced amino acid sequence had 99% identity to that of cuhivar " Fudingdabai" , among which there were 3 loci of amino acid substitution. The expression of cold inducible zinc finger protein in " Xiaoxueya" ,sas significantly lower than in " Fudingdabai". It is considered that the mutation in gene sequence resulted in the sensitivity of cultivar "Xiaoxueya" to low temperature through low expression and mutation of the cold inducible zinc finger protein.%分析了低温诱导型新梢白化茶树品种“小雪芽”叶片低温诱导锌指蛋白基因cDNA序列。该序列长度为698bp,与大豆锌指蛋白mRNA同源性为83%,与蓖麻锌指蛋白mRNA同源性为82%;具有可编码230个氨基酸的开放阅读框。与“福鼎大白茶”冷诱导锌指蛋白eDNA序列相比,该eDNA序列在50—51位上核苷酸AT缺失,第143位的A被置换为G,第654位T缺失。其翻译的蛋白质氨基酸序列与“福鼎大白茶”同源性达到99%,但有3个位点的氨基酸变异。该基因表达丰度明显低于“福鼎大白茶”。研究认为,基因结构差异,引起表达强度和蛋白质氨基酸序列的差异,可能是引起“小雪芽”品种对低温敏感的重要因素。

  9. Use of zinc-finger nucleases to knock out the WAS gene in K562 cells: a human cellular model for Wiskott-Aldrich syndrome

    Directory of Open Access Journals (Sweden)

    Miguel G. Toscano

    2013-03-01

    Mutations in the WAS gene cause Wiskott-Aldrich syndrome (WAS, which is characterized by eczema, immunodeficiency and microthrombocytopenia. Although the role of WASP in lymphocytes and myeloid cells is well characterized, its role on megakaryocyte (MK development is poorly understood. In order to develop a human cellular model that mimics the megakaryocytic-derived defects observed in WAS patients we used K562 cells, a well-known model for study of megakaryocytic development. We knocked out the WAS gene in K562 cells using a zinc-finger nuclease (ZFN pair targeting the WAS intron 1 and a homologous donor DNA that disrupted WASP expression. Knockout of WASP on K562 cells (K562WASKO cells resulted in several megakaryocytic-related defects such as morphological alterations, lower expression of CD41ɑ, lower increments in F-actin polymerization upon stimulation, reduced CD43 expression and increased phosphatidylserine exposure. All these defects have been previously described either in WAS-knockout mice or in WAS patients, validating K562WASKO as a cell model for WAS. However, K562WASPKO cells showed also increased basal F-actin and adhesion, increased expression of CD61 and reduced expression of TGFβ and Factor VIII, defects that have never been described before for WAS-deficient cells. Interestingly, these phenotypic alterations correlate with different roles for WASP in megakaryocytic differentiation. All phenotypic alterations observed in K562WASKO cells were alleviated upon expression of WAS following lentiviral transduction, confirming the role of WASP in these phenotypes. In summary, in this work we have validated a human cellular model, K562WASPKO, that mimics the megakaryocytic-related defects found in WAS-knockout mice and have found evidences for a role of WASP as regulator of megakaryocytic differentiation. We propose the use of K562WASPKO cells as a tool to study the molecular mechanisms involved in the megakaryocytic-related defects observed in WAS

  10. Zinc-Finger Nuclease Knockout of Dual-Specificity Protein Phosphatase-5 Enhances the Myogenic Response and Autoregulation of Cerebral Blood Flow in FHH.1BN Rats

    Science.gov (United States)

    Fan, Fan; Geurts, Aron M.; Pabbidi, Mallikarjuna R.; Smith, Stanley V.; Harder, David R.; Jacob, Howard; Roman, Richard J.

    2014-01-01

    We recently reported that the myogenic responses of the renal afferent arteriole (Af-Art) and middle cerebral artery (MCA) and autoregulation of renal and cerebral blood flow (RBF and CBF) were impaired in Fawn Hooded hypertensive (FHH) rats and were restored in a FHH.1BN congenic strain in which a small segment of chromosome 1 from the Brown Norway (BN) containing 15 genes including dual-specificity protein phosphatase-5 (Dusp5) were transferred into the FHH genetic background. We identified 4 single nucleotide polymorphisms in the Dusp5 gene in FHH as compared with BN rats, two of which altered CpG sites and another that caused a G155R mutation. To determine whether Dusp5 contributes to the impaired myogenic response in FHH rats, we created a Dusp5 knockout (KO) rat in the FHH.1BN genetic background using a zinc-finger nuclease that introduced an 11 bp frame-shift deletion and a premature stop codon at AA121. The expression of Dusp5 was decreased and the levels of its substrates, phosphorylated ERK1/2 (p-ERK1/2), were enhanced in the KO rats. The diameter of the MCA decreased to a greater extent in Dusp5 KO rats than in FHH.1BN and FHH rats when the perfusion pressure was increased from 40 to 140 mmHg. CBF increased markedly in FHH rats when MAP was increased from 100 to 160 mmHg, and CBF was better autoregulated in the Dusp5 KO and FHH.1BN rats. The expression of Dusp5 was higher at the mRNA level but not at the protein level and the levels of p-ERK1/2 and p-PKC were lower in cerebral microvessels and brain tissue isolated from FHH than in FHH.1BN rats. These results indicate that Dusp5 modulates myogenic reactivity in the cerebral circulation and support the view that a mutation in Dusp5 may enhance Dusp5 activity and contribute to the impaired myogenic response in FHH rats. PMID:25397684

  11. Zinc-finger nuclease knockout of dual-specificity protein phosphatase-5 enhances the myogenic response and autoregulation of cerebral blood flow in FHH.1BN rats.

    Directory of Open Access Journals (Sweden)

    Fan Fan

    Full Text Available We recently reported that the myogenic responses of the renal afferent arteriole (Af-Art and middle cerebral artery (MCA and autoregulation of renal and cerebral blood flow (RBF and CBF were impaired in Fawn Hooded hypertensive (FHH rats and were restored in a FHH.1BN congenic strain in which a small segment of chromosome 1 from the Brown Norway (BN containing 15 genes including dual-specificity protein phosphatase-5 (Dusp5 were transferred into the FHH genetic background. We identified 4 single nucleotide polymorphisms in the Dusp5 gene in FHH as compared with BN rats, two of which altered CpG sites and another that caused a G155R mutation. To determine whether Dusp5 contributes to the impaired myogenic response in FHH rats, we created a Dusp5 knockout (KO rat in the FHH.1BN genetic background using a zinc-finger nuclease that introduced an 11 bp frame-shift deletion and a premature stop codon at AA121. The expression of Dusp5 was decreased and the levels of its substrates, phosphorylated ERK1/2 (p-ERK1/2, were enhanced in the KO rats. The diameter of the MCA decreased to a greater extent in Dusp5 KO rats than in FHH.1BN and FHH rats when the perfusion pressure was increased from 40 to 140 mmHg. CBF increased markedly in FHH rats when MAP was increased from 100 to 160 mmHg, and CBF was better autoregulated in the Dusp5 KO and FHH.1BN rats. The expression of Dusp5 was higher at the mRNA level but not at the protein level and the levels of p-ERK1/2 and p-PKC were lower in cerebral microvessels and brain tissue isolated from FHH than in FHH.1BN rats. These results indicate that Dusp5 modulates myogenic reactivity in the cerebral circulation and support the view that a mutation in Dusp5 may enhance Dusp5 activity and contribute to the impaired myogenic response in FHH rats.

  12. Zinc finger nuclease mediated knockout of ADP-dependent glucokinase in cancer cell lines: effects on cell survival and mitochondrial oxidative metabolism.

    Directory of Open Access Journals (Sweden)

    Susan Richter

    Full Text Available Zinc finger nucleases (ZFN are powerful tools for editing genes in cells. Here we use ZFNs to interrogate the biological function of ADPGK, which encodes an ADP-dependent glucokinase (ADPGK, in human tumour cell lines. The hypothesis we tested is that ADPGK utilises ADP to phosphorylate glucose under conditions where ATP becomes limiting, such as hypoxia. We characterised two ZFN knockout clones in each of two lines (H460 and HCT116. All four clones had frameshift mutations in all alleles at the target site in exon 1 of ADPGK, and were ADPGK-null by immunoblotting. ADPGK knockout had little or no effect on cell proliferation, but compromised the ability of H460 cells to survive siRNA silencing of hexokinase-2 under oxic conditions, with clonogenic survival falling from 21±3% for the parental line to 6.4±0.8% (p = 0.002 and 4.3±0.8% (p = 0.001 for the two knockouts. A similar increased sensitivity to clonogenic cell killing was observed under anoxia. No such changes were found when ADPGK was knocked out in HCT116 cells, for which the parental line was less sensitive than H460 to anoxia and to hexokinase-2 silencing. While knockout of ADPGK in HCT116 cells caused few changes in global gene expression, knockout of ADPGK in H460 cells caused notable up-regulation of mRNAs encoding cell adhesion proteins. Surprisingly, we could discern no consistent effect on glycolysis as measured by glucose consumption or lactate formation under anoxia, or extracellular acidification rate (Seahorse XF analyser under oxic conditions in a variety of media. However, oxygen consumption rates were generally lower in the ADPGK knockouts, in some cases markedly so. Collectively, the results demonstrate that ADPGK can contribute to tumour cell survival under conditions of high glycolytic dependence, but the phenotype resulting from knockout of ADPGK is cell line dependent and appears to be unrelated to priming of glycolysis in these lines.

  13. Zinc Finger Nuclease Mediated Knockout of ADP-Dependent Glucokinase in Cancer Cell Lines: Effects on Cell Survival and Mitochondrial Oxidative Metabolism

    Science.gov (United States)

    Richter, Susan; Morrison, Shona; Connor, Tim; Su, Jiechuang; Print, Cristin G.; Ronimus, Ron S.; McGee, Sean L.; Wilson, William R.

    2013-01-01

    Zinc finger nucleases (ZFN) are powerful tools for editing genes in cells. Here we use ZFNs to interrogate the biological function of ADPGK, which encodes an ADP-dependent glucokinase (ADPGK), in human tumour cell lines. The hypothesis we tested is that ADPGK utilises ADP to phosphorylate glucose under conditions where ATP becomes limiting, such as hypoxia. We characterised two ZFN knockout clones in each of two lines (H460 and HCT116). All four clones had frameshift mutations in all alleles at the target site in exon 1 of ADPGK, and were ADPGK-null by immunoblotting. ADPGK knockout had little or no effect on cell proliferation, but compromised the ability of H460 cells to survive siRNA silencing of hexokinase-2 under oxic conditions, with clonogenic survival falling from 21±3% for the parental line to 6.4±0.8% (p = 0.002) and 4.3±0.8% (p = 0.001) for the two knockouts. A similar increased sensitivity to clonogenic cell killing was observed under anoxia. No such changes were found when ADPGK was knocked out in HCT116 cells, for which the parental line was less sensitive than H460 to anoxia and to hexokinase-2 silencing. While knockout of ADPGK in HCT116 cells caused few changes in global gene expression, knockout of ADPGK in H460 cells caused notable up-regulation of mRNAs encoding cell adhesion proteins. Surprisingly, we could discern no consistent effect on glycolysis as measured by glucose consumption or lactate formation under anoxia, or extracellular acidification rate (Seahorse XF analyser) under oxic conditions in a variety of media. However, oxygen consumption rates were generally lower in the ADPGK knockouts, in some cases markedly so. Collectively, the results demonstrate that ADPGK can contribute to tumour cell survival under conditions of high glycolytic dependence, but the phenotype resulting from knockout of ADPGK is cell line dependent and appears to be unrelated to priming of glycolysis in these lines. PMID:23799003

  14. MiRNA-205 modulates cellular invasion and migration via regulating zinc finger E-box binding homeobox 2 expression in esophageal squamous cell carcinoma cells

    Directory of Open Access Journals (Sweden)

    Yamashita Shunichi

    2011-03-01

    Full Text Available Abstract Background Esophageal squamous cell carcinoma (ESCC is often diagnosed at later stages until they are incurable. MicroRNA (miR is a small, non-coding RNA that negatively regulates gene expression mainly via translational repression. Accumulating evidence indicates that deregulation of miR is associated with human malignancies including ESCC. The aim of this study was to identify miR that could be specifically expressed and exert distinct biological actions in ESCC. Methods Total RNA was extracted from ESCC cell lines, OE21 and TE10, and a non-malignant human esophageal squamous cell line, Het-1A, and subjected to microarray analysis. Expression levels of miR that showed significant differences between the 2 ESCC and Het-1A cells based on the comprehensive analysis were analyzed by the quantitative reverse transcriptase (RT-PCR method. Then, functional analyses, including cellular proliferation, apoptosis and Matrigel invasion and the wound healing assay, for the specific miR were conducted. Using ESCC tumor samples and paired surrounding non-cancerous tissue obtained endoscopically, the association with histopathological differentiation was examined with quantitative RT-PCR. Results Based on the miR microarray analysis, there were 14 miRs that showed significant differences (more than 2-fold in expression between the 2 ESCC cells and non-malignant Het-1A. Among the significantly altered miRs, miR-205 expression levels were exclusively higher in 5 ESCC cell lines examined than any other types of malignant cell lines and Het-1A. Thus, miR-205 could be a specific miR in ESCC. Modulation of miR-205 expression by transfection with its precursor or anti-miR-205 inhibitor did not affect ESCC cell proliferation and apoptosis, but miR-205 was found to be involved in cell invasion and migration. Western blot revealed that knockdown of miR-205 expression in ESCC cells substantially enhanced expression of zinc finger E-box binding homeobox 2

  15. Satellite Boreal Measurements over Alaska and Canada During June-July 2004: Simultaneous Measurements of Upper Tropospheric CO, C2H6, HCN, CH3Cl, CH4, C2H2, CH2OH, HCOOH, OCS, and SF6 Mixing Ratios

    Science.gov (United States)

    Rinsland, Curtis P.; Dufour, Gaelle; Boone, Chris D.; Bernath, Peter F.; Chiou, Linda; Coheur, Pierre-Francois; Turquety, Solene; Clerbaux, Cathy

    2007-01-01

    Simultaneous ACE (Atmospheric Chemistry Experiment) upper tropospheric CO, C2H6, HCN, CH3Cl, CH4 , C2H2 , CH30H, HCOOH, and OCS measurements show plumes up to 185 ppbv (10 (exp -9) per unit volume) for CO, 1.36 ppbv for C2H6, 755 pptv (10(exp -12) per unit volume) for HCN, 1.12 ppbv for CH3C1, 1.82 ppmv, (10(exp -6) per unit volume) for CH4, 0.178 ppbv for C2H2, 3.89 ppbv for CH30H, 0.843 ppbv for HCOOH, and 0.48 ppbv for OCS in western Canada and Alaska at 50 deg N-68 deg N latitude between 29 June and 23 July 2004. Enhancement ratios and emission factors for HCOOH, CH30H, HCN, C2H6, and OCS relative to CO at 250-350 hPa are inferred from measurements of young plumes compared with lower mixing ratios assumed to represent background conditions based on a CO emission factor derived from boreal measurements. Results are generally consistent with the limited data reported for various vegetative types and emission phases measured in extratropical forests including boreal forests. The low correlation between fire product emission mixing ratios and the S176 mixing ratio is consistent with no significant SF6 emissions from the biomass fires.

  16. The zinc finger protein PtaZFP2 negatively controls stem growth and gene expression responsiveness to external mechanical loads in poplar.

    Science.gov (United States)

    Martin, Ludovic; Decourteix, Mélanie; Badel, Eric; Huguet, Stéphanie; Moulia, Bruno; Julien, Jean-Louis; Leblanc-Fournier, Nathalie

    2014-07-01

    Mechanical cues are essential signals regulating plant growth and development. In response to wind, trees develop a thigmomorphogenetic response characterized by a reduction in longitudinal growth, an increase in diameter growth, and changes in mechanical properties. The molecular mechanisms behind these processes are poorly understood. In poplar, PtaZFP2, a C2H2 transcription factor, is rapidly up-regulated after stem bending. To investigate the function of PtaZFP2, we analyzed PtaZFP2-overexpressing poplars (Populus tremula × Populus alba). To unravel the genes downstream PtaZFP2, a transcriptomic analysis was performed. PtaZFP2-overexpressing poplars showed longitudinal and cambial growth reductions together with an increase in the tangent and hardening plastic moduli. The regulation level of mechanoresponsive genes was much weaker after stem bending in PtaZFP2-overexpressing poplars than in wild-type plants, showing that PtaZFP2 negatively modulates plant responsiveness to mechanical stimulation. Microarray analysis revealed a high proportion of down-regulated genes in PtaZFP2-overexpressing poplars. Among these genes, several were also shown to be regulated by mechanical stimulation. Our results confirmed the important role of PtaZFP2 during plant acclimation to mechanical load, in particular through a negative control of plant molecular responsiveness. This desensitization process could modulate the amplitude and duration of the plant response during recurrent stimuli.

  17. Finger Stiffness

    OpenAIRE

    Oosterhoff, Thijs C.H.; Nota, Sjoerd P. F. T.; Ring, David

    2014-01-01

    Background Finger stiffness varies substantially in patients with hand and upper extremity illness and can be notably more than expected for a given pathophysiology. In prior studies, pain intensity and magnitude of disability consistently correlate with coping strategies such as catastrophic thinking and kinesiophobia, which can be characterized as overprotectiveness. In this retrospective study we address the primary research question whether patients with finger stiffness are more often ov...

  18. The kinetics and product state distributions from gas-phase reactions of small atomic and molecular cations with C2H4, C2H3F, 1,1-C2H2F2, C2HF3 and C2F4.

    Science.gov (United States)

    Parkes, Michael A; Simpson, Matthew J; Mikhailov, Victor; Tuckett, Richard P

    2014-02-28

    The reactions of twenty one gas-phase cations with C2H3F, 1,1-C2H2F2, C2HF3 and C2F4 have been studied in a selected ion flow tube at 298 K. The cations are both atomic and molecular with recombination energies in the range 6-22 eV, and the kinetics and branching ratios into product ions are revealed for all the reactions. These data, together with that from an earlier study of reactions of C(x)F(y)(+) with these four fluorinated ethenes (J. Phys. Chem. A., 2012, 116, 8119), are compared with the reactions of these ions with C2H4, where available. Nearly all the reactions have a rate coefficient close to the collisional value calculated by either Langevin or modified average dipole orientation theories. The products of the reactions of N(+) and N2(+) with C2H4 are found to be anomalous, compared to their reactions with the four fluorinated ethenes. The branching ratios into product cations are compared with those from a high resolution (ca. 0.002 eV) photoionisation (hν = 10-22 eV) study of C2H3F, 1,1-C2H2F2, C2HF3 and C2F4 (Phys. Chem. Chem. Phys., 2012, 14, 3935) in order to gauge the importance of electron transfer in ion-molecule reactions. The higher the recombination energy of the cation, the better the agreement between the two sets of product branching ratios. Where there is disagreement at lower recombination energies, it appears that there is more fragmentation of the products in the photoionisation experiment compared to the ion-molecule reactions. PMID:24418988

  19. Finger Stiffness.

    Science.gov (United States)

    Oosterhoff, Thijs C H; Nota, Sjoerd P F T; Ring, David

    2015-06-01

    Background Finger stiffness varies substantially in patients with hand and upper extremity illness and can be notably more than expected for a given pathophysiology. In prior studies, pain intensity and magnitude of disability consistently correlate with coping strategies such as catastrophic thinking and kinesiophobia, which can be characterized as overprotectiveness. In this retrospective study we address the primary research question whether patients with finger stiffness are more often overprotective when the primary pathology is outside the hand (e.g. distal radius fracture) than when it is located within the hand. Methods In an orthopaedic hand surgery department 160 patients diagnosed with more finger stiffness than expected for a given pathophysiology or time point of recovery between December 2006 and September 2012 were analyzed to compare the proportion of patients characterized as overprotective for differences by site of pathology: (1) inside the hand, (2) outside the hand, and (3) psychiatric etiology (e.g. clenched fist). Results Among 160 subjects with more finger stiffness than expected, 132 (82 %) were characterized as overprotective including 88 of 108 (81 %) with pathology in the hand, 39 of 44 (89 %) with pathology outside the hand, and 5 of 8 (63 %) with psychiatric etiology. These differences were not significant. Conclusions Overprotectiveness is common in patients with more finger stiffness than expected regardless the site and type of primary pathology. It seems worthwhile to recognize and treat maladaptive coping strategies early during recovery to limit impairment, symptoms, and disability. PMID:26078497

  20. Nucleus Accumbens 1, a Pox virus and Zinc finger/Bric-a-brac Tramtrack Broad protein binds to TAR DNA-binding protein 43 and has a potential role in Amyotrophic Lateral Sclerosis.

    Science.gov (United States)

    Scofield, M D; Korutla, L; Jackson, T G; Kalivas, P W; Mackler, S A

    2012-12-27

    Protein degradation is a critical component of cellular maintenance. The intracellular translocation and targeting of the Ubiquitin Proteasome System (UPS) differentially coordinates a protein's half-life and thereby its function. Nucleus Accumbens 1 (NAC1), a member of the Pox virus and Zinc finger/Bric-a-brac Tramtrack Broad complex (POZ/BTB) family of proteins, participates in the coordinated proteolysis of synaptic proteins by mediating recruitment of the UPS to dendritic spines. Here we report a novel interaction between NAC1 and TAR DNA-binding protein 43 (TDP-43), a protein identified as the primary component of ubiquitinated protein aggregates found in patients with Amyotrophic Lateral Sclerosis (ALS). In vitro translated full-length TDP-43 associated with both the POZ/BTB domain and the non-POZ/BTB domain of NAC1 in GST pulldown assays. Other POZ/BTB proteins (including zinc finger POZ/BTB proteins and atypical POZ/BTB proteins) showed weak interactions with TDP-43. In addition, NAC1 and TDP-43 were present in the same immunocomplexes in different regions of mouse brain and spinal cord. In primary spinal cord cultures, TDP-43 expression was mainly nuclear, whereas NAC1 was both nuclear and cytoplasmic. In order to mimic ALS-like toxicity in the spinal cord culture system, we elevated extracellular glutamate levels resulting in the selective loss of motor neurons. Using this model, it was found that glutamate toxicity elicited a dose-dependent translocation of TDP-43 out of the nucleus of cholinergic neurons and increased the co-localization of NAC1 and TDP-43. These findings suggest that NAC1 may function to link TDP-43 to the proteasome; thereby, facilitating the post-translational modifications of TDP-43 that lead to the development of ALS.

  1. Cloning and Molecular Characterization of a RING Zinc-Finger Gene of Hevea brasiliensis%巴西橡胶树环锌指蛋白基因克隆及其分子特性

    Institute of Scientific and Technical Information of China (English)

    朱家红; 李辉亮; 屠发志; 田维敏; 彭世清

    2006-01-01

    Using the information about the sequence from a differentially expressed clone (designated as HbSSH10)encodes a protein specifying a cysteine-rich sequence containing a putative "RING finger" or "C3HC4" consensus motif that was cloned recently by the subtractive hybridization between latex and leaves from rubber tree (Hevea brasiliensis). A full-length cDNA encoding C3HC4 type zinc-finger protein was isolated and characterized from rubber tree. Sequence analysis revealed that the ORFs of HbRZF encode 156 amino acid residues with a total predicted molecular mass of 17.2 kD,HbRZF protein having a putative "RING finger" segment (amino acid residues 100-144). The deduced amino acid sequences of HbRZF showed high identities of 48%,52% and 50% to those of the ring zinc protein from Poncirus trifoliata, Arabidopsis thaliana, Thellungiella halophila. The result of Northern blot analysis indicated that the transcripts of the HbRZF were expressed more in the latex than in the leaves, whereas little expression was detected in roots and flowers. The transcription of HbRZF was induced by jasmonic acid, whereas ethylene had little effect.%在先前的研究中通过抑制缩减杂交获得了一个在巴西橡胶树胶乳中特异表达的片段(HbSSH10),该片段含有"RING finger"或"C3HC4"保守序列.根据HbSSH10的序列信息设计引物并通过3'-RACE和5'-RACE的方法,获得了一个全长的cDNA(HbRZF).该cDNA含有589个核苷酸,含有完整的阅读框架,编码156个氨基酸.从它推导出的氨基酸序列中含有"RING finger"或"C3HC4"保守区(氨基酸100~144).该氨基酸序列与Poncirus trifoliata、Arabidopsis thaliana和Thellungiella halophila的环锌指蛋白的同源性分别为48%、52%和50%.Northern杂交分析表明HbRZF在胶乳中大量表达,在叶片中微量表达,而在根和花中几乎没有表达.茉莉酸处理可以诱导胶乳中HbRZF的表达,而乙烯对胶乳中HbRZF的表达基本上没有影响.

  2. The C3H-type zinc finger protein GDS1/C3H42 is a nuclear-speckle-localized protein that is essential for normal growth and development in Arabidopsis.

    Science.gov (United States)

    Kim, Dae Won; Jeon, Su Jeong; Hwang, Sung Min; Hong, Jong Chan; Bahk, Jeong Dong

    2016-09-01

    Eukaryotic C3H-type zinc finger proteins (Znfs) comprise a large family of regulatory proteins involved in many aspects of plant stress response, growth and development. However, compared to mammalian, only a few plant Znfs have been functionally characterized. Here, T-DNA inserted gds1 (growth, development and splicing 1) mutant, displayed abnormal growth throughout the lifecycle owing to the reduction of cell size and number. Inverse PCR analysis revealed that the abnormal growth was caused by the disruption of At3g47120, which encodes a C3H42 protein belonging to the C-X7-C-X5-C-X3-H class of the Znf family. GDS1 was ubiquitously transcribed, but shows high levels of expression in young seedling and unexpanded new leaves. In gds1, the transcripts of many growth- and development-related genes were down-regulated, and the auxin response was dramatically reduced. A fluorescence-based assay revealed that the GDS1 protein was localized to the nucleus, prominently in the speckle compartments. Its arginine/serine dipeptide-rich-like (RS-like) domain was essential for nuclear localization. In addition, the SR1, SRm102 and U1-70K components of the U1 spliceosome interacted with GDS1 in the nuclear speckle compartments. Taken together, these suggest that GDS1, a nuclear-speckle-associated Znf, might play a significant role in splicing during plant growth and development. PMID:27457991

  3. Research Advance on Zinc-finger Nucleases Technology in Mammalian Transgenesis%锌指核酸酶技术在动物转基因中的研究进展

    Institute of Scientific and Technical Information of China (English)

    袁玉国; 彭秋玲

    2014-01-01

    锌指核酸酶(zinc finger nucleases,ZFNs)技术是近年来发展起来的一种基因修饰技术,利用ZFNs可特异识别靶位点DNA序列并使其断裂.与传统基于同源重组的基因打靶相比,ZFNs能使打靶效率提高100000倍,并使基因断裂与突变修复达到同样效果.ZFNs已经应用于多种生物,包括昆虫、两栖类生物、线虫、植物、多种动物和人类细胞.这些应用提高了人们对复杂生理系统的理解,使ZFNs成为一种生产转基因动物、细胞系和植物的有力工具,并在治疗人类疾病中发挥重要作用.文章对ZFNs技术原理及其应用于哺乳类动物转基因的研究进行了回顾和展望.

  4. Herpes simplex virus induces the marked up-regulation of the zinc finger transcriptional factor INSM1, which modulates the expression and localization of the immediate early protein ICP0

    Directory of Open Access Journals (Sweden)

    Kimura Hiroshi

    2011-05-01

    Full Text Available Abstract Background Herpes simplex viruses (HSVs rapidly shut off macromolecular synthesis in host cells. In contrast, global microarray analyses have shown that HSV infection markedly up-regulates a number of host cell genes that may play important roles in HSV-host cell interactions. To understand the regulatory mechanisms involved, we initiated studies focusing on the zinc finger transcription factor insulinoma-associated 1 (INSM1, a host cell protein markedly up-regulated by HSV infection. Results INSM1 gene expression in HSV-1-infected normal human epidermal keratinocytes increased at least 400-fold 9 h after infection; INSM1 promoter activity was also markedly stimulated. Expression and subcellular localization of the immediate early HSV protein ICP0 was affected by INSM1 expression, and chromatin immunoprecipitation (ChIP assays revealed binding of INSM1 to the ICP0 promoter. Moreover, the role of INSM1 in HSV-1 infection was further clarified by inhibition of HSV-1 replication by INSM1-specific siRNA. Conclusions The results suggest that INSM1 up-regulation plays a positive role in HSV-1 replication, probably by binding to the ICP0 promoter.

  5. The mouse nac1 gene, encoding a cocaine-regulated Bric-a-brac Tramtrac Broad complex/Pox virus and Zinc finger protein, is regulated by AP1.

    Science.gov (United States)

    Mackler, S A; Homan, Y X; Korutla, L; Conti, A C; Blendy, J A

    2003-01-01

    NAC1 cDNA was identified as a novel transcript induced in the nucleus accumbens from rats chronically treated with cocaine. NAC1 is a member of the Bric-a-brac Tramtrac Broad complex/Pox virus and Zinc finger family of transcription factors and has been shown by overexpression studies to prevent the development of behavioral sensitization resulting from repeated cocaine treatment. This paper reports the cloning and characterization of the corresponding gene. The mouse Nac1 gene consist of six exons, with exon 2 containing an alternative splice donor, providing a molecular explanation of the splice variants observed in mouse and rat. Transcripts of Nac1 were ubiquitously detected in different mouse tissues with prominent expression in the brain. The mouse Nac1 gene was localized to chromosome 8, suggesting a highly plausible candidate gene to explain differences in cocaine-induced behaviors between C57BL6/J and DBA/2J mice that had previously been mapped to the area. In addition, a functional AP1 binding site has been identified in an intron 1 enhancer of the Nac1 gene that plays an essential role in the activation of the gene in differentiation of neuroblastoma cells. Co-transfection with c-jun and c-fos expression plasmids, which encode the two subunits of AP1, activated the wild type Nac1 intron 1 enhancer two-fold over basal, nearly at the level of NAC1 enhancer activity seen in differentiated N2A cells. Mutation of the AP1 site completely abrogated all activation of the NAC1 enhancer in differentiated N2A cells. Activation of immediate early genes such as c-fos and c-jun following chronic drug treatments has been well characterized. The present data describe one potential regulatory cascade involving these transcription factors and activation of NAC1. Identification of drug induced alterations in gene expression is key to understanding the types of molecular adaptations underlying addiction.

  6. An A20/AN1-zinc-finger domain containing protein gene in tea is differentially expressed during winter dormancy and in response to abiotic stress and plant growth regulators

    Directory of Open Access Journals (Sweden)

    Asosii Paul

    2015-03-01

    Full Text Available The present manuscript describes cloning and expression characterization of A20/AN1-zinc-finger domain containing protein (CsZfp gene in an evergreen tree tea [Camellia sinensis (L. O. Kuntze] in response to winter dormancy (WD, abiotic stresses (polyethylene glycol, hydrogen peroxide, and sodium chloride and plant growth regulators [abscisic acid (ABA, and gibberellic acid (GA3]. CsZfp encoded a putative protein of 173 amino acids with a calculated molecular weight of 18.44 kDa, an isoelectric point (pI of 6.50 and grand average of hydropathicity (GRAVY value of −0.334. The gene did not have an intron, and belonged to a multi-gene family. During the period of active growth (PAG, CsZfp showed maximum expression in root and fruit as compared to leaf, floral bud and stem. Interaction studies between temperature and plant growth regulators on the expression of CsZfp showed that ABA upregulated CsZfp expression at growth temperature (GT; 25 °C but had no effect at low temperature (LT; 4 °C. In response to GA3, upregulation was observed at LT but not at GT. Further, the expression was not modulated by LT either in the tissue harvested during PAG or during WD. It was interesting to record that the expression of CsZfp was upregulated by hydrogen peroxide and sodium chloride, whereas it was non-responsive to polyethylene glycol. The possible role of CsZfp in playing key but differential roles in tea to various abiotic stresses is discussed.

  7. 锌指蛋白185对胶质瘤细胞增殖影响的研究%The effect of zinc finger protein 185 on the proliferation of human glioma cells

    Institute of Scientific and Technical Information of China (English)

    陆斌; 郑全辉

    2015-01-01

    目的:探讨锌指蛋白ZNF185在人脑胶质瘤细胞增殖中的作用。方法标本取自2011年1月—2013年12月于唐山市工人医院就诊,经病理确诊为胶质瘤患者的肿瘤组织,以瘤旁组织作对照。提取不同组织总蛋白, Western-blot检测ZNF185的表达。提取瘤旁组织总RNA,反转录扩增ZNF185编码序列并克隆至pEGFPC2质粒,构建ZNF185表达载体。采用Lipofactamine2000将ZNF185表达载体转染人胶质瘤细胞系SF767,以转染pEGFPC2空载体细胞作为对照。采用流式细胞仪检测细胞周期变化;MTT法检测细胞增殖活性。结果与瘤旁组织相比, ZNF185在人脑胶质瘤组织中表达显著降低(P<0.01);转染ZNF185表达载体的胶质瘤细胞与对照细胞相比ZNF185的表达显著增加(P<0.01);过表达ZNF185导致SF767细胞G0~G1期细胞比例显著增加(P<0.05),而S期细胞比例减少(P<0.05)。同时,过表达ZNF185也导致SF767细胞的增殖速度显著降低(P<0.05)。结论ZNF185在人脑胶质瘤细胞中发挥抑制细胞增殖的作用。%Objective To explore the role of zinc finger protein (ZNF)185 in the proliferation of human glioma cells. Methods Human glioma tissues and tumor adjacent tissues were obtained from glioma patients diagnosed pathologically in Tangshan Gongren Hospital from January 2011 to December 2013. Total protein was extracted from different tissues. The ZNF185 expression was detected by Western-blot assay. Total RNA was extracted from tumor adjacent tissues. ZNF 185 cod⁃ing sequence was obtained by RT-PCR and inserted into pEGFPC2 plasmid to construct the ZNF185 expression vector. Li⁃pofactamine2000 was used to transfect the ZNF185 expression vector to human glioma cell SF767. pEGFPC2 blank vector transfected SF767 cells were used as control. Changes of cell cycle were analyzed by flow cytometry, and cell proliferation was analyzed by MTT assay. Results The expression of ZNF185 decreased

  8. Gold finger formation studied by high-resolution mass spectrometry and in silico methods

    NARCIS (Netherlands)

    Laskay, Ü.A.; Garino, C.; Tsybin, Y.O.; Salassa, L.; Casini, A.

    2015-01-01

    High-resolution mass spectrometry and quantum mechanics/molecular mechanics studies were employed for characterizing the formation of two gold finger (GF) domains from the reaction of zinc fingers (ZF) with gold complexes. The influence of both the gold oxidation state and the ZF coordination sphere

  9. Multiple Fingers - One Gestalt.

    Science.gov (United States)

    Lezkan, Alexandra; Manuel, Steven G; Colgate, J Edward; Klatzky, Roberta L; Peshkin, Michael A; Drewing, Knut

    2016-01-01

    The Gestalt theory of perception offered principles by which distributed visual sensations are combined into a structured experience ("Gestalt"). We demonstrate conditions whereby haptic sensations at two fingertips are integrated in the perception of a single object. When virtual bumps were presented simultaneously to the right hand's thumb and index finger during lateral arm movements, participants reported perceiving a single bump. A discrimination task measured the bump's perceived location and perceptual reliability (assessed by differential thresholds) for four finger configurations, which varied in their adherence to the Gestalt principles of proximity (small versus large finger separation) and synchrony (virtual spring to link movements of the two fingers versus no spring). According to models of integration, reliability should increase with the degree to which multi-finger cues integrate into a unified percept. Differential thresholds were smaller in the virtual-spring condition (synchrony) than when fingers were unlinked. Additionally, in the condition with reduced synchrony, greater proximity led to lower differential thresholds. Thus, with greater adherence to Gestalt principles, thresholds approached values predicted for optimal integration. We conclude that the Gestalt principles of synchrony and proximity apply to haptic perception of surface properties and that these principles can interact to promote multi-finger integration.

  10. Simulation of nanoparticle coagulation in radio-frequency C2H2/Ar microdischarges

    Science.gov (United States)

    Xiang-Mei, Liu; Qi-Nan, Li; Rui, Li

    2016-06-01

    The nanoparticle coagulation is investigated by using a couple of fluid models and aerosol dynamics model in argon with a 5% molecular acetylene admixture rf microdischarges, with the total input gas flow rate of 400 sccm. It co-exists with a homogeneous, secondary electron-dominated low temperature γ-mode glow discharges. The heat transfer equation and flow equation for neutral gas are taken into account. We mainly focused on investigations of the nanoparticle properties in atmospheric pressure microdischarges, and discussed the influences of pressure, electrode spacing, and applied voltage on the plasma density and nanoparticle density profiles. The results show that the characteristics of microdischarges are quite different from those of low pressure radio-frequency discharges. First, the nanoparticle density in the bulk plasma in microdischarges is much larger than that of low pressure discharges. Second, the nanoparticle density of 10 nm experiences an exponential increase as soon as the applied voltage increases, especially in the presheath. Finally, as the electrode spacing increases, the nanoparticle density decreased instead of increasing. Project supported by the Natural Science Foundation of Heilongjiang Province, China (Grant Nos. A2015011 and A2015010), the Postdoctoral Scientific Research Development Fund of Heilongjiang Province, China (Grant No. LBH-Q14159), the Program for Young Teachers Scientific Research in Qiqihar University (Grant No. 2014k-Z11), the National Natural Science Foundation of China (Grant No. 11404180), and the University Nursing Program for Yong Scholars with Creative Talents in Heilongjiang Province, China (Grant No. UNPYSCT-2015095).

  11. Demonstrated in vitro activity of nitrogenase (C2H2) rhizosphere microorganisms in sugarcane

    International Nuclear Information System (INIS)

    The sugar cane (Saccharum officinarum L.) and rice (Oryza sativa var K 017) production improvement is an urgency for tropical developing countries. In order to investigate the associative N2 fixing activity by rhizospheric microorganisms, experiments were made on Sugar cane and rice cultivated in Zaire. In vitro evidence of the Acetylene Reduction Activity (ARA) was obtained from root pe ices of sugar cane and rice seedlings (six weeks old) growing on synthetic medium supplemented with rhizospheric soil dilutions. The determined ARA was higher in root cane samples than in rice seedling cultures.

  12. Arsenite binding-induced zinc loss from PARP-1 is equivalent to zinc deficiency in reducing PARP-1 activity, leading to inhibition of DNA repair

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Xi; Zhou, Xixi [Department of Pharmaceutical Sciences, College of Pharmacy, University of New Mexico Health Sciences Center, Albuquerque, NM 87131 (United States); Du, Libo [Center for Molecular Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190 (China); Liu, Wenlan [Department of Pharmaceutical Sciences, College of Pharmacy, University of New Mexico Health Sciences Center, Albuquerque, NM 87131 (United States); Liu, Yang [Center for Molecular Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190 (China); Hudson, Laurie G. [Department of Pharmaceutical Sciences, College of Pharmacy, University of New Mexico Health Sciences Center, Albuquerque, NM 87131 (United States); Liu, Ke Jian, E-mail: kliu@salud.unm.edu [Department of Pharmaceutical Sciences, College of Pharmacy, University of New Mexico Health Sciences Center, Albuquerque, NM 87131 (United States)

    2014-01-15

    Inhibition of DNA repair is a recognized mechanism for arsenic enhancement of ultraviolet radiation-induced DNA damage and carcinogenesis. Poly(ADP-ribose) polymerase-1 (PARP-1), a zinc finger DNA repair protein, has been identified as a sensitive molecular target for arsenic. The zinc finger domains of PARP-1 protein function as a critical structure in DNA recognition and binding. Since cellular poly(ADP-ribosyl)ation capacity has been positively correlated with zinc status in cells, we hypothesize that arsenite binding-induced zinc loss from PARP-1 is equivalent to zinc deficiency in reducing PARP-1 activity, leading to inhibition of DNA repair. To test this hypothesis, we compared the effects of arsenite exposure with zinc deficiency, created by using the membrane-permeable zinc chelator TPEN, on 8-OHdG formation, PARP-1 activity and zinc binding to PARP-1 in HaCat cells. Our results show that arsenite exposure and zinc deficiency had similar effects on PARP-1 protein, whereas supplemental zinc reversed these effects. To investigate the molecular mechanism of zinc loss induced by arsenite, ICP-AES, near UV spectroscopy, fluorescence, and circular dichroism spectroscopy were utilized to examine arsenite binding and occupation of a peptide representing the first zinc finger of PARP-1. We found that arsenite binding as well as zinc loss altered the conformation of zinc finger structure which functionally leads to PARP-1 inhibition. These findings suggest that arsenite binding to PARP-1 protein created similar adverse biological effects as zinc deficiency, which establishes the molecular mechanism for zinc supplementation as a potentially effective treatment to reverse the detrimental outcomes of arsenic exposure. - Highlights: • Arsenite binding is equivalent to zinc deficiency in reducing PARP-1 function. • Zinc reverses arsenic inhibition of PARP-1 activity and enhancement of DNA damage. • Arsenite binding and zinc loss alter the conformation of zinc finger

  13. Arsenite binding-induced zinc loss from PARP-1 is equivalent to zinc deficiency in reducing PARP-1 activity, leading to inhibition of DNA repair

    International Nuclear Information System (INIS)

    Inhibition of DNA repair is a recognized mechanism for arsenic enhancement of ultraviolet radiation-induced DNA damage and carcinogenesis. Poly(ADP-ribose) polymerase-1 (PARP-1), a zinc finger DNA repair protein, has been identified as a sensitive molecular target for arsenic. The zinc finger domains of PARP-1 protein function as a critical structure in DNA recognition and binding. Since cellular poly(ADP-ribosyl)ation capacity has been positively correlated with zinc status in cells, we hypothesize that arsenite binding-induced zinc loss from PARP-1 is equivalent to zinc deficiency in reducing PARP-1 activity, leading to inhibition of DNA repair. To test this hypothesis, we compared the effects of arsenite exposure with zinc deficiency, created by using the membrane-permeable zinc chelator TPEN, on 8-OHdG formation, PARP-1 activity and zinc binding to PARP-1 in HaCat cells. Our results show that arsenite exposure and zinc deficiency had similar effects on PARP-1 protein, whereas supplemental zinc reversed these effects. To investigate the molecular mechanism of zinc loss induced by arsenite, ICP-AES, near UV spectroscopy, fluorescence, and circular dichroism spectroscopy were utilized to examine arsenite binding and occupation of a peptide representing the first zinc finger of PARP-1. We found that arsenite binding as well as zinc loss altered the conformation of zinc finger structure which functionally leads to PARP-1 inhibition. These findings suggest that arsenite binding to PARP-1 protein created similar adverse biological effects as zinc deficiency, which establishes the molecular mechanism for zinc supplementation as a potentially effective treatment to reverse the detrimental outcomes of arsenic exposure. - Highlights: • Arsenite binding is equivalent to zinc deficiency in reducing PARP-1 function. • Zinc reverses arsenic inhibition of PARP-1 activity and enhancement of DNA damage. • Arsenite binding and zinc loss alter the conformation of zinc finger

  14. Spiral viscous fingering.

    Science.gov (United States)

    Nagatsu, Yuichiro; Hayashi, Atsushi; Kato, Yoshihito; Tada, Yutaka

    2006-11-01

    When a less-viscous fluid displaces a more-viscous fluid in a radial Hele-Shaw cell, viscous fingering pattern is believed to develop in a radial direction. We performed experiments on viscous fingering in a radial Hele-Shaw cell when a polymer solution, a sodium polyacrylate (SPA) solution is used as the more-viscous fluid and the trivalent iron (Fe^3+) solution is as the less-viscous fluid. The experiment was done by varying the concentration of Fe^3+, cFe3+. We have found that viscous fingering pattern develops spirally when cFe3+ is larger than a threshold value, while the pattern develops in a radial direction for small cFe3+. We confirmed from different experiments that an instantaneous chemical reaction takes place between SPA solution and Fe^3+ solution. The chemical reaction produces precipitation and significantly reduces the viscosity of the SPA solution. The quantity of the precipitation is increased with cFe3+. We will make a discussion on the relationship between the formation of spiral viscous fingering and the chemical reaction taking place between the two fluids.

  15. Finger cold induced vasodilation

    NARCIS (Netherlands)

    Daanen, H.A.M.

    2007-01-01

    There are indications that subjects with a reduced finger CIVD response are more prone to get local cold injuries, but more epidemiological research is needed to establish a firm relationship. Although it was observed that an early CIVD onset was associated with initially superior manual performance

  16. X-Ray Exam: Finger

    Science.gov (United States)

    ... Tropical Delight: Melon Smoothie Pregnant? Your Baby's Growth X-Ray Exam: Finger KidsHealth > For Parents > X-Ray Exam: Finger Print A A A Text Size ... español Radiografía: dedo What It Is A finger X-ray is a safe and painless test that uses ...

  17. Design of lanthanide fingers: compact lanthanide-binding metalloproteins.

    Science.gov (United States)

    am Ende, Christopher W; Meng, Hai Yun; Ye, Mao; Pandey, Anil K; Zondlo, Neal J

    2010-08-16

    Lanthanides have interesting chemical properties; these include luminescent, magnetic, and catalytic functions. Toward the development of proteins incorporating novel functions, we have designed a new lanthanide-binding motif, lanthanide fingers. These were designed based on the Zif268 zinc finger, which exhibits a beta beta alpha structural motif. Lanthanide fingers utilize an Asp(2)Glu(2) metal-coordination environment to bind lanthanides through a tetracarboxylate peptide ligand. The iterative design of a general lanthanide-binding peptide incorporated the following key elements: 1) residues with high alpha-helix and beta-sheet propensities in the respective secondary structures; 2) an optimized big box alpha-helix N-cap; 3) a Schellman alpha-helix C-cap motif; and 4) an optional D-Pro-Ser type II' beta-turn in the beta-hairpin. The peptides were characterized for lanthanide binding by circular dichroism (CD), NMR, and fluorescence spectroscopy. In all instances, stabilization of the peptide secondary structures resulted in an increase in metal affinity. The optimized protein design was a 25-residue peptide that was a general lanthanide-binding motif; this binds all lanthanides examined in a competitive aqueous environment, with a dissociation constant of 9.3 microM for binding Er(3+). CD spectra of the peptide-lanthanide complexes are similar to those of zinc fingers and other beta beta alpha proteins. Metal binding involves residues from the N-terminal beta-hairpin and the C terminal alpha-helical segments of the peptide. NMR data indicated that metal binding induced a global change in the peptide structure. The D-Pro-Ser type II' beta-turn motif could be replaced by Thr-Ile to generate genetically encodable lanthanide fingers. Replacement of the central Phe with Trp generated genetically encodable lanthanide fingers that exhibited terbium luminescence greater than that of an EF-hand peptide.

  18. Finger Forces in Clarinet Playing

    Science.gov (United States)

    Hofmann, Alex; Goebl, Werner

    2016-01-01

    Clarinettists close and open multiple tone holes to alter the pitch of the tones. Their fingering technique must be fast, precise, and coordinated with the tongue articulation. In this empirical study, finger force profiles and tongue techniques of clarinet students (N = 17) and professional clarinettists (N = 6) were investigated under controlled performance conditions. First, in an expressive-performance task, eight selected excerpts from the first Weber Concerto were performed. These excerpts were chosen to fit in a 2 × 2 × 2 design (register: low–high; tempo: slow–fast, dynamics: soft–loud). There was an additional condition controlled by the experimenter, which determined the expression levels (low–high) of the performers. Second, a technical-exercise task, an isochronous 23-tone melody was designed that required different effectors to produce the sequence (finger-only, tongue-only, combined tongue-finger actions). The melody was performed in three tempo conditions (slow, medium, fast) in a synchronization-continuation paradigm. Participants played on a sensor-equipped Viennese clarinet, which tracked finger forces and reed oscillations simultaneously. From the data, average finger force (Fmean) and peak force (Fmax) were calculated. The overall finger forces were low (Fmean = 1.17 N, Fmax = 3.05 N) compared to those on other musical instruments (e.g., guitar). Participants applied the largest finger forces during the high expression level performance conditions (Fmean = 1.21 N). For the technical exercise task, timing and articulation information were extracted from the reed signal. Here, the timing precision of the fingers deteriorated the timing precision of the tongue for combined tongue-finger actions, especially for faster tempi. Although individual finger force profiles were overlapping, the group of professional players applied less finger force overall (Fmean = 0.54 N). Such sensor instruments provide useful insights into player

  19. Viscous fingering patterns in ferrofluids

    OpenAIRE

    Widom, Michael; Miranda, Jose A.

    1998-01-01

    Viscous fingering occurs in the flow of two immiscible, viscous fluids between the plates of a Hele-Shaw cell. Due to pressure gradients or gravity, the initially planar interface separating the two fluids undergoes a Saffman-Taylor instability and develops finger-like structures. When one of the fluids is a ferrofluid and a perpendicular magnetic field is applied, the labyrinthine instability supplements the usual viscous fingering instability, resulting in visually striking, complex pattern...

  20. Robotic Finger Assembly

    Science.gov (United States)

    Ihrke, Chris A. (Inventor); Bridgwater, Lyndon (Inventor); Diftler, Myron A. (Inventor); Linn, Douglas Martin (Inventor); Platt, Robert J., Jr. (Inventor); Hargrave, Brian (Inventor); Askew, Scott R. (Inventor); Valvo, Michael C. (Inventor)

    2014-01-01

    A robotic hand includes a finger with first, second, and third phalanges. A first joint rotatably connects the first phalange to a base structure. A second joint rotatably connects the first phalange to the second phalange. A third joint rotatably connects the third phalange to the second phalange. The second joint and the third joint are kinematically linked such that the position of the third phalange with respect to the second phalange is determined by the position of the second phalange with respect to the first phalange.

  1. Finger Mathematics for EMR Children.

    Science.gov (United States)

    Ogletree, Earl J.; Chavez, Maria

    An approach to teaching mildly retarded children math skills using finger calculation is discussed. Drills progress from using one to two hands and doing multiplication and division. The appropriateness of finger calculation with children in the sensory motor and preoperational stages of development is noted, and the approach's ability to enhance…

  2. Emotional Communication in Finger Braille

    Directory of Open Access Journals (Sweden)

    Yasuhiro Matsuda

    2010-01-01

    Full Text Available We describe analyses of the features of emotions (neutral, joy, sadness, and anger expressed by Finger Braille interpreters and subsequently examine the effectiveness of emotional expression and emotional communication between people unskilled in Finger Braille. The goal is to develop a Finger Braille system to teach emotional expression and a system to recognize emotion. The results indicate the following features of emotional expression by interpreters. The durations of the code of joy were significantly shorter than the durations of the other emotions, the durations of the code of sadness were significantly longer, and the finger loads of anger were significantly larger. The features of emotional expression by unskilled subjects were very similar to those of the interpreters, and the coincidence ratio of emotional communication was 75.1%. Therefore, it was confirmed that people unskilled in Finger Braille can express and communicate emotions using this communication medium.

  3. Mixing methods, tasting fingers

    DEFF Research Database (Denmark)

    Mann, Anna; Mol, Annemarie; Satalkar, Priya;

    2011-01-01

    This article reports on an ethnographic experiment. Four finger eating experts and three novices sat down for a hot meal and ate with their hands. Drawing on the technique of playing with the familiar and the strange, our aim was not to explain our responses, but to articulate them. As we seek wo...... available for articulation. But is it okay? Will we be allowed to mess with textbook biology in this way and interfere, not just with anthropological theory, but with the English language itself?...... hard to name. Pleasure and embarrassment, food-­like vitality, erotic titillation, the satisfaction or discomfort that follow a meal-we suggest that these may all be included in "tasting." Thus teasing the language alters what speakers and eaters may sense and say. It complements the repertoires...

  4. 锌指核酸酶技术制备肌肉生长抑制素基因敲除的五指山小型猪成纤维细胞%Production of myostatin gene knockout Wuzhishan miniature pig fibroblasts with zinc-finger nucleases

    Institute of Scientific and Technical Information of China (English)

    曹随忠; 岳成鹤; 李西睿; 冯冲; 龙川; 潘登科

    2013-01-01

    Disruption of myostatin (MSTN) gene in pigs may improve porcine lean meat percentage (LMP), and create an animal model for certain human diseases. Using zinc-finger nucleases (ZFNs) technology, MSTN gene was deleted in Wuzhishan miniature pig fibroblasts by transfection of either ZFNs plasmids or ZFNs mRNA in high efficiency. Strikingly, ZFNs encoding mRNA could produce MSTN+/-and MSTN-/- cell colonies with single or double allele deletion by single transfection. Sequencing results demonstrated that 92.18% of the mutations were short fragment deletions or insertions (≤10 bp). Prediction of amino acids sequences indicated that more than half of the mutations cause premature transla-tional-termination codon. MSTN+/+, MSTN+/-, and MSTK-/- cell colonies were used as nuclear donor for somatic cell nuclear transfer (SCNT), and developmental potential of SCNT embryos were measured by the blastocyst rate. The results revealed no significant difference in development competence among the three kinds of reconstructed embryos (14.29% vs. 19.64% vs. 16.13%), which provides the possibility of making myostatin knock out pigs in the future.%敲除猪肌肉生长抑制素(Myostatin,MSTN)基因可能提高猪瘦肉率,MSTN 基因敲除猪也可作为相关疾病的动物模型.文章利用锌指核酸酶(Zinc-finger nucleases,ZFNs)技术敲除五指山小型猪胎儿成纤维细胞MSTN基因,为制备MSTN 基因敲除猪奠定基础.ZFNs 质粒或编码ZFNs 的mRNA 均能高效敲除MSTN 基因,使用ZFNs mRNA 能直接得到MSTN+/-和MSTN-/-两种基因型的细胞克隆.DNA 序列测定与分析发现,细胞克隆的突变类型多为ZFNs 作用靶位点处不大于10 bp 的碱基插入或缺失(92.18 %); 氨基酸预测发现,突变型MSTN 基因的终止密码子常常提前出现.将MSTN 基因敲除的细胞进行体细胞核移植(Somatic cell nuclear transfer,SCNT)发现,胚胎体外早期发育潜力与野生型无显著差异,表明这些细胞可用于后续MSTN 基因敲除猪的制备.

  5. Covering the Dorsal Finger Defect with Reverse Cross Finger Flap

    Directory of Open Access Journals (Sweden)

    Kaan Gurbuz

    2014-12-01

    Full Text Available Reconstruction of finger extensor zone defects with or without tendon gaps still remains a challenge for surgeons. Although surgical treatments may differ, and range from the use of local, regional, to free flaps, the outcomes for all cases are not satisfactory. In this case report, we present a case of a 3rd finger extensor side crush injury including a defect of Dd (Digit Dorsal 1, Dd2 and Dd3 defects of extensor zones with tendon gap. Tendon gap was reconstructed using m. palmaris longus tendon graft and the defect was covered with reversed cross-finger flap (random pattern with good cosmetic and excellent functional results.

  6. Steadily translating parabolic dissolution fingers

    CERN Document Server

    Kondratiuk, Paweł

    2015-01-01

    Dissolution fingers (or wormholes) are formed during the dissolution of a porous rock as a result of nonlinear feedbacks between the flow, transport and chemical reactions at pore surfaces. We analyze the shapes and growth velocities of such fingers within the thin-front approximation, in which the reaction is assumed to take place instantaneously with the reactants fully consumed at the dissolution front. We concentrate on the case when the main flow is driven by the constant pressure gradient far from the finger, and the permeability contrast between the inside and the outside of the finger is finite. Using Ivantsov ansatz and conformal transformations we find the family of steadily translating fingers characterized by a parabolic shape. We derive the reactant concentration field and the pressure field inside and outside of the fingers and show that the flow within them is uniform. The advancement velocity of the finger is shown to be inversely proportional to its radius of curvature in the small P\\'{e}clet...

  7. Zinc Deficiency in Humans and its Amelioration

    OpenAIRE

    Yashbir Singh Shivay

    2015-01-01

    Zinc (Zn) deficiency in humans has recently received considerable attention. Global mortality in children under 5 years of age in 2004 due to Zn deficiency was estimated at 4,53,207 as against 6,66,771 for vitamin A deficiency; 20,854 for iron deficiency and 3,619 for iodine deficiency. In humans 2800-3000 proteins contain Zn prosthetic group and Zn is an integral component of zinc finger prints that regulate DNA transcription. Zinc is a Type-2 nutrient, which means that its concentration in ...

  8. Finger Forces in Clarinet Playing

    Directory of Open Access Journals (Sweden)

    Alex Hofmann

    2016-08-01

    Full Text Available Clarinettists close and open multiple tone holes to alter the pitch of the tones. Their fingering technique must be fast, precise, and coordinated with the tongue articulation. In this empirical study, finger force profiles and tongue techniques of clarinet students (N = 17 and professional clarinettists (N = 6 were investigated under controlled performance conditions. First, in an expressive-performance task, eight selected excerpts from the first Weber Concerto were performed. These excerpts were chosen to fit in a 2 x 2 x 2 design (register: low--high; tempo: slow--fast, dynamics: soft--loud. There was an additional condition controlled by the experimenter, which determined the expression levels (low--high of the performers. Second, a technical-exercise task, an isochronous 23-tone melody was designed that required different effectors to produce the sequence (finger-only, tongue-only, combined tongue-finger actions. The melody was performed in three tempo conditions (slow, medium, fast in a synchronization-continuation paradigm. Participants played on a sensor-equipped Viennese clarinet, which tracked finger forces and reed oscillations simultaneously. From the data, average finger force (Fmean and peak force (Fmax were calculated. The overall finger forces were low (Fmean = 1.17 N, Fmax = 3.05 N compared to those on other musical instruments (e.g. guitar. Participants applied the largest finger forces during the high expression level performance conditions (Fmean = 1.21 N.For the technical exercise task, timing and articulation information were extracted from the reed signal. Here, the timing precision of the fingers deteriorated the timing precision of the tongue for combined tongue-finger actions, especially for faster tempi. Although individual finger force profiles were overlapping, the group of professional players applied less finger force overall (Fmean = 0.54 N. Such sensor instruments provide useful insights into player

  9. Finger Forces in Clarinet Playing.

    Science.gov (United States)

    Hofmann, Alex; Goebl, Werner

    2016-01-01

    Clarinettists close and open multiple tone holes to alter the pitch of the tones. Their fingering technique must be fast, precise, and coordinated with the tongue articulation. In this empirical study, finger force profiles and tongue techniques of clarinet students (N = 17) and professional clarinettists (N = 6) were investigated under controlled performance conditions. First, in an expressive-performance task, eight selected excerpts from the first Weber Concerto were performed. These excerpts were chosen to fit in a 2 × 2 × 2 design (register: low-high; tempo: slow-fast, dynamics: soft-loud). There was an additional condition controlled by the experimenter, which determined the expression levels (low-high) of the performers. Second, a technical-exercise task, an isochronous 23-tone melody was designed that required different effectors to produce the sequence (finger-only, tongue-only, combined tongue-finger actions). The melody was performed in three tempo conditions (slow, medium, fast) in a synchronization-continuation paradigm. Participants played on a sensor-equipped Viennese clarinet, which tracked finger forces and reed oscillations simultaneously. From the data, average finger force (F mean ) and peak force (F max ) were calculated. The overall finger forces were low (F mean = 1.17 N, F max = 3.05 N) compared to those on other musical instruments (e.g., guitar). Participants applied the largest finger forces during the high expression level performance conditions (F mean = 1.21 N). For the technical exercise task, timing and articulation information were extracted from the reed signal. Here, the timing precision of the fingers deteriorated the timing precision of the tongue for combined tongue-finger actions, especially for faster tempi. Although individual finger force profiles were overlapping, the group of professional players applied less finger force overall (F mean = 0.54 N). Such sensor instruments provide useful insights into player

  10. Finger Forces in Clarinet Playing.

    Science.gov (United States)

    Hofmann, Alex; Goebl, Werner

    2016-01-01

    Clarinettists close and open multiple tone holes to alter the pitch of the tones. Their fingering technique must be fast, precise, and coordinated with the tongue articulation. In this empirical study, finger force profiles and tongue techniques of clarinet students (N = 17) and professional clarinettists (N = 6) were investigated under controlled performance conditions. First, in an expressive-performance task, eight selected excerpts from the first Weber Concerto were performed. These excerpts were chosen to fit in a 2 × 2 × 2 design (register: low-high; tempo: slow-fast, dynamics: soft-loud). There was an additional condition controlled by the experimenter, which determined the expression levels (low-high) of the performers. Second, a technical-exercise task, an isochronous 23-tone melody was designed that required different effectors to produce the sequence (finger-only, tongue-only, combined tongue-finger actions). The melody was performed in three tempo conditions (slow, medium, fast) in a synchronization-continuation paradigm. Participants played on a sensor-equipped Viennese clarinet, which tracked finger forces and reed oscillations simultaneously. From the data, average finger force (F mean ) and peak force (F max ) were calculated. The overall finger forces were low (F mean = 1.17 N, F max = 3.05 N) compared to those on other musical instruments (e.g., guitar). Participants applied the largest finger forces during the high expression level performance conditions (F mean = 1.21 N). For the technical exercise task, timing and articulation information were extracted from the reed signal. Here, the timing precision of the fingers deteriorated the timing precision of the tongue for combined tongue-finger actions, especially for faster tempi. Although individual finger force profiles were overlapping, the group of professional players applied less finger force overall (F mean = 0.54 N). Such sensor instruments provide useful insights into player

  11. The Response of a Putative Maize Zinc-Finger Protein Gene ZmAN14 in Transgenic Tabacco to Abiotic Stress%玉米锌指蛋白基因ZmAN14过表达转基因烟草对非生物胁迫的响应

    Institute of Scientific and Technical Information of China (English)

    宣宁; 柳絮; 张华; 陈高; 刘国霞; 边斐; 姚方印

    2015-01-01

    [Objective]ZmAN14is a member of the maize A20/AN1 zinc finger protein gene family. This gene family of rice is involved in the response to abiotic stress. The expression ofZmAN14inthe maize inbred line H21 and transgenic ZmAN14tobacco under abiotic stress was analyzed. The results will provide novel information for the comprehensive analysis of the functional and molecular mechanisms of maizeZmAN14and the entire gene family.[Method] Sequence analysis ofZmAN14was performed to confirm that this gene is a member of the maize A20/AN1 zinc finger protein gene family. The experiment was conducted by using maize inbred line H2. At the three-leaf stage, the seedlings were treated with multiple abiotic stress or induced by ABA. After treatment for 0, 1, 3, 6, 12 and 24 h, the whole plants were harvested. In the meantime, the roots, stems, leaves, coleptiles, pistil, stamen, silks and bract leaves were harvested from different growing periods of maize. Real-time quantitative fluorescence PCR was used to analyze the expression profile in different tissues and abiotic stress response ofZmAN14and its ABA-induced expression profile. Thecis-acting element of the promoter area was cloned to perform the comparative analysis. TheZmAN14coding sequence was cloned onto the GFP expressing vector pMDC85. A subcellular localization method was used to verify the localization of the ZmAN14 protein in the cell. TheZmAN14coding sequence was cloned onto the GAL4 DNA-binding domain vector. This vector was used to transform yeast. The transformed yeast was spotted onto the defect medium, and the resulted transcriptional activation activity was analyzed. TheZmAN14coding region was ligated to the plant expression vector p1300-221 to create an overexpression vector. The overexpression vector was transformed into tobacco. T2homozygous transgenic lines with highZmAN14 mRNA expression level were used to conduct salt-, drought- and ABA induced experiments, and its response to abiotic stress was

  12. Domain Modeling: NP_065914.2 [SAHG[Archive

    Lifescience Database Archive (English)

    Full Text Available NP_065914.2 chr8 Solution structure of the 6th C2H2 zinc finger of human Zinc finge...r protein 406 p2elva_ chr8/NP_065914.2/NP_065914.2_holo_398-430.pdb blast 406C,408I,409C,410E,411R,422H,425V,426H _ZN 0 ...

  13. Viscous fingering of miscible slices

    CERN Document Server

    De Wit, A; Martin, M; Wit, Anne De; Bertho, Yann; Martin, Michel

    2005-01-01

    Viscous fingering of a miscible high viscosity slice of fluid displaced by a lower viscosity fluid is studied in porous media by direct numerical simulations of Darcy's law coupled to the evolution equation for the concentration of a solute controlling the viscosity of miscible solutions. In contrast with fingering between two semi-infinite regions, fingering of finite slices is a transient phenomenon due to the decrease in time of the viscosity ratio across the interface induced by fingering and dispersion processes. We show that fingering contributes transiently to the broadening of the peak in time by increasing its variance. A quantitative analysis of the asymptotic contribution of fingering to this variance is conducted as a function of the four relevant parameters of the problem i.e. the log-mobility ratio R, the length of the slice l, the Peclet number Pe and the ratio between transverse and axial dispersion coefficients $\\epsilon$. Relevance of the results is discussed in relation with transport of vi...

  14. EST Table: CA946165 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CA946165 KI000253R 10/09/28 42 %/154 aa ref|XP_001952754.1| PREDICTED: similar to Z...el KRAB box and zinc finger, C2H2 type domain containing protein [Tribolium castaneum] CK539334 L9 ...

  15. Hypermethylated in cancer 1 (HIC1) recruits polycomb repressive complex 2 (PRC2) to a subset of its target genes through interaction with human polycomb-like (hPCL) proteins

    DEFF Research Database (Denmark)

    Boulay, Gaylor; Dubuissez, Marion; Van Rechem, Capucine;

    2012-01-01

    Hic1-deficient mice die perinatally and exhibit gross developmental defects throughout the second half of development. HIC1 encodes a transcriptional repressor with five C(2)H(2) zinc fingers mediating sequence-specific DNA binding and two repression domains: an N-terminal BTB/POZ domain and a central...

  16. Interaction of finger enslaving and error compensation in multiple finger force production

    OpenAIRE

    Martin, Joel R.; Mark L. Latash; Zatsiorsky, Vladimir M.

    2008-01-01

    Previous studies have documented two patterns of finger interaction during multi-finger pressing tasks, enslaving and error compensation, which do not agree with each other. Enslaving is characterized by positive correlation between instructed (master) and non-instructed (slave) finger(s) while error compensation can be described as a pattern of negative correlation between master and slave fingers. We hypothesize that pattern of finger interaction, enslaving or compensation, depends on the i...

  17. Mesofluidic controlled robotic or prosthetic finger

    Energy Technology Data Exchange (ETDEWEB)

    Lind, Randall F; Jansen, John F; Love, Lonnie J

    2013-11-19

    A mesofluidic powered robotic and/or prosthetic finger joint includes a first finger section having at least one mesofluidic actuator in fluid communication with a first actuator, a second mesofluidic actuator in fluid communication with a second actuator and a second prosthetic finger section pivotally connected to the first finger section by a joint pivot, wherein the first actuator pivotally cooperates with the second finger to provide a first mechanical advantage relative to the joint point and wherein the second actuator pivotally cooperates with the second finger section to provide a second mechanical advantage relative to the joint point.

  18. The SABS finger ring service

    International Nuclear Information System (INIS)

    All radiation workers are required by law to wear personal dosimeters to measure the radiation dose they receive in the course of their work. However, there was a need to measure the radiation dose received by the fingers and hands, because they are much closer to the source of radiation than the rest of the body. In April 1982 a finger ring service was instituted on a routine basis. A thermoluminescent dosimeter (TLD) is used, which is suitable for both X-rays and gamma rays. Materials that are used for thermoluminescent dosimeters includes LiF:Mg, CaF2:Dy and CaSO4:Dy

  19. Association analysis of zinc finger protein 804A gene with bipolar disorders and its subtypes in Han Chinese%锌指蛋白804A基因与中国汉族双相障碍及其亚型的关联分析

    Institute of Scientific and Technical Information of China (English)

    张晨; 汪作为; 吴志国; 洪武; 陈俊; 彭代辉; 方贻儒

    2015-01-01

    Objective To investigate the potential association between zinc finger protein 804A (ZNF804A) rs1344706 polymorphism and bipolar disorder as well as its subtypes.Methods A total of 304 bipolar Ⅰ disorder,442 bipolar Ⅱ disorder and 762 healthy controls subjects were recruited from Division of Mood Disorders,Shanghai Mental Health Center,Shanghai Jiao Tong University School of Medicine,from November 2006 to August 2014.Rs1344706 was genotyped by SNaPshot genotyping assay.Results In comparison between the patients with bipolar disorder and controls,there were significant differences in the genotypic (TT:210(28.2%) vs.175(23.0%),TG:378(50.7%) vs.399(52.4%),GG:158 (21.2%) vs.188(24.7%)) and allelic (T:798(53.5%) vs.749(49.1%),G:694(46.5%) vs.775(50.9%)) distributions (x2=6.18,P=0.04,x2=5.68,P=0.017,respectively).The frequency of T allele in patients with bipolar disorder was higher than that in controls (OR=1.19,95%CI 1.03-1.37).After stratified by diagnosed subtypes,there were significant differences in the genotypic and allelic distribution between patients with bipolar Ⅰ disorder and controls (x2=7.57,P=0.02,x2=7.22,P=0.007).The frequency of T allele in patients with bipolar Ⅰ disorder was higher than those in controls (OR=1.30,95% CI 1.07-1.56).In comparison between patients with bipolar Ⅱ disorder and controls,there was no significant difference in the distributions of genotype and allele.Conclusion There is positive association between rs1344706 polymorphism and bipolar Ⅰ disorder in a Chinese Han population.ZNF804A gene may confer susceptibility to bipolar Ⅰ disorder in Han Chinese.%目的 探讨位于锌指蛋白804A(zinc finger protein 804A,ZNF804A)内含子2区的多态性位点rs 1344706与双相障碍及其亚型的关系.方法 收集2006年11月至2014年8月在上海交通大学医学院附属精神卫生中心心境障碍科就诊的双相障碍患者746例(患者组),根据DSM-Ⅳ中双相障碍诊断亚型

  20. 27 CFR 9.34 - Finger Lakes.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Finger Lakes. 9.34 Section... Lakes. (a) Name. The name of the viticultural area described in this section is “Finger Lakes.” (b) Approved maps. The appropriate maps for determining the boundaries of the Finger Lakes viticultural...

  1. Multi-finger interaction during involuntary and voluntary single finger force changes

    OpenAIRE

    J R Martin; Zatsiorsky, V.M.; Latash, M.L.

    2010-01-01

    Two types of finger interaction are characterized by positive co-variation (enslaving) or negative co-variation (error compensation) of finger forces. Enslaving reflects mechanical and neural connections among fingers, while error compensation results from synergic control of fingers to stabilize their net output. Involuntary and voluntary force changes by a finger were used to explore these patterns. We hypothesized that synergic mechanisms will dominate during involuntary force changes, whi...

  2. Does finger sense predict addition performance?

    Science.gov (United States)

    Newman, Sharlene D

    2016-05-01

    The impact of fingers on numerical and mathematical cognition has received a great deal of attention recently. However, the precise role that fingers play in numerical cognition is unknown. The current study explores the relationship between finger sense, arithmetic and general cognitive ability. Seventy-six children between the ages of 5 and 12 participated in the study. The results of stepwise multiple regression analyses demonstrated that while general cognitive ability including language processing was a predictor of addition performance, finger sense was not. The impact of age on the relationship between finger sense, and addition was further examined. The participants were separated into two groups based on age. The results showed that finger gnosia score impacted addition performance in the older group but not the younger group. These results appear to support the hypothesis that fingers provide a scaffold for calculation and that if that scaffold is not properly built, it has continued differential consequences to mathematical cognition. PMID:26993292

  3. FINGER-VEIN RECOGNITION SYSTEMS

    Directory of Open Access Journals (Sweden)

    A.Haritha Deepthi

    2015-10-01

    Full Text Available As the Person‟s/Organization‟s Private information‟s are becoming very easy to access, the demand for a Simple, Convenient, Efficient, and a highly Securable Authentication System has been increased. In considering these requirements for data Protection, Biometrics, which uses human physiological or behavioral system for personal Identification has been found as a solution for these difficulties. However most of the biometric systems have high complexity in both time and space. So we are going to use a Real time Finger-Vein recognition System for authentication purposes. In this paper we had implemented the Finger Vein Recognition concept using MATLAB R2013a. The features used are Lacunarity Distance, Blanket Dimension distance. This has more accuracy when compared to conventional methods.

  4. Finger Indexed Sets: New Approaches

    CERN Document Server

    Sioutas, Spyros

    2008-01-01

    In the particular case we have insertions/deletions at the tail of a given set S of $n$ one-dimensional elements, we present a simpler and more concrete algorithm than that presented in [Anderson, 2007] achieving the same (but also amortized) upper bound of $O(\\sqrt{logd/loglogd})$ for finger searching queries, where $d$ is the number of sorted keys between the finger element and the target element we are looking for. Furthermore, in general case we have insertions/deletions anywhere we present a new randomized algorithm achieving the same expected time bounds. Even the new solutions achieve the optimal bounds in amortized or expected case, the advantage of simplicity is of great importance due to practical merits we gain.

  5. Finger clubbing in cystic fibrosis.

    OpenAIRE

    Pitts-Tucker, T J; Miller, M G; Littlewood, J M

    1986-01-01

    Finger clubbing was measured in 73 of 105 patients with cystic fibrosis undergoing full assessment. The sign correlated well with the chest x ray score and indices of pulmonary function and infection but not with weight, height, age, liver function, or degree of fat malabsorption. The presence of clubbing suggests appreciable pulmonary involvement. Most probably its progression indicates a deterioration in pulmonary state. In both instances increased efforts should be made to treat the infect...

  6. Current status of ultrasonography of the finger

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Seun Ah; Kim, Baek Hyun [Dept. of Radiology, Korea University Ansan Hospital, Korea University College of Medicine, Ansan (Korea, Republic of); Kim, Seon Jeong [Dept. of Radiology, Myongji Hospital, Seonam University College of Medicine, Goyang (Korea, Republic of); Kim, Ji Na [Dept. of Radiology, Chungnam National University Hospital, Chungnam National University School of Medicine, Daejeon (Korea, Republic of); Park, Sun Young [Dept. of Radiology, Hallym University Sacred Heart Hospital, Hallym University College of Medicine, Anyang (Korea, Republic of); Choi, Kyung Hee [Incheon Baek Hospital, Incheon (Korea, Republic of)

    2016-03-15

    The recent development of advanced high-resolution transducers has enabled the fast, easy, and dynamic ultrasonographic evaluation of small, superficial structures such as the finger. In order to best exploit these advances, it is important to understand the normal anatomy and the basic pathologies of the finger, as exemplified by the following conditions involving the dorsal, volar, and lateral sections of the finger: sagittal band injuries, mallet finger, and Boutonnière deformity (dorsal aspect); flexor tendon tears, trigger finger, and volar plate injuries (volar aspect); gamekeeper’s thumb (Stener lesions) and other collateral ligament tears (lateral aspect); and other lesions. This review provides a basis for understanding the ultrasonography of the finger and will therefore be useful for radiologists.

  7. Domain Modeling: NP_005072.2 [SAHG[Archive

    Lifescience Database Archive (English)

    Full Text Available NP_005072.2 chr2 Solution structure of the N-terminal C2H2 type zinc-binding domain... of the Zinc finger protein 64, isoforms 1 and 2 c2dmda_ chr2/NP_005072.2/NP_005072.2_holo_380-476.pdb psi-blast 383C,386C,388Y,399H,404H,412C,415C,428H,429K,447A,449Q,450E _ZN 0 ...

  8. Trigger Finger: Adult and Pediatric Treatment Strategies.

    Science.gov (United States)

    Giugale, Juan M; Fowler, John R

    2015-10-01

    Trigger fingers are common tendinopathies representing a stenosing flexor tenosynovitis of the fingers. Adult trigger finger can be treated nonsurgically using activity modification, splinting, and/or corticosteroid injections. Surgical treatment options include percutaneous A1 pulley release and open A1 pulley release. Excision of a slip of the flexor digitorum superficialis is reserved for patients with persistent triggering despite A1 release or patients with persistent flexion contracture. Pediatric trigger thumb is treated with open A1 pulley release. Pediatric trigger finger is treated with release of the A1 pulley with excision of a slip or all of the flexor digitorum superficialis if triggering persists. PMID:26410644

  9. Fluid Mixing from Viscous Fingering

    CERN Document Server

    Jha, Birendra; Juanes, Ruben

    2010-01-01

    We study, by means of numerical simulation, the mixing of two fluids of different viscosities in advection-dominated flows in a porous medium. It is well known that when a less viscous fluid displaces a more viscous fluid, the displacement front is unstable and leads to the formation of a pattern known as viscous fingering. We present a high-resolution simulation approach that is stable for arbitrary viscosity ratios, and study mixing under different configurations with viscosity contrasts up to M = 400. We observe, in agreement with lab experiments, that for high-M displacements, the growth of new fingers follows the trace of previous ones. This channeling effect, which is a result of the nonlocal coupling through the pressure field, greatly reduces mixing. A two-equation mixing model using the scalar variance and its dissipation rate is derived from the advection-diffusion equation. It provides a measure of effective diffusivity due to convective and diffusive mixing processes. Our analysis predicts the opt...

  10. Reversed cross finger subcutaneous flap: A rapid way to cover finger defects

    Directory of Open Access Journals (Sweden)

    Fejjal Nawfal

    2008-01-01

    Full Text Available Adequate coverage of dorsal finger wounds is often a challenge. The reversed cross finger subcutaneous flap to cover defects on the dorsum of phalanx constitutes an excellent option for coverage of wounds over the middle and distal phalanges of the index, middle, ring, and small fingers. It′s an easy flap and represents our first choice to cover those defects.

  11. Axon reflexes in human cold exposed fingers

    NARCIS (Netherlands)

    Daanen, H.A.M.; Ducharme, M.B.

    2000-01-01

    Exposure of fingers to severe cold induces cold induced vasodilation (CIVD). The mechanism of CIVD is still debated. The original theory states that an axon reflex causes CIVD. To test this hypothesis, axon reflexes were evoked by electrical stimulation of the middle fingers of hands immersed in wat

  12. Surgical Treatment of Trigger Finger: Open Release

    Directory of Open Access Journals (Sweden)

    Firat Ozan

    2016-01-01

    Full Text Available In this study, open A1 pulley release results were evaluated in patients with a trigger finger diagnosis. 45 patients (29 females, 16 males, mean age 50.7 ± 11.9; range (24-79, 45 trigger fingers were released via open surgical technique. On the 25 of 45 cases were involved in the right hand and 16 of them were at the thumb, 2 at index, 6 at the middle and 1 at ring finger. Similarly, at the left hand, 15 of 20 cases were at the thumb, 1 at the index finger, 2 at middle finger and 2 at ring finger. Average follow-up time was 10.2 ± 2.7 (range, 6-15 months. Comorbidities in patients were; diabetes mellitus at 6 cases (13.3%, hypertension at 11 cases (24.4%, hyperthyroidism at 2 cases (4.4%, dyslipidemia at 2 cases (4.4% and lastly 2 cases had carpal tunnel syndrome operation. The mean time between the onset of symptoms to surgery was 6.9 ± 4.8 (range, 2-24 months. Patient satisfaction was very good in 34 cases (75.4% and good in 11 (24.6% patients. The distance between the pulpa of the operated finger and the palm was normal in every case postoperatively. We have not encountered any postoperative complications. We can recommend that; A1 pulley release via open incision is an effective and reliable method in trigger finger surgery.

  13. Zinc'ing down RNA polymerase I

    OpenAIRE

    Chanfreau, Guillaume F.

    2013-01-01

    Most RNA polymerases contain zinc, yet the precise function of zinc and its influence of polymerases stability are unknown. A recent study provides evidence that zinc levels control the stability of RNA polymerase I in vivo and that the enzyme might serve as a zinc reservoir for other proteins.

  14. Symptomatic zinc deficiency in experimental zinc deprivation.

    OpenAIRE

    Taylor, C M; Goode, H F; Aggett, P J; Bremner, I; Walker, B E; Kelleher, J.

    1992-01-01

    An evaluation of indices of poor zinc status was undertaken in five male subjects in whom dietary zinc intake was reduced from 85 mumol d-1 in an initial phase of the study to 14 mumol d-1. One of the subjects developed features consistent with zinc deficiency after receiving the low zinc diet for 12 days. These features included retroauricular acneform macullo-papular lesions on the face, neck, and shoulders and reductions in plasma zinc, red blood cell zinc, neutrophil zinc and plasma alkal...

  15. Layer Formation in Sedimentary Fingering Convection

    CERN Document Server

    Reali, J F; Alsinan, A; Meiburg, E

    2016-01-01

    When particles settle through a stable temperature or salinity gradient they can drive an instability known as sedimentary fingering convection. This phenomenon is thought to occur beneath sediment-rich river plumes in lakes and oceans, in the context of marine snow where decaying organic materials serve as the suspended particles, or in the atmosphere in the presence of aerosols or volcanic ash. Laboratory experiments of Houk and Green (1973) and Green (1987) have shown sedimentary fingering convection to be similar to the more commonly known thermohaline fingering convection in many ways. Here, we study the phenomenon using 3D direct numerical simulations. We find evidence for layer formation in sedimentary fingering convection in regions of parameter space where it does not occur for non-sedimentary systems. This is due to two complementary effects. Sedimentation affects the turbulent fluxes and broadens the region of parameter space unstable to the $\\gamma$-instability (Radko 2003) to include systems at l...

  16. Double dislocation of finger interphalangeal joints

    OpenAIRE

    Jahangiri, Saqib Aziz; Mestha, Prabhakar; McNally, Scarlett

    2012-01-01

    A 62-year-old, right-hand-dominant man who had dementia and lived in an Elderly Mentally Infirm (EMI) nursing home was admitted through Accident & Emergency (A&E) department following unwitnessed injury to the left little finger.

  17. Fingering Convection in Red Giants Revisited

    CERN Document Server

    Wachlin, F C; Althaus, L G

    2014-01-01

    Fingering (thermohaline) convection has been invoked for several years as a possible extra-mixing which could occur in Red Giant stars due to the modification of the chemical composition induced by nuclear reactions in the hydrogen burning zone. Recent studies show however that this mixing is not sufficient to account for the needed surface abundances. A new prescription for fingering convection, based on 3D numerical simulations has recently been proposed (BGS). The resulting mixing coefficient is larger than the ones previously given in the literature. We compute models using this new coefficient and compare them to previous studies. We use the LPCODE stellar evolution code with the GNA generalized version of the mixing length theory to compute Red Giant models and we introduce fingering convection using the BGS prescription. The results show that, although the fingering zone now reaches the outer dynamical convective zone, the efficiency of the mixing is not enough to account for the observations. The fing...

  18. Surface tension and dynamics of fingering patterns

    OpenAIRE

    Magdaleno, F. X.; Casademunt, J.

    1997-01-01

    We study the minimal class of exact solutions of the Saffman-Taylor problem with zero surface tension, which contains the physical fixed points of the regularized (non-zero surface tension) problem. New fixed points are found and the basin of attraction of the Saffman-Taylor finger is determined within that class. Specific features of the physics of finger competition are identified and quantitatively defined, which are absent in the zero surface tension case. This has dramatic consequences f...

  19. Histopathology of tenosynovium in trigger fingers.

    Science.gov (United States)

    Uchihashi, Kazuyoshi; Tsuruta, Toshiyuki; Mine, Hiroko; Aoki, Shigehisa; Nishijima-Matsunobu, Aki; Yamamoto, Mihoko; Kuraoka, Akio; Toda, Shuji

    2014-06-01

    Stenosing flexor tenosynovitis, trigger finger, is a common clinical disorder causing painful locking or contracture of the involved digits, and most instances are idiopathic. This problem is generally caused by a size mismatch between the swollen flexor tendon and the thickened first annular pulley. Although hypertrophic pulleys have been histologically and ultrasonographically detected, little is known about the histopathology of the tenosynovium covering the tendons of trigger fingers. We identified chondrocytoid cells that produced hyaluronic acid in 23 (61%) fingers and hypocellular collagen matrix in 32 (84%) fingers around the tenosynovium among 38 specimens of tenosynovium from patients with trigger fingers. These chondrocytoid cells expressed the synovial B cell marker CD44, but not the chondrocyte marker S-100 protein. The incidence of these findings was much higher than that of conventional findings of synovitis, such as inflammatory infiltrate (37%), increased vascularity (37%), hyperplasia of synovial lining cells (21%), or fibrin exudation (5%). We discovered the following distinctive histopathological features of trigger finger: hyaluronic acid-producing chondrocytoid cells originated from fibroblastic synovial B cells, and a hypocellular collagen matrix surrounding the tenosynovium. Thus, an edematous extracellular matrix with active hyaluronic acid synthesis might increase pressure under the pulley and contribute to the progression of stenosis. PMID:24965110

  20. Finger Search in the Implicit Model

    DEFF Research Database (Denmark)

    Brodal, Gerth Stølting; Nielsen, Jesper Asbjørn Sindahl; Truelsen, Jakob

    2012-01-01

    the finger to another element is Ω(q− 1(logn)), where t is the rank distance between the query element and the finger. We present an optimal implicit static structure matching this lower bound. We furthermore present a near optimal implicit dynamic structure supporting search, change-finger, insert......We address the problem of creating a dictionary with the finger search property in the strict implicit model, where no information is stored between operations, except the array of elements. We show that for any implicit dictionary supporting finger searches in q(t) = Ω(logt) time, the time to move......, and delete in times $\\mathcal{O}(q(t))$, $\\mathcal{O}(q^{-1}(\\log n)\\log n)$, $\\mathcal{O}(\\log n)$, and $\\mathcal{O}(\\log n)$, respectively, for any q(t) = Ω(logt). Finally we show that the search operation must take Ω(logn) time for the special case where the finger is always changed to the element...

  1. Highly transparent front electrodes with metal fingers for p-i-n thin-film silicon solar cells

    Directory of Open Access Journals (Sweden)

    Moulin Etienne

    2015-01-01

    Full Text Available The optical and electrical properties of transparent conductive oxides (TCOs, traditionally used in thin-film silicon (TF-Si solar cells as front-electrode materials, are interlinked, such that an increase in TCO transparency is generally achieved at the cost of reduced lateral conductance. Combining a highly transparent TCO front electrode of moderate conductance with metal fingers to support charge collection is a well-established technique in wafer-based technologies or for TF-Si solar cells in the substrate (n-i-p configuration. Here, we extend this concept to TF-Si solar cells in the superstrate (p-i-n configuration. The metal fingers are used in conjunction with a millimeter-scale textured foil, attached to the glass superstrate, which provides an antireflective and retroreflective effect; the latter effect mitigates the shadowing losses induced by the metal fingers. As a result, a substantial increase in power conversion efficiency, from 8.7% to 9.1%, is achieved for 1-μm-thick microcrystalline silicon solar cells deposited on a highly transparent thermally treated aluminum-doped zinc oxide layer combined with silver fingers, compared to cells deposited on a state-of-the-art zinc oxide layer.

  2. Integrated criteria document Zinc

    NARCIS (Netherlands)

    Cleven RFMJ; Janus JA; Annema JA; Slooff W

    1993-01-01

    This report contains information on zinc and zinc compounds concerning standards, emissions, exposure levels and effect levels. It includes a risk evaluation and presents proposals for maximum permissible concentrations of zinc in the environment. This study indicates that the concentration of zinc

  3. Ultrafast High-Resolution Mass Spectrometric Finger Pore Imaging in Latent Finger Prints

    Science.gov (United States)

    Elsner, Christian; Abel, Bernd

    2014-11-01

    Latent finger prints (LFPs) are deposits of sweat components in ridge and groove patterns, left after human fingers contact with a surface. Being important targets in biometry and forensic investigations they contain more information than topological patterns. With laser desorption mass spectrometry imaging (LD-MSI) we record `three-dimensional' finger prints with additional chemical information as the third dimension. Here we show the potential of fast finger pore imaging (FPI) in latent finger prints employing LD-MSI without a classical matrix in a high- spatial resolution mode. Thin films of gold rapidly sputtered on top of the sample are used for desorption. FPI employing an optical image for rapid spatial orientation and guiding of the desorption laser enables the rapid analysis of individual finger pores, and the chemical composition of their excretions. With this approach we rapidly detect metabolites, drugs, and characteristic excretions from the inside of the human organism by a minimally-invasive strategy, and distinguish them from chemicals in contact with fingers without any labeling. The fast finger pore imaging, analysis, and screening approach opens the door for a vast number of novel applications in such different fields as forensics, doping and medication control, therapy, as well as rapid profiling of individuals.

  4. Design of a 2-Finger Hand Exoskeleton for Finger Stiffness Measurements

    Directory of Open Access Journals (Sweden)

    Angelo Emanuele Fiorilla

    2009-01-01

    Full Text Available Recent studies of human arm movements have suggested that the control of stiffness may be important both for maintaining stability and for achieving differences in movement accuracy. Several studies in the robotic field demonstrated that grasp stiffness is useful for modelling and controlling manipulators but, even though it is accredited that having models of the human finger impedance would be very desirable for the control of anthropomorphous robot's hands, relatively few studies have focused on finger and hand stiffness. To allow the measurement of such entities at the finger level, an appropriate device capable of applying fast force transients while at the same time be able to monitor the finger movements is required. The work presented in this paper is a very detailed report about the design of a new hand exoskeleton system that will be used in our future works to investigate the finger stiffness range in different grasping postures and conditions.

  5. Saffman-Taylor fingers with kinetic undercooling

    KAUST Repository

    Gardiner, Bennett P. J.

    2015-02-23

    © 2015 American Physical Society. The mathematical model of a steadily propagating Saffman-Taylor finger in a Hele-Shaw channel has applications to two-dimensional interacting streamer discharges which are aligned in a periodic array. In the streamer context, the relevant regularization on the interface is not provided by surface tension but instead has been postulated to involve a mechanism equivalent to kinetic undercooling, which acts to penalize high velocities and prevent blow-up of the unregularized solution. Previous asymptotic results for the Hele-Shaw finger problem with kinetic undercooling suggest that for a given value of the kinetic undercooling parameter, there is a discrete set of possible finger shapes, each analytic at the nose and occupying a different fraction of the channel width. In the limit in which the kinetic undercooling parameter vanishes, the fraction for each family approaches 1/2, suggesting that this "selection" of 1/2 by kinetic undercooling is qualitatively similar to the well-known analog with surface tension. We treat the numerical problem of computing these Saffman-Taylor fingers with kinetic undercooling, which turns out to be more subtle than the analog with surface tension, since kinetic undercooling permits finger shapes which are corner-free but not analytic. We provide numerical evidence for the selection mechanism by setting up a problem with both kinetic undercooling and surface tension and numerically taking the limit that the surface tension vanishes.

  6. Perceiving fingers in single-digit arithmetic problems

    Directory of Open Access Journals (Sweden)

    Ilaria eBerteletti

    2015-03-01

    Full Text Available In this study, we investigate in children the neural underpinnings of finger representation and finger movement involved in single-digit arithmetic problems. Evidence suggests that finger representation and finger-based strategies play an important role in learning and understanding arithmetic. Because different operations rely on different networks, we compared activation for subtraction and multiplication problems in independently localized finger somatosensory and motor areas and tested whether activation was related to skill. Brain activations from children between 8 and 13 years of age revealed that only subtraction problems significantly activated finger motor areas, suggesting reliance on finger-based strategies. In addition, larger subtraction problems yielded greater somatosensory activation than smaller problems, suggesting a greater reliance on finger representation for larger numerical values. Interestingly, better performance in subtraction problems was associated with lower activation in the finger somatosensory area. Our results support the importance of fine-grained finger representation in arithmetical skill and are the first neurological evidence for a functional role of the somatosensory finger area in proficient arithmetical problem solving, in particular for those problems requiring quantity manipulation. From an educational perspective, these results encourage investigating whether different finger-based strategies facilitate arithmetical understanding and encourage educational practices aiming at integrating finger representation and finger-based strategies as a tool for instilling stronger numerical sense.

  7. Domain Modeling: NP_003414.2 [SAHG[Archive

    Lifescience Database Archive (English)

    Full Text Available NP_003414.2 chr19 Solution structure of C2H2 type Zinc finger domain 345 in Zinc fi...nger protein 278 c2yt9a_ chr19/NP_003414.2/NP_003414.2_holo_617-702.pdb blast 624C,626E,627C,628G,629K,640H,644H,652C,654E,655C,668H,672H,680C,682E,683C,684G,696H,699I,700H _ZN 0 ...

  8. Experiment list: SRX100521 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available to the mouse Bcl11a/Evi9 protein. Functions as a myeloid and B-cell proto-oncogen...e Bcl11a/Evi9 protein. Functions as a myeloid and B-cell proto-oncogene. May play important roles in leukemo...n=This gene encodes a C2H2 type zinc-finger protein by its similarity to the mous

  9. Fingering instability in combustion: an extended view.

    Science.gov (United States)

    Zik, O; Moses, E

    1999-07-01

    We detail the experimental situation concerning the fingering instability that occurs when a solid fuel is forced to burn against a horizontal oxidizing wind. The instability appears when the Rayleigh number for convection is below criticality. The focus is on the developed fingering state. We present direct measurements of the depletion of oxygen by the front as well as new results that connect heat losses to the characteristic scale of the instability. In addition, we detail the experimental system, elaborate (qualitatively and quantitatively) on the results that were previously presented, and discuss new observations. We also show that the same phenomenological model applies to electrochemical deposition.

  10. Autonomous control of multi-fingered hand

    Institute of Scientific and Technical Information of China (English)

    JIANG Li; LIU Hong

    2006-01-01

    This paper describes a novel autonomous control strategy of multi-fingered hand based on a modular control system of dexterous manipulation. A simple proportional-integral-derivative(PID) position control with friction compensation, which requires few friction parameters, is used to realize accurate and smooth trajectory tracking in pregrasp phase. In grasp and manipulation phases, an event-driven switcher is adopted to determine the switching between unconstrained position control and constrained torque control, and an improved explicit integral force control strategy is implemented to realize simultaneously stable contact transition and accurate force tracking. Experimental results have verified the effectiveness of the proposed autonomous control strategy of multi-fingered hand.

  11. Trace elements in human physiology and pathology: zinc and metallothioneins.

    Science.gov (United States)

    Tapiero, Haim; Tew, Kenneth D

    2003-11-01

    Zinc is one of the most abundant nutritionally essential elements in the human body. It is found in all body tissues with 85% of the whole body zinc in muscle and bone, 11% in the skin and the liver and the remaining in all the other tissues. In multicellular organisms, virtually all zinc is intracellular, 30-40% is located in the nucleus, 50% in the cytoplasm, organelles and specialized vesicles (for digestive enzymes or hormone storage) and the remainder in the cell membrane. Zinc intake ranges from 107 to 231 micromol/d depending on the source, and human zinc requirement is estimated at 15 mg/d. Zinc has been shown to be essential to the structure and function of a large number of macromolecules and for over 300 enzymic reactions. It has both catalytic and structural roles in enzymes, while in zinc finger motifs, it provides a scaffold that organizes protein sub-domains for the interaction with either DNA or other proteins. It is critical for the function of a number of metalloproteins, inducing members of oxido-reductase, hydrolase ligase, lyase family and has co-activating functions with copper in superoxide dismutase or phospholipase C. The zinc ion (Zn(++)) does not participate in redox reactions, which makes it a stable ion in a biological medium whose potential is in constant flux. Zinc ions are hydrophilic and do not cross cell membranes by passive diffusion. In general, transport has been described as having both saturable and non-saturable components, depending on the Zn(II) concentrations involved. Zinc ions exist primarily in the form of complexes with proteins and nucleic acids and participate in all aspects of intermediary metabolism, transmission and regulation of the expression of genetic information, storage, synthesis and action of peptide hormones and structural maintenance of chromatin and biomembranes. PMID:14652165

  12. Zinc: the neglected nutrient.

    Science.gov (United States)

    Shambaugh, G E

    1989-03-01

    Zinc was first recognized as essential for animals at the University of Illinois School of Agriculture in 1916, when it was found that zinc-deficient baby pigs were runty, developed dermatitis on their legs, and were sterile. Zinc deficiency was first recognized in man by Dr. Ananda Prasad of Detroit 26 years ago when he measured serum and hair zinc levels in young male Egyptian dwarfs who had failed to mature and were small in stature. By simply adding zinc to their regular diet, they grew in height and became sexually mature. It is now recognized that dwarfism in males is frequent around the Mediterranean, where wheat is the staple of life and has been grown for 4,000 years on the same soil, thereby resulting in the depletion of zinc. Professor Robert Henkin first suggested that zinc deficiency might cause hearing-nerve impairment. Assay of the soft tissues of the cochlea and vestibule revealed a zinc level higher than that of any other part of the body. Previously, the eye was considered to have the highest level of zinc of any organ. To diagnose zinc deficiency clinically, we use serum zinc assays made at the Mayo Clinic Trace Element Laboratory. With zinc supplementation in patients who are marginally zinc deficient, there has been improvement in tinnitus and sensorineural hearing loss in about one-third of elderly adults. We believe zinc deficiency is one causation of presbycusis; by recognizing and correcting it, a progressive hearing loss can be arrested. PMID:2786676

  13. Zinc: the neglected nutrient.

    Science.gov (United States)

    Shambaugh, G E

    1989-03-01

    Zinc was first recognized as essential for animals at the University of Illinois School of Agriculture in 1916, when it was found that zinc-deficient baby pigs were runty, developed dermatitis on their legs, and were sterile. Zinc deficiency was first recognized in man by Dr. Ananda Prasad of Detroit 26 years ago when he measured serum and hair zinc levels in young male Egyptian dwarfs who had failed to mature and were small in stature. By simply adding zinc to their regular diet, they grew in height and became sexually mature. It is now recognized that dwarfism in males is frequent around the Mediterranean, where wheat is the staple of life and has been grown for 4,000 years on the same soil, thereby resulting in the depletion of zinc. Professor Robert Henkin first suggested that zinc deficiency might cause hearing-nerve impairment. Assay of the soft tissues of the cochlea and vestibule revealed a zinc level higher than that of any other part of the body. Previously, the eye was considered to have the highest level of zinc of any organ. To diagnose zinc deficiency clinically, we use serum zinc assays made at the Mayo Clinic Trace Element Laboratory. With zinc supplementation in patients who are marginally zinc deficient, there has been improvement in tinnitus and sensorineural hearing loss in about one-third of elderly adults. We believe zinc deficiency is one causation of presbycusis; by recognizing and correcting it, a progressive hearing loss can be arrested.

  14. ANALYSIS WITH MSC ADAMS OF A 5-FINGER AND 3-PHALANX /FINGER UNDER-ACTUATEDMECHANICAL HAND

    Directory of Open Access Journals (Sweden)

    Gheorghe POPESCU

    2013-05-01

    Full Text Available This paper studies the analysis with MSC ADAMS of a 5-fingered and 3-phalanx/finger underactuatedmechanical hand, designed by the author to work on industrial robots. Moreover, in order to increasegrasping safety in the automated handling process, the author has fitted each finger with a locking sequence inthe final phase of grasping. Thus, the mechanism of mechanical hand is considered to be a mechanical systemand is treated like a set of rigid bodies connected by mechanical linkages and elastic elements. To model andsimulate this mechanism with MSC ADAMS programme, the author covered the following stages: constructionof the model, testing-simulation, validation, finishing, parameterization, and optimization

  15. Is zinc a neuromodulator?

    Science.gov (United States)

    Kay, Alan R; Tóth, Katalin

    2008-01-01

    The vesicles of certain glutamatergic terminals in the mammalian forebrain are replete with ionic zinc. It is believed that during synaptic transmission zinc is released, binds to receptors on the pre- or postsynaptic membranes, and hence acts as a neuromodulator. Although exogenous zinc modulates a wide variety of channels, whether synaptic zinc transits across the synaptic cleft and alters the response of channels has been difficult to establish. We will review the evidence for zinc as a neuromodulator and propose diagnostic criteria for establishing whether it is indeed one. Moreover, we will delineate alternative ways in which zinc might act at synapses.

  16. Reactivity of Cys4 Zinc Finger Domains with Gold(III) Complexes : Insights into the Formation of "Gold Fingers"

    NARCIS (Netherlands)

    Jacques, Aurélie; Lebrun, Colette; Casini, Angela; Kieffer, Isabelle; Proux, Olivier; Latour, Jean-Marc; Sénèque, Olivier

    2015-01-01

    Gold(I) complexes such as auranofin or aurothiomalate have been used as therapeutic agents for the treatment of rheumatoid arthritis for several decades. Several gold(I) and gold(III) complexes have also shown in vitro anticancer properties against human cancer cell lines, including cell lines resis

  17. The PHD finger of human UHRF1 reveals a new subgroup of unmethylated histone H3 tail readers.

    Directory of Open Access Journals (Sweden)

    Nada Lallous

    Full Text Available The human UHRF1 protein (ubiquitin-like containing PHD and RING finger domains 1 has emerged as a potential cancer target due to its implication in cell cycle regulation, maintenance of DNA methylation after replication and heterochromatin formation. UHRF1 functions as an adaptor protein that binds to histones and recruits histone modifying enzymes, like HDAC1 or G9a, which exert their action on chromatin. In this work, we show the binding specificity of the PHD finger of human UHRF1 (huUHRF1-PHD towards unmodified histone H3 N-terminal tail using native gel electrophoresis and isothermal titration calorimetry. We report the molecular basis of this interaction by determining the crystal structure of huUHRF1-PHD in complex with the histone H3 N-terminal tail. The structure reveals a new mode of histone recognition involving an extra conserved zinc finger preceding the conventional PHD finger region. This additional zinc finger forms part of a large surface cavity that accommodates the side chain of the histone H3 lysine K4 (H3K4 regardless of its methylation state. Mutation of Q330, which specifically interacts with H3K4, to alanine has no effect on the binding, suggesting a loose interaction between huUHRF1-PHD and H3K4. On the other hand, the recognition appears to rely on histone H3R2, which fits snugly into a groove on the protein and makes tight interactions with the conserved aspartates D334 and D337. Indeed, a mutation of the former aspartate disrupts the formation of the complex, while mutating the latter decreases the binding affinity nine-fold.

  18. Effective Length Design of Humanoid Robot Fingers Using Biomimetic Optimization

    Directory of Open Access Journals (Sweden)

    Byoung-Ho Kim

    2015-10-01

    Full Text Available In this study, we propose an effective design method for the phalangeal parameters and the total size of humanoid robot fingers based on a biomimetic optimization. For the optimization, an interphalangeal joint coordination parameter and the length constraints inherent in human fingers are considered from a biomimetic perspective. A reasonable grasp formulation is also taken into account from the viewpoint of power grasping, where the grasp space of a humanoid robot finger is importantly considered to determine the phalangeal length parameters. The usefulness of the devised biomimetic optimization method is shown through the design examples of various humanoid robot fingers. In fact, the optimization-based finger design method enables us to determine effectively the proper phalangeal size of humanoid robot fingers for human-like object handling tasks. In addition, we discuss its contribution to the structural configuration and coordinated motion of a humanoid robot finger, and address its practical availability in terms of effective finger design.

  19. Acceleration Workspace of Cooperating Multi-Finger Robot Systems

    Institute of Scientific and Technical Information of China (English)

    Hyungwon Shim; Jihong Lee

    2008-01-01

    We present a mathematical method for acceleration workspace analysis of cooperating multi-finger robot systems using a model of point-contact with friction. A new unified formulation from dynamic equations of cooperating multi-finger robots is derived considering the force and acceleration relationships between the fingers and the object to be handled. From the dynamic equation, maximum translational and rotational acceleration bounds of an object are calculated under given constraints of contact conditions, configurations of fingers, and bounds on the torques of joint actuators for each finger. Here, the rotational acceleration bounds can be applied as an important manipulability index when the multi-finger robot grasps an object. To verify the proposed method, we used a set of case studies with a simple multi-finger mechanism system. The achievable acceleration boundary in task space can be obtained successfully with the proposed method and the acceleration boundary depends on the configurations of fingers.

  20. Clubbed fingers: the claws we lost?

    NARCIS (Netherlands)

    Brouwers, A.A.M.; Vermeij-Keers, C.; Zoelen, E.J.J. van; Gooren, L.J.G.

    2004-01-01

    Clubbed digits resemble the human embryonic fingers and toes, which took like the digits of a claw. Clubbed digits, thus, may represent the return of the embryonic claw and may even represent the claws man has lost during evolution, if ontogenesis realty recapitulates phylogenesis. We put forward th

  1. Viscous fingering with partial miscible fluids

    Science.gov (United States)

    Fu, Xiaojing; Cueto-Felgueroso, Luis; Juanes, Ruben

    2015-11-01

    When a less viscous fluid displaces a more viscous fluid, the contrast in viscosity destabilizes the interface between the two fluids, leading to the formation of fingers. Studies of viscous fingering have focused on fluids that are either fully miscible or perfectly immiscible. In practice, however, the miscibility of two fluids can change appreciably with temperature and pressure, and often falls into the case of partial miscibility, where two fluids have limited solubility in each other. Following our recent work for miscible (Jha et al., PRL 2011, 2013) and immiscible systems (Cueto-Felgueroso and Juanes, PRL 2012, JFM 2014), here we propose a phase-field model for fluid-fluid displacements in a Hele-Shaw cell, when the two fluids have limited (but nonzero) solubility in one another. Partial miscibility is characterized through the design of thermodynamic free energy of the two-fluid system. We elucidate the key dimensionless groups that control the behavior of the system. We present high-resolution numerical simulations of the model applied to the viscous fingering problem. On one hand, we demonstrate the effect of partial miscibility on the hydrodynamic instability. On the other, we elucidate the role of the degree of fingering on the rate of mutual fluid dissolution.

  2. Coriolis effects on fingering patterns under rotation.

    Science.gov (United States)

    Alvarez-Lacalle, Enrique; Gadêlha, Hermes; Miranda, José A

    2008-08-01

    The development of immiscible viscous fingering patterns in a rotating Hele-Shaw cell is investigated. We focus on understanding how the time evolution and the resulting morphologies are affected by the action of the Coriolis force. The problem is approached analytically and numerically by employing a vortex sheet formalism. The vortex sheet strength and a linear dispersion relation are derived analytically, revealing that the most relevant Coriolis force contribution comes from the normal component of the averaged interfacial velocity. It is shown that this normal velocity, uniquely due to the presence of the Coriolis force, is responsible for the complex-valued nature of the linear dispersion relation making the linear phases vary with time. Fully nonlinear stages are studied through intensive numerical simulations. A suggestive interplay between inertial and viscous effects is found, which modifies the dynamics, leading to different pattern-forming structures. The inertial Coriolis contribution plays a characteristic role: it generates a phase drift by deviating the fingers in the sense opposite to the actual rotation of the cell. However, the direction and intensity of finger bending is predominantly determined by viscous effects, being sensitive to changes in the magnitude and sign of the viscosity contrast. The finger competition behavior at advanced time stages is also discussed.

  3. Finger-jointed beams in bending

    DEFF Research Database (Denmark)

    Andreasen, Lotte; Hoffmeyer, Preben

    1997-01-01

    An investigation of the dynamic and static fatique of finger-jointed beams in bending was carried out. Results were obtained for five different frequencies from static loading to a load cycle period of two minutes. A total of seven series were long-term tested and five series were short-term tested...

  4. Finger cold-induced vasodilation : a review

    NARCIS (Netherlands)

    Daanen, H.A.M.

    2003-01-01

    Cold-induced vasodilation (CIVD) in the finger tips generally occurs 5-10 min after the start of local cold exposure of the extremities. This phenomenon is believed to reduce the risk of local cold injuries. However, CIVD is almost absent during hypothermia, when survival of the organism takes prece

  5. Transdermal anaesthesia for percutaneous trigger finger release.

    Science.gov (United States)

    Yiannakopoulos, Christos K; Ignatiadis, Ioannis A

    2006-01-01

    The purpose of this study was to evaluate the safety and efficiency of transdermal anaesthesia using eutectic mixture of lidocaine and prilocaine (EMLA) in patients undergoing percutaneous trigger finger release and to compare it with lidocaine infiltration. In this prospective, randomised study percutaneous release of the A1 annular pulley was performed to treat stenosing tenosynovitis (trigger finger syndrome) in 50 patients (50 fingers). The procedure was performed either under transdermal anaesthesia using EMLA applied transcutaneously 120 minutes prior to the operation (Group A, n = 25) or using local infiltration anaesthesia using lidocaine (Group B, n = 25). Pain experienced during administration of anaesthesia and during the operation was assessed using a 10-point Visual Analogue Pain Scale (VAPS), while all patients rated the effectiveness of anaesthesia with a 5-point scale. There were no significant differences between the two groups in the VAPS during the operation (1.33 +/- 0.52 versus 1.59 +/- 0.87) and the satisfaction scores (4.6 +/- 0.2 versus 4.4 +/- 0.3). The VAPS score during the administration of anaesthesia was statistically significantly less in the EMLA group (0 versus 5.96 +/- 2.41). All patients were satisfied with the final result of the operation. Percutaneous trigger finger release can be performed as an office procedure with the use of EMLA avoiding the use of injectable local infiltration anaesthesia. PMID:17405199

  6. Fingering phenomena during grain-grain displacement

    Science.gov (United States)

    Mello, Nathália M. P.; Paiva, Humberto A.; Combe, G.; Atman, A. P. F.

    2016-05-01

    Spontaneous formation of fingered patterns during the displacement of dense granular assemblies was experimentally reported few years ago, in a radial Hele-Shaw cell. Here, by means of discrete element simulations, we have recovered the experimental findings and extended the original study to explore the control parameters space. In particular, using assemblies of grains with different geometries (monodisperse, bidisperse, or polydisperse), we measured the macroscopic stress tensor in the samples in order to confirm some conjectures proposed in analogy with Saffman-Taylor viscous fingering phenomena for immiscible fluids. Considering an axial setup which allows to control the discharge of grains and to follow the trajectory and the pressure gradient along the displacing interface, we have applied the Darcy law for laminar flow in fluids in order to measure an "effective viscosity" for each assembly combination, in an attempt to mimic variation of the viscosity ratio between the injected/displaced fluids in the Saffman-Taylor experiment. The results corroborate the analogy with the viscous fluids displacement, with the bidisperse assembly corresponding to the less viscous geometry. But, differently to fluid case, granular fingers only develop for a specific combination of displaced/injected geometries, and we have demonstrated that it is always related with the formation of a force chain network along the finger direction.

  7. Ten Trigger Fingers in an Adult Man : A Case Report

    OpenAIRE

    Lee, Young-Keun; Kam, Byung-Sup; Lee, Kwang-Won; Kim, Whoan-Jeang; Choy, Won-Sik

    2007-01-01

    Trigger finger is a common disease particularly in the middle aged women. A very rare case in which an adult man had 10 trigger fingers was experienced. He was treated with local steroid injections in both thumbs, but trigger finger disease has been aggravated in every digit of both hands. We performed an early operative treatment. Three months after the operation, the patient could perform his work without discomfort in his hands and showed normal range of motion in all fingers.

  8. A palmitoylated RING finger ubiquitin ligase and its homologue in the brain membranes.

    Science.gov (United States)

    Araki, Kazuaki; Kawamura, Meiko; Suzuki, Toshiaki; Matsuda, Noriyuki; Kanbe, Daiji; Ishii, Kyoko; Ichikawa, Tomio; Kumanishi, Toshiro; Chiba, Tomoki; Tanaka, Keiji; Nawa, Hiroyuki

    2003-08-01

    Ubiquitin (Ub) ligation is implicated in active protein metabolism and subcellular trafficking and its impairment is involved in various neurologic diseases. In rat brain, we identified two novel Ub ligases, Momo and Sakura, carrying double zinc finger motif and RING finger domain. Momo expression is enriched in the brain gray matter and testis, and Sakura expression is more widely detected in the brain white matter as well as in many peripheral organs. Both proteins associate with the cell membranes of neuronal and/or glial cells. We examined their Ub ligase activity in vivo and in vitro using viral expression vectors carrying myc-tagged Momo and Sakura. Overexpression of either Momo or Sakura in mixed cortical cultures increased total polyubiquitination levels. In vitro ubiquitination assay revealed that the combination of Momo and UbcH4 and H5c, or of Sakura and UbcH4, H5c and H6 is required for the reaction. Deletion mutagenesis suggested that the E3 Ub ligase activity of Momo and Sakura depended on their C-terminal domains containing RING finger structure, while their N-terminal domains influenced their membrane association. In agreement, Sakura associating with the membrane was specifically palmitoylated. Although the molecular targets of their Ub ligation remain to be identified, these findings imply a novel function of the palmitoylated E3 Ub ligase(s).

  9. Bacitracin zinc overdose

    Science.gov (United States)

    Bacitracin zinc is a medicine that is used on cuts and other skin wounds to help prevent infection. Bacitracin ... medicine that kills germs. Small amounts of bacitracin zinc are dissolved in petroleum jelly to create antibiotic ...

  10. Zinc oxide overdose

    Science.gov (United States)

    Zinc oxide is an ingredient in many products. Some of these are certain creams and ointments used ... prevent or treat minor skin burns and irritation. Zinc oxide overdose occurs when someone eats one of ...

  11. Error compensation during finger force production after oneand four-finger voluntarily fatiguing exercise

    OpenAIRE

    Kruger, Eric S.; Hoopes, Josh A.; Cordial, Rory J.; Li, Sheng

    2007-01-01

    The effect of muscle fatigue on error compensation strategies during multi-finger ramp force production tasks was investigated. Thirteen young, healthy subjects were instructed to produce a total force with four fingers of the right hand to accurately match a visually displayed template. The template consisted of a 3-s waiting period, a 3-s ramp force production (from 0 to 30% maximal voluntary contraction, MVC), and a 3-s constant force production. A series of twelve ramp trials was performe...

  12. Robot-assisted Guitar Hero for finger rehabilitation after stroke.

    Science.gov (United States)

    Taheri, Hossein; Rowe, Justin B; Gardner, David; Chan, Vicky; Reinkensmeyer, David J; Wolbrecht, Eric T

    2012-01-01

    This paper describes the design and testing of a robotic device for finger therapy after stroke: FINGER (Finger Individuating Grasp Exercise Robot). FINGER makes use of stacked single degree-of-freedom mechanisms to assist subjects in moving individual fingers in a naturalistic grasping pattern through much of their full range of motion. The device has a high bandwidth of control (-3dB at approximately 8 Hz) and is backdriveable. These characteristics make it capable of assisting in grasping tasks that require precise timing. We therefore used FINGER to assist individuals with a stroke (n= 8) and without impairment (n= 4) in playing a game similar to Guitar Hero©. The subjects attempted to move their fingers to target positions at times specified by notes that were graphically streamed to popular music. We show here that by automatically adjusting the robot gains, it is possible to use FINGER to modulate the subject's success rate at the game, across a range of impairment levels. Modulating success rates did not alter the stroke subject's effort, although the unimpaired subjects exerted more force when they were made less successful. We also present a novel measure of finger individuation that can be assessed as individuals play Guitar Hero with FINGER. The results demonstrate the ability of FINGER to provide controlled levels of assistance during an engaging computer game, and to quantify finger individuation after stroke. PMID:23366783

  13. Robot-assisted Guitar Hero for finger rehabilitation after stroke.

    Science.gov (United States)

    Taheri, Hossein; Rowe, Justin B; Gardner, David; Chan, Vicky; Reinkensmeyer, David J; Wolbrecht, Eric T

    2012-01-01

    This paper describes the design and testing of a robotic device for finger therapy after stroke: FINGER (Finger Individuating Grasp Exercise Robot). FINGER makes use of stacked single degree-of-freedom mechanisms to assist subjects in moving individual fingers in a naturalistic grasping pattern through much of their full range of motion. The device has a high bandwidth of control (-3dB at approximately 8 Hz) and is backdriveable. These characteristics make it capable of assisting in grasping tasks that require precise timing. We therefore used FINGER to assist individuals with a stroke (n= 8) and without impairment (n= 4) in playing a game similar to Guitar Hero©. The subjects attempted to move their fingers to target positions at times specified by notes that were graphically streamed to popular music. We show here that by automatically adjusting the robot gains, it is possible to use FINGER to modulate the subject's success rate at the game, across a range of impairment levels. Modulating success rates did not alter the stroke subject's effort, although the unimpaired subjects exerted more force when they were made less successful. We also present a novel measure of finger individuation that can be assessed as individuals play Guitar Hero with FINGER. The results demonstrate the ability of FINGER to provide controlled levels of assistance during an engaging computer game, and to quantify finger individuation after stroke.

  14. Zinc and gastrointestinal disease

    Institute of Scientific and Technical Information of China (English)

    Sonja; Skrovanek; Katherine; DiGuilio; Robert; Bailey; William; Huntington; Ryan; Urbas; Barani; Mayilvaganan; Giancarlo; Mercogliano; James; M; Mullin

    2014-01-01

    This review is a current summary of the role that both zinc deficiency and zinc supplementation can play in the etiology and therapy of a wide range of gastrointestinal diseases. The recent literature describing zinc action on gastrointestinal epithelial tight junctions and epithelial barrier function is described. Zinc enhancement of gastrointestinal epithelial barrier function may figure prominently in its potential therapeutic action in several gastrointestinal diseases.

  15. ZINC ABSORPTION BY INFANTS

    Science.gov (United States)

    Zinc is a vital mineral in human nutrition, and rare cases of overt zinc deficiency are well described in term and preterm infants. A variety of methods have been developed to assess zinc absorption, retention, and balance in humans, either using mass (metabolic) balance or stable isotope-based METH...

  16. Dynamic analysis of C/C composite finger seal

    Institute of Scientific and Technical Information of China (English)

    Chen Guoding; Wang Li’na; Yu Qiangpeng; Su Hua

    2014-01-01

    A seal device as an important component of aeroengines has decisive influence on per-formance, reliability, and working life of aeroengines. With the development of aeroengines, demands on the performance characteristics of seal devices are made strictly. Finger seal as a novel kind of sealing device, recently attracts more and more attentions in academic circles and engineer-ing fields at home and abroad. Research on finger seals has been extensively developed, especially on leakage and wear performances under dynamic conditions. However, it is a pity that the work on finger seals has been limited with a single approach that is improving the performance by structural optimization;in addition, the technology of dynamic analysis on finger seals is weak. Aiming at the problems mentioned above, a distributed mass equivalent dynamic model of finger seals considering the coupling effect of overlaid laminates is established in the present paper, the dynamic perfor-mance of 2.5 dimension C/C composite finger seal is analyzed with the model, and then the effects of fiber bundle density and fiber bundle preparation direction on finger seal’s dynamic performance are discussed, as well as compared with those of Co-based alloy finger seal. The current work is about dynamic analysis of finger seals and application of C/C composite in this paper may have much academic significance and many engineering values for improving research level of finger seal dynamics and exploring feasibility of C/C composite being used for finger seals.

  17. Dynamic analysis of C/C composite finger seal

    Directory of Open Access Journals (Sweden)

    Chen Guoding

    2014-06-01

    Full Text Available A seal device as an important component of aeroengines has decisive influence on performance, reliability, and working life of aeroengines. With the development of aeroengines, demands on the performance characteristics of seal devices are made strictly. Finger seal as a novel kind of sealing device, recently attracts more and more attentions in academic circles and engineering fields at home and abroad. Research on finger seals has been extensively developed, especially on leakage and wear performances under dynamic conditions. However, it is a pity that the work on finger seals has been limited with a single approach that is improving the performance by structural optimization; in addition, the technology of dynamic analysis on finger seals is weak. Aiming at the problems mentioned above, a distributed mass equivalent dynamic model of finger seals considering the coupling effect of overlaid laminates is established in the present paper, the dynamic performance of 2.5 dimension C/C composite finger seal is analyzed with the model, and then the effects of fiber bundle density and fiber bundle preparation direction on finger seal’s dynamic performance are discussed, as well as compared with those of Co-based alloy finger seal. The current work is about dynamic analysis of finger seals and application of C/C composite in this paper may have much academic significance and many engineering values for improving research level of finger seal dynamics and exploring feasibility of C/C composite being used for finger seals.

  18. [Zinc and gastrointestinal disorders].

    Science.gov (United States)

    Higashimura, Yasuki; Takagi, Tomohisa; Naito, Yuji

    2016-07-01

    Zinc, an essential trace element, affects immune responses, skin metabolism, hormone composition, and some sensory function, so that the deficiency presents various symptoms such as immunodeficiency and taste obstacle. Further, the zinc deficiency also considers as a risk of various diseases. Recent reports demonstrated that -20% of the Japanese population was marginally zinc deficiency, and over 25% of the global population is at high risk of zinc deficiency. In gastrointestinal disorders, zinc plays an important role in the healing of mucosal and epithelial damage. In fact, polaprezinc, a chelate compound of zinc and L-carnosine, has been used for the treatment of gastric ulcer and gastritis. We describe here the therapeutic effect of zinc on gastrointestinal disorders. PMID:27455800

  19. Nylon-muscle-actuated robotic finger

    Science.gov (United States)

    Wu, Lianjun; Jung de Andrade, Monica; Rome, Richard S.; Haines, Carter; Lima, Marcio D.; Baughman, Ray H.; Tadesse, Yonas

    2015-04-01

    This paper describes the design and experimental analysis of novel artificial muscles, made of twisted and coiled nylon fibers, for powering a biomimetic robotic hand. The design is based on circulating hot and cold water to actuate the artificial muscles and obtain fast finger movements. The actuation system consists of a spring and a coiled muscle within a compliant silicone tube. The silicone tube provides a watertight, expansible compartment within which the coiled muscle contracts when heated and expands when cooled. The fabrication and characterization of the actuating system are discussed in detail. The performance of the coiled muscle fiber in embedded conditions and the related characteristics of the actuated robotic finger are described.

  20. Angiolipoma of index finger: A case report

    Directory of Open Access Journals (Sweden)

    Muzaffer Durmus

    2016-04-01

    Full Text Available Angiolipomas are usually found in the upper extremities, shoulder and back. They are seldom found in the hands, face and lower extremities. They usually occur as painful soft tissue masses or they may compress the neighboring structures (e.g. nerves depending on the size and location. In this report we present an angiolipoma case located in the finger and discuss related recent cases described in the literature. [Hand Microsurg 2016; 5(1.000: 22-25

  1. Ferritin, finger clubbing, and lung disease.

    OpenAIRE

    Shneerson, J M; Jones, B M

    1981-01-01

    The serum ferritin concentration has been determined by an immunoradiometric assay in 90 subjects with a variety of pulmonary diseases. No association between ferritin concentrations and finger clubbing has been found in any of the diseases studied. Ferritin levels were significantly raised in the subjects with bronchial carcinoma, but were not useful in monitoring recurrence of the tumour. Pulmonary artery and pulmonary vein ferritin concentrations were similar to systemic venous concentrati...

  2. A Color Based Touchless Finger Mouse

    OpenAIRE

    Kah-Meng Kwong; Chin-Poo Lee; Kian-Ming Lim; Siew-Chin Chong; Siti Fatimah Abdul Razak

    2012-01-01

    People work with computers almost anytime, everywhere  in the current trend. However, continuously controlling a computer with mouse for a long time might cause much strains to people’s wrist. This work proposes a touchless finger mouse using webcam. A marker with different colours representing different actions is used. The webcam will capture the information on the marker and trigger the associated actions. This prototype is proven to be able to perform most of the actions a normal mouser c...

  3. Viscous fingering in volatile thin films

    OpenAIRE

    Agam, Oded

    2008-01-01

    A thin water film on a cleaved mica substrate undergoes a first order phase transition between two values of film thickness. By inducing a finite evaporation rate of the water, the interface between the two phases develops a fingering instability similar to that observed in the Saffman-Taylor problem. We draw the connection between the two problems, and construct solutions describing the dynamics of evaporation in this system.

  4. Angiolipoma of index finger: A case report

    OpenAIRE

    Muzaffer Durmus; Nuri Yigit; Abdul Kerim Yapici

    2016-01-01

    Angiolipomas are usually found in the upper extremities, shoulder and back. They are seldom found in the hands, face and lower extremities. They usually occur as painful soft tissue masses or they may compress the neighboring structures (e.g. nerves) depending on the size and location. In this report we present an angiolipoma case located in the finger and discuss related recent cases described in the literature. [Hand Microsurg 2016; 5(1.000): 22-25

  5. Saffman-Taylor fingers with kinetic undercooling

    CERN Document Server

    Gardiner, Bennett P J; Dallaston, Michael C; Moroney, Timothy J

    2015-01-01

    The mathematical model of a steadily propagating Saffman-Taylor finger in a Hele-Shaw channel has applications to two-dimensional interacting streamer discharges which are aligned in a periodic array. In the streamer context, the relevant regularisation on the interface is not provided by surface tension, but instead has been postulated to involve a mechanism equivalent to kinetic undercooling, which acts to penalise high velocities and prevent blow-up of the unregularised solution. Previous asymptotic results for the Hele-Shaw finger problem with kinetic undercooling suggest that for a given value of the kinetic undercooling parameter, there is a discrete set of possible finger shapes, each analytic at the nose and occupying a different fraction of the channel width. In the limit in which the kinetic undercooling parameter vanishes, the fraction for each family approaches 1/2, suggesting that this 'selection' of 1/2 by kinetic undercooling is qualitatively similar to the well-known analogue with surface tens...

  6. Pacifier Use, Finger Sucking, and Infant Sleep.

    Science.gov (United States)

    Butler, Rachel; Moore, Melisa; Mindell, Jodi A

    2016-01-01

    Few studies to date have investigated the relationship between pacifier use or finger sucking and infant sleep. One hundred and four mothers of infants (ages 0-11 months) completed the Brief Infant Sleep Questionnaire (BISQ). Infants who engaged in finger sucking had fewer night wakings and longer stretches of nighttime sleep, although less daytime sleep. There were no significant differences in sleep patterns between pacifier users and infants who did not engage in nonnutritive sucking. Furthermore, no significant differences were found across groups for sleep ecology, including parental involvement at bedtime and following night wakings. Finally, infants were consistently able to retrieve their pacifiers independently by 7 months of age, although this did not appear to be associated with sleep outcomes. Results suggest that when parents are deciding whether to give their infant a pacifier, sleep may not be a critical factor. In contrast, parents of finger and thumb suckers should be reassured that this nonnutritive sucking is beneficial to sleep, at least in the first year of life. PMID:26548755

  7. Visual Foraging With Fingers and Eye Gaze.

    Science.gov (United States)

    Jóhannesson, Ómar I; Thornton, Ian M; Smith, Irene J; Chetverikov, Andrey; Kristjánsson, Árni

    2016-03-01

    A popular model of the function of selective visual attention involves search where a single target is to be found among distractors. For many scenarios, a more realistic model involves search for multiple targets of various types, since natural tasks typically do not involve a single target. Here we present results from a novel multiple-target foraging paradigm. We compare finger foraging where observers cancel a set of predesignated targets by tapping them, to gaze foraging where observers cancel items by fixating them for 100 ms. During finger foraging, for most observers, there was a large difference between foraging based on a single feature, where observers switch easily between target types, and foraging based on a conjunction of features where observers tended to stick to one target type. The pattern was notably different during gaze foraging where these condition differences were smaller. Two conclusions follow: (a) The fact that a sizeable number of observers (in particular during gaze foraging) had little trouble switching between different target types raises challenges for many prominent theoretical accounts of visual attention and working memory. (b) While caveats must be noted for the comparison of gaze and finger foraging, the results suggest that selection mechanisms for gaze and pointing have different operational constraints. PMID:27433323

  8. Nano structured porous anodized aluminium oxide by using C2H2O4 for electronic applications: Study of the cell potential effects on formation of porous alumina

    International Nuclear Information System (INIS)

    In this research, a nano porous anodized aluminium oxide AAO thin film was successfully grown onto oxide layer on silicon substrate. The anodization of Si/ SiO2/ Al substrate was conducted in a vigorous stirring oxalic acid bath solution. The rate of growth, morphology and also the kinetic study of the AAO thin film were investigated. The resulting array, pores structure and pores density of AAO strongly depends on an applied voltage of the anodizing process. (author)

  9. A Study on the Effects of Carrier Gases on the Structure and Morphology of Carbon Nanotubes Prepared by Pyrolysis of Ferrocene and C2H2 Mixture

    Institute of Scientific and Technical Information of China (English)

    Wanliang Mi; Jerry Yuesheng Lin; Qian Mao; Yongdan Li; Baoquan Zhang

    2005-01-01

    Carbon nanotubes (CNTs) were prepared using different carrier gases,with ferrocene as the catalyst precusor and acetylene as the carbon source. The effects of ammonia and nitrogen as carrier gases on the structure and morphology of CNTs were investigated. Transmission electron microscope (TEM),high-resolution electron microscope (HRTEM),scanning electron microscope (SEM) and X-ray diffraction (XRD) were employed to characterize the products and the catalyst. Experiment results show that the CNTs grown in N2 gas exhibited cylindrical and tubular structure,while a bamboo-like structure was observed for the CNTs grown in NH3 gas. Moreover,vertically aligned CNTs were obtained on an Al2O3 disk when NH3 was used as the carrier gas. The carrier gas also exerted influence on the shape of the catalyst. Based on the theory of active centers of catalysis and combined with the particle shape of the catalyst,a growth model for the vertically aligned CNTs on the substrate is given.

  10. The influence of C2H2 and dust formation on the time dependence of metastable argon density in pulsed plasmas

    OpenAIRE

    Stefanovic, Ilija; Sadeghi, Nader; Winter, Jörg

    2010-01-01

    Abstract Diode laser absorption at 772.38 nm is used to measure the time resolved density of Ar*(3 P 2) metastable atoms in a capacitively coupled radio-frequency (RF) discharge running in argon/acetylene mixture at 0.1 mbar. The RF power is pulsed at 100 Hz and the density of Ar*(3 P 2) atoms in the 5 ms ON time and in the afterglow are recorded. Different plasma conditions, namely: 1) pure argon, 2) argon + 7% acetylene before powder formation, 3) argon + 7% acetylene after dust particle...

  11. A philosophical theory on human communication and modern physics: e(,2)c(,2)H('2)T energy-exchange and consciousness-change toward humanism, healing, and transformation

    Science.gov (United States)

    Jenkins-Tate, Marnishia Laverne

    This dissertation addresses the need for a body of human communication theory that can be useful toward advancing personal and social transformation. Of the humanistic genre, it suggests that there is a need to promote humanism, healing, and personal transformation in the non-clinical settings of everyday living. Three questions guide the effort. First, it asks: what kind of human communication theory might describe some of the underlying dynamics of human interaction, while also suggesting ways to improve the quality of interactions of any related philosophical theory be grounded by some scientific discipline? Then finally, it asks: how might these proposed concepts be captured in a manner that can be useful to human beings in everyday human interaction? Extending the work of modern physics to the realm of human communication, the theory integrates conceptual aspects of quantum theory, relativity theory, communication accommodation theory, and various nonverbal communication theory. Then, it proposes the philosophical framework for a new body of theory which it calls the energy-exchange theory of human communication. Treating human beings as living forms of matter, it suggests that ``energy'' is the life-force that sustains all human beings, and that ``consciousness'' is that qualitative level of development at which energy manifests itself in the human experience. It proposes that human beings have the capacity to exchange energy and influence consciousness during the human communication process, and that these interactions can advance humanism, healing, and transformation-which it proposes are the higher states and levels of human consciousness. Thus, this research effort sought to know and to describe a phenomenon that is the interactive human being; and to suggest useful ways that this volitional being can know and transform itself through human interaction. With verisimilitude as a driving factor in describing human beings as communicators, the research is ontology- centered. It suggests that human beings are most notably creatures of feeling and soul, and that it is through interaction involving these dimensions that one can best come to know the human being. Accordingly, the research employs a hermeneutic phenomenological approach toward participative inquiry and experiential knowing. With practicality as a driving factor in suggesting ways that the human phenomenon can know and transform itself, this work utilizes a pragmatic approach to theory- building. Pragmatism suggests that all theories are approximations that ultimately should be judged on their abilities to truthfully describe phenomena and to solve human problems. Thus, having undergone an inductive progression from data to theory-building, now this work must go from theory to data, such that next steps should involve formal assessment of its verisimilitude and pragmatism based on feedback gained through field research on various persons who may apply the theory and its model to everyday living. (Abstract shortened by UMI.)

  12. Is the Reaction of C3N(-) with C2H2 a Possible Process for Chain Elongation in Titan's Ionosphere?

    Science.gov (United States)

    Lindén, Fredrik; Alcaraz, Christian; Ascenzi, Daniela; Guillemin, Jean-Claude; Koch, Leopold; Lopes, Allan; Polášek, Miroslav; Romanzin, Claire; Žabka, Jan; Zymak, Illia; Geppert, Wolf D

    2016-07-14

    The reaction of C3N(-) with acetylene was studied using three different experimental setups, a triple quadrupole mass spectrometer (Trento), a tandem quadrupole mass spectrometer (Prague), and the "CERISES" guided ion beam apparatus at Orsay. The process is of astrophysical interest because it can function as a chain elongation mechanism to produce larger anions that have been detected in Titan's ionosphere by the Cassini Plasma Spectrometer. Three major products of primary processes, C2H(-), CN(-), and C5N(-), have been identified, whereby the production of the cyanide anion is probably partly due to collisional induced dissociation. The formations of all these products show considerable reaction thresholds and also display comparatively small cross sections. Also, no strong signals of anionic products for collision energies lower than 1 eV have been observed. Ab initio calculations have been performed to identify possible pathways leading to the observed products of the title reaction and to elucidate the thermodynamics of these processes. Although the productions of CN(-) and C5N(-) are exoergic, all reaction pathways have considerable barriers. Overall, the results of these computations are in agreement with the observed reaction thresholds. Due to the existence of considerable reaction energy barriers and the small observed cross sections, the title reaction is not very likely to play a major role in the buildup of large anions in cold environments like the interstellar medium or planetary and satellite ionospheres. PMID:27135984

  13. Small-scale behavior of single gravity-driven fingers in an initially dry fracture

    Energy Technology Data Exchange (ETDEWEB)

    Nicholl, M.J.; Glass, R.J.; Nguyen, H.A.

    1992-12-31

    Experiments investigating the behavior of individual, gravity-driven fingers in an initially dry, rough-walled analog fracture are presented. Fingers were initiated from constant flow to a point source. Finger structure is described in detail; specific phenomena observed include: desaturation behind the finger-tip, variation in finger path, intermittent flow structures, finger-tip bifurcation, and formation of dendritic sub-fingers. Measurements were made of finger-tip velocity, finger width, and finger-tip length. Non-dimensional forms of the measured variables are analyzed relative to the independent parameters, flow rate and gravitational gradient.

  14. Small-scale behavior of single gravity-driven fingers in an initially dry fracture

    International Nuclear Information System (INIS)

    Experiments investigating the behavior of individual, gravity-driven fingers in an initially dry, rough-walled analog fracture are presented. Fingers were initiated from constant flow to a point source. Finger structure is described in detail; specific phenomena observed include: desaturation behind the finger-tip, variation in finger path, intermittent flow structures, finger-tip bifurcation, and formation of dendritic sub-fingers. Measurements were made of finger-tip velocity, finger width, and finger-tip length. Non-dimensional forms of the measured variables are analyzed relative to the independent parameters, flow rate and gravitational gradient

  15. Viscous fingering with partially miscible fluids

    Science.gov (United States)

    Fu, X.; Cueto-Felgueroso, L.; Juanes, R.

    2015-12-01

    When a less viscous fluid displaces a more viscous fluid, the contrast in viscosity destabilizes the interface between the two fluids, leading to the formation of fingers. Experimental and numerical studies of viscous fingering have focused on fluids that are either fully miscible (e.g. water and glycerol) or perfectly immiscible (e.g. water and oil). In practice, however, the miscibility of two fluids can change appreciably with temperature and pressure, and often falls into the case of partial miscibility, where two fluids have limited solubility in each other (e.g. CO2 and water). Following our recent work for miscible systems (Jha et al., PRL 2011, 2013) and immiscible systems (Cueto-Felgueroso and Juanes, PRL 2012, JFM 2014), here we propose a phase-field model for fluid-fluid displacements in a porous medium, when the two fluids have limited (but nonzero) solubility in one another. In our model, partial miscibility is characterized through the design of the thermodynamic free energy of the two-fluid system. We express the model in dimensionless form and elucidate the key dimensionless groups that control the behavior of the system. We present high-resolution numerical simulations of the model applied to the viscous fingering problem. On one hand, we demonstrate the effect of partial miscibility on the hydrodynamic instability. On the other, we elucidate the role of the degree of fingering on the rate of mutual fluid dissolution. Figure caption: final snapshots in simulations of viscous fingering with a two-fluid system mimicking that of CO2 and water. The colormap corresponds to the concentration of CO2. A band of less viscous gas phase rich in CO2 (red) displaces through the more viscous liquid phase that is undersaturated with CO2 (blue). At the fluid interface, an exchange of CO2 occurs as a result of local chemical potentials that drives the system towards thermodynamic equilibrium. This results in a shrinkage of gas phase as well as a local increase in

  16. The role of fingers in number processing in young children.

    Science.gov (United States)

    Lafay, Anne; Thevenot, Catherine; Castel, Caroline; Fayol, Michel

    2013-01-01

    The aim of the present study was to investigate the relationship between finger counting and numerical processing in 4-7-year-old children. Children were assessed on a variety of numerical tasks and we examined the correlations between their rates of success and their frequency of finger use in a counting task. We showed that children's performance on finger pattern comparison and identification tasks did not correlate with the frequency of finger use. However, this last variable correlated with the percentages of correct responses in an enumeration task (i.e., Give-N task), even when the age of children was entered as a covariate in the analysis. Despite this correlation, we showed that some children who never used their fingers in the counting task were able to perform optimally in the enumeration task. Overall, our results support the conclusion that finger counting is useful but not necessary to develop accurate symbolic numerical skills. Moreover, our results suggest that the use of fingers in a counting task is related to the ability of children in a dynamic enumeration task but not to static tasks involving recognition or comparison of finger patterns. Therefore, it could be that the link between fingers and numbers remain circumscribed to counting tasks and do not extent to static finger montring situations. PMID:23908643

  17. The role of fingers in number processing in young children

    Science.gov (United States)

    Lafay, Anne; Thevenot, Catherine; Castel, Caroline; Fayol, Michel

    2013-01-01

    The aim of the present study was to investigate the relationship between finger counting and numerical processing in 4–7-year-old children. Children were assessed on a variety of numerical tasks and we examined the correlations between their rates of success and their frequency of finger use in a counting task. We showed that children's performance on finger pattern comparison and identification tasks did not correlate with the frequency of finger use. However, this last variable correlated with the percentages of correct responses in an enumeration task (i.e., Give-N task), even when the age of children was entered as a covariate in the analysis. Despite this correlation, we showed that some children who never used their fingers in the counting task were able to perform optimally in the enumeration task. Overall, our results support the conclusion that finger counting is useful but not necessary to develop accurate symbolic numerical skills. Moreover, our results suggest that the use of fingers in a counting task is related to the ability of children in a dynamic enumeration task but not to static tasks involving recognition or comparison of finger patterns. Therefore, it could be that the link between fingers and numbers remain circumscribed to counting tasks and do not extent to static finger montring situations. PMID:23908643

  18. The role of fingers in number processing in young children

    Directory of Open Access Journals (Sweden)

    Anne eLafay

    2013-07-01

    Full Text Available The aim of the present study was to investigate the relationship between finger counting and numerical processing in 4- to 7-year-old children. Children were assessed on a variety of numerical tasks and we examined the correlations between their rates of success and their frequency of finger use in a counting task. We showed that children’s performance on finger pattern comparison and identification tasks did not correlate with the frequency of finger use. However, this last variable correlated with the percentages of correct responses in an enumeration task (i.e., Give-N task, even when the age of children was entered as a covariate in the analysis. Despite this correlation, we showed that some children who never used their fingers in the counting task were able to perform optimally in the enumeration task. Overall, our results support the conclusion that finger counting is useful but not necessary to develop accurate symbolic numerical skills. Moreover, our results suggest that the use of fingers in a counting task is related to the ability of children in a dynamic enumeration task but not to static tasks involving recognition or comparison of finger patterns. Therefore, it could be that the link between fingers and numbers remain circumscribed to counting tasks and do not extent to static finger montring situations.

  19. VISIONS FOR FOOTWEAR TIP SHAPE ACCORDING TO THE CONFIGURATION FINGER

    Directory of Open Access Journals (Sweden)

    MALCOCI Marina

    2015-05-01

    Full Text Available Compatibility between the consumer and the interior leg permanent footwear raises a number of issues. And any new form of footwear is time for a new silhouette last. Fashion is a factor in determining the shape of the last significant role. The most important influence on fashion in footwear that has at one time is found in peak shape. During registered a variety of forms leading to the last, for example, pointed, oval, round, square, asymmetrical, curved, trapezoidal, etc. Each has added a tip top recommended. The paper analyzes the morphofunctional characteristic, namely, finger configuration. The configuration of the fingers is determined from the positions of all the fingers of one another, as are six variants. Analysis of the shape and configuration of the arm fingers allow us to make the following recommendations to consumers: people showing finger configuration as in variant V and VI are advised not to wear pointy shoes because of the limited movement of the foot, which favors the diversion finger I exterior and deformed finger V; persons who fall within I-IV variant can procure pointy shoes; a round-tipped shoes, square, curved or asymmetric may be purchased by any consumer regardless of the configuration of the fingers; shoes with cut edge must be present only in garderopa people in variant I and II; consumers whose configuration is like finger-VI and III variants are awkwardly shaped fingers can buy shoes closed in the previous summer, but of different perforations or overlapping strips.

  20. C3HC4-type RING finger protein NbZFP1 is involved in growth and fruit development in Nicotiana benthamiana.

    Directory of Open Access Journals (Sweden)

    Wenxian Wu

    Full Text Available C3HC4-type RING finger proteins constitute a large family in the plant kingdom and play important roles in various physiological processes of plant life. In this study, a C3HC4-type zinc finger gene was isolated from Nicotiana benthamiana. Sequence analysis indicated that the gene encodes a 24-kDa protein with 191 amino acids containing one typical C3HC4-type zinc finger domain; this gene was named NbZFP1. Transient expression of pGDG-NbZFP1 demonstrated that NbZFP1 was localized to the chloroplast, especially in the chloroplasts of cells surrounding leaf stomata. Virus-induced gene silencing (VIGS analysis indicated that silencing of NbZFP1 hampered fruit development, although the height of the plants was normal. An overexpression construct was then designed and transferred into Nicotiana benthamiana, and PCR and Southern blot showed that the NbZFP1 gene was successfully integrated into the Nicotiana benthamiana genome. The transgenic lines showed typical compactness, with a short internode length and sturdy stems. This is the first report describing the function of a C3HC4-type RING finger protein in tobacco.

  1. Improved zinc electrode and rechargeable zinc-air battery

    Science.gov (United States)

    Ross, P.N. Jr.

    1988-06-21

    The invention comprises an improved rechargeable zinc-air cell/battery having recirculating alkaline electrolyte and a zinc electrode comprising a porous foam support material which carries the active zinc electrode material. 5 figs.

  2. Mutational analysis of a predicted zinc-binding motif in the 26-kilodalton protein encoded by the vaccinia virus A2L gene: correlation of zinc binding with late transcriptional transactivation activity.

    OpenAIRE

    Keck, J G; Feigenbaum, F; B. Moss

    1993-01-01

    Transient transfection assays indicated that A2L is one of three vaccinia virus intermediate genes that are required for the transcriptional transactivation of viral late genes. We have expressed the A2L open reading frame in Escherichia coli and shown by blotting experiments that the 26-kDa protein binds zinc, a property predicted by the presence of a CX2CX13CX2C zinc finger motif. The specificity for zinc binding was demonstrated by competition with other metals. The role of the sequence mo...

  3. Experiments of periodic forcing of Saffman-Taylor fingers

    Science.gov (United States)

    Torralba, M.; Ortín, J.; Hernández-Machado, A.; Poiré, E. Corvera

    2008-03-01

    We report on an experimental study of long normal Saffman-Taylor fingers subject to periodic forcing. The sides of the finger develop a low amplitude, long wavelength instability. We discuss the finger response in stationary and nonstationary situations, as well as the dynamics towards the stationary states. The response frequency of the instability increases with forcing frequency at low forcing frequencies, while, remarkably, it becomes independent of forcing frequency at large forcing frequencies. This implies a process of wavelength selection. These observations are in good agreement with previous numerical results reported in [Ledesma-Aguilar , Phys. Rev. E 71, 016312 (2005)]. We also study the average value of the finger width, and its fluctuations, as a function of forcing frequency. The average finger width is always smaller than the width of the steady-state finger. Fluctuations have a nonmonotonic behavior with a maximum at a particular frequency.

  4. Fingering instabilities in Newtonian and non-Newtonian fluids

    Science.gov (United States)

    Kennedy, Kristi E.

    Fingering has been studied in different fluid systems. Viscous fingering, which is driven by a difference in viscosity between fluids, has been studied by both experiments and numerical simulations. We used a single fluid with a temperature-dependent viscosity and studied the instability for a range of inlet pressures and viscosity ratios. The spreading and fingering of a fluid drop subjected to a centrifugal force, known as spin coating, has also been studied for a range of drop volumes and rotation speeds, both for a Newtonian and a non-Newtonian fluid. Experiments on viscous fingering with a single fluid, glycerine, show that an instability occurs at the boundary separating hot and cold fluid. The results indicate that the instability is similar to that which occurs between two miscible fluids. Fingering only occurs for high enough values of the inlet pressure and viscosity ratio. The wavelength of the fingering pattern is found to be proportional to the cell width for the two smallest cell widths used. The fingering patterns seen in the simulations are very similar to the experimental patterns, although there are some quantitative differences. In particular, the wavelength of the instability is seen to depend only weakly on the cell width. The spreading of silicone oil, a Newtonian fluid, during spin coating follows the time dependence predicted theoretically, although with a shift in the scaled time variable. Once the radius of the spreading silicone oil drop becomes large enough, fingers form around the perimeter of the drop for all experimental conditions studied. The number of fingers and the growth rate of the fingers are in agreement with theoretical predictions. Fingers are also observed to form for high enough drop volumes and rotation speeds during the spinning of a non-Newtonian fluid drop, Carbopol, which possesses a yield stress. In this case the fingering is a localized effect, occuring once the stress on the drop exceeds the yield stress, rather

  5. Effective Length Design of Humanoid Robot Fingers Using Biomimetic Optimization

    OpenAIRE

    Byoung-Ho Kim

    2015-01-01

    In this study, we propose an effective design method for the phalangeal parameters and the total size of humanoid robot fingers based on a biomimetic optimization. For the optimization, an interphalangeal joint coordination parameter and the length constraints inherent in human fingers are considered from a biomimetic perspective. A reasonable grasp formulation is also taken into account from the viewpoint of power grasping, where the grasp space of a humanoid robot finger is importantly cons...

  6. Forced ventilation increases variability of isometric finger forces

    OpenAIRE

    Li, Sheng; Yasuda, Nobuo

    2006-01-01

    The study aimed to assess the effects of forced ventilation on variability of the index finger force at the submaximal levels. Fourteen (6 men, 8 women) healthy subjects were instructed to perform self-initiated forced inspiration and forced expiration, the Valsalva maneuver and normal breathing while sustaining 15%, 30%, and 45% of maximal voluntary contraction (MVC) by the index finger. Standard deviation (S.D.) of finger forces increased significantly with the level of force. At each level...

  7. Finteraction : Finger Interaction with Mobile Phone

    OpenAIRE

    Jenabi, Mahsa; Reiterer, Harald

    2008-01-01

    Touch interaction with mobile phones enable users to have a more natural interaction with the device, since touch is a natural way of direct accessing an ob ject of interest. But one disadvantage is occlusion; i.e., the user loses a high percentage of information presented on the small screen of the mobile phone during the interaction using his big finger . On the other hand, interaction enhanced with accelerometers can react according to the device movements, e.g., while the user is tilting...

  8. A Color Based Touchless Finger Mouse

    Directory of Open Access Journals (Sweden)

    Kah-Meng Kwong

    2012-01-01

    Full Text Available People work with computers almost anytime, everywhere  in the current trend. However, continuously controlling a computer with mouse for a long time might cause much strains to people’s wrist. This work proposes a touchless finger mouse using webcam. A marker with different colours representing different actions is used. The webcam will capture the information on the marker and trigger the associated actions. This prototype is proven to be able to perform most of the actions a normal mouser can perform.

  9. Chelators for investigating zinc metalloneurochemistry.

    Science.gov (United States)

    Radford, Robert J; Lippard, Stephen J

    2013-04-01

    The physiology and pathology of mobile zinc signaling has become an important topic in metalloneurochemistry. To study the action of mobile zinc effectively, specialized tools are required that probe the temporal and positional changes of zinc ions within live tissue and cells. In the present article we describe the design and implementation of selective zinc chelators as antagonists to interrogate the function of mobile zinc, with an emphasis on the pools of vesicular zinc in the terminals of hippocampal mossy fiber buttons.

  10. Recovery of zinc and manganese, and other metals (Fe, Cu, Ni, Co, Cd, Cr, Na, K) from Zn-MnO2 and Zn-C waste batteries: Hydroxyl and carbonate co-precipitation from solution after reducing acidic leaching with use of oxalic acid

    Science.gov (United States)

    Sobianowska-Turek, A.; Szczepaniak, W.; Maciejewski, P.; Gawlik-Kobylińska, M.

    2016-09-01

    The article discusses the current situation of the spent batteries and portable accumulators management. It reviews recycling technologies of the spent batteries and portable accumulators which are used in the manufacturing installations in the world. Also, it presents the authors' research results on the reductive acidic leaching of waste material of the zinc-carbon batteries (Zn-C) and zinc-manganese batteries (alkaline Zn-MnO2) delivered by a company dealing with mechanical treatment of this type of waste stream. The research data proved that the reductive acidic leaching (H2SO4 + C2H2O4) of the battery's black mass allows to recover 85.0% of zinc and 100% of manganese. Moreover, it was found that after the reductive acidic leaching it is possible to recover nearly 100% of manganese, iron, cadmium, and chromium, 98.0% of cobalt, 95.5% of zinc, and 85.0% of copper and nickel from the solution with carbonate method. On the basis of the results, it is possible to assume that the carbonate method can be used for the preparation of manganese-zinc ferrite.

  11. Effect of Finger Joint on Flexural Strength of Teak Wood

    Directory of Open Access Journals (Sweden)

    Bharatesh A. Danawade

    2014-01-01

    Full Text Available This paper presents the flexural properties of rectangular Burma teak wood beam without finger joint and with finger joint. Finger joints enable full utilization of wood. Finger jointing technique is also used to eliminate wood defects which weaken the strength of wood. This paper considers finger joint as defined defect and its effect on the flexural strength is determined. Teakwood is hard and heavy, seasons rapidly and has good durability. The specimens were studied under three point bending test. Both edge wise and flat wise tests were carried out. It is observed that Burma teakwood beam without finger joint is stronger than beams with finger joints. Because of finger jointing the flexural strength reduces. It can be concluded that the strength loss can improved upon by selecting suitable geometry of finger joint and a suitable adhesive. It is recognized that further studies are necessary on jointing techniques of wood and type of adhesive so as to equal the flexural strength properties of clear teak wood beams.

  12. Spatial Circular Granulation Method Based on Multimodal Finger Feature

    Directory of Open Access Journals (Sweden)

    Jinfeng Yang

    2016-01-01

    Full Text Available Finger-based personal identification has become an active research topic in recent years because of its high user acceptance and convenience. How to reliably and effectively fuse the multimodal finger features together, however, has still been a challenging problem in practice. In this paper, viewing the finger trait as the combination of a fingerprint, finger vein, and finger-knuckle-print, a new multimodal finger feature recognition scheme is proposed based on granular computing. First, the ridge texture features of FP, FV, and FKP are extracted using Gabor Ordinal Measures (GOM. Second, combining the three-modal GOM feature maps in a color-based manner, we then constitute the original feature object set of a finger. To represent finger features effectively, they are granulated at three levels of feature granules (FGs in a bottom-up manner based on spatial circular granulation. In order to test the performance of the multilevel FGs, a top-down matching method is proposed. Experimental results show that the proposed method achieves higher accuracy recognition rate in finger feature recognition.

  13. The design and development of a finger joint simulator.

    Science.gov (United States)

    Joyce, Thomas J

    2016-05-01

    Artificial finger joints lack the long-term clinical success seen with hip and knee prostheses. In part, this can be explained by the challenges of rheumatoid arthritis, a progressive disease which attacks surrounding tissues as well as the joint itself. Therefore, the natural finger joints' biomechanics are adversely affected, and consequently, this imbalance due to subluxing forces further challenges any prosthesis. Many different designs of finger prosthesis have been offered over a period of greater than 50 years. Most of these designs have failed, and it is likely that many of these failures could have been identified had the prostheses been appropriately tested prior to implantation into patients. While finger joint simulators have been designed, arguably only those from a single centre have been able to reproduce clinical-type failures of the finger prostheses tested in them. This article describes the design and development of a finger simulator at Durham University, UK. It explains and justifies the engineering decisions made and thus the evolution of the finger simulator. In vitro results and their linkage to clinical-type failures are outlined to help to show the effectiveness of the simulator. Failures of finger implants in vivo continue to occur, and the need for appropriate in vitro testing of finger prostheses remains strong. PMID:26833697

  14. Acute finger injuries: part I. Tendons and ligaments.

    Science.gov (United States)

    Leggit, Jeffrey C; Meko, Christian J

    2006-03-01

    Improper diagnosis and treatment of finger injuries can cause deformity and dysfunction over time. A basic understanding of the complex anatomy of the finger and of common tendon and ligament injury mechanisms can help physicians properly diagnose and treat finger injuries. Evaluation includes a general musculoskeletal examination as well as radiography (oblique, anteroposterior, and true lateral views). Splinting and taping are effective treatments for tendon and ligament injuries. Treatment should restrict the motion of injured structures while allowing uninjured joints to remain mobile. Although family physicians are usually the first to evaluate patients with finger injuries, it is important to recognize when a referral is needed to ensure optimal outcomes.

  15. Extrinsic versus intrinsic hand muscle dominance in finger flexion.

    Science.gov (United States)

    Al-Sukaini, A; Singh, H P; Dias, J J

    2016-05-01

    This study aims to identify the patterns of dominance of extrinsic or intrinsic muscles in finger flexion during initiation of finger curl and mid-finger flexion. We recorded 82 hands of healthy individuals (18-74 years) while flexing their fingers and tracked the finger joint angles of the little finger using video motion tracking. A total of 57 hands (69.5%) were classified as extrinsic dominant, where the finger flexion was initiated and maintained at proximal interphalangeal and distal interphalangeal joints. A total of 25 (30.5%) were classified as intrinsic dominant, where the finger flexion was initiated and maintained at the metacarpophalangeal joint. The distribution of age, sex, dominance, handedness and body mass index was similar in the two groups. This knowledge may allow clinicians to develop more efficient rehabilitation regimes, since intrinsic dominant individuals would not initiate extrinsic muscle contraction till later in finger flexion, and might therefore be allowed limited early active motion. For extrinsic dominant individuals, by contrast, initial contraction of extrinsic muscles would place increased stress on the tendon repair site if early motion were permitted. PMID:26744509

  16. The design and development of a finger joint simulator.

    Science.gov (United States)

    Joyce, Thomas J

    2016-05-01

    Artificial finger joints lack the long-term clinical success seen with hip and knee prostheses. In part, this can be explained by the challenges of rheumatoid arthritis, a progressive disease which attacks surrounding tissues as well as the joint itself. Therefore, the natural finger joints' biomechanics are adversely affected, and consequently, this imbalance due to subluxing forces further challenges any prosthesis. Many different designs of finger prosthesis have been offered over a period of greater than 50 years. Most of these designs have failed, and it is likely that many of these failures could have been identified had the prostheses been appropriately tested prior to implantation into patients. While finger joint simulators have been designed, arguably only those from a single centre have been able to reproduce clinical-type failures of the finger prostheses tested in them. This article describes the design and development of a finger simulator at Durham University, UK. It explains and justifies the engineering decisions made and thus the evolution of the finger simulator. In vitro results and their linkage to clinical-type failures are outlined to help to show the effectiveness of the simulator. Failures of finger implants in vivo continue to occur, and the need for appropriate in vitro testing of finger prostheses remains strong.

  17. Zinc deficiency and eating disorders.

    Science.gov (United States)

    Humphries, L; Vivian, B; Stuart, M; McClain, C J

    1989-12-01

    Decreased food intake, a cyclic pattern of eating, and weight loss are major manifestations of zinc deficiency. In this study, zinc status was evaluated in 62 patients with bulimia and 24 patients with anorexia nervosa. Forty percent of patients with bulimia and 54% of those with anorexia nervosa had biochemical evidence of zinc deficiency. The authors suggest that for a variety of reasons, such as lower dietary intake of zinc, impaired zinc absorption, vomiting, diarrhea, and binging on low-zinc foods, patients with eating disorders may develop zinc deficiency. This acquired zinc deficiency could then add to the chronicity of altered eating behavior in those patients. PMID:2600063

  18. Lipid Gymnastics: Tethers and Fingers in membrane

    Science.gov (United States)

    Tayebi, Lobat; Miller, Gregory; Parikh, Atul

    2009-03-01

    A significant body of evidence now links local mesoscopic structure (e.g., shape and composition) of the cell membrane with its function; the mechanisms by which cellular membranes adopt the specific shapes remain poorly understood. Among all the different structures adopted by cellular membranes, the tubular shape is one of the most surprising one. While their formation is typically attributed to the reorganization of membrane cytoskeleton, many exceptions exist. We report the instantaneous formation of tubular membrane mesophases following the hydration under specific thermal conditions. The shapes emerge in a bimodal way where we have two distinct diameter ranges for tubes, ˜20μm and ˜1μm, namely fat fingers and narrow tethers. We study the roughening of hydrated drops of 3 lipids in 3 different spontaneous curvatures at various temp. and ionic strength to figure out the dominant effect in selection of tethers and fingers. Dynamics of the tubes are of particular interest where we observe four distinct steps of birth, coiling, uncoiling and retraction with different lifetime on different thermal condition. These dynamics appear to reflect interplay between membrane elasticity, surface adhesion, and thermal or hydrodynamic gradient.

  19. Teleoperation of Robonaut Using Finger Tracking

    Science.gov (United States)

    Champoux, Rachel G.; Luo, Victor

    2012-01-01

    With the advent of new finger tracking systems, the idea of a more expressive and intuitive user interface is being explored and implemented. One practical application for this new kind of interface is that of teleoperating a robot. For humanoid robots, a finger tracking interface is required due to the level of complexity in a human-like hand, where a joystick isn't accurate. Moreover, for some tasks, using one's own hands allows the user to communicate their intentions more effectively than other input. The purpose of this project was to develop a natural user interface for someone to teleoperate a robot that is elsewhere. Specifically, this was designed to control Robonaut on the international space station to do tasks too dangerous and/or too trivial for human astronauts. This interface was developed by integrating and modifying 3Gear's software, which includes a library of gestures and the ability to track hands. The end result is an interface in which the user can manipulate objects in real time in the user interface. then, the information is relayed to a simulator, the stand in for Robonaut, at a slight delay.

  20. Finger-like voids induced by viscous fingering during phase inversion of alumina/PES/NMP suspensions

    KAUST Repository

    Wang, Bo

    2012-07-01

    The formation mechanism of phase-inversion ceramic hollow fibre membranes has not been well understood. In this paper, we report on the formation of finger-like macrovoids during non-solvent-induced phase inversion of alumina/PES/NMP suspensions. A membrane structure without such finger-like macrovoids was observed when the suspension was slowly immersed into pure ethanol or a mixture of 70. wt% NMP and 30. wt% water, whereas finger-like macrovoids occurred when the suspension was slid into the non-solvents at higher speeds. We found that the formation process of finger-like macrovoids could be fully or partially reversed when nascent membranes were taken out from water shortly after immersion, depending on the duration of the immersion. Splitting of the fingers during the formation of the macrovoids was also observed during the phase inversion of two alumina/PES/NMP suspensions. These experimental observations were not predicted by current theories of finger-like macrovoid formation in polymer membranes, but appear to mimic the well-known viscous fingering phenomenon. We therefore propose that in the phase inversion of ceramic suspensions, the viscous fingering phenomenon is an important mechanism in the formation of finger-like voids. © 2012 Elsevier B.V.

  1. Analysis of zinc borates

    International Nuclear Information System (INIS)

    Methods for analyzing zinc borates: B2O3 determination in the presence of zinc ions and determination of zinc in the presence of borates are developed. Distributing effect of zinc in alcalometrical determination of B2O3 is removed using either its binding cationite KU-2 in H-form in hydrochloric acid medium or using complexone 3 masking. In the first case the results are underestimated, in the second one - are overestimated. When analyzing Zn the complexonometrical titration with sodium teraborate is carried out. Borate ions don't affect the accuracy of determination. Zinc borate samples of 0.1-0.15 g in dimesion are recommended according to the method suggested

  2. Finger curvature movement recognition interface technique using SEMG signals

    Directory of Open Access Journals (Sweden)

    Y. Itoh

    2007-08-01

    Full Text Available Purpose: Until recently, keyboard has been used as the primary input method for machinery operation system. But in recent years, numerous methods related to direct input interface have been developed. One of them is to measure the surface electric potential that generates on the skin surface during muscle contraction. Based on this fact, hand finger operation can also be recognized with the help of the surface muscle electric potential. The purpose of this study is to identify the hand finger operation using surface electromyogram (SEMG during crookedness state of the finger.Design/methodology/approach: Two electrodes (Ag-AgCl electrode were sticked randomly on the forearm muscles and the intensity of EMG signals at different muscles were measured for each crooked finger. Then depending on the intensity of the obtained electric potentials, a position was located and considered to have participated most actively during the crookedness state of that finger. Thus five locations on the forearm muscles were identified for five different fingers. Moreover, four different types of crookedness states were considered for each finger.Findings: In this experimental study, the electric current that generates on the skin during muscle activity was measured for different hand finger operations. As a result, it is found that there is a specified position related to the maximum intensity of EMG signals for each finger. Practical implications: This paper cleared that the amount of crookedness of each finger can also be recognized with the help of surface EMG. It could be used as a machine interface technology in the field of welfare equipments, robot hand operation, virtual reality, etc.Originality/value: The objective of this research project was to develop the method of recognizing the hand finger operation and their crookedness states from surface electromyogram (SEMG.

  3. Zinc supplementation alters airway inflammation and airway hyperresponsiveness to a common allergen

    Directory of Open Access Journals (Sweden)

    Morgan Carrie I

    2011-12-01

    Full Text Available Abstract Background Zinc supplementation can modulate immunity through inhibition of NF-κB, a transcription factor that controls many immune response genes. Thus, we sought to examine the mechanism by which zinc supplementation tempers the response to a common allergen and determine its effect on allergic airway inflammation. Methods Mice were injected with zinc gluconate prior to German cockroach (GC feces (frass exposure and airway inflammation was assessed. Primary bone marrow-derived neutrophils and DMSO-differentiated HL-60 cells were used to assess the role of zinc gluconate on tumor necrosis factor (TNFα expression. NF-κB:DNA binding and IKK activity were assessed by EMSA and in vitro kinase assay. Protein levels of A20, RIP1 and TRAF6 were assessed by Western blot analysis. Establishment of allergic airway inflammation with GC frass was followed by administration of zinc gluconate. Airway hyperresponsiveness, serum IgE levels, eosinophilia and Th2 cytokine production were assessed. Results Administration of zinc gluconate prior to allergen exposure resulted in significantly decreased neutrophil infiltration and TNFα cytokine release into the airways. This correlated with decreased NF-κB activity in the whole lung. Treatment with zinc gluconate significantly decreased GC frass-mediated TNFα production from bone-marrow derived neutrophils and HL-60 cells. We confirmed zinc-mediated decreases in NF-κB:DNA binding and IKK activity in HL-60 cells. A20, a natural inhibitor of NF-κB and a zinc-fingered protein, is a potential target of zinc. Zinc treatment did not alter A20 levels in the short term, but resulted in the degradation of RIP1, an important upstream activator of IKK. TRAF6 protein levels were unaffected. To determine the application for zinc as a therapeutic for asthma, we administered zinc following the establishment of allergic airway inflammation in a murine model. Zinc supplementation decreased airway hyperresponsiveness

  4. Zinc transporters that regulate vacuolar zinc storage in Saccharomyces cerevisiae

    OpenAIRE

    MacDiarmid, Colin W; Gaither, L.Alex; Eide, David

    2000-01-01

    All cells regulate their intracellular zinc levels. In yeast, zinc uptake is mediated by Zrt1p and Zrt2p, which belong to the ZIP family of metal transporters. Under zinc limitation, ZRT1 and ZRT2 transcription is induced by the Zap1p transcriptional activator. We describe here a new component of zinc homeostasis, vacuolar zinc storage, that is also regulated by Zap1p. Zinc-replete cells accumulate zinc in the vacuole via the Zrc1p and Cot1p transporters. Our results indicate that another zin...

  5. Melorheostosis of the index finger: a case report

    OpenAIRE

    Quarles, Ellen K.; Richardson, Michael L.

    2015-01-01

    We describe a case of melorheostosis involving multiple bones of the left index finger. This patient presented with a many-year history of a finger mass, and demonstrated the typical radiographic findings of melorheostosis on radiographs and computed tomography (CT) of the hand. Following excisional biopsy and bone grafting, this patient is doing well and undergoing physical therapy.

  6. A force feedback master finger in exoskeleton type

    Institute of Scientific and Technical Information of China (English)

    Fang Honggen; Liu Hong; Xie Zongwu

    2010-01-01

    In order to eliminate the drawbacks of conventional force feedback gloves,a new type of master finger has been developed.By utilizing three"four-bar mechanism joint"in series and wire coupling mechanism,the master finger transmission ratio is kept exactly 1:1.4:1 in the whole movement range and it can make active motions in both extension and flexion directions.Additionally,to assttre faster data transmission and near zero delay in the master-slave operation,a digital signal processing/field programmable gate array(DSP/FPGA-FPGA)structure with 200μs cycle time is designed.The operating modes of the master finger can be contact or non-contact,which depends on the motion states of a slave finger,free motion or constrained motion.The position control employed in non-contact mode ensures unconstrained motion and the force control adopted in contact mode guarantees natural contact sensation.To evaluate the performances of the master finger,an experiment between the master finger and a DLR/HIT dexterous finger is conducted.The results demonstrate that this new type master finger can augment telepresence.

  7. Evaluation of the finger wrinkling test : a pilot study

    NARCIS (Netherlands)

    van Barneveld, S.; van der Palen, J.; van Putten, M. J. A. M.

    2010-01-01

    Tilt table testing mainly evaluates the systemic cardiovascular part of the autonomic nervous system, while it is assumed that the finger wrinkling test assesses the peripheral part of the autonomic nervous system. In this study we explored whether the finger wrinkling test could be a useful test fo

  8. Detection of finger gesture using singular spectrum transformation

    International Nuclear Information System (INIS)

    The purpose of this study is to detect finger movement using a singular spectrum transformation method. Human gesture recognition is essential for realizing natural user interfaces. However, constructing a robust, easily installable interface is extremely difficult. Our proposed method uses singular spectrum transformation to classify finger movements. This method robustly classifies gestures and behavior

  9. 21 CFR 888.3230 - Finger joint polymer constrained prosthesis.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Finger joint polymer constrained prosthesis. 888... constrained prosthesis. (a) Identification. A finger joint polymer constrained prosthesis is a device intended... generic type of device includes prostheses that consist of a single flexible across-the-joint...

  10. Zinc sulphate in rheumatoid arthritis

    OpenAIRE

    Mattingly, P. C.; Mowat, A G

    1982-01-01

    To assess the antirheumatic activity of zinc sulphate, 27 patients with active rheumatoid arthritis took part in a 6-month, randomised, double-blind, between-group trial of oral zinc sulphate versus placebo. Twelve patients on zinc and 9 on placebo completed the trial, but no significant antirheumatic activity of zinc sulphate was demonstrated.

  11. Geometric approach to viscous fingering on a cone

    International Nuclear Information System (INIS)

    We study fluid flow and the formation of viscous fingering patterns on a two-dimensional conical background space, defined as the conical Hele-Shaw cell. We approach the problem geometrically and study how the nontrivial topological structure of the conical cell affects the evolution of the interface separating two viscous fluids. We perform a perturbative weakly nonlinear analysis of the problem and derive a mode-coupling differential equation which describes fluid-fluid interface behaviour. Our nonlinear study predicts the formation of fingering structures in which fingers of different lengths compete and split at their tips. The shape of the emerging patterns show a significant sensitivity to variations in the cell's topological features, which can be monitored by changing the cone opening angle. We find that for increasingly larger values of the opening angle, finger competition is inhibited while finger tip-splitting is enhanced

  12. A new algorithmic approach for fingers detection and identification

    Science.gov (United States)

    Mubashar Khan, Arslan; Umar, Waqas; Choudhary, Taimoor; Hussain, Fawad; Haroon Yousaf, Muhammad

    2013-03-01

    Gesture recognition is concerned with the goal of interpreting human gestures through mathematical algorithms. Gestures can originate from any bodily motion or state but commonly originate from the face or hand. Hand gesture detection in a real time environment, where the time and memory are important issues, is a critical operation. Hand gesture recognition largely depends on the accurate detection of the fingers. This paper presents a new algorithmic approach to detect and identify fingers of human hand. The proposed algorithm does not depend upon the prior knowledge of the scene. It detects the active fingers and Metacarpophalangeal (MCP) of the inactive fingers from an already detected hand. Dynamic thresholding technique and connected component labeling scheme are employed for background elimination and hand detection respectively. Algorithm proposed a new approach for finger identification in real time environment keeping the memory and time constraint as low as possible.

  13. Surface Tension and Fingering of Miscible Interfaces

    Science.gov (United States)

    Abib, Mohammed; Liu, Jian-Bang; Ronney, Paul D.

    1999-01-01

    Experiments on miscible, buoyantly unstable reaction-diffusion fronts and non-reacting displacement fronts in Hele-Shaw cells show a fingering-type instability whose wavelengths (lambda*) are consistent with an interfacial tension (sigma) at the front caused by the change in chemical composition, even though the solutions are miscible in all proportions. In conjunction with the Saffman-Taylor model, the relation sigma = K/tau, where tau is the interface thickness and K approximately equal 4 +/- 2 x 10(exp -6) dyne, enables prediction of our measured values of lambda* as well as results from prior experiments on miscible interfaces. These results indicate that even for miscible fluids, surface tension is generally a more significant factor than diffusion in interfacial stability and flow characteristics.

  14. The Silent Language of Fingers and Hands-The Role of Finger and Hand Motions in Non-verbal Communication

    Institute of Scientific and Technical Information of China (English)

    周樱

    2013-01-01

    This paper engages in the discussion about nonverbal intercultural communication from the perspective of hand and finger gestures, probing into the functions of non-verbal cues in cross-cultural interaction, talking over two different attitudes to⁃ward gestures, and giving a detailed analysis about the silent language of some specific gestures with fingers and hands in different cultures.

  15. Zinc metabolism in thyroid disease.

    OpenAIRE

    Nishi, Y.; Kawate, R.; Usui, T

    1980-01-01

    This study was designed to evaluate the zinc metabolism in adults of both sexes with thyroid disease. Plasma and erythrocyte zinc concentration and urinary zinc excretion were investigated. The mean concentration of plasma zinc in hypothyroid patients and in euthyroid patients, previously either hyperthyroid or hypothyroid, was lower than that of control subjects, whereas no statistically significant differences were observed in plasma zinc values between hyperthyroid patients and control sub...

  16. Zinc bioavailability and homeostasis1234

    OpenAIRE

    Hambidge, K Michael; Miller, Leland V; Westcott, Jamie E; Sheng, Xiaoyang; Krebs, Nancy F.

    2010-01-01

    Zinc has earned recognition recently as a micronutrient of outstanding and diverse biological, clinical, and global public health importance. Regulation of absorption by zinc transporters in the enterocyte, together with saturation kinetics of the absorption process into and across the enterocyte, are the principal means by which whole-body zinc homeostasis is maintained. Several physiologic factors, most notably the quantity of zinc ingested, determine the quantity of zinc absorbed and the e...

  17. Zinc in diet

    Science.gov (United States)

    Symptoms of zinc deficiency include: Frequent infections Hypogonadism in males Loss of hair Poor appetite Problems with the sense of taste Problems with the sense of smell Skin sores Slow growth Trouble seeing ...

  18. Zinc electrode and rechargeable zinc-air battery

    Science.gov (United States)

    Ross, Jr., Philip N.

    1989-01-01

    An improved zinc electrode is disclosed for a rechargeable zinc-air battery comprising an outer frame and a porous foam electrode support within the frame which is treated prior to the deposition of zinc thereon to inhibit the formation of zinc dendrites on the external surface thereof. The outer frame is provided with passageways for circulating an alkaline electrolyte through the treated zinc-coated porous foam. A novel rechargeable zinc-air battery system is also disclosed which utilizes the improved zinc electrode and further includes an alkaline electrolyte within said battery circulating through the passageways in the zinc electrode and an external electrolyte circulation means which has an electrolyte reservoir external to the battery case including filter means to filter solids out of the electrolyte as it circulates to the external reservoir and pump means for recirculating electrolyte from the external reservoir to the zinc electrode.

  19. AAV-Mediated Delivery of Zinc Finger Nucleases Targeting Hepatitis B Virus Inhibits Active Replication

    OpenAIRE

    Weber, Nicholas D.; Daniel Stone; Ruth Hall Sedlak; De Silva Feelixge, Harshana S.; Pavitra Roychoudhury; Schiffer, Joshua T.; Martine Aubert; Jerome, Keith R.

    2014-01-01

    Despite an existing effective vaccine, hepatitis B virus (HBV) remains a major public health concern. There are effective suppressive therapies for HBV, but they remain expensive and inaccessible to many, and not all patients respond well. Furthermore, HBV can persist as genomic covalently closed circular DNA (cccDNA) that remains in hepatocytes even during otherwise effective therapy and facilitates rebound in patients after treatment has stopped. Therefore, the need for an effective treatme...

  20. Mining the O-glycoproteome using zinc-finger nuclease-glycoengineered SimpleCell lines

    DEFF Research Database (Denmark)

    Steentoft, Catharina; Vakhrushev, Sergey Y; Vester-Christensen, Malene B;

    2011-01-01

    targeting to truncate the O-glycan elongation pathway in human cells, generating stable 'SimpleCell' lines with homogenous O-glycosylation. Three SimpleCell lines expressing only truncated GalNAca or NeuAca2-6GalNAca O-glycans were produced, allowing straightforward isolation and sequencing of GalNAc O......-glycopeptides from total cell lysates using lectin chromatography and nanoflow liquid chromatography-mass spectrometry (nLC-MS/MS) with electron transfer dissociation fragmentation. We identified >100 O-glycoproteins with >350 O-glycan sites (the great majority previously unidentified), including a GalNAc O...

  1. Brittle Cornea Syndrome Associated with a Missense Mutation in the Zinc-Finger 469 Gene

    DEFF Research Database (Denmark)

    Christensen, Anne Elisabeth; Knappskog, Per Morten; Midtbø, Marit;

    2010-01-01

    Purpose: To investigate the diverse clinical manifestations, identify the causative mutation and explain the association with red hair in a family with brittle cornea syndrome (BCS). Methods: Eight family members in three generations underwent ophthalmic, dental, and general medical examination...... mapping with SNP markers, DNA sequencing, and MC1R genotyping. Results: At 42 and 48 years of age, respectively, both affected individuals were blind due to retinal detachment and secondary glaucoma. They had extremely thin and bulging corneas, velvety skin, chestnut colored hair, scoliosis, reduced BMD......, dental anomalies, hearing loss and minor cardiac defects. The morphologies of the skin biopsies were normal except that in some areas slightly thinner collagen fibrils were seen in one of the affected individuals. Molecular genetic analysis revealed a novel missense mutation of ZNF469, c.10016G...

  2. Expression of Zinc Finger Protein 804A (ZNF804A) in the brain

    DEFF Research Database (Denmark)

    Benedikz, Eirikur

    Schizophrenia is a severe psychiatric disorder with lifetime prevalence between 0.5 and 1%. The disease is characterized by delusions, hallucinations, altered cognition, emotional reactivity and disorganized behavior. Research increasingly suggests that schizophrenia is a subtle disorder of brain...

  3. Glycoengineering of Human Cell Lines Using Zinc Finger Nuclease Gene Targeting

    DEFF Research Database (Denmark)

    Steentoft, Catharina; Bennett, Eric Paul; Clausen, Henrik

    2013-01-01

    Lectin affinity chromatography is a powerful technique for isolation of glycoproteins carrying a specific glycan structure of interest. However, the enormous diversity of glycans present on the cell surface, as well as on individual proteins, makes it difficult to isolate an entire glycoproteome ...

  4. The zinc finger protein ZAT11 modulates paraquat-induced programmed cell death in Arabidopsis thaliana

    NARCIS (Netherlands)

    Qureshi, Muhammad Kamran; Sujeeth, Neerakkal; Gechev, Tsanko S.; Hille, Jacques; Liu, J.-H.

    2013-01-01

    Plants use programmed cell death (PCD) as a tool in their growth and development. PCD is also involved in defense against different kinds of stresses including pathogen attack. In both types of PCD, reactive oxygen species (ROS) play an important role. ROS is not only a toxic by-product but also act

  5. E(var)3-9 of Drosophila melanogaster Encodes a Zinc Finger Protein

    OpenAIRE

    Weiler, Karen S.

    2007-01-01

    The importance of a gene's natural chromatin environment for its normal expression is poignantly illustrated when a change in chromosome position results in variable gene repression, such as is observed in position effect variegation (PEV) when the Drosophila melanogaster white (w) gene is juxtaposed with heterochromatin. The Enhancer of variegation 3-9 [E(var)3-9] gene was one of over a hundred loci identified in screens for mutations that dominantly modify PEV. Haploinsufficiency for E(var)...

  6. Re-engineering the zinc fingers of PRDM9 reverses hybrid sterility in mice.

    Science.gov (United States)

    Davies, Benjamin; Hatton, Edouard; Altemose, Nicolas; Hussin, Julie G; Pratto, Florencia; Zhang, Gang; Hinch, Anjali Gupta; Moralli, Daniela; Biggs, Daniel; Diaz, Rebeca; Preece, Chris; Li, Ran; Bitoun, Emmanuelle; Brick, Kevin; Green, Catherine M; Camerini-Otero, R Daniel; Myers, Simon R; Donnelly, Peter

    2016-02-11

    The DNA-binding protein PRDM9 directs positioning of the double-strand breaks (DSBs) that initiate meiotic recombination in mice and humans. Prdm9 is the only mammalian speciation gene yet identified and is responsible for sterility phenotypes in male hybrids of certain mouse subspecies. To investigate PRDM9 binding and its role in fertility and meiotic recombination, we humanized the DNA-binding domain of PRDM9 in C57BL/6 mice. This change repositions DSB hotspots and completely restores fertility in male hybrids. Here we show that alteration of one Prdm9 allele impacts the behaviour of DSBs controlled by the other allele at chromosome-wide scales. These effects correlate strongly with the degree to which each PRDM9 variant binds both homologues at the DSB sites it controls. Furthermore, higher genome-wide levels of such 'symmetric' PRDM9 binding associate with increasing fertility measures, and comparisons of individual hotspots suggest binding symmetry plays a downstream role in the recombination process. These findings reveal that subspecies-specific degradation of PRDM9 binding sites by meiotic drive, which steadily increases asymmetric PRDM9 binding, has impacts beyond simply changing hotspot positions, and strongly support a direct involvement in hybrid infertility. Because such meiotic drive occurs across mammals, PRDM9 may play a wider, yet transient, role in the early stages of speciation.

  7. The retinoblastoma-interacting zinc-finger protein RIZ is a downstream effector of estrogen action

    Science.gov (United States)

    Abbondanza, Ciro; Medici, Nicola; Nigro, Vincenzo; Rossi, Valentina; Gallo, Luigi; Piluso, Giulio; Belsito, Angela; Roscigno, Annarita; Bontempo, Paola; Puca, Annibale A.; Molinari, Anna Maria; Moncharmont, Bruno; Puca, Giovanni A.

    2000-01-01

    Co-immunoprecipitation experiments in cell extract from cultured cells or target tissues indicated that estrogen receptor was complexed with the retinoblastoma binding protein RIZ in a ligand-dependent manner. Mapping of interaction sites indicated that in both proteins the same regions and motifs responsible for the interaction of transcriptional co-activator and nuclear receptors were involved. In cultured cells, estradiol induced a redistribution of RIZ protein within the nucleus and in the cytoplasm. A similar effect was produced in vivo, in prepuberal rat endometrium, by administration of a physiological dose of estradiol. Therefore, RIZ protein could be a specific effector of estrogen action downstream of the hormone-receptor interaction, presumably involved in proliferation control. PMID:10706618

  8. Highly efficient modification of beta-lactoglobulin (BLG) gene via zinc-finger nucleases in cattle

    Institute of Scientific and Technical Information of China (English)

    Shengli Yu; Junjie Luo; Zhiyuan Song; Fangrong Ding; Yunping Dai; Ning Li

    2011-01-01

    Dear Editor,Gene targeting is in widespread use as a gold standard for determining the function of genes in mice and human embryonic stem cells [1].However,the poor efficiency of this technology has hindered its application to domestic animals,for which embryonic stem cells are not available.Although gene-targeted large domestic animals have been produced successfully by combination of homologous recombination-based targeting strategy and cloning [2-4],the efficiency is very low and,more importantly,the disruption of the targeted gene is usually mono-allelic.It thus takes a long time to obtain a null mutant.

  9. Cooperation of the BTB-Zinc finger protein, Abrupt, with cytoskeletal regulators in Drosophila epithelial tumorigenesis

    Directory of Open Access Journals (Sweden)

    Nezaket Turkel

    2015-08-01

    Full Text Available The deregulation of cell polarity or cytoskeletal regulators is a common occurrence in human epithelial cancers. Moreover, there is accumulating evidence in human epithelial cancer that BTB-ZF genes, such as Bcl6 and ZBTB7A, are oncogenic. From our previous studies in the vinegar fly, Drosophila melanogaster, we have identified a cooperative interaction between a mutation in the apico-basal cell polarity regulator Scribble (Scrib and overexpression of the BTB-ZF protein Abrupt (Ab. Herein, we show that co-expression of ab with actin cytoskeletal regulators, RhoGEF2 or Src64B, in the developing eye-antennal epithelial tissue results in the formation of overgrown amorphous tumours, whereas ab and DRac1 co-expression leads to non-cell autonomous overgrowth. Together with ab, these genes affect the expression of differentiation genes, resulting in tumours locked in a progenitor cell fate. Finally, we show that the expression of two mammalian genes related to ab, Bcl6 and ZBTB7A, which are oncogenes in mammalian epithelial cancers, significantly correlate with the upregulation of cytoskeletal genes or downregulation of apico-basal cell polarity neoplastic tumour suppressor genes in colorectal, lung and other human epithelial cancers. Altogether, this analysis has revealed that upregulation of cytoskeletal regulators cooperate with Abrupt in Drosophila epithelial tumorigenesis, and that high expression of human BTB-ZF genes, Bcl6 and ZBTB7A, shows significant correlations with cytoskeletal and cell polarity gene expression in specific epithelial tumour types. This highlights the need for further investigation of the cooperation between these genes in mammalian systems.

  10. Tangential finger forces use mechanical advantage during static grasping.

    Science.gov (United States)

    Slota, Gregory P; Latash, Mark L; Zatsiorsky, Vladimir M

    2012-02-01

    When grasping and manipulating objects, the central controller utilizes the mechanical advantage of the normal forces of the fingers for torque production. Whether the same is valid for tangential forces is unknown. The main purpose of this study was to determine the patterns of finger tangential forces and the use of mechanical advantage as a control mechanism when dealing with objects of nonuniform finger positioning. A complementary goal was to explore the interaction of mechanical advantage (moment arm) and the role a finger has as a torque agonist/antagonist with respect to external torques (±0.4 N m). Five 6-df force/torque transducers measured finger forces while subjects held a prism handle (6 cm width × 9 cm height) with and without a single finger displaced 2 cm (handle width). The effect of increasing the tangential moment arm was significant (p forces (in >70% of trials) and hence creating greater moments. Thus, the data provides evidence that the grasping system as a rule utilizes mechanical advantage for generating tangential forces. The increase in tangential force was independent of whether the finger was acting as a torque agonist or antagonist, revealing their effects to be additive.

  11. Torque control of underactuated tendon-driven fingers

    Directory of Open Access Journals (Sweden)

    M. E. Abdallah

    2011-02-01

    Full Text Available Given an underactuated tendon-driven finger, the finger posture is underdetermined and can move freely ("flop" in a region of slack tendons. This work shows that such an underactuated finger can be operated in tendon force control (rather than position control with effective performance. The force control eliminates the indeterminate slack while commanding a parameterized space of desired torques. The torque will either push the finger to the joint limits or wrap around an external object with variable torque – behavior that is sufficient for primarily gripping fingers. In addition, introducing asymmetric joint radii to the design allows the finger to command an expanded range of joint torques and to scan an expanded set of external surfaces. This study is motivated by the design and control of the secondary fingers of the NASA-GM R2 humanoid hand.

    This paper was presented at the IFToMM/ASME International Workshop on Underactuated Grasping (UG2010, 19 August 2010, Montréal, Canada.

  12. Design Considerations for an Underactuated Robotic Finger Mechanism

    Institute of Scientific and Technical Information of China (English)

    YAO Shuangji; CECCARELLI Marco; ZHAN Qiang; ARBONE Giuseppe; LU Zhen

    2009-01-01

    A design approach is presented in this paper for underactuation in robotic finger mechanisms. The characters of underactuated finger mechanisms are introduced as based on linkage and spring systems. The feature of self-adaptive enveloping grasp by underactuated finger mechanisms is discussed with feasible in grasping unknown objects. The design problem of robotic fingers is analyzed by looking at many aspects for an optimal functionality. Design problems and requirements for underactuated mechanisms are formulated as related to human-like robotic fingers. In particular, characteristics of finger mechanisms are analyzed and optimality criteria are summarized with the aim to formulate a general design algorithm. A general multi-objective optimization design approach is applied as based on a suitable optimization problem by using suitable expressions of optimality criteria. An example is illustrated as an improvement of finger mechanism in Laboratory of Robotics and Mechatronics (LARM) Hand. Results of design outputs and grasp simulations are reported with the aim to show the practical feasibility of the proposed concepts and computations.

  13. Control of zinc transfer between thionein, metallothionein, and zinc proteins

    OpenAIRE

    Jacob, Claus; Maret, Wolfgang; Vallee, Bert L.

    1998-01-01

    Metallothionein (MT), despite its high metal binding constant (KZn = 3.2 × 1013 M−1 at pH 7.4), can transfer zinc to the apoforms of zinc enzymes that have inherently lower stability constants. To gain insight into this paradox, we have studied zinc transfer between zinc enzymes and MT. Zinc can be transferred in both directions—i.e., from the enzymes to thionein (the apoform of MT) and from MT to the apoenzymes. Agents that mediate or enhance zinc transfer have be...

  14. Reduction of arsenite-enhanced ultraviolet radiation-induced DNA damage by supplemental zinc

    Energy Technology Data Exchange (ETDEWEB)

    Cooper, Karen L.; King, Brenee S.; Sandoval, Monica M.; Liu, Ke Jian; Hudson, Laurie G., E-mail: lhudson@salud.unm.edu

    2013-06-01

    Arsenic is a recognized human carcinogen and there is evidence that arsenic augments the carcinogenicity of DNA damaging agents such as ultraviolet radiation (UVR) thereby acting as a co-carcinogen. Inhibition of DNA repair is one proposed mechanism to account for the co-carcinogenic actions of arsenic. We and others find that arsenite interferes with the function of certain zinc finger DNA repair proteins. Furthermore, we reported that zinc reverses the effects of arsenite in cultured cells and a DNA repair target protein, poly (ADP-ribose) polymerase-1. In order to determine whether zinc ameliorates the effects of arsenite on UVR-induced DNA damage in human keratinocytes and in an in vivo model, normal human epidermal keratinocytes and SKH-1 hairless mice were exposed to arsenite, zinc or both before solar-simulated (ss) UVR exposure. Poly (ADP-ribose) polymerase activity, DNA damage and mutation frequencies at the Hprt locus were measured in each treatment group in normal human keratinocytes. DNA damage was assessed in vivo by immunohistochemical staining of skin sections isolated from SKH-1 hairless mice. Cell-based findings demonstrate that ssUVR-induced DNA damage and mutagenesis are enhanced by arsenite, and supplemental zinc partially reverses the arsenite effect. In vivo studies confirm that zinc supplementation decreases arsenite-enhanced DNA damage in response to ssUVR exposure. From these data we can conclude that zinc offsets the impact of arsenic on ssUVR-stimulated DNA damage in cells and in vivo suggesting that zinc supplementation may provide a strategy to improve DNA repair capacity in arsenic exposed human populations. - Highlights: • Low levels of arsenite enhance UV-induced DNA damage in human keratinocytes. • UV-initiated HPRT mutation frequency is enhanced by arsenite. • Zinc supplementation offsets DNA damage and mutation frequency enhanced by arsenite. • Zinc-dependent reduction of arsenite enhanced DNA damage is confirmed in vivo.

  15. CH_4 and C_2H_2 Detection System with Non-dispersed Infrared Technique Based on GFC%基于GFC的非分散红外CH_4和C_2H_2气体检测系统

    Institute of Scientific and Technical Information of China (English)

    胡艳玲; 王玉田; 尚明丽; 杨程

    2009-01-01

    基于甲烷气体和乙炔气体近红外吸收的机理,研究了一种高灵敏度非分散红外系统,同时测量甲烷气体和乙炔气体浓度.系统采用折反式吸收池作为气室,结合先进的气体滤波相关技术和相关检测技术,实现了对微弱光谱信号的调制和检测,最后实现对浓度的反演,从而实现了甲烷和乙炔气体实时测量.试验结果表明:系统具有10~(-5)的测量分辨力.

  16. Finger Movement Based Wearable Communication & Navigation Aid for partially disabled

    Directory of Open Access Journals (Sweden)

    Jobin Jose

    2016-02-01

    Full Text Available The FMBWCN Aid is a portable and wearable multi-purpose system for the partially disabled. The Aid is a glove-based system in which different trained finger movements leads to different modes of operation of the system such as APR Voice Module and Wheelchair Controller Module. The finger movements (bending movement leads to the flexion of the flex sensor attached to the glove. Flexion leads to change in resistance of the flex sensors, which will be recorded via a microcontroller (Arduino Uno and different sets of movement of the fingers will lead to different modules of operation as specified in the default settings of the FMBWCN Aid.

  17. The social and economic consequences of finger amputations.

    Science.gov (United States)

    Hovgaard, C; Angermann, P; Hovgaard, D

    1994-06-01

    120 patients with amputation of at least 1 of the 4 ulnar fingers were admitted to hospital. In none was replantation considered to be possible because of serious damage to the soft tissues and bone. 12 (3-18) years after the accident 80 percent of the patients assessed their condition as good or fair, even those with proximal amputation or loss of 2 or 3 fingers. Our observations do not support replantation when only one of the second-to-fifth fingers have been amputated.

  18. Narrow fingers in the Saffman-Taylor instability

    Science.gov (United States)

    Couder, Y.; Gerard, N.; Rabaud, M.

    1986-12-01

    Saffman-Taylor fingers with a relative width much smaller than the classical limit lambda = 0.5 are found when a small isolated bubble is located at their tip. These solutions are members of a family found by Saffman and Taylor (1958) neglecting superficial tension. Recent theories have shown that when capillary forces are taken into account an unphysical cusplike singularity would appear at the tip of all the fingers with lambda less than 0.5. Conversely, here the replacement of the tip by a small bubble makes these solutions possible. At large velocity these fingers show dendritic instability.

  19. Treatment of zinc deficiency without zinc fortification

    Institute of Scientific and Technical Information of China (English)

    Donald OBERLEAS; Barbara F. HARLAND

    2008-01-01

    Zinc (Zn) deficiency in animals became of interest until the 1950s. In this paper, progresses in researches on physi-ology of Zn deficiency in animals, phytate effect on bioavailability of Zn, and role of phytase in healing Zn deficiency of animals were reviewed. Several studies demonstrated that Zn is recycled via the pancreas; the problem of Zn deficiency was controlled by Zn homeostasis. The endogenous secretion of Zn is considered as an important factor influencing Zn deficiency, and the critical molar ratio is 10. Phytate (inositol hexaphosphate) constituted up to 90% of the organically bound phosphorus in seeds. Great improvement has been made in recent years on isolating and measuring phytate, and its structure is clear. Phytate is considered to reduce Zn bioavailability in animal. Phytase is the enzyme that hydrolyzes phytate and is present in yeast, rye bran, wheat bran, barley, triticale, and many bacteria and fungi. Zinc nutrition and bioavailability can be enhanced by addition of phytase to animal feeds. Therefore, using phytase as supplements, the most prevalent Zn deficiency in animals may be effectively corrected without the mining and smelting of several tons of zinc daily needed to correct this deficiency by fortification worldwide.

  20. The bioavailability of four zinc oxide sources and zinc sulphate in broiler chickens

    OpenAIRE

    Veldkamp, T; Diepen, van, F.N.J.; Bikker, P.

    2014-01-01

    Zinc is an essential trace element for all farm animal species. It is commonly included in animal diets as zinc oxide, zinc sulphate or organically bound zinc. Umicore Zinc Chemicals developed zinc oxide products with different mean particle sizes. Umicore Zinc Chemicals requested Wageningen UR Livestock Research to determine the bioavailability of four zinc oxide sources and zinc sulphate in broiler chickens. A precise estimate of the bioavailability of zinc sources is required both for fulf...

  1. Examiner's finger-mounted fetal tissue oximetry

    Science.gov (United States)

    Kanayama, Naohiro; Niwayama, Masatsugu

    2014-06-01

    The best way to assess fetal condition is to observe the oxygen status of the fetus (as well as to assess the condition of infants, children, and adults). Previously, several fetal oximeters have been developed; however, no instrument has been utilized in clinical practice because of the low-capturing rate of the fetal oxygen saturation. To overcome the problem, we developed a doctor's finger-mounted fetal tissue oximeter, whose sensor volume is one hundredth of the conventional one. Additionally, we prepared transparent gloves. The calculation algorithm of the hemoglobin concentration was derived from the light propagation analysis based on the transport theory. We measured neonatal and fetal oxygen saturation (StO2) with the new tissue oximeter. Neonatal StO was measured at any position of the head regardless of amount of hair. Neonatal StO was found to be around 77%. Fetal StO was detected in every position of the fetal head during labor regardless of the presence of labor pain. Fetal StO without labor pain was around 70% in the first stage of labor and around 60% in the second stage of labor. We concluded that our new concept of fetal tissue oximetry would be useful for detecting fetal StO in any condition of the fetus.

  2. Finger Length Ratios in Serbian Transsexuals

    Directory of Open Access Journals (Sweden)

    Svetlana Vujović

    2014-01-01

    Full Text Available Atypical prenatal hormone exposure could be a factor in the development of transsexualism. There is evidence that the 2nd and 4th digit ratio (2D : 4D associates negatively with prenatal testosterone and positively with estrogens. The aim was to assess the difference in 2D : 4D between female to male transsexuals (FMT and male to female transsexuals (MFT and controls. We examined 42 MFT, 38 FMT, and 45 control males and 48 control females. Precise measurements were made by X-rays at the ventral surface of both hands from the basal crease of the digit to the tip using vernier calliper. Control male and female patients had larger 2D : 4D of the right hand when compared to the left hand. Control male’s left hand ratio was lower than in control female’s left hand. There was no difference in 2D : 4D between MFT and control males. MFT showed similar 2D : 4D of the right hand with control women indicating possible influencing factor in embryogenesis and consequently finger length changes. FMT showed the lowest 2D : 4D of the left hand when compared to the control males and females. Results of our study go in favour of the biological aetiology of transsexualism.

  3. Possible Gigantic Variations on the Width of Viscoelastic Fingers

    CERN Document Server

    Poire, E C; Poire, Eugenia Corvera

    2002-01-01

    We analyze the effect of frequency on the width of a single finger displacing a viscoelastic fluid. We derive a generalized Darcy's law in the frequency domain for a linear viscoelastic fluid flowing in a Hele Shaw cell. This leads to an analytic expression for the dynamic permeability that has maxima which are several orders of magnitude larger than the static permeability. We then follow an argument of de Gennes to obtain the smallest possible finger width when viscoelasticity is important. Using this, and a conservation law, we obtain a lowest bound for the width of a single finger displacing a viscoelastic fluid. Our results indicate that when a small amplitude signal of the frequency that maximizes the permeability is overimposed to a constant pressure drop, gigantic variations are obtained for the finger width.

  4. Mallet finger deformities treated by buried purse-string suture

    Institute of Scientific and Technical Information of China (English)

    YAO Jian-min; ZHAO Feng-jing

    2011-01-01

    Objective: To investigate the efficacy of buried purse-string suture in the treatment of mallet finger deformities. Methods: From February 2009 to February 2010, 12 patients with closed non-fracture mallet fingers were treated by buried purse-string suture. The rupture tendons were sutured by purse-string suture with an atraumatic needle, and the knots were buried under subcutaneous tissue. External fixator was used at the extension position of the finger every night within three weeks after operation. Results: All patients were followed up for 6-12 months, mean 7 months. According to the Patel's evaluation criteria, 2 cases (17%) obtained excellent results, 7 good (58%), 2 fair (17%) and 1 poor (8%). The overall rate of the cases with excellent and good outcomes was 75%. Conclusion: Buried purse-string suture is an easy and effective way to treat mallet finger deformities, with no serious postoperative complications or no need for reoperation.

  5. Online Credit Card Transaction Using Finger Print Recognition

    Directory of Open Access Journals (Sweden)

    S.Sivasubramanian

    2010-10-01

    Full Text Available Internet shopping, a strong alternative to traditional “go, see, touch and buy” shopping, has been one of the mostly used facilities of the Internet. Security in online payment systems has been a wide research area since the early days of the Internet and several approaches have been devised by various Organizations. But, none of the system overcome the weakness in those system. Several online shopping systems serve internet users all around the world and enable people to get the products they need with a small effort. This paper proposes a new solution that combines finger print recognition with online credit card transactions. Here the proposed system provides more security then existing system with finger print recognitionbecause of finger print is unique. Here no need to remember the more passwords, your finger is your password.

  6. [Raynaud's phenomenon and other circulatory disorders of the fingers].

    Science.gov (United States)

    Mahler, Felix

    2014-02-26

    Raynaud's phenomenon (RP) is defined as attacks of blanking, subsequent cyanosis and rubeosis of fingers due to vasospasms in response to cold or emotional stimuli. Primary RP has no known underlying cause and occurs mainly in young and otherwise healthy women. Secondary RP goes along with various causes such as connective tissue diseases, toxic substances, drugs, physical trauma or organic finger artery occlusions, and occurs at any age and in both genders. Related affections are acrocyanosis and finger artery occlusions either due to arteriosclerosis or vasculitis. Also spontaneous finger hematoma may provoke an episode of RP. Therapeutically strict cold protection and avoidance of possible noxa is recommended besides the treatment of underlying diseases. No standard vasoactive drug has proven ideal for RP due to side effects. In cases with rest pain or ulcerations the same principles are applied as in ischemic diseases with no possibility for revascularization.

  7. Zinc and its deficiency diseases.

    Science.gov (United States)

    Evans, G W

    1986-01-01

    The pervasive role of zinc in the metabolic function of the body results from its function as a cofactor of a multitude of enzymes. Zinc is found in every tissue in the body, and because zinc metalloenzymes are found in every known class of enzymes, the metal has a function in every conceivable type of biochemical pathway. Symptoms resulting from zinc deficiency are as diverse as the enzymes with which the metal is associated. If chronic, severe, and untreated, zinc deficiency can be fatal. Less drastic symptoms include infections, hypogonadism, weight loss, emotional disturbance, dermatitis, alopecia, impaired taste acuity, night blindness, poor appetite, delayed wound healing, and elevated blood ammonia levels. Many symptoms of zinc deficiency result from poor diet consumption, but often the most severe symptoms result from other factors including excessive alcohol use, liver diseases, malabsorption syndromes, renal disease, enteral or parenteral alimentation, administration of sulfhydryl-containing drugs, and sickle cell disease. The most severe symptoms of zinc deficiency occur in young children affected with the autosomal-recessive trait, acrodermatitis enteropathica. This disease results in decreased synthesis of picolinic acid which causes an impaired ability to utilize zinc from common food. Because simple laboratory analyses are often not reliable in determining zinc nutriture of a patient, those symptoms caused by suspected zinc deficiency are best verified by the oral administration of zinc dipicolinate. This zinc compound is efficacious and safe and would provide an accurate means of identifying symptoms that do result from zinc deficiency. PMID:3514057

  8. Finger Movement Based Wearable Communication & Navigation Aid for partially disabled

    OpenAIRE

    Jobin Jose; Kundan Joshi

    2016-01-01

    The FMBWCN Aid is a portable and wearable multi-purpose system for the partially disabled. The Aid is a glove-based system in which different trained finger movements leads to different modes of operation of the system such as APR Voice Module and Wheelchair Controller Module. The finger movements (bending movement) leads to the flexion of the flex sensor attached to the glove. Flexion leads to change in resistance of the flex sensors, which will be recorded via a microcontroller ...

  9. Finger blood content, light transmission, and pulse oximetry errors.

    Science.gov (United States)

    Craft, T M; Lawson, R A; Young, J D

    1992-01-01

    The changes in light emitting diode current necessary to maintain a constant level of light incident upon a photodetector were measured in 20 volunteers at the two wavelengths employed by pulse oximeters. Three states of finger blood content were assessed; exsanguinated, hyperaemic, and normal. The changes in light emitting diode current with changes in finger blood content were small and are not thought to represent a significant source of error in saturation as measured by pulse oximetry. PMID:1536406

  10. Topological Considerations for Tuning and Fingering Stringed Instruments

    OpenAIRE

    Allen, Terry; Goudeseune, Camille

    2011-01-01

    We present a formal language for assigning pitches to strings for fingered multi-string instruments, particularly the six-string guitar. Given the instrument's tuning (the strings' open pitches) and the compass of the fingers of the hand stopping the strings, the formalism yields a framework for simultaneously optimizing three things: the mapping of pitches to strings, the choice of instrument tuning, and the key of the composition. Final optimization relies on heuristics idiomatic to the tun...

  11. Viscous and Gravitational Fingering in Multiphase Compositional and Compressible Flow

    OpenAIRE

    Moortgat, Joachim

    2016-01-01

    Viscous and gravitational fingering refer to flow instabilities in porous media that are triggered by adverse mobility or density ratios, respectively. These instabilities have been studied extensively in the past for 1) single-phase flow (e.g., contaminant transport in groundwater, first-contact-miscible displacement of oil by gas in hydrocarbon production), and 2) multi-phase immiscible and incompressible flow (e.g., water-alternating-gas (WAG) injection in oil reservoirs). Fingering in mul...

  12. Finger blood content, light transmission, and pulse oximetry errors.

    Science.gov (United States)

    Craft, T M; Lawson, R A; Young, J D

    1992-01-01

    The changes in light emitting diode current necessary to maintain a constant level of light incident upon a photodetector were measured in 20 volunteers at the two wavelengths employed by pulse oximeters. Three states of finger blood content were assessed; exsanguinated, hyperaemic, and normal. The changes in light emitting diode current with changes in finger blood content were small and are not thought to represent a significant source of error in saturation as measured by pulse oximetry.

  13. Unveiling the Biometric Potential of Finger-Based ECG Signals

    OpenAIRE

    André Lourenço; Hugo Silva; Ana Fred

    2011-01-01

    The ECG signal has been shown to contain relevant information for human identification. Even though results validate the potential of these signals, data acquisition methods and apparatus explored so far compromise user acceptability, requiring the acquisition of ECG at the chest. In this paper, we propose a finger-based ECG biometric system, that uses signals collected at the fingers, through a minimally intrusive 1-lead ECG setup recurring to Ag/AgCl electrodes without gel as interface with...

  14. Finger avulsion injuries: A report of four cases

    Directory of Open Access Journals (Sweden)

    Fejjal N

    2008-01-01

    Full Text Available Injury that occurs to a finger wearing a ring though rare can have grave consequences. It is a preventable injury which has a peculiar mode of trauma that is usually occupational. Injury ranges from simple contusion to degloving of soft tissues to traumatic amputation. We hereby report our experience of four cases of finger avulsion injuries due to a ring and discuss their variable clinical presentation and individualized management.

  15. Stress Distribution in Implant Retained Finger Prosthesis: A Finite Element Study

    OpenAIRE

    Amornvit, Pokpong; Rokaya, Dinesh; Keawcharoen, Konrawee; Thongpulsawasdi, Nimit

    2013-01-01

    Background: Finger amputation may result from congenital cause, trauma, infection and tumours. The finger amputation may be rehabilitated with dental implant-retained finger prosthesis. The success of implant-retained finger prosthesis is determined by the implant loading. The type of the force is a determining factor in implant loading.

  16. Enhancement of finger motion range with compliant anthropomorphic joint design.

    Science.gov (United States)

    Çulha, Utku; Iida, Fumiya

    2016-04-01

    Robotic researchers have been greatly inspired by the human hand in the search to design and build adaptive robotic hands. Especially, joints have received a lot of attention upon their role in maintaining the passive compliance that gives the fingers flexibility and extendible motion ranges. Passive compliance, which is the tendency to be employed in motion under the influence of an external force, is the result of the stiffness and the geometrical constraints of the joints that define the direction of the motion. Based on its building elements, human finger joints have multi-directional passive compliance which means that they can move in multiple axis of motion under external force. However, due to their complex anatomy, only simplified biomechanical designs based on physiological analysis are preferred in present day robotics. To imitate the human joints, these designs either use fixed degree of freedom mechanisms which substantially limit the motion axes of compliance, or soft materials that can deform in many directions but hinder the fingers' force exertion capacities. In order to find a solution that lies between these two design approaches, we are using anatomically correct finger bones, elastic ligaments and antagonistic tendons to build anthropomorphic joints with multi-directional passive compliance and strong force exertion capabilities. We use interactions between an index finger and a thumb to show that our joints allow the extension of the range of motion of the fingers up to 245% and gripping size to 63% which can be beneficial for mechanical adaptation in gripping larger objects. PMID:26891473

  17. Fluidic Channels Produced by Electro Hydrodynamic Viscous Fingering

    Science.gov (United States)

    Behler, Kristopher; Wetzel, Eric

    2010-03-01

    Viscous fingering is a term describing fingerlike extensions of liquid from a column of low viscosity liquid that has been injected into a more viscous liquid. The modification of viscous fingering, known as electro hydrodynamic viscous fingering (EHVF), utilizes large electrical potentials of 10-60 kV. The fingers see a reduction in size and increase in branching behavior due to the potential applied to the system. The resulting finely structured patterns are analogous to biological systems such as blood vessels and the lymphatic system. In this study silicone oils and water were studied in thin channel Hele-Shaw cells. The interfacial tension was optimized by altering the surfactant concentration in the silicone oils. EHVF of liquid filled packed beds consisting of beads and silicone oils showed retardation of the relaxation of the fingers after the voltage was turned off. Decreased relaxation provides a means to solidify patterns into a curable material, such as polydimethylsiloxane (PDMS). After the water is evacuated from the fingers, the cured materials then possess hollow channels that can be refilled and emptied, thus creating an artificial circulatory system.

  18. Elastic fingering in rotating Hele-Shaw flows

    KAUST Repository

    Carvalho, Gabriel D.

    2014-05-21

    The centrifugally driven viscous fingering problem arises when two immiscible fluids of different densities flow in a rotating Hele-Shaw cell. In this conventional setting an interplay between capillary and centrifugal forces makes the fluid-fluid interface unstable, leading to the formation of fingered structures that compete dynamically and reach different lengths. In this context, it is known that finger competition is very sensitive to changes in the viscosity contrast between the fluids. We study a variant of such a rotating flow problem where the fluids react and produce a gellike phase at their separating boundary. This interface is assumed to be elastic, presenting a curvature-dependent bending rigidity. A perturbative weakly nonlinear approach is used to investigate how the elastic nature of the interface affects finger competition events. Our results unveil a very different dynamic scenario, in which finger length variability is not regulated by the viscosity contrast, but rather determined by two controlling quantities: a characteristic radius and a rigidity fraction parameter. By properly tuning these quantities one can describe a whole range of finger competition behaviors even if the viscosity contrast is kept unchanged. © 2014 American Physical Society.

  19. Biomechanical Analysis of Force Distribution in Human Finger Extensor Mechanisms

    Directory of Open Access Journals (Sweden)

    Dan Hu

    2014-01-01

    Full Text Available The complexities of the function and structure of human fingers have long been recognised. The in vivo forces in the human finger tendon network during different activities are critical information for clinical diagnosis, surgical treatment, prosthetic finger design, and biomimetic hand development. In this study, we propose a novel method for in vivo force estimation for the finger tendon network by combining a three-dimensional motion analysis technique and a novel biomechanical tendon network model. The extensor mechanism of a human index finger is represented by an interconnected tendinous network moving around the phalanx’s dorsum. A novel analytical approach based on the “Principle of Minimum Total Potential Energy” is used to calculate the forces and deformations throughout the tendon network of the extensor mechanism when subjected to an external load and with the finger posture defined by measurement data. The predicted deformations and forces in the tendon network are in broad agreement with the results obtained by previous experimental in vitro studies. The proposed methodology provides a promising tool for investigating the biomechanical function of complex interconnected tendon networks in vivo.

  20. Integrating optical finger motion tracking with surface touch events

    Directory of Open Access Journals (Sweden)

    Jennifer eMacRitchie

    2015-06-01

    Full Text Available This paper presents a method of integrating two contrasting sensor systems for studying human interaction with a mechanical system, using piano performance as the case study. Piano technique requires both precise small-scale motion of fingers on the key surfaces and planned large-scale movement of the hands and arms. Where studies of performance often focus on one of these scales in isolation, this paper investigates the relationship between them. Two sensor systems were installed on an acoustic grand piano: a monocular high-speed camera tracking the position of painted markers on the hands, and capacitive touch sensors attach to the key surfaces which measure the location of finger-key contacts. This paper highlights a method of fusing the data from these systems, including temporal and spatial alignment, segmentation into notes and automatic fingering annotation. Three case studies demonstrate the utility of the multi-sensor data: analysis of finger flexion or extension based on touch and camera marker location, timing analysis of finger-key contact preceding and following key presses, and characterisation of individual finger movements in the transitions between successive key presses. Piano performance is the focus of this paper, but the sensor method could equally apply to other fine motor control scenarios, with applications to human-computer interaction.