Genome-wide dynamics of alternative polyadenylation in rice

PubMed Central

Fu, Haihui; Yang, Dewei; Su, Wenyue; Ma, Liuyin; Shen, Yingjia; Ji, Guoli; Ye, Xinfu; Wu, Xiaohui

2016-01-01

Alternative polyadenylation (APA), in which a transcript uses one of the poly(A) sites to define its 3′-end, is a common regulatory mechanism in eukaryotic gene expression. However, the potential of APA in determining crop agronomic traits remains elusive. This study systematically tallied poly(A) sites of 14 different rice tissues and developmental stages using the poly(A) tag sequencing (PAT-seq) approach. The results indicate significant involvement of APA in developmental and quantitative trait loci (QTL) gene expression. About 48% of all expressed genes use APA to generate transcriptomic and proteomic diversity. Some genes switch APA sites, allowing differentially expressed genes to use alternate 3′ UTRs. Interestingly, APA in mature pollen is distinct where differential expression levels of a set of poly(A) factors and different distributions of APA sites are found, indicating a unique mRNA 3′-end formation regulation during gametophyte development. Equally interesting, statistical analyses showed that QTL tends to use APA for regulation of gene expression of many agronomic traits, suggesting a potential important role of APA in rice production. These results provide thus far the most comprehensive and high-resolution resource for advanced analysis of APA in crops and shed light on how APA is associated with trait formation in eukaryotes. PMID:27733415

The poly(rC)-binding protein αCP2 is a noncanonical factor in X. laevis cytoplasmic polyadenylation

PubMed Central

Vishnu, Melanie R.; Sumaroka, Marina; Klein, Peter S.; Liebhaber, Stephen A.

2011-01-01

Post-transcriptional control of mRNA stability and translation is central to multiple developmental pathways. This control can be linked to cytoplasmic polyadenylation in certain settings. In maturing Xenopus oocytes, specific mRNAs are targeted for polyadenylation via recruitment of the Cytoplasmic Polyadenylation Element (CPE) binding protein (CPEB) to CPE(s) within the 3′ UTR. Cytoplasmic polyadenylation is also critical to early embryonic events, although corresponding determinants are less defined. Here, we demonstrate that the Xenopus ortholog of the poly(rC) binding protein αCP2 can recruit cytoplasmic poly(A) polymerase activity to mRNAs in Xenopus post-fertilization embryos, and that this recruitment relies on cis sequences recognized by αCP2. We find that the hα-globin 3′ UTR, a validated mammalian αCP2 target, constitutes an effective target for cytoplasmic polyadenylation in Xenopus embryos, but not during Xenopus oocyte maturation. We further demonstrate that the cytoplasmic polyadenylation activity is dependent on the action of the C-rich αCP-binding site in conjunction with the adjacent AAUAAA. Consistent with its ability to target mRNA for poly(A) addition, we find that XαCP2 associates with core components of the Xenopus cytoplasmic polyadenylation complex, including the cytoplasmic poly(A) polymerase XGLD2. Furthermore, we observe that the C-rich αCP-binding site can robustly enhance the activity of a weak canonical oocyte maturation CPE in early embryos, possibly via a direct interaction between XαCP2 and CPEB1. These studies establish XαCP2 as a novel cytoplasmic polyadenylation trans factor, indicate that C-rich sequences can function as noncanonical cytoplasmic polyadenylation elements, and expand our understanding of the complexities underlying cytoplasmic polyadenylation in specific developmental settings. PMID:21444632

αCP Poly(C) Binding Proteins Act as Global Regulators of Alternative Polyadenylation

PubMed Central

Ji, Xinjun; Wan, Ji; Vishnu, Melanie

2013-01-01

We have previously demonstrated that the KH-domain protein αCP binds to a 3′ untranslated region (3′UTR) C-rich motif of the nascent human alpha-globin (hα-globin) transcript and enhances the efficiency of 3′ processing. Here we assess the genome-wide impact of αCP RNA-protein (RNP) complexes on 3′ processing with a specific focus on its role in alternative polyadenylation (APA) site utilization. The major isoforms of αCP were acutely depleted from a human hematopoietic cell line, and the impact on mRNA representation and poly(A) site utilization was determined by direct RNA sequencing (DRS). Bioinformatic analysis revealed 357 significant alterations in poly(A) site utilization that could be specifically linked to the αCP depletion. These APA events correlated strongly with the presence of C-rich sequences in close proximity to the impacted poly(A) addition sites. The most significant linkage was the presence of a C-rich motif within a window 30 to 40 bases 5′ to poly(A) signals (AAUAAA) that were repressed upon αCP depletion. This linkage is consistent with a general role for αCPs as enhancers of 3′ processing. These findings predict a role for αCPs in posttranscriptional control pathways that can alter the coding potential and/or levels of expression of subsets of mRNAs in the mammalian transcriptome. PMID:23629627

Compilation of mRNA Polyadenylation Signals in Arabidopsis Revealed a New Signal Element and Potential Secondary Structures1[w

PubMed Central

Loke, Johnny C.; Stahlberg, Eric A.; Strenski, David G.; Haas, Brian J.; Wood, Paul Chris; Li, Qingshun Quinn

2005-01-01

Using a novel program, SignalSleuth, and a database containing authenticated polyadenylation [poly(A)] sites, we analyzed the composition of mRNA poly(A) signals in Arabidopsis (Arabidopsis thaliana), and reevaluated previously described cis-elements within the 3′-untranslated (UTR) regions, including near upstream elements and far upstream elements. As predicted, there are absences of high-consensus signal patterns. The AAUAAA signal topped the near upstream elements patterns and was found within the predicted location to only approximately 10% of 3′-UTRs. More importantly, we identified a new set, named cleavage elements, of poly(A) signals flanking both sides of the cleavage site. These cis-elements were not previously revealed by conventional mutagenesis and are contemplated as a cluster of signals for cleavage site recognition. Moreover, a single-nucleotide profile scan on the 3′-UTR regions unveiled a distinct arrangement of alternate stretches of U and A nucleotides, which led to a prediction of the formation of secondary structures. Using an RNA secondary structure prediction program, mFold, we identified three main types of secondary structures on the sequences analyzed. Surprisingly, these observed secondary structures were all interrupted in previously constructed mutations in these regions. These results will enable us to revise the current model of plant poly(A) signals and to develop tools to predict 3′-ends for gene annotation. PMID:15965016

Structure of an Rrp6-RNA exosome complex bound to poly(A) RNA

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wasmuth, Elizabeth V.; Januszyk, Kurt; Lima, Christopher D.

The eukaryotic RNA exosome processes and degrades RNA by directing substrates to the distributive or processive 3' to 5' exoribonuclease activities of Rrp6 or Rrp44, respectively. The non-catalytic nine-subunit exosome core (Exo9) features a prominent central channel. Although RNA can pass through the channel to engage Rrp44, it is not clear how RNA is directed to Rrp6 or whether Rrp6 uses the central channel. Here we report a 3.3 Å crystal structure of a ten-subunit RNA exosome complex from Saccharomyces cerevisiae composed of the Exo9 core and Rrp6 bound to single-stranded poly(A) RNA. The Rrp6 catalytic domain rests on topmore » of the Exo9 S1/KH ring above the central channel, the RNA 3' end is anchored in the Rrp6 active site, and the remaining RNA traverses the S1/KH ring in an opposite orientation to that observed in a structure of a Rrp44-containing exosome complex. Solution studies with human and yeast RNA exosome complexes suggest that the RNA path to Rrp6 is conserved and dependent on the integrity of the S1/KH ring. Although path selection to Rrp6 or Rrp44 is stochastic in vitro, the fate of a particular RNA may be determined in vivo by the manner in which cofactors present RNA to the RNA exosome.« less

E2F mediates enhanced alternative polyadenylation in proliferation.

PubMed

Elkon, Ran; Drost, Jarno; van Haaften, Gijs; Jenal, Mathias; Schrier, Mariette; Oude Vrielink, Joachim A F; Agami, Reuven

2012-07-02

The majority of mammalian genes contain multiple poly(A) sites in their 3' UTRs. Alternative cleavage and polyadenylation are emerging as an important layer of gene regulation as they generate transcript isoforms that differ in their 3' UTRs, thereby modulating genes' response to 3' UTR-mediated regulation. Enhanced cleavage at 3' UTR proximal poly(A) sites resulting in global 3' UTR shortening was recently linked to proliferation and cancer. However, mechanisms that regulate this enhanced alternative polyadenylation are unknown. Here, we explored, on a transcriptome-wide scale, alternative polyadenylation events associated with cellular proliferation and neoplastic transformation. We applied a deep-sequencing technique for identification and quantification of poly(A) sites to two human cellular models, each examined under proliferative, arrested and transformed states. In both cell systems we observed global 3' UTR shortening associated with proliferation, a link that was markedly stronger than the association with transformation. Furthermore, we found that proliferation is also associated with enhanced cleavage at intronic poly(A) sites. Last, we found that the expression level of the set of genes that encode for 3'-end processing proteins is globally elevated in proliferation, and that E2F transcription factors contribute to this regulation. Our results comprehensively identify alternative polyadenylation events associated with cellular proliferation and transformation, and demonstrate that the enhanced alternative polyadenylation in proliferative conditions results not only in global 3' UTR shortening but also in enhanced premature cleavage in introns. Our results also indicate that E2F-mediated co-transcriptional regulation of 3'-end processing genes is one of the mechanisms that links enhanced alternative polyadenylation to proliferation.

PAPERCLIP identifies microRNA targets and a role of CstF64/64tau in promoting non-canonical poly(A) site usage

PubMed Central

Hwang, Hun-Way; Park, Christopher Y.; Goodarzi, Hani; Fak, John J.; Mele, Aldo; Moore, Michael J.; Saito, Yuhki; Darnell, Robert B.

2016-01-01

Accurate and precise annotation of the 3′ untranslated regions (3′ UTRs) is critical in understanding how mRNAs are regulated by microRNAs (miRNAs) and RNA-binding proteins (RBPs). Here we describe a method, PAPERCLIP (Poly(A) binding Protein-mediated mRNA 3′ End Retrieval by CrossLinking ImmunoPrecipitation), which shows high specificity for the mRNA 3′ ends and compares favorably to existing 3′ end mapping methods. PAPERCLIP uncovers a previously unrecognized role of CstF64/64tau in promoting the usage of a selected group of non-canonical poly(A) sites, the majority of them containing a downstream GUKKU motif. Furthermore, in mouse brain, PAPERCLIP discovers extended 3′ UTR sequences harboring functional miRNA binding sites and reveals developmentally regulated APA shifts including one in Atp2b2 that is evolutionarily conserved in human and results in a gain of a functional binding site of miR-137. PAPERCLIP provides a powerful tool to decipher post-transcriptional regulation of mRNAs through APA in vivo. PMID:27050522

E2F mediates enhanced alternative polyadenylation in proliferation

PubMed Central

2012-01-01

Background The majority of mammalian genes contain multiple poly(A) sites in their 3' UTRs. Alternative cleavage and polyadenylation are emerging as an important layer of gene regulation as they generate transcript isoforms that differ in their 3' UTRs, thereby modulating genes' response to 3' UTR-mediated regulation. Enhanced cleavage at 3' UTR proximal poly(A) sites resulting in global 3' UTR shortening was recently linked to proliferation and cancer. However, mechanisms that regulate this enhanced alternative polyadenylation are unknown. Results Here, we explored, on a transcriptome-wide scale, alternative polyadenylation events associated with cellular proliferation and neoplastic transformation. We applied a deep-sequencing technique for identification and quantification of poly(A) sites to two human cellular models, each examined under proliferative, arrested and transformed states. In both cell systems we observed global 3' UTR shortening associated with proliferation, a link that was markedly stronger than the association with transformation. Furthermore, we found that proliferation is also associated with enhanced cleavage at intronic poly(A) sites. Last, we found that the expression level of the set of genes that encode for 3'-end processing proteins is globally elevated in proliferation, and that E2F transcription factors contribute to this regulation. Conclusions Our results comprehensively identify alternative polyadenylation events associated with cellular proliferation and transformation, and demonstrate that the enhanced alternative polyadenylation in proliferative conditions results not only in global 3' UTR shortening but also in enhanced premature cleavage in introns. Our results also indicate that E2F-mediated co-transcriptional regulation of 3'-end processing genes is one of the mechanisms that links enhanced alternative polyadenylation to proliferation. PMID:22747694

Alternative polyadenylation of tumor suppressor genes in small intestinal neuroendocrine tumors.

PubMed

Rehfeld, Anders; Plass, Mireya; Døssing, Kristina; Knigge, Ulrich; Kjær, Andreas; Krogh, Anders; Friis-Hansen, Lennart

2014-01-01

The tumorigenesis of small intestinal neuroendocrine tumors (SI-NETs) is poorly understood. Recent studies have associated alternative polyadenylation (APA) with proliferation, cell transformation, and cancer. Polyadenylation is the process in which the pre-messenger RNA is cleaved at a polyA site and a polyA tail is added. Genes with two or more polyA sites can undergo APA. This produces two or more distinct mRNA isoforms with different 3' untranslated regions. Additionally, APA can also produce mRNAs containing different 3'-terminal coding regions. Therefore, APA alters both the repertoire and the expression level of proteins. Here, we used high-throughput sequencing data to map polyA sites and characterize polyadenylation genome-wide in three SI-NETs and a reference sample. In the tumors, 16 genes showed significant changes of APA pattern, which lead to either the 3' truncation of mRNA coding regions or 3' untranslated regions. Among these, 11 genes had been previously associated with cancer, with 4 genes being known tumor suppressors: DCC, PDZD2, MAGI1, and DACT2. We validated the APA in three out of three cases with quantitative real-time-PCR. Our findings suggest that changes of APA pattern in these 16 genes could be involved in the tumorigenesis of SI-NETs. Furthermore, they also point to APA as a new target for both diagnostic and treatment of SI-NETs. The identified genes with APA specific to the SI-NETs could be further tested as diagnostic markers and drug targets for disease prevention and treatment.

Alternative Polyadenylation of Tumor Suppressor Genes in Small Intestinal Neuroendocrine Tumors

PubMed Central

Rehfeld, Anders; Plass, Mireya; Døssing, Kristina; Knigge, Ulrich; Kjær, Andreas; Krogh, Anders; Friis-Hansen, Lennart

2014-01-01

The tumorigenesis of small intestinal neuroendocrine tumors (SI-NETs) is poorly understood. Recent studies have associated alternative polyadenylation (APA) with proliferation, cell transformation, and cancer. Polyadenylation is the process in which the pre-messenger RNA is cleaved at a polyA site and a polyA tail is added. Genes with two or more polyA sites can undergo APA. This produces two or more distinct mRNA isoforms with different 3′ untranslated regions. Additionally, APA can also produce mRNAs containing different 3′-terminal coding regions. Therefore, APA alters both the repertoire and the expression level of proteins. Here, we used high-throughput sequencing data to map polyA sites and characterize polyadenylation genome-wide in three SI-NETs and a reference sample. In the tumors, 16 genes showed significant changes of APA pattern, which lead to either the 3′ truncation of mRNA coding regions or 3′ untranslated regions. Among these, 11 genes had been previously associated with cancer, with 4 genes being known tumor suppressors: DCC, PDZD2, MAGI1, and DACT2. We validated the APA in three out of three cases with quantitative real-time-PCR. Our findings suggest that changes of APA pattern in these 16 genes could be involved in the tumorigenesis of SI-NETs. Furthermore, they also point to APA as a new target for both diagnostic and treatment of SI-NETs. The identified genes with APA specific to the SI-NETs could be further tested as diagnostic markers and drug targets for disease prevention and treatment. PMID:24782827

Specific Tandem 3'UTR Patterns and Gene Expression Profiles in Mouse Thy1+ Germline Stem Cells

PubMed Central

Lin, Zhuoheng; Feng, Xuyang; Jiang, Xue; Songyang, Zhou; Huang, Junjiu

2015-01-01

A recently developed strategy of sequencing alternative polyadenylation (APA) sites (SAPAS) with second-generation sequencing technology can be used to explore complete genome-wide patterns of tandem APA sites and global gene expression profiles. spermatogonial stem cells (SSCs) maintain long-term reproductive abilities in male mammals. The detailed mechanisms by which SSCs self-renew and generate mature spermatozoa are not clear. To understand the specific alternative polyadenylation pattern and global gene expression profile of male germline stem cells (GSCs, mainly referred to SSCs here), we isolated and purified mouse Thy1+ cells from testis by magnetic-activated cell sorting (MACS) and then used the SAPAS method for analysis, using pluripotent embryonic stem cells (ESCs) and differentiated mouse embryonic fibroblast cells (MEFs) as controls. As a result, we obtained 99,944 poly(A) sites, approximately 40% of which were newly detected in our experiments. These poly(A) sites originated from three mouse cell types and covered 17,499 genes, including 831 long non-coding RNA (lncRNA) genes. We observed that GSCs tend to have shorter 3'UTR lengths while MEFs tend towards longer 3'UTR lengths. We also identified 1337 genes that were highly expressed in GSCs, and these genes were highly consistent with the functional characteristics of GSCs. Our detailed bioinformatics analysis identified APA site-switching events at 3'UTRs and many new specifically expressed genes in GSCs, which we experimentally confirmed. Furthermore, qRT-PCR was performed to validate several events of the 334 genes with distal-to-proximal poly(A) switch in GSCs. Consistently APA reporter assay confirmed the total 3'UTR shortening in GSCs compared to MEFs. We also analyzed the cis elements around the proximal poly(A) site preferentially used in GSCs and found C-rich elements may contribute to this regulation. Overall, our results identified the expression level and polyadenylation site profiles and these data provide new insights into the processes potentially involved in the GSC life cycle and spermatogenesis. PMID:26713853

3′ UTR lengthening as a novel mechanism in regulating cellular senescence

PubMed Central

Chen, Meng; Lyu, Guoliang; Han, Miao; Nie, Hongbo; Shen, Ting; Chen, Wei; Niu, Yichi; Song, Yifan; Li, Xueping; Li, Huan; Chen, Xinyu; Wang, Ziyue; Xia, Zheng; Li, Wei; Tian, Xiao-Li; Ding, Chen; Gu, Jun; Zheng, Yufang; Liu, Xinhua; Hu, Jinfeng; Wei, Gang; Tao, Wei

2018-01-01

Cellular senescence has been viewed as a tumor suppression mechanism and also as a contributor to individual aging. Widespread shortening of 3′ untranslated regions (3′ UTRs) in messenger RNAs (mRNAs) by alternative polyadenylation (APA) has recently been discovered in cancer cells. However, the role of APA in the process of cellular senescence remains elusive. Here, we found that hundreds of genes in senescent cells tended to use distal poly(A) (pA) sites, leading to a global lengthening of 3′ UTRs and reduced gene expression. Genes that harbor longer 3′ UTRs in senescent cells were enriched in senescence-related pathways. Rras2, a member of the Ras superfamily that participates in multiple signal transduction pathways, preferred longer 3′ UTR usage and exhibited decreased expression in senescent cells. Depletion of Rras2 promoted senescence, while rescue of Rras2 reversed senescence-associated phenotypes. Mechanistically, splicing factor TRA2B bound to a core “AGAA” motif located in the alternative 3′ UTR of Rras2, thereby reducing the RRAS2 protein level and causing senescence. Both proximal and distal poly(A) signals showed strong sequence conservation, highlighting the vital role of APA regulation during evolution. Our results revealed APA as a novel mechanism in regulating cellular senescence. PMID:29440281

Promoter-Terminator Gene Loops Affect Alternative 3'-End Processing in Yeast.

PubMed

Lamas-Maceiras, Mónica; Singh, Badri Nath; Hampsey, Michael; Freire-Picos, María A

2016-04-22

Many eukaryotic genes undergo alternative 3'-end poly(A)-site selection producing transcript isoforms with 3'-UTRs of different lengths and post-transcriptional fates. Gene loops are dynamic structures that juxtapose the 3'-ends of genes with their promoters. Several functions have been attributed to looping, including memory of recent transcriptional activity and polarity of transcription initiation. In this study, we investigated the relationship between gene loops and alternative poly(A)-site. Using the KlCYC1 gene of the yeast Kluyveromyces lactis, which includes a single promoter and two poly(A) sites separated by 394 nucleotides, we demonstrate in two yeast species the formation of alternative gene loops (L1 and L2) that juxtapose the KlCYC1 promoter with either proximal or distal 3'-end processing sites, resulting in the synthesis of short and long forms of KlCYC1 mRNA. Furthermore, synthesis of short and long mRNAs and formation of the L1 and L2 loops are growth phase-dependent. Chromatin immunoprecipitation experiments revealed that the Ssu72 RNA polymerase II carboxyl-terminal domain phosphatase, a critical determinant of looping, peaks in early log phase at the proximal poly(A) site, but as growth phase advances, it extends to the distal site. These results define a cause-and-effect relationship between gene loops and alternative poly(A) site selection that responds to different physiological signals manifested by RNA polymerase II carboxyl-terminal domain phosphorylation status. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

In silico Analysis of 3′-End-Processing Signals in Aspergillus oryzae Using Expressed Sequence Tags and Genomic Sequencing Data

PubMed Central

Tanaka, Mizuki; Sakai, Yoshifumi; Yamada, Osamu; Shintani, Takahiro; Gomi, Katsuya

2011-01-01

To investigate 3′-end-processing signals in Aspergillus oryzae, we created a nucleotide sequence data set of the 3′-untranslated region (3′ UTR) plus 100 nucleotides (nt) sequence downstream of the poly(A) site using A. oryzae expressed sequence tags and genomic sequencing data. This data set comprised 1065 sequences derived from 1042 unique genes. The average 3′ UTR length in A. oryzae was 241 nt, which is greater than that in yeast but similar to that in plants. The 3′ UTR and 100 nt sequence downstream of the poly(A) site is notably U-rich, while the region located 15–30 nt upstream of the poly(A) site is markedly A-rich. The most frequently found hexanucleotide in this A-rich region is AAUGAA, although this sequence accounts for only 6% of all transcripts. These data suggested that A. oryzae has no highly conserved sequence element equivalent to AAUAAA, a mammalian polyadenylation signal. We identified that putative 3′-end-processing signals in A. oryzae, while less well conserved than those in mammals, comprised four sequence elements: the furthest upstream U-rich element, A-rich sequence, cleavage site, and downstream U-rich element flanking the cleavage site. Although these putative 3′-end-processing signals are similar to those in yeast and plants, some notable differences exist between them. PMID:21586533

Novel mechanism of conjoined gene formation in the human genome.

PubMed

Kim, Ryong Nam; Kim, Aeri; Choi, Sang-Haeng; Kim, Dae-Soo; Nam, Seong-Hyeuk; Kim, Dae-Won; Kim, Dong-Wook; Kang, Aram; Kim, Min-Young; Park, Kun-Hyang; Yoon, Byoung-Ha; Lee, Kang Seon; Park, Hong-Seog

2012-03-01

Recently, conjoined genes (CGs) have emerged as important genetic factors necessary for understanding the human genome. However, their formation mechanism and precise structures have remained mysterious. Based on a detailed structural analysis of 57 human CG transcript variants (CGTVs, discovered in this study) and all (833) known CGs in the human genome, we discovered that the poly(A) signal site from the upstream parent gene region is completely removed via the skipping or truncation of the final exon; consequently, CG transcription is terminated at the poly(A) signal site of the downstream parent gene. This result led us to propose a novel mechanism of CG formation: the complete removal of the poly(A) signal site from the upstream parent gene is a prerequisite for the CG transcriptional machinery to continue transcribing uninterrupted into the intergenic region and downstream parent gene. The removal of the poly(A) signal sequence from the upstream gene region appears to be caused by a deletion or truncation mutation in the human genome rather than post-transcriptional trans-splicing events. With respect to the characteristics of CG sequence structures, we found that intergenic regions are hot spots for novel exon creation during CGTV formation and that exons farther from the intergenic regions are more highly conserved in the CGTVs. Interestingly, many novel exons newly created within the intergenic and intragenic regions originated from transposable element sequences. Additionally, the CGTVs showed tumor tissue-biased expression. In conclusion, our study provides novel insights into the CG formation mechanism and expands the present concepts of the genetic structural landscape, gene regulation, and gene formation mechanisms in the human genome.

Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro

PubMed Central

Jablonski, Joseph; Clementz, Mark; Ryan, Kevin; Valente, Susana T.

2014-01-01

The 3’ end of mammalian mRNAs is not formed by abrupt termination of transcription by RNA polymerase II (RNPII). Instead, RNPII synthesizes precursor mRNA beyond the end of mature RNAs, and an active process of endonuclease activity is required at a specific site. Cleavage of the precursor RNA normally occurs 10-30 nt downstream from the consensus polyA site (AAUAAA) after the CA dinucleotides. Proteins from the cleavage complex, a multifactorial protein complex of approximately 800 kDa, accomplish this specific nuclease activity. Specific RNA sequences upstream and downstream of the polyA site control the recruitment of the cleavage complex. Immediately after cleavage, pre-mRNAs are polyadenylated by the polyA polymerase (PAP) to produce mature stable RNA messages. Processing of the 3’ end of an RNA transcript may be studied using cellular nuclear extracts with specific radiolabeled RNA substrates. In sum, a long 32P-labeled uncleaved precursor RNA is incubated with nuclear extracts in vitro, and cleavage is assessed by gel electrophoresis and autoradiography. When proper cleavage occurs, a shorter 5’ cleaved product is detected and quantified. Here, we describe the cleavage assay in detail using, as an example, the 3’ end processing of HIV-1 mRNAs. PMID:24835792

Poly(A) polymerase contains multiple functional domains.

PubMed Central

Raabe, T; Murthy, K G; Manley, J L

1994-01-01

Poly(A) polymerase (PAP) contains regions of similarity with several known protein domains. Through site-directed mutagenesis, we provide evidence that PAP contains a functional ribonucleoprotein-type RNA binding domain (RBD) that is responsible for primer binding, making it the only known polymerase to contain such a domain. The RBD is adjacent to, and probably overlaps with, an apparent catalytic region responsible for polymerization. Despite the presence of sequence similarities, this catalytic domain appears to be distinct from the conserved polymerase module found in a large number of RNA-dependent polymerases. PAP contains two nuclear localization signals (NLSs) in its C terminus, each by itself similar to the consensus bipartite NLS found in many nuclear proteins. Mutagenesis experiments indicate that both signals, which are separated by nearly 140 residues, play important roles in directing PAP exclusively to the nucleus. Surprisingly, basic amino acids in the N-terminal-most NLS are also essential for AAUAAA-dependent polyadenylation but not for nonspecific poly(A) synthesis, suggesting that this region of PAP is involved in interactions both with nuclear targeting proteins and with nuclear polyadenylation factors. The serine/threonine-rich C terminus is multiply phosphorylated, including at sites affected by mutations in either NLS. Images PMID:8164653

Poly(A) code analyses reveal key determinants for tissue-specific mRNA alternative polyadenylation

PubMed Central

Weng, Lingjie; Li, Yi; Xie, Xiaohui; Shi, Yongsheng

2016-01-01

mRNA alternative polyadenylation (APA) is a critical mechanism for post-transcriptional gene regulation and is often regulated in a tissue- and/or developmental stage-specific manner. An ultimate goal for the APA field has been to be able to computationally predict APA profiles under different physiological or pathological conditions. As a first step toward this goal, we have assembled a poly(A) code for predicting tissue-specific poly(A) sites (PASs). Based on a compendium of over 600 features that have known or potential roles in PAS selection, we have generated and refined a machine-learning algorithm using multiple high-throughput sequencing-based data sets of tissue-specific and constitutive PASs. This code can predict tissue-specific PASs with >85% accuracy. Importantly, by analyzing the prediction performance based on different RNA features, we found that PAS context, including the distance between alternative PASs and the relative position of a PAS within the gene, is a key feature for determining the susceptibility of a PAS to tissue-specific regulation. Our poly(A) code provides a useful tool for not only predicting tissue-specific APA regulation, but also for studying its underlying molecular mechanisms. PMID:27095026

Genome-Wide Association Study of a Validated Case Definition of Gulf War Illness in a Population-Representative Sample

DTIC Science & Technology

2013-09-01

sequence dataset. All procedures were performed by personnel in the IIMT UT Southwestern Genomics and Microarray Core using standard protocols. More... sequencing run, samples were demultiplexed using standard algorithms in the Genomics and Microarray Core and processed into individual sample Illumina single... Sequencing (RNA-Seq), using Illumina’s multiplexing mRNA-Seq to generate full sequence libraries from the poly-A tailed RNA to a read depth of 30

UV-VIS extinction spectra of gold particle coated by oligonucleotide shell

NASA Astrophysics Data System (ADS)

Bogatyrev, Vladimir A.; Vrublevsky, Stanislav A.; Trachuk, Lyubov A.; Khlebtsov, Nikolai G.

2005-06-01

We describe synthesis process of an oligonucleotide-functionalized colloidal gold marker CG-l5-T28, its optical properties and interaction with poly(A) in solution and on a solid-phase substrate. The marker is a complex of 15 nm diameter colloidal gold nanoparticles with covalently attached 5'-thiolated 28-base oligothymidine macromolecules. A positive hybridization reaction of the marker with poly(A) is observed by solid-phase analysis on hanging a spot color (from red to blue ) or on appearance of a red dye in dot-blot test as compared to control experiments with poly(U) target. The principles of spectrophotometric monitoring all stages of the marker preparation and application of spectrophotometry to detection of the polynucleotide hybridization in vitro are described. Experimental data were compared with theoretical calculations based on Mie theory for 2-layer model of gold core in polymeric shell with imaginary part of refractive index that typical for the real absorption spectra of NA. To explain the aggregation of CG-15-T28 caused by interaction with poly(A) in solution, we suggest a new model differing from a standard model of cross-linker binding.

A comprehensive analysis of 3′ end sequencing data sets reveals novel polyadenylation signals and the repressive role of heterogeneous ribonucleoprotein C on cleavage and polyadenylation

PubMed Central

Gruber, Andreas J.; Schmidt, Ralf; Gruber, Andreas R.; Martin, Georges; Ghosh, Souvik; Belmadani, Manuel; Keller, Walter

2016-01-01

Alternative polyadenylation (APA) is a general mechanism of transcript diversification in mammals, which has been recently linked to proliferative states and cancer. Different 3′ untranslated region (3′ UTR) isoforms interact with different RNA-binding proteins (RBPs), which modify the stability, translation, and subcellular localization of the corresponding transcripts. Although the heterogeneity of pre-mRNA 3′ end processing has been established with high-throughput approaches, the mechanisms that underlie systematic changes in 3′ UTR lengths remain to be characterized. Through a uniform analysis of a large number of 3′ end sequencing data sets, we have uncovered 18 signals, six of which are novel, whose positioning with respect to pre-mRNA cleavage sites indicates a role in pre-mRNA 3′ end processing in both mouse and human. With 3′ end sequencing we have demonstrated that the heterogeneous ribonucleoprotein C (HNRNPC), which binds the poly(U) motif whose frequency also peaks in the vicinity of polyadenylation (poly(A)) sites, has a genome-wide effect on poly(A) site usage. HNRNPC-regulated 3′ UTRs are enriched in ELAV-like RBP 1 (ELAVL1) binding sites and include those of the CD47 gene, which participate in the recently discovered mechanism of 3′ UTR–dependent protein localization (UDPL). Our study thus establishes an up-to-date, high-confidence catalog of 3′ end processing sites and poly(A) signals, and it uncovers an important role of HNRNPC in regulating 3′ end processing. It further suggests that U-rich elements mediate interactions with multiple RBPs that regulate different stages in a transcript's life cycle. PMID:27382025

Human Fip1 is a subunit of CPSF that binds to U-rich RNA elements and stimulates poly(A) polymerase.

PubMed

Kaufmann, Isabelle; Martin, Georges; Friedlein, Arno; Langen, Hanno; Keller, Walter

2004-02-11

In mammals, polyadenylation of mRNA precursors (pre-mRNAs) by poly(A) polymerase (PAP) depends on cleavage and polyadenylation specificity factor (CPSF). CPSF is a multisubunit complex that binds to the canonical AAUAAA hexamer and to U-rich upstream sequence elements on the pre-mRNA, thereby stimulating the otherwise weakly active and nonspecific polymerase to elongate efficiently RNAs containing a poly(A) signal. Based on sequence similarity to the Saccharomyces cerevisiae polyadenylation factor Fip1p, we have identified human Fip1 (hFip1) and found that the protein is an integral subunit of CPSF. hFip1 interacts with PAP and has an arginine-rich RNA-binding motif that preferentially binds to U-rich sequence elements on the pre-mRNA. Recombinant hFip1 is sufficient to stimulate the in vitro polyadenylation activity of PAP in a U-rich element-dependent manner. hFip1, CPSF160 and PAP form a ternary complex in vitro, suggesting that hFip1 and CPSF160 act together in poly(A) site recognition and in cooperative recruitment of PAP to the RNA. These results show that hFip1 significantly contributes to CPSF-mediated stimulation of PAP activity.

Human Fip1 is a subunit of CPSF that binds to U-rich RNA elements and stimulates poly(A) polymerase

PubMed Central

Kaufmann, Isabelle; Martin, Georges; Friedlein, Arno; Langen, Hanno; Keller, Walter

2004-01-01

In mammals, polyadenylation of mRNA precursors (pre-mRNAs) by poly(A) polymerase (PAP) depends on cleavage and polyadenylation specificity factor (CPSF). CPSF is a multisubunit complex that binds to the canonical AAUAAA hexamer and to U-rich upstream sequence elements on the pre-mRNA, thereby stimulating the otherwise weakly active and nonspecific polymerase to elongate efficiently RNAs containing a poly(A) signal. Based on sequence similarity to the Saccharomyces cerevisiae polyadenylation factor Fip1p, we have identified human Fip1 (hFip1) and found that the protein is an integral subunit of CPSF. hFip1 interacts with PAP and has an arginine-rich RNA-binding motif that preferentially binds to U-rich sequence elements on the pre-mRNA. Recombinant hFip1 is sufficient to stimulate the in vitro polyadenylation activity of PAP in a U-rich element-dependent manner. hFip1, CPSF160 and PAP form a ternary complex in vitro, suggesting that hFip1 and CPSF160 act together in poly(A) site recognition and in cooperative recruitment of PAP to the RNA. These results show that hFip1 significantly contributes to CPSF-mediated stimulation of PAP activity. PMID:14749727

Changes in RNA polymerase II progression influence somatic hypermutation of Ig-related genes by AID

PubMed Central

Kodgire, Prashant; Mukkawar, Priyanka; Ratnam, Sarayu; Martin, Terence E.

2013-01-01

Somatic hypermutation (SHM) of Ig genes is initiated by the activation-induced cytidine deaminase (AID), and requires target gene transcription. We previously proposed that AID may associate with the RNA polymerase II (Pol). Here, to determine aspects of the transcription process required for SHM, we knocked-in a transcription terminator into an Ig gene variable region in DT40 chicken B cell line. We found that the human β-globin terminator was an efficient inhibitor of downstream transcription in these cells. The terminator reduced mutations downstream of the poly(A) signal, suggesting that the process of transcription is essential for efficient SHM and that AID has better access to its target when Pol is in the elongating rather than terminating mode. Mutations upstream of the poly(A) site were almost doubled in the active terminator clones compared with an inactivated terminator, and this region showed more single-stranded DNA, indicating that Pol pausing assists SHM. Moreover, the nontranscribed DNA strand was the preferred SHM target upstream of the active terminator. Pol pausing during poly(A) site recognition may facilitate persistence of negative supercoils, exposing the coding single strand and possibly allowing the nascent RNA intermittent reannealing with the template strand, for prolonged access of AID. PMID:23752228

siRNAs targeted to certain polyadenylation sites promote specific, RISC-independent degradation of messenger RNAs.

PubMed

Vickers, Timothy A; Crooke, Stanley T

2012-07-01

While most siRNAs induce sequence-specific target mRNA cleavage and degradation in a process mediated by Ago2/RNA-induced silencing complex (RISC), certain siRNAs have also been demonstrated to direct target RNA reduction through deadenylation and subsequent degradation of target transcripts in a process which involves Ago1/RISC and P-bodies. In the current study, we present data suggesting that a third class of siRNA exist, which are capable of promoting target RNA reduction that is independent of both Ago and RISC. These siRNAs bind the target messenger RNA at the polyA signal and are capable of redirecting a small amount of polyadenylation to downstream polyA sites when present, however, the majority of the activity appears to be due to inhibition of polyadenylation or deadenylation of the transcript, followed by exosomal degradation of the immature mRNA.

Back to basics: the untreated rabbit reticulocyte lysate as a competitive system to recapitulate cap/poly(A) synergy and the selective advantage of IRES-driven translation.

PubMed

Soto Rifo, Ricardo; Ricci, Emiliano P; Décimo, Didier; Moncorgé, Olivier; Ohlmann, Théophile

2007-01-01

Translation of most eukaryotic mRNAs involves the synergistic action between the 5' cap structure and the 3' poly(A) tail at the initiation step. The poly(A) tail has also been shown to stimulate translation of picornavirus internal ribosome entry sites (IRES)-directed translation. These effects have been attributed principally to interactions between eIF4G and poly(A)-binding protein (PABP) but also to the participation of PABP in other steps during translation initiation. As the rabbit reticulocyte lysate (RRL) does not recapitulate this cap/poly(A) synergy, several systems based on cellular cell-free extracts have been developed to study the effects of poly(A) tail in vitro but they generally exhibit low translational efficiency. Here, we describe that the non-nuclease-treated RRL (untreated RRL) is able to recapitulate the effects of poly(A) tail on translation in vitro. In this system, translation of a capped/polyadenylated RNA was specifically inhibited by either Paip2 or poly(rA), whereas translation directed by HCV IRES remained unaffected. Moreover, cleavage of eIF4G by FMDV L protease strongly stimulated translation directed by the EMCV IRES, thus recapitulating the competitive advantage that the proteolytic processing of eIF4G confers to IRES-driven RNAs.

Spectroscopic study of a DNA brush synthesized in situ by surface initiated enzymatic polymerization.

PubMed

Khan, M Nuruzzaman; Tjong, Vinalia; Chilkoti, Ashutosh; Zharnikov, Michael

2013-08-29

We used a combination of synchrotron-based X-ray photoelectron spectroscopy (XPS) and angle-resolved near-edge X-ray absorption fine structure (NEXAFS) spectroscopy to study the chemical integrity, purity, and possible internal alignment of single-strand (ss) adenine deoxynucleotide (poly(A)) DNA brushes. The brushes were synthesized by surface-initiated enzymatic polymerization (SIEP) on a 25-mer of adenine self-assembled monolayer (SAM) on gold (A25-SH), wherein the terminal 3'-OH of the A25-SH serve as the initiation sites for SIEP of poly(A). XPS and NEXAFS spectra of poly(A) brushes were found to be almost identical to those of A25-SH initiator, with no unambiguous traces of contamination. Apart from the well-defined chemical integrity and contamination-free character, the brushes were found to have a high degree of orientational order, with an upright orientation of individual strands, despite their large thickness up to ~55 nm, that corresponds to a chain length of at least several hundred nucleotides for individual ssDNA molecules. The orientational order exhibited by these poly(A) DNA brushes, mediated presumably by base stacking, was found to be independent of the brush thickness as long as the packing density was high enough. The well-defined character and orientational ordering of the ssDNA brushes make them a potentially promising system for different applications.

PABP is not essential for microRNA-mediated translational repression and deadenylation in vitro

PubMed Central

Fukaya, Takashi; Tomari, Yukihide

2011-01-01

MicroRNAs silence their complementary target genes via formation of the RNA-induced silencing complex (RISC) that contains an Argonaute (Ago) protein at its core. It was previously proposed that GW182, an Ago-associating protein, directly binds to poly(A)-binding protein (PABP) and interferes with its function, leading to silencing of the target mRNAs. Here we show that Drosophila Ago1-RISC induces silencing via two independent pathways: shortening of the poly(A) tail and pure repression of translation. Our data suggest that although PABP generally modulates poly(A) length and translation efficiency, neither PABP function nor GW182–PABP interaction is a prerequisite for these two silencing pathways. Instead, we propose that each of the multiple functional domains within GW182 has a potential for silencing, and yet they need to act together in the context of full-length GW182 to exert maximal silencing. PMID:22117217

Complex and dynamic landscape of RNA polyadenylation revealed by PAS-Seq

PubMed Central

Shepard, Peter J.; Choi, Eun-A; Lu, Jente; Flanagan, Lisa A.; Hertel, Klemens J.; Shi, Yongsheng

2011-01-01

Alternative polyadenylation (APA) of mRNAs has emerged as an important mechanism for post-transcriptional gene regulation in higher eukaryotes. Although microarrays have recently been used to characterize APA globally, they have a number of serious limitations that prevents comprehensive and highly quantitative analysis. To better characterize APA and its regulation, we have developed a deep sequencing-based method called Poly(A) Site Sequencing (PAS-Seq) for quantitatively profiling RNA polyadenylation at the transcriptome level. PAS-Seq not only accurately and comprehensively identifies poly(A) junctions in mRNAs and noncoding RNAs, but also provides quantitative information on the relative abundance of polyadenylated RNAs. PAS-Seq analyses of human and mouse transcriptomes showed that 40%–50% of all expressed genes produce alternatively polyadenylated mRNAs. Furthermore, our study detected evolutionarily conserved polyadenylation of histone mRNAs and revealed novel features of mitochondrial RNA polyadenylation. Finally, PAS-Seq analyses of mouse embryonic stem (ES) cells, neural stem/progenitor (NSP) cells, and neurons not only identified more poly(A) sites than what was found in the entire mouse EST database, but also detected significant changes in the global APA profile that lead to lengthening of 3′ untranslated regions (UTR) in many mRNAs during stem cell differentiation. Together, our PAS-Seq analyses revealed a complex landscape of RNA polyadenylation in mammalian cells and the dynamic regulation of APA during stem cell differentiation. PMID:21343387

Evidence that Poly(A) Binding Protein C1 Binds Nuclear Pre-mRNA Poly(A) Tails

PubMed Central

Hosoda, Nao; Lejeune, Fabrice; Maquat, Lynne E.

2006-01-01

In mammalian cells, poly(A) binding protein C1 (PABP C1) has well-known roles in mRNA translation and decay in the cytoplasm. However, PABPC1 also shuttles in and out of the nucleus, and its nuclear function is unknown. Here, we show that PABPC1, like the major nuclear poly(A) binding protein PABPN1, associates with nuclear pre-mRNAs that are polyadenylated and intron containing. PABPC1 does not bind nonpolyadenylated histone mRNA, indicating that the interaction of PABPC1 with pre-mRNA requires a poly(A) tail. Consistent with this conclusion, UV cross-linking results obtained using intact cells reveal that PABPC1 binds directly to pre-mRNA poly(A) tails in vivo. We also show that PABPC1 immunopurifies with poly(A) polymerase, suggesting that PABPC1 is acquired by polyadenylated transcripts during poly(A) tail synthesis. Our findings demonstrate that PABPC1 associates with polyadenylated transcripts earlier in mammalian mRNA biogenesis than previously thought and offer insights into the mechanism by which PABPC1 is recruited to newly synthesized poly(A). Our results are discussed in the context of pre-mRNA processing and stability and mRNA trafficking and the pioneer round of translation. PMID:16581783

Inhibition of Poly(A)-binding protein with a synthetic RNA mimic reduces pain sensitization in mice.

PubMed

Barragán-Iglesias, Paulino; Lou, Tzu-Fang; Bhat, Vandita D; Megat, Salim; Burton, Michael D; Price, Theodore J; Campbell, Zachary T

2018-01-02

Nociceptors rely on cap-dependent translation to rapidly induce protein synthesis in response to pro-inflammatory signals. Comparatively little is known regarding the role of the regulatory factors bound to the 3' end of mRNA in nociceptor sensitization. Poly(A)-binding protein (PABP) stimulates translation initiation by bridging the Poly(A) tail to the eukaryotic initiation factor 4F complex associated with the mRNA cap. Here, we use unbiased assessment of PABP binding specificity to generate a chemically modified RNA-based competitive inhibitor of PABP. The resulting RNA mimic, which we designated as the Poly(A) SPOT-ON, is more stable than unmodified RNA and binds PABP with high affinity and selectivity in vitro. We show that injection of the Poly(A) SPOT-ON at the site of an injury can attenuate behavioral response to pain. Collectively, these results suggest that PABP is integral for nociceptive plasticity. The general strategy described here provides a broad new source of mechanism-based inhibitors for RNA-binding proteins and is applicable for in vivo studies.

The 5'-poly(A) leader of poxvirus mRNA confers a translational advantage that can be achieved in cells with impaired cap-dependent translation

PubMed Central

Dhungel, Pragyesh; Cao, Shuai

2017-01-01

The poly(A) leader at the 5’-untranslated region (5’-UTR) is an unusually striking feature of all poxvirus mRNAs transcribed after viral DNA replication (post-replicative mRNAs). These poly(A) leaders are non-templated and of heterogeneous lengths; and their function during poxvirus infection remains a long-standing question. Here, we discovered that a 5’-poly(A) leader conferred a selective translational advantage to mRNA in poxvirus-infected cells. A constitutive and uninterrupted 5’-poly(A) leader with 12 residues was optimal. Because the most frequent lengths of the 5’-poly(A) leaders are 8–12 residues, the result suggests that the poly(A) leader has been evolutionarily optimized to boost poxvirus protein production. A 5’-poly(A) leader also could increase protein production in the bacteriophage T7 promoter-based expression system of vaccinia virus, the prototypic member of poxviruses. Interestingly, although vaccinia virus post-replicative mRNAs do have 5’- methylated guanosine caps and can use cap-dependent translation, in vaccinia virus-infected cells, mRNA with a 5’-poly(A) leader could also be efficiently translated in cells with impaired cap-dependent translation. However, the translation was not mediated through an internal ribosome entry site (IRES). These results point to a fundamental mechanism poxvirus uses to efficiently translate its post-replicative mRNAs. PMID:28854224

The Geographic Distribution of Human Y Chromosome Variation

PubMed Central

Hammer, M. F.; Spurdle, A. B.; Karafet, T.; Bonner, M. R.; Wood, E. T.; Novelletto, A.; Malaspina, P.; Mitchell, R. J.; Horai, S.; Jenkins, T.; Zegura, S. L.

1997-01-01

We examined variation on the nonrecombining portion of the human Y chromosome to investigate human evolution during the last 200,000 years. The Y-specific polymorphic sites included the Y Alu insertional polymorphism or ``YAP'' element (DYS287), the poly(A) tail associated with the YAP element, three point mutations in close association with the YAP insertion site, an A-G polymorphic transition (DYS271), and a tetranucleotide microsatellite (DYS19). Global variation at the five bi-allelic sites (DYS271, DYS287, and the three point mutations) gave rise to five ``YAP haplotypes'' in 60 populations from Africa, Europe, Asia, Australasia, and the New World (n = 1500). Combining the multi-allelic variation at the microsatellite loci (poly(A) tail and DYS19) with the YAP haplotypes resulted in a total of 27 ``combination haplotypes''. All five of the YAP haplotypes and 21 of the 27 combination haplotypes were found in African populations, which had greater haplotype diversity than did populations from other geographical locations. Only subsets of the five YAP haplotypes were found outside of Africa. Patterns of observed variation were compatible with a variety of hypotheses, including multiple human migrations and range expansions. PMID:9055088

Characterization of stress-responsive lncRNAs in Arabidopsis thaliana by integrating expression, epigenetic and structural features.

PubMed

Di, Chao; Yuan, Jiapei; Wu, Yue; Li, Jingrui; Lin, Huixin; Hu, Long; Zhang, Ting; Qi, Yijun; Gerstein, Mark B; Guo, Yan; Lu, Zhi John

2014-12-01

Recently, in addition to poly(A)+ long non-coding RNAs (lncRNAs), many lncRNAs without poly(A) tails, have been characterized in mammals. However, the non-polyA lncRNAs and their conserved motifs, especially those associated with environmental stresses, have not been fully investigated in plant genomes. We performed poly(A)- RNA-seq for seedlings of Arabidopsis thaliana under four stress conditions, and predicted lncRNA transcripts. We classified the lncRNAs into three confidence levels according to their expression patterns, epigenetic signatures and RNA secondary structures. Then, we further classified the lncRNAs to poly(A)+ and poly(A)- transcripts. Compared with poly(A)+ lncRNAs and coding genes, we found that poly(A)- lncRNAs tend to have shorter transcripts and lower expression levels, and they show significant expression specificity in response to stresses. In addition, their differential expression is significantly enriched in drought condition and depleted in heat condition. Overall, we identified 245 poly(A)+ and 58 poly(A)- lncRNAs that are differentially expressed under various stress stimuli. The differential expression was validated by qRT-PCR, and the signaling pathways involved were supported by specific binding of transcription factors (TFs), phytochrome-interacting factor 4 (PIF4) and PIF5. Moreover, we found many conserved sequence and structural motifs of lncRNAs from different functional groups (e.g. a UUC motif responding to salt and a AU-rich stem-loop responding to cold), indicated that the conserved elements might be responsible for the stress-responsive functions of lncRNAs. © 2014 The Authors The Plant Journal © 2014 John Wiley & Sons Ltd.

Polyadenylation state microarray (PASTA) analysis.

PubMed

Beilharz, Traude H; Preiss, Thomas

2011-01-01

Nearly all eukaryotic mRNAs terminate in a poly(A) tail that serves important roles in mRNA utilization. In the cytoplasm, the poly(A) tail promotes both mRNA stability and translation, and these functions are frequently regulated through changes in tail length. To identify the scope of poly(A) tail length control in a transcriptome, we developed the polyadenylation state microarray (PASTA) method. It involves the purification of mRNA based on poly(A) tail length using thermal elution from poly(U) sepharose, followed by microarray analysis of the resulting fractions. In this chapter we detail our PASTA approach and describe some methods for bulk and mRNA-specific poly(A) tail length measurements of use to monitor the procedure and independently verify the microarray data.

Sequence variation between 462 human individuals fine-tunes functional sites of RNA processing

NASA Astrophysics Data System (ADS)

Ferreira, Pedro G.; Oti, Martin; Barann, Matthias; Wieland, Thomas; Ezquina, Suzana; Friedländer, Marc R.; Rivas, Manuel A.; Esteve-Codina, Anna; Estivill, Xavier; Guigó, Roderic; Dermitzakis, Emmanouil; Antonarakis, Stylianos; Meitinger, Thomas; Strom, Tim M.; Palotie, Aarno; François Deleuze, Jean; Sudbrak, Ralf; Lerach, Hans; Gut, Ivo; Syvänen, Ann-Christine; Gyllensten, Ulf; Schreiber, Stefan; Rosenstiel, Philip; Brunner, Han; Veltman, Joris; Hoen, Peter A. C. T.; Jan van Ommen, Gert; Carracedo, Angel; Brazma, Alvis; Flicek, Paul; Cambon-Thomsen, Anne; Mangion, Jonathan; Bentley, David; Hamosh, Ada; Rosenstiel, Philip; Strom, Tim M.; Lappalainen, Tuuli; Guigó, Roderic; Sammeth, Michael

2016-09-01

Recent advances in the cost-efficiency of sequencing technologies enabled the combined DNA- and RNA-sequencing of human individuals at the population-scale, making genome-wide investigations of the inter-individual genetic impact on gene expression viable. Employing mRNA-sequencing data from the Geuvadis Project and genome sequencing data from the 1000 Genomes Project we show that the computational analysis of DNA sequences around splice sites and poly-A signals is able to explain several observations in the phenotype data. In contrast to widespread assessments of statistically significant associations between DNA polymorphisms and quantitative traits, we developed a computational tool to pinpoint the molecular mechanisms by which genetic markers drive variation in RNA-processing, cataloguing and classifying alleles that change the affinity of core RNA elements to their recognizing factors. The in silico models we employ further suggest RNA editing can moonlight as a splicing-modulator, albeit less frequently than genomic sequence diversity. Beyond existing annotations, we demonstrate that the ultra-high resolution of RNA-Seq combined from 462 individuals also provides evidence for thousands of bona fide novel elements of RNA processing—alternative splice sites, introns, and cleavage sites—which are often rare and lowly expressed but in other characteristics similar to their annotated counterparts.

Journey into Problem Solving: A Gift from Polya

ERIC Educational Resources Information Center

Lederman, Eric

2009-01-01

In "How to Solve It", accomplished mathematician and skilled communicator George Polya describes a four-step universal solving technique designed to help students develop mathematical problem-solving skills. By providing a glimpse at the grace with which experts solve problems, Polya provides definable methods that are not exclusive to…

In vivo analysis of polyadenylation in prokaryotes.

PubMed

Mohanty, Bijoy K; Kushner, Sidney R

2014-01-01

Polyadenylation at the 3' ends of mRNAs, tRNAs, rRNAs, and sRNAs plays important roles in RNA metabolism in both prokaryotes and eukaryotes. However, the nature of poly(A) tails in prokaryotes is distinct compared to their eukaryotic counterparts. Specifically, depending on the organism, eukaryotic poly(A) tails average between 50 and >200 nt and can easily be isolated by several techniques involving oligo(dT)-dependent cDNA amplification. In contrast, the bulk of the poly(A) tails present on prokaryotic transcripts is relatively short (<10 nt) and is difficult to characterize using similar techniques. This chapter describes methods that can circumvent these problems. For example, we discuss how to isolate total RNA and characterize its overall polyadenylation status employing a poly(A) sizing assay. Furthermore, we describe a technique involving RNase H treatment of total RNA followed by northern analysis in order to distinguish length of poly(A) tails on various types of transcripts. Finally, we outline a useful procedure to clone the poly(A) tails of specific transcripts using 5'-3' end-ligated RNA, which is independent of oligo(dT)-dependent cDNA amplification. These approaches are particularly helpful in analyzing transcripts with either short or long poly(A) tails both in prokaryotes and eukaryotes.

The poly(A) tail length of casein mRNA in the lactating mammary gland changes depending upon the accumulation and removal of milk.

PubMed Central

Kuraishi, T; Sun, Y; Aoki, F; Imakawa, K; Sakai, S

2000-01-01

The length of casein mRNA from the lactating mouse mammary gland, as assessed on Northern blots, is shorter after weaning, but is elongated following the removal of milk. In order to investigate this phenomenon, the molecular structures of beta- and gamma-casein mRNAs were analysed. The coding and non-coding regions of the two forms were the same length, but the long form of casein mRNA had a longer poly(A) tail than the short form (P<0.05). In order to examine the stability of casein mRNA under identical conditions, casein mRNAs with the long and short poly(A) tails were incubated in the rabbit reticulocyte lysate (RRL) cell-free translation system. Casein mRNA with the long poly(A) tail had a longer half-life than that with the short tail (P<0.05). The beta- and gamma-casein mRNAs were first degraded into 0.92 and 0.81 kb fragments respectively. With undegraded mRNA, the poly(A) tail shortening by exoribonuclease was not observed until the end of the incubation. Northern blot analysis showed that casein mRNA with the long poly(A) tail was protected efficiently from endoribonucleases. We conclude that the length of the poly(A) tail of casein mRNA in the lactating mammary gland changes depending upon the accumulation and removal of the gland's milk, and we show that the longer poly(A) tail potentially protects the mRNA from degradation by endoribonucleases. PMID:10749689

The human myelin oligodendrocyte glycoprotein (MOG) gene: Complete nucleotide sequence and structural characterization

DOE Office of Scientific and Technical Information (OSTI.GOV)

Paule Roth, M.; Malfroy, L.; Offer, C.

1995-07-20

Human myelin oligodendrocyte glycoprotein (MOG), a myelin component of the central nervous system, is a candidate target antigen for autoimmune-mediated demyelination. We have isolated and sequenced part of a cosmid clone that contains the entire human MOG gene. The primary nuclear transcript, extending from the putative start of transcription to the site of poly(A) addition, is 15,561 nucleotides in length. The human MOG gene contains 8 exons, separated by 7 introns; canonical intron/exon boundary sites are observed at each junction. The introns vary in size from 242 to 6484 bp and contain numerous repetitive DNA elements, including 14 Alu sequencesmore » within 3 introns. Another Alu element is located in the 3{prime}-untranslated region of the gene. Alu sequences were classified with respect to subfamily assignment. Seven hundred sixty-three nucleotides 5{prime} of the transcription start and 1214 nucleotides 3{prime} of the poly(A) addition sites were also sequenced. The 5{prime}-flanking region revealed the presence of several consensus sequences that could be relevant in the transcription of the MOG gene, in particular binding sites in common with other myelin gene promoters. Two polymorphic intragenic dinucleotide (CA){sub n} and tetranucleotide (TAAA){sub n} repeats were identified and may provide genetic marker tools for association and linkage studies. 50 refs., 3 figs., 3 tabs.« less

Spontaneous and engineered deletions in the 3' noncoding region of tick-borne encephalitis virus: construction of highly attenuated mutants of a flavivirus.

PubMed

Mandl, C W; Holzmann, H; Meixner, T; Rauscher, S; Stadler, P F; Allison, S L; Heinz, F X

1998-03-01

The flavivirus genome is a positive-strand RNA molecule containing a single long open reading frame flanked by noncoding regions (NCR) that mediate crucial processes of the viral life cycle. The 3' NCR of tick-borne encephalitis (TBE) virus can be divided into a variable region that is highly heterogeneous in length among strains of TBE virus and in certain cases includes an internal poly(A) tract and a 3'-terminal conserved core element that is believed to fold as a whole into a well-defined secondary structure. We have now investigated the genetic stability of the TBE virus 3' NCR and its influence on viral growth properties and virulence. We observed spontaneous deletions in the variable region during growth of TBE virus in cell culture and in mice. These deletions varied in size and location but always included the internal poly(A) element of the TBE virus 3' NCR and never extended into the conserved 3'-terminal core element. Subsequently, we constructed specific deletion mutants by using infectious cDNA clones with the entire variable region and increasing segments of the core element removed. A virus mutant lacking the entire variable region was indistinguishable from wild-type virus with respect to cell culture growth properties and virulence in the mouse model. In contrast, even small extensions of the deletion into the core element led to significant biological effects. Deletions extending to nucleotides 10826, 10847, and 10870 caused distinct attenuation in mice without measurable reduction of cell culture growth properties, which, however, were significantly restricted when the deletion was extended to nucleotide 10919. An even larger deletion (to nucleotide 10994) abolished viral viability. In spite of their high degree of attenuation, these mutants efficiently induced protective immune responses even at low inoculation doses. Thus, 3'-NCR deletions represent a useful technique for achieving stable attenuation of flaviviruses that can be included in the rational design of novel flavivirus live vaccines.

Budding yeast telomerase RNA transcription termination is dictated by the Nrd1/Nab3 non-coding RNA termination pathway

PubMed Central

Noël, Jean-François; Larose, Stéphanie; Abou Elela, Sherif; Wellinger, Raymund J.

2012-01-01

The RNA component of budding yeast telomerase (Tlc1) occurs in two forms, a non-polyadenylated form found in functional telomerase and a rare polyadenylated version with unknown function. Previous work suggested that the functional Tlc1 polyA− RNA is processed from the polyA+ form, but the mechanisms regulating its transcription termination and 3′-end formation remained unclear. Here we examined transcription termination of Tlc1 RNA in the sequences 3′ of the TLC1 gene and relate it to telomere maintenance. Strikingly, disruption of all probable or cryptic polyadenylation signals near the 3′-end blocked the accumulation of the previously reported polyA+ RNA without affecting the level, function or specific 3′ nucleotide of the mature polyA− form. A genetic approach analysing TLC1 3′-end sequences revealed that transcription terminates upstream of the polyadenylation sites. Furthermore, the results also demonstrate that the function of this Tlc1 terminator depends on the Nrd1/Nab3 transcription termination pathway. The data thus show that transcription termination of the budding yeast telomerase RNA occurs as that of snRNAs and Tlc1 functions in telomere maintenance are not strictly dependent on a polyadenylated precursor, even if the polyA+ form can serve as intermediate in a redundant termination/maturation pathway. PMID:22379137

Distinct regulation of alternative polyadenylation and gene expression by nuclear poly(A) polymerases

PubMed Central

Li, Wencheng; Laishram, Rakesh S.; Hoque, Mainul; Ji, Zhe

2017-01-01

Abstract Polyadenylation of nascent RNA by poly(A) polymerase (PAP) is important for 3′ end maturation of almost all eukaryotic mRNAs. Most mammalian genes harbor multiple polyadenylation sites (PASs), leading to expression of alternative polyadenylation (APA) isoforms with distinct functions. How poly(A) polymerases may regulate PAS usage and hence gene expression is poorly understood. Here, we show that the nuclear canonical (PAPα and PAPγ) and non-canonical (Star-PAP) PAPs play diverse roles in PAS selection and gene expression. Deficiencies in the PAPs resulted in perturbations of gene expression, with Star-PAP impacting lowly expressed mRNAs and long-noncoding RNAs to the greatest extent. Importantly, different PASs of a gene are distinctly regulated by different PAPs, leading to widespread relative expression changes of APA isoforms. The location and surrounding sequence motifs of a PAS appear to differentiate its regulation by the PAPs. We show Star-PAP-specific PAS usage regulates the expression of the eukaryotic translation initiation factor EIF4A1, the tumor suppressor gene PTEN and the long non-coding RNA NEAT1. The Star-PAP-mediated APA of PTEN is essential for DNA damage-induced increase of PTEN protein levels. Together, our results reveal a PAS-guided and PAP-mediated paradigm for gene expression in response to cellular signaling cues. PMID:28911096

The Apc5 Subunit of the Anaphase-Promoting Complex/Cyclosome Interacts with Poly(A) Binding Protein and Represses Internal Ribosome Entry Site-Mediated Translation

PubMed Central

Koloteva-Levine, Nadejda; Pinchasi, Dalia; Pereman, Idan; Zur, Amit; Brandeis, Michael; Elroy-Stein, Orna

2004-01-01

The anaphase-promoting complex/cyclosome (APC/C) is a multisubunit ubiquitin ligase that mediates the proteolysis of cell cycle proteins in mitosis and G1. We used a yeast three-hybrid screen to identify proteins that interact with the internal ribosome entry site (IRES) of platelet-derived growth factor 2 mRNA. Surprisingly, this screen identified Apc5, although it does not harbor a classical RNA binding domain. We found that Apc5 binds the poly(A) binding protein (PABP), which directly binds the IRES element. PABP was found to enhance IRES-mediated translation, whereas Apc5 overexpression counteracted this effect. In addition to its association with the APC/C complex, Apc5 binds much heavier complexes and cosediments with the ribosomal fraction. In contrast to Apc3, which is associated only with the APC/C and remains intact during differentiation, Apc5 is degraded upon megakaryocytic differentiation in correlation with IRES activation. Expression of Apc5 in differentiated cells abolished IRES activation. This is the first report implying an additional role for an APC/C subunit, apart from its being part of the APC/C complex. PMID:15082755

Comprehensive profiling of rhizome-associated alternative splicing and alternative polyadenylation in moso bamboo (Phyllostachys edulis).

PubMed

Wang, Taotao; Wang, Huiyuan; Cai, Dawei; Gao, Yubang; Zhang, Hangxiao; Wang, Yongsheng; Lin, Chentao; Ma, Liuyin; Gu, Lianfeng

2017-08-01

Moso bamboo (Phyllostachys edulis) represents one of the fastest-spreading plants in the world, due in part to its well-developed rhizome system. However, the post-transcriptional mechanism for the development of the rhizome system in bamboo has not been comprehensively studied. We therefore used a combination of single-molecule long-read sequencing technology and polyadenylation site sequencing (PAS-seq) to re-annotate the bamboo genome, and identify genome-wide alternative splicing (AS) and alternative polyadenylation (APA) in the rhizome system. In total, 145 522 mapped full-length non-chimeric (FLNC) reads were analyzed, resulting in the correction of 2241 mis-annotated genes and the identification of 8091 previously unannotated loci. Notably, more than 42 280 distinct splicing isoforms were derived from 128 667 intron-containing full-length FLNC reads, including a large number of AS events associated with rhizome systems. In addition, we characterized 25 069 polyadenylation sites from 11 450 genes, 6311 of which have APA sites. Further analysis of intronic polyadenylation revealed that LTR/Gypsy and LTR/Copia were two major transposable elements within the intronic polyadenylation region. Furthermore, this study provided a quantitative atlas of poly(A) usage. Several hundred differential poly(A) sites in the rhizome-root system were identified. Taken together, these results suggest that post-transcriptional regulation may potentially have a vital role in the underground rhizome-root system. © 2017 The Authors The Plant Journal © 2017 John Wiley & Sons Ltd.

A nonradioactive assay for poly(a)-specific ribonuclease activity by methylene blue colorimetry.

PubMed

Cheng, Yuan; Liu, Wei-Feng; Yan, Yong-Bin; Zhou, Hai-Meng

2006-01-01

A simple nonradioactive assay, which was based on the specific shift of the absorbance maximum of methylene blue induced by its intercalation into poly(A) molecules, was developed for poly(A)-specific ribonuclease (PARN). A good linear relationship was found between the absorbance at 662 nm and the poly(A) concentration. The assay conditions, including the concentration of methylene blue, the incubation temperature and time, and the poly(A) concentration were evaluated and optimized.

RNA binding protein and binding site useful for expression of recombinant molecules

DOEpatents

Mayfield, Stephen P.

2006-10-17

The present invention relates to a gene expression system in eukaryotic and prokaryotic cells, preferably plant cells and intact plants. In particular, the invention relates to an expression system having a RB47 binding site upstream of a translation initiation site for regulation of translation mediated by binding of RB47 protein, a member of the poly(A) binding protein family. Regulation is further effected by RB60, a protein disulfide isomerase. The expression system is capable of functioning in the nuclear/cytoplasm of cells and in the chloroplast of plants. Translation regulation of a desired molecule is enhanced approximately 100 fold over that obtained without RB47 binding site activation.

RNA binding protein and binding site useful for expression of recombinant molecules

DOEpatents

Mayfield, Stephen

2000-01-01

The present invention relates to a gene expression system in eukaryotic and prokaryotic cells, preferably plant cells and intact plants. In particular, the invention relates to an expression system having a RB47 binding site upstream of a translation initiation site for regulation of translation mediated by binding of RB47 protein, a member of the poly(A) binding protein family. Regulation is further effected by RB60, a protein disulfide isomerase. The expression system is capable of functioning in the nuclear/cytoplasm of cells and in the chloroplast of plants. Translation regulation of a desired molecule is enhanced approximately 100 fold over that obtained without RB47 binding site activation.

Expression of eukaryotic polypeptides in chloroplasts

DOEpatents

Mayfield, Stephen P.

2013-06-04

The present invention relates to a gene expression system in eukaryotic and prokaryotic cells, preferably plant cells and intact plants. In particular, the invention relates to an expression system having a RB47 binding site upstream of a translation initiation site for regulation of translation mediated by binding of RB47 protein, a member of the poly(A) binding protein family. Regulation is further effected by RB60, a protein disulfide isomerase. The expression system is capable of functioning in the nuclear/cytoplasm of cells and in the chloroplast of plants. Translation regulation of a desired molecule is enhanced approximately 100 fold over that obtained without RB47 binding site activation.

Optimization of Streptomyces bacteriophage phi C31 integrase system to prevent post integrative gene silencing in pulmonary type II cells.

PubMed

Aneja, Manish Kumar; Geiger, Johannes; Imker, Rabea; Uzgun, Senta; Kormann, Michael; Hasenpusch, Guenther; Maucksch, Christof; Rudolph, Carsten

2009-12-31

phi C31 integrase has emerged as a potent tool for achieving long-term gene expression in different tissues. The present study aimed at optimizing elements of phi C31 integrase system for alveolar type II cells. Luciferase and beta-galactosidase activities were measured at different time points post transfection. 5-Aza-2'deoxycytidine (AZA) and trichostatin A (TSA) were used to inhibit DNA methyltransferase and histone deacetylase complex (HDAC) respectively. In A549 cells, expression of the integrase using a CMV promoter resulted in highest integrase activity, whereas in MLE12 cells, both CAG and CMV promoter were equally effective. Effect of polyA site was observed only in A549 cells, where replacement of SV40 polyA by bovine growth hormone (BGH) polyA site resulted in an enhancement of integrase activity. Addition of a C-terminal SV40 nuclear localization signal (NLS) did not result in any significant increase in integrase activity. Long-term expression studies with AZA and TSA, provided evidence for post-integrative gene silencing. In MLE12 cells, both DNA methylases and HDACs played a significant role in silencing, whereas in A549 cells, it could be attributed majorly to HDAC activity. Donor plasmids comprising cellular promoters ubiquitin B (UBB), ubiquitin C (UCC) and elongation factor 1 alpha (EF1 alpha) in an improved backbone prevented post-integrative gene silencing. In contrast to A549 and MLE12 cells, no silencing could be observed in human bronchial epithelial cells, BEAS-2B. Donor plasmid coding for murine erythropoietin under the EF1 alpha promoter when combined with phi C31 integrase resulted in higher long-term erythropoietin expression and subsequently higher hematocrit levels in mice after intravenous delivery to the lungs. These results provide evidence for cell specific post integrative gene silencing with C31 integrase and demonstrate the pivotal role of donor plasmid in long-term expression attained with this system.

Interaction of phenazinium dyes with double-stranded poly(A): Spectroscopy and isothermal titration calorimetry studies

NASA Astrophysics Data System (ADS)

Khan, Asma Yasmeen; Saha, Baishakhi; Kumar, Gopinatha Suresh

2014-10-01

A comprehensive study on the binding of phenazinium dyes viz. janus green B, indoine blue, safranine O and phenosafranine with double stranded poly(A) using various spectroscopic and calorimetric techniques is presented. A higher binding of janus green B and indoine blue over safranine O and phenosafranine to poly(A) was observed from all experiments. Intercalative mode of binding of the dyes was inferred from fluorescence polarization anisotropy, iodide quenching and viscosity experiments. Circular dichroism study revealed significant perturbation of the secondary structure of poly(A) on binding of these dyes. Results from isothermal titration calorimetry experiments suggested that the binding was predominantly entropy driven with a minor contribution of enthalpy to the standard molar Gibbs energy. The results presented here may open new opportunities in the application of these dyes as RNA targeted therapeutic agents.

Structural basis of UGUA recognition by the Nudix protein CFIm25 and implications for a regulatory role in mRNA 3′ processing

PubMed Central

Yang, Qin; Gilmartin, Gregory M.; Doublié, Sylvie

2010-01-01

Human Cleavage Factor Im (CFIm) is an essential component of the pre-mRNA 3′ processing complex that functions in the regulation of poly(A) site selection through the recognition of UGUA sequences upstream of the poly(A) site. Although the highly conserved 25 kDa subunit (CFIm25) of the CFIm complex possesses a characteristic α/β/α Nudix fold, CFIm25 has no detectable hydrolase activity. Here we report the crystal structures of the human CFIm25 homodimer in complex with UGUAAA and UUGUAU RNA sequences. CFIm25 is the first Nudix protein to be reported to bind RNA in a sequence-specific manner. The UGUA sequence contributes to binding specificity through an intramolecular G:A Watson–Crick/sugar-edge base interaction, an unusual pairing previously found to be involved in the binding specificity of the SAM-III riboswitch. The structures, together with mutational data, suggest a novel mechanism for the simultaneous sequence-specific recognition of two UGUA elements within the pre-mRNA. Furthermore, the mutually exclusive binding of RNA and the signaling molecule Ap4A (diadenosine tetraphosphate) by CFIm25 suggests a potential role for small molecules in the regulation of mRNA 3′ processing. PMID:20479262

Structural basis of UGUA recognition by the Nudix protein CFI(m)25 and implications for a regulatory role in mRNA 3' processing.

PubMed

Yang, Qin; Gilmartin, Gregory M; Doublié, Sylvie

2010-06-01

Human Cleavage Factor Im (CFI(m)) is an essential component of the pre-mRNA 3' processing complex that functions in the regulation of poly(A) site selection through the recognition of UGUA sequences upstream of the poly(A) site. Although the highly conserved 25 kDa subunit (CFI(m)25) of the CFI(m) complex possesses a characteristic alpha/beta/alpha Nudix fold, CFI(m)25 has no detectable hydrolase activity. Here we report the crystal structures of the human CFI(m)25 homodimer in complex with UGUAAA and UUGUAU RNA sequences. CFI(m)25 is the first Nudix protein to be reported to bind RNA in a sequence-specific manner. The UGUA sequence contributes to binding specificity through an intramolecular G:A Watson-Crick/sugar-edge base interaction, an unusual pairing previously found to be involved in the binding specificity of the SAM-III riboswitch. The structures, together with mutational data, suggest a novel mechanism for the simultaneous sequence-specific recognition of two UGUA elements within the pre-mRNA. Furthermore, the mutually exclusive binding of RNA and the signaling molecule Ap(4)A (diadenosine tetraphosphate) by CFI(m)25 suggests a potential role for small molecules in the regulation of mRNA 3' processing.

hnRNP L controls HPV16 RNA polyadenylation and splicing in an Akt kinase-dependent manner

PubMed Central

Kajitani, Naoko; Glahder, Jacob; Wu, Chengjun; Yu, Haoran; Nilsson, Kersti

2017-01-01

Abstract Inhibition of the Akt kinase activates HPV16 late gene expression by reducing HPV16 early polyadenylation and by activating HPV16 late L1 mRNA splicing. We identified ‘hot spots’ for RNA binding proteins at the early polyA signal and at splice sites on HPV16 late mRNAs. We observed that hnRNP L was associated with sequences at all HPV16 late splice sites and at the early polyA signal. Akt kinase inhibition resulted in hnRNP L dephosphorylation and reduced association of hnRNP L with HPV16 mRNAs. This was accompanied by an increased binding of U2AF65 and Sam68 to HPV16 mRNAs. Furthermore, siRNA knock-down of hnRNP L or Akt induced HPV16 gene expression. Treatment of HPV16 immortalized keratinocytes with Akt kinase inhibitor reduced hnRNP L binding to HPV16 mRNAs and induced HPV16 L1 mRNA production. Finally, deletion of the hnRNP L binding sites in HPV16 subgenomic expression plasmids resulted in activation of HPV16 late gene expression. In conclusion, the Akt kinase inhibits HPV16 late gene expression at the level of RNA processing by controlling the RNA-binding protein hnRNP L. We speculate that Akt kinase activity upholds an intracellular milieu that favours HPV16 early gene expression and suppresses HPV16 late gene expression. PMID:28934469

Influence of nucleotide modifications at the C2’ position on the Hoogsteen base-paired parallel-stranded duplex of poly(A) RNA

PubMed Central

Copp, William; Denisov, Alexey Y.; Xie, Jingwei; Noronha, Anne M.; Liczner, Christopher; Safaee, Nozhat

2017-01-01

Abstract Polyadenylate (poly(A)) has the ability to form a parallel duplex with Hoogsteen adenine:adenine base pairs at low pH or in the presence of ammonium ions. In order to evaluate the potential of this structural motif for nucleic acid-based nanodevices, we characterized the effects on duplex stability of substitutions of the ribose sugar with 2′-deoxyribose, 2′-O-methyl-ribose, 2′-deoxy-2′-fluoro-ribose, arabinose and 2′-deoxy-2′-fluoro-arabinose. Deoxyribose substitutions destabilized the poly(A) duplex both at low pH and in the presence of ammonium ions: no duplex formation could be detected with poly(A) DNA oligomers. Other sugar C2’ modifications gave a variety of effects. Arabinose and 2′-deoxy-2′-fluoro-arabinose nucleotides strongly destabilized poly(A) duplex formation. In contrast, 2′-O-methyl and 2′-deoxy-2′-fluoro-ribo modifications were stabilizing either at pH 4 or in the presence of ammonium ions. The differential effect suggests they could be used to design molecules selectively responsive to pH or ammonium ions. To understand the destabilization by deoxyribose, we determined the structures of poly(A) duplexes with a single DNA residue by nuclear magnetic resonance spectroscopy and X-ray crystallography. The structures revealed minor structural perturbations suggesting that the combination of sugar pucker propensity, hydrogen bonding, pKa shifts and changes in hydration determine duplex stability. PMID:28973475

Influence of nucleotide modifications at the C2' position on the Hoogsteen base-paired parallel-stranded duplex of poly(A) RNA.

PubMed

Copp, William; Denisov, Alexey Y; Xie, Jingwei; Noronha, Anne M; Liczner, Christopher; Safaee, Nozhat; Wilds, Christopher J; Gehring, Kalle

2017-09-29

Polyadenylate (poly(A)) has the ability to form a parallel duplex with Hoogsteen adenine:adenine base pairs at low pH or in the presence of ammonium ions. In order to evaluate the potential of this structural motif for nucleic acid-based nanodevices, we characterized the effects on duplex stability of substitutions of the ribose sugar with 2'-deoxyribose, 2'-O-methyl-ribose, 2'-deoxy-2'-fluoro-ribose, arabinose and 2'-deoxy-2'-fluoro-arabinose. Deoxyribose substitutions destabilized the poly(A) duplex both at low pH and in the presence of ammonium ions: no duplex formation could be detected with poly(A) DNA oligomers. Other sugar C2' modifications gave a variety of effects. Arabinose and 2'-deoxy-2'-fluoro-arabinose nucleotides strongly destabilized poly(A) duplex formation. In contrast, 2'-O-methyl and 2'-deoxy-2'-fluoro-ribo modifications were stabilizing either at pH 4 or in the presence of ammonium ions. The differential effect suggests they could be used to design molecules selectively responsive to pH or ammonium ions. To understand the destabilization by deoxyribose, we determined the structures of poly(A) duplexes with a single DNA residue by nuclear magnetic resonance spectroscopy and X-ray crystallography. The structures revealed minor structural perturbations suggesting that the combination of sugar pucker propensity, hydrogen bonding, pKa shifts and changes in hydration determine duplex stability. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

Mechanisms and consequences of alternative polyadenylation

PubMed Central

Di Giammartino, Dafne Campigli; Nishida, Kensei; Manley, James L.

2011-01-01

Summary Alternative polyadenylation (APA) is emerging as a widespread mechanism used to control gene expression. Like alternative splicing, usage of alternative poly(A) sites allows a single gene to encode multiple mRNA transcripts. In some cases, this changes the mRNA coding potential; in other cases, the code remains unchanged but the 3’UTR length is altered, influencing the fate of mRNAs in several ways, for example, by altering the availability of RNA binding protein sites and microRNA binding sites. The mechansims governing both global and gene-specific APA are only starting to be deciphered. Here we review what is known about these mechanisms and the functional consequences of alternative polyadenlyation. PMID:21925375

Histone-poly(A) hybrid molecules as tools to block nuclear pores.

PubMed

Cremer, G; Wojtech, E; Kalbas, M; Agutter, P S; Prochnow, D

1995-04-01

Histone-poly(A) hybrid molecules were used for transport experiments with resealed nuclear envelopes and after attachment of a cleavable cross-linker (SASD) to identify nuclear proteins. In contrast to histones, the hybrid molecules cannot be accumulated in resealed nuclear envelopes, and in contrast to poly(A), the export of hybrids from preloaded nuclear envelopes is completely impaired. The experiments strongly confirm the existence of poly(A) as an export signal in mRNA which counteracts the nuclear location signals (NLS) in histones. The contradicting transport signals in the hybrid molecules impair translocation through the nuclear pore complex. The failure to accumulate hybrid molecules into resealed nuclear envelopes results from the covalent attachment of polyadenylic acid to histones in a strict 1:1 molar ratio. This was demonstrated in control transport experiments where radiolabeled histones were simply mixed with nonlabeled poly(A) or radiolabeled poly(A) mixed with nonlabeled histones. In comparison, control uptake experiments with histones covalently linked to a single UMP-mononucleotide are strongly enhanced. Such controls exclude the conceivable possibility of a simple masking of the nuclear location signal in the histones by the covalent attached poly(A) moiety. Photoreactive histone-poly(A) hybrid analogs serve to identify nuclear envelope proteins--presumably in the nuclear pore--with molecular weights of 110, 80, and 71.4 kDa.

Fluorescence in-situ hybridization method reveals that carboxyl-terminal fragments of transactive response DNA-binding protein-43 truncated at the amino acid residue 218 reduce poly(A)+ RNA expression.

PubMed

Higashi, Shinji; Watanabe, Ryohei; Arai, Tetsuaki

2018-07-04

Transactive response (TAR) DNA-binding protein 43 (TDP-43) has emerged as an important contributor to amyotrophic lateral sclerosis and frontotemporal lobar degeneration. To understand the association of TDP-43 with complex RNA processing in disease pathogenesis, we performed fluorescence in-situ hybridization using HeLa cells transfected with a series of deleted TDP-43 constructs and investigated the effect of truncation of TDP-43 on the expression of poly(A) RNA. Endogenous and overexpressed full-length TDP-43 localized to the perichromatin region and interchromatin space adjacent to poly(A) RNA. Deleted variants of TDP-43 containing RNA recognition motif 1 and truncating N-terminal region induced cytoplasmic inclusions in which poly(A) RNA was recruited. Carboxyl-terminal TDP-43 truncated at residue 202 or 218 was distributed in the cytoplasm as punctate structures. Carboxyl-terminal TDP-43 truncated at residue 218, but not at 202, significantly decreased poly(A) RNA expression by ∼24% compared with the level in control cells. Our results suggest that the disturbance of RNA metabolism induced by pathogenic fragments plays central roles in the pathogenesis of amyotrophic lateral sclerosis and frontotemporal lobar degeneration.

Transcript Lifetime Is Balanced between Stabilizing Stem-Loop Structures and Degradation-Promoting Polyadenylation in Plant Mitochondria

PubMed Central

Kuhn, Josef; Tengler, Ulrike; Binder, Stefan

2001-01-01

To determine the influence of posttranscriptional modifications on 3′ end processing and RNA stability in plant mitochondria, pea atp9 and Oenothera atp1 transcripts were investigated for the presence and function of 3′ nonencoded nucleotides. A 3′ rapid amplification of cDNA ends approach initiated at oligo(dT)-adapter primers finds the expected poly(A) tails predominantly attached within the second stem or downstream of the double stem-loop structures at sites of previously mapped 3′ ends. Functional studies in a pea mitochondrial in vitro processing system reveal a rapid removal of the poly(A) tails up to termini at the stem-loop structure but little if any influence on further degradation of the RNA. In contrast 3′ poly(A) tracts at RNAs without such stem-loop structures significantly promote total degradation in vitro. To determine the in vivo identity of 3′ nonencoded nucleotides more accurately, pea atp9 transcripts were analyzed by a direct anchor primer ligation-reverse transcriptase PCR approach. This analysis identified maximally 3-nucleotide-long nonencoded extensions most frequently of adenosines combined with cytidines. Processing assays with substrates containing homopolymer stretches of different lengths showed that 10 or more adenosines accelerate RNA processivity, while 3 adenosines have no impact on RNA life span. Thus polyadenylation can generally stimulate the decay of RNAs, but processivity of degradation is almost annihilated by the stabilizing effect of the stem-loop structures. These antagonistic actions thus result in the efficient formation of 3′ processed and stable transcripts. PMID:11154261

[Polyadenylated RNA and mRNA export factors in extrachromosomal nuclear domains of vitellogenic oocytes of the insect Tenebrio molitor].

PubMed

Bogoliubov, D S; Kiselev, A M; Shabel'nikov, S V; Parfenov, V N

2012-01-01

The nucleus ofvitellogenic oocytes of the yellow mealworm, Tenebrio molitor, contains a karyosphere that consists of the condensed chromatin embedded in an extrachromosomal fibrogranular material. Numerous nuclear bodies located freely in the nucleoplasm are also observed. Amongst these bodies, counterparts of nuclear speckles (= interchromatin granule clusters, IGCs) can be identified by the presence of the marker protein SC35. Microinjections of fluorescently tagged methyloligoribonucleotide probes 2'-O-Me(U)22, complementary to poly(A) tails of RNAs, revealed poly(A)+ RNA in the vast majority of IGCs. We found that all T. molitor oocyte IGCs contain heterogeneous ribonucleoprotein (hnRNP) core protein Al that localizes to IGCs in an RNA-dependent manner. The extrachromosomal material of the karyosphere and a part of nucleoplasmic IGCs also contain the adapter protein Aly that is known to provide a link between pre-mRNA splicing and mRNA export. The essential mRNA export factor/receptor NXF1 was observed to colocalize with Aly. In nucleoplasmic IGCs, NXF1 was found to localize in an RNA-dependent manner whereas it is RNA-independently located in the extrachromosomal material of the karyosphere. We believe our data suggest on a role of the nucleoplasmic IGCs in mRNA biogenesis and retention in a road to nuclear export.

A deadenylase assay by size-exclusion chromatography.

PubMed

He, Guang-Jun; Yan, Yong-Bin

2012-01-01

The shortening of the 3'-end poly(A) tail, also called deadenylation, is crucial to the regulation of mRNA processing, transportation, translation and degradation. The deadenylation process is achieved by deadenylases, which specifically catalyze the removal of the poly(A) tail at the 3'-end of eukaryotic mRNAs and release 5'-AMP as the product. To achieve their physiological functions, all deadenylases have numerous binding partners that may regulate their catalytic properties or recruit them into various protein complexes. To study the effects of various partners, it is important to develop new deadenylase assay that can be applied either in vivo or in vitro. In this research, we developed the deadenylase assay by the size-exclusion chromatography (SEC) method. The SEC analysis indicated that the poly(A) or oligo(A) substrate and the product AMP could be successfully separated and quantified. The enzymatic parameters of deadenylase could be obtained by quantifying the AMP generation. When using the commercial poly(A) as the substrate, a biphasic catalytic process was observed, which might correlate to the two distinct states of poly(A) in the commercial samples. Different lots of commercial poly(A) had dissimilar size distributions and were dissimilar in response to the degradation of deadenylase. The deadenylation pattern, processive or distributive, could also be investigated using the SEC assay by monitoring the status of the substrate and the generation kinetics of AMP and A2. The SEC assay was applicable to both simple samples using the purified enzyme and complex enzyme reaction conditions such as using protein mixtures or crude cell extracts as samples. The influence of solutes with absorption at 254 nm could be successfully eliminated by constructing the different SEC profiles.

Human mRNA polyadenylate binding protein: evolutionary conservation of a nucleic acid binding motif.

PubMed Central

Grange, T; de Sa, C M; Oddos, J; Pictet, R

1987-01-01

We have isolated a full length cDNA (cDNA) coding for the human poly(A) binding protein. The cDNA derived 73 kd basic translation product has the same Mr, isoelectric point and peptidic map as the poly(A) binding protein. DNA sequence analysis reveals a 70,244 dalton protein. The N terminal part, highly homologous to the yeast poly(A) binding protein, is sufficient for poly(A) binding activity. This domain consists of a four-fold repeated unit of approximately 80 amino acids present in other nucleic acid binding proteins. In the C terminal part there is, as in the yeast protein, a sequence of approximately 150 amino acids, rich in proline, alanine and glutamine which together account for 48% of the residues. A 2,9 kb mRNA corresponding to this cDNA has been detected in several vertebrate cell types and in Drosophila melanogaster at every developmental stage including oogenesis. Images PMID:2885805

Equivalent Colorings with "Maple"

ERIC Educational Resources Information Center

Cecil, David R.; Wang, Rongdong

2005-01-01

Many counting problems can be modeled as "colorings" and solved by considering symmetries and Polya's cycle index polynomial. This paper presents a "Maple 7" program link http://users.tamuk.edu/kfdrc00/ that, given Polya's cycle index polynomial, determines all possible associated colorings and their partitioning into equivalence classes. These…

Distinct solvent- and temperature-dependent packing arrangements of anti-parallel β-sheet polyalanines studied with solid-state 13C NMR and MD simulation.

PubMed

Kametani, Shunsuke; Tasei, Yugo; Nishimura, Akio; Asakura, Tetsuo

2017-08-09

Polyalanine (polyA) sequences are well known as the simplest sequence that naturally forms anti-parallel β-sheets and constitute a key element in the structure of spider and wild silkworm silk fibers. We have carried out a systematic analysis of the packing of anti-parallel β-sheets for (Ala) n , n = 5, 6, 7 and 12, using primarily 13 C solid-state NMR and MD simulation. HFIP and TFA are frequently used as the dope solvents for recombinant silks, and polyA was solidified from both HFIP and TFA solutions by drying. An analysis of Ala Cβ peaks in the 13 C CP/MAS NMR spectra indicated that polyA from HFIP was mainly rectangular but polyA from TFA was mainly staggered. The transition from the rectangular to the staggered arrangement in (Ala) 6 was observed for the first time from the change in the Ala Cβ peak through heat treatment at 200 °C for 4 h. The removal of the bound water was confirmed by thermal analysis. This transition could be reproduced by MD simulation of (Ala) 6 molecules at 200 °C after removal of the bound water molecules. In this way, the origin of the stability of the different packing arrangements of polyA was clarified.

Evaluation of two main RNA-seq approaches for gene quantification in clinical RNA sequencing: polyA+ selection versus rRNA depletion.

PubMed

Zhao, Shanrong; Zhang, Ying; Gamini, Ramya; Zhang, Baohong; von Schack, David

2018-03-19

To allow efficient transcript/gene detection, highly abundant ribosomal RNAs (rRNA) are generally removed from total RNA either by positive polyA+ selection or by rRNA depletion (negative selection) before sequencing. Comparisons between the two methods have been carried out by various groups, but the assessments have relied largely on non-clinical samples. In this study, we evaluated these two RNA sequencing approaches using human blood and colon tissue samples. Our analyses showed that rRNA depletion captured more unique transcriptome features, whereas polyA+ selection outperformed rRNA depletion with higher exonic coverage and better accuracy of gene quantification. For blood- and colon-derived RNAs, we found that 220% and 50% more reads, respectively, would have to be sequenced to achieve the same level of exonic coverage in the rRNA depletion method compared with the polyA+ selection method. Therefore, in most cases we strongly recommend polyA+ selection over rRNA depletion for gene quantification in clinical RNA sequencing. Our evaluation revealed that a small number of lncRNAs and small RNAs made up a large fraction of the reads in the rRNA depletion RNA sequencing data. Thus, we recommend that these RNAs are specifically depleted to improve the sequencing depth of the remaining RNAs.

Transcriptome dynamics through alternative polyadenylation in developmental and environmental responses in plants revealed by deep sequencing

PubMed Central

Shen, Yingjia; Venu, R.C.; Nobuta, Kan; Wu, Xiaohui; Notibala, Varun; Demirci, Caghan; Meyers, Blake C.; Wang, Guo-Liang; Ji, Guoli; Li, Qingshun Q.

2011-01-01

Polyadenylation sites mark the ends of mRNA transcripts. Alternative polyadenylation (APA) may alter sequence elements and/or the coding capacity of transcripts, a mechanism that has been demonstrated to regulate gene expression and transcriptome diversity. To study the role of APA in transcriptome dynamics, we analyzed a large-scale data set of RNA “tags” that signify poly(A) sites and expression levels of mRNA. These tags were derived from a wide range of tissues and developmental stages that were mutated or exposed to environmental treatments, and generated using digital gene expression (DGE)–based protocols of the massively parallel signature sequencing (MPSS-DGE) and the Illumina sequencing-by-synthesis (SBS-DGE) sequencing platforms. The data offer a global view of APA and how it contributes to transcriptome dynamics. Upon analysis of these data, we found that ∼60% of Arabidopsis genes have multiple poly(A) sites. Likewise, ∼47% and 82% of rice genes use APA, supported by MPSS-DGE and SBS-DGE tags, respectively. In both species, ∼49%–66% of APA events were mapped upstream of annotated stop codons. Interestingly, 10% of the transcriptomes are made up of APA transcripts that are differentially distributed among developmental stages and in tissues responding to environmental stresses, providing an additional level of transcriptome dynamics. Examples of pollen-specific APA switching and salicylic acid treatment-specific APA clearly demonstrated such dynamics. The significance of these APAs is more evident in the 3034 genes that have conserved APA events between rice and Arabidopsis. PMID:21813626

Teaching and Learning. A Problem-Solving Focus.

ERIC Educational Resources Information Center

Curcio, Frances R., Ed.

This book is dedicated to George Polya, who focused on problem solving as the means for teaching and learning mathematics. The first chapter is a reprint of his article "On Learning, Teaching, and Learning Teaching." Then, G. L. Alexanderson paints a portrait of "George Polya, Teacher," including some anecdotes that exemplify…

Modified nucleoside dependent Watson-Crick and wobble codon binding by tRNALysUUU species.

PubMed

Yarian, C; Marszalek, M; Sochacka, E; Malkiewicz, A; Guenther, R; Miskiewicz, A; Agris, P F

2000-11-07

Nucleoside modifications are important to the structure of all tRNAs and are critical to the function of some tRNA species. The transcript of human tRNA(Lys3)(UUU) with a UUU anticodon, and the corresponding anticodon stem and loop domain (ASL(Lys3)(UUU)), are unable to bind to poly-A programmed ribosomes. To determine if specific anticodon domain modified nucleosides of tRNA(Lys) species would restore ribosomal binding and also affect thermal stability, we chemically synthesized ASL(Lys) heptadecamers and site-specifically incorporated the anticodon domain modified nucleosides pseudouridine (Psi(39)), 5-methylaminomethyluridine (mnm(5)U(34)) and N6-threonylcarbamoyl-adenosine (t(6)A(37)). Incorporation of t(6)A(37) and mnm(5)U(34) contributed structure to the anticodon loop, apparent by increases in DeltaS, and significantly enhanced the ability of ASL(Lys3)(UUU) to bind poly-A programmed ribosomes. Neither ASL(Lys3)(UUU)-t(6)A(37) nor ASL(Lys3)(UUU)-mnm(5)U(34) bound AAG programmed ribosomes. Only the presence of both t(6)A(37) and mnm(5)U(34) enabled ASL(Lys3)(UUU) to bind AAG programmed ribosomes, as well as increased its affinity for poly-A programmed ribosomes to the level of native Escherichia coli tRNA(Lys). The completely unmodified anticodon stem and loop of human tRNA(Lys1,2)(CUU) with a wobble position-34 C bound AAG, but did not wobble to AAA, even when the ASL was modified with t(6)A(37). The data suggest that tRNA(Lys)(UUU) species require anticodon domain modifications in the loop to impart an ordered structure to the anticodon for ribosomal binding to AAA and require a combination of modified nucleosides to bind AAG.

Reviving Polya's "Look Back" in a Singapore School

ERIC Educational Resources Information Center

Leong, Yew Hoong; Tay, Eng Guan; Toh, Tin Lam; Quek, Khiok Seng; Dindyal, Jaguthsing

2011-01-01

This study is based on the stance that Polya's "Look Back," though understudied, remains relevant to Mathematics curricula that place emphasis on problem solving. Although the Singapore Mathematics curriculum adopts the goal of teaching Look Back, research about how it is carried out in actual classroom practice is rare. In our project,…

An Appropriate Prompts System Based on the Polya Method for Mathematical Problem-Solving

ERIC Educational Resources Information Center

Lee, Chien I.

2017-01-01

Current mathematics education emphasizes techniques, formulas, and procedures, neglecting the importance of understanding, presentation, and reasoning. This turns students into passive listeners that are well-practiced only in using formulas that they do not understand. We therefore adopted the Polya problem-solving method to provide students with…

Evidence for a Complex Class of Nonadenylated mRNA in Drosophila

PubMed Central

Zimmerman, J. Lynn; Fouts, David L.; Manning, Jerry E.

1980-01-01

The amount, by mass, of poly(A+) mRNA present in the polyribosomes of third-instar larvae of Drosophila melanogaster, and the relative contribution of the poly(A+) mRNA to the sequence complexity of total polysomal RNA, has been determined. Selective removal of poly(A+) mRNA from total polysomal RNA by use of either oligo-dT-cellulose, or poly(U)-sepharose affinity chromatography, revealed that only 0.15% of the mass of the polysomal RNA was present as poly(A+) mRNA. The present study shows that this RNA hybridized at saturation with 3.3% of the single-copy DNA in the Drosophila genome. After correction for asymmetric transcription and reactability of the DNA, 7.4% of the single-copy DNA in the Drosophila genome is represented in larval poly(A+) mRNA. This corresponds to 6.73 x 106 nucleotides of mRNA coding sequences, or approximately 5,384 diverse RNA sequences of average size 1,250 nucleotides. However, total polysomal RNA hybridizes at saturation to 10.9% of the single-copy DNA sequences. After correcting this value for asymmetric transcription and tracer DNA reactability, 24% of the single-copy DNA in Drosophila is represented in total polysomal RNA. This corresponds to 2.18 x 107 nucleotides of RNA coding sequences or 17,440 diverse RNA molecules of size 1,250 nucleotides. This value is 3.2 times greater than that observed for poly(A+) mRNA, and indicates that ≃69% of the polysomal RNA sequence complexity is contributed by nonadenylated RNA. Furthermore, if the number of different structural genes represented in total polysomal RNA is ≃1.7 x 104, then the number of genes expressed in third-instar larvae exceeds the number of chromomeres in Drosophila by about a factor of three. This numerology indicates that the number of chromomeres observed in polytene chromosomes does not reflect the number of structural gene sequences in the Drosophila genome. PMID:6777246

Relooking "Look Back": A Student's Attempt at Problem Solving Using Polya's Model

ERIC Educational Resources Information Center

Leong, Yew Hoong; Toh, Tin Lam; Tay, Eng Guan; Quek, Khiok Seng; Dindyal, Jaguthsing

2012-01-01

Against the backdrop of half a century of research in mathematics problem solving, Polya's last stage is especially conspicuous--by the scarcity of research on it! Much of the research focused on the first three stages (J.M. Francisco and C.A. Maher, "Conditions for promoting reasoning in problem solving: Insights from a longitudinal…

Mathematics in the Making: Mapping Verbal Discourse in Polya's "Let Us Teach Guessing" Lesson

ERIC Educational Resources Information Center

Truxaw, Mary P.; DeFranco, Thomas C.

2007-01-01

This paper describes a detailed analysis of verbal discourse within an exemplary mathematics lesson--that is, George Polya teaching in the Mathematics Association of America [MAA] video classic, "Let Us Teach Guessing" (1966). The results of the analysis reveal an inductive model of teaching that represents recursive cycles rather than linear…

Effect of Polya Problem-Solving Model on Senior Secondary School Students' Performance in Current Electricity

ERIC Educational Resources Information Center

Olaniyan, Ademola Olatide; Omosewo, Esther O.; Nwankwo, Levi I.

2015-01-01

This study was designed to investigate the Effect of Polya Problem-Solving Model on Senior School Students' Performance in Current Electricity. It was a quasi experimental study of non- randomized, non equivalent pre-test post-test control group design. Three research questions were answered and corresponding three research hypotheses were tested…

The Effects of Polya's Heuristic and Diary Writing on Children's Problem Solving

ERIC Educational Resources Information Center

Hensberry, Karina K. R.; Jacobbe, Tim

2012-01-01

This paper presents the results of a study that aimed at increasing students' problem-solving skills. Polya's (1985) heuristic for problem solving was used and students were required to articulate their thought processes through the use of a structured diary. The diary prompted students to answer questions designed to engage them in the phases of…

An Improved Estimation Using Polya-Gamma Augmentation for Bayesian Structural Equation Models with Dichotomous Variables

ERIC Educational Resources Information Center

Kim, Seohyun; Lu, Zhenqiu; Cohen, Allan S.

2018-01-01

Bayesian algorithms have been used successfully in the social and behavioral sciences to analyze dichotomous data particularly with complex structural equation models. In this study, we investigate the use of the Polya-Gamma data augmentation method with Gibbs sampling to improve estimation of structural equation models with dichotomous variables.…

Poly(A) tail length regulates PABPC1 expression to tune translation in the heart.

PubMed

Chorghade, Sandip; Seimetz, Joseph; Emmons, Russell; Yang, Jing; Bresson, Stefan M; Lisio, Michael De; Parise, Gianni; Conrad, Nicholas K; Kalsotra, Auinash

2017-06-27

The rate of protein synthesis in the adult heart is one of the lowest in mammalian tissues, but it increases substantially in response to stress and hypertrophic stimuli through largely obscure mechanisms. Here, we demonstrate that regulated expression of cytosolic poly(A)-binding protein 1 (PABPC1) modulates protein synthetic capacity of the mammalian heart. We uncover a poly(A) tail-based regulatory mechanism that dynamically controls PABPC1 protein synthesis in cardiomyocytes and thereby titrates cellular translation in response to developmental and hypertrophic cues. Our findings identify PABPC1 as a direct regulator of cardiac hypertrophy and define a new paradigm of gene regulation in the heart, where controlled changes in poly(A) tail length influence mRNA translation.

Aberrant termination of reproduction-related TMEM30C transcripts in the hominoids.

PubMed

Osada, Naoki; Hashimoto, Katsuyuki; Hirai, Momoki; Kusuda, Jun

2007-05-01

Finding genetic novelties that may contribute to human-specific physiology and diseases is a key issue of current biomedical studies. TMEM30C is a gene containing two transmembrane (TM) domains and homologous to the yeast CDC50 family, which is related to polarized cell division. It is conserved among mammals along with two other paralogs, TMEM30A and TMEM30B. We found that TMEM30C is expressed specifically in the testis of mammals, in contrast to the relatively wide expression distributions of the other paralogs. While macaques expressed two alternative splicing isoforms which include one or two TM domains, humans and chimpanzees predominantly expressed truncated transcripts because of the mutations in the splicing and/or poly(A) signal sites. The major transcript in humans harbored non-stop ORF (open reading frame) while the chimpanzee counterpart encoded a protein with one TM domain. The difference was due to the 1-bp indel upstream of the poly(A) signal site. In addition, both the hominoids expressed minor transcripts encoding short proteins with one TM domain. Phylogenetic analysis has showed the acceleration of amino acid substitution after the human and chimpanzee divergence, which may have been caused by a recent relaxation in functional constraints or positive selection on TMEM30C. Elucidating the precise reproductive function of TMEM30C in mammals will be important to the foundation of divergence in higher primates at a molecular level.

Symbolic and Verbal Representation Process of Student in Solving Mathematics Problem Based Polya's Stages

ERIC Educational Resources Information Center

Anwar, Rahmad Bustanul; Rahmawati, Dwi

2017-01-01

The purpose of this research was to reveal how the construction process of symbolic representation and verbal representation made by students in problem solving. The construction process in this study referred to the problem-solving stage by Polya covering; 1) understanding the problem, 2) devising a plan, 3) carrying out the plan, and 4) looking…

Students' Errors in Solving the Permutation and Combination Problems Based on Problem Solving Steps of Polya

ERIC Educational Resources Information Center

Sukoriyanto; Nusantara, Toto; Subanji; Chandra, Tjang Daniel

2016-01-01

This article was written based on the results of a study evaluating students' errors in problem solving of permutation and combination in terms of problem solving steps according to Polya. Twenty-five students were asked to do four problems related to permutation and combination. The research results showed that the students still did a mistake in…

Changes in Gene Expression during Tomato Fruit Ripening 1

PubMed Central

Biggs, M. Scott; Harriman, Robert W.; Handa, Avtar K.

1986-01-01

Total proteins from pericarp tissue of different chronological ages from normally ripening tomato (Lycopersicon esculentum Mill. cv Rutgers) fruits and from fruits of the isogenic ripening-impaired mutants rin, nor, and Nr were extracted and separated by sodium dodecylsulfate-polyacrylamide gel electrophoresis. Analysis of the stained bands revealed increases in 5 polypeptides (94, 44, 34, 20, and 12 kilodaltons), decreases in 12 polypeptides (106, 98, 88, 76, 64, 52, 48, 45, 36, 28, 25, and 15 kilodaltons), and fluctuations in 5 polypeptides (85, 60, 26, 21, and 16 kilodaltons) as normal ripening proceeded. Several polypeptides present in ripening normal pericarp exhibited very low or undetectable levels in developing mutant pericarp. Total RNAs extracted from various stages of Rutgers pericarp and from 60 to 65 days old rin, nor, and Nr pericarp were fractionated into poly(A)+ and poly(A)− RNAs. Peak levels of total RNA, poly(A)+ RNA, and poly(A)+ RNA as percent of total RNA occurred between the mature green to breaker stages of normal pericarp. In vitro translation of poly(A)+ RNAs from normal pericarp in rabbit reticulocyte lysates revealed increases in mRNAs for 9 polypeptides (116, 89, 70, 42, 38, 33, 31, 29, and 26 kilodaltons), decreases in mRNAs for 2 polypeptides (41 and 35 kilodaltons), and fluctuations in mRNAs for 5 polypeptides (156, 53, 39, 30, and 14 kilodaltons) during normal ripening. Analysis of two-dimensional separation of in vitro translated polypeptides from poly(A)+ RNAs isolated from different developmental stages revealed even more extensive changes in mRNA populations during ripening. In addition, a polygalacturonase precursor (54 kilodaltons) was immunoprecipitated from breaker, turning, red ripe, and 65 days old Nr in vitro translation products. Images Fig. 1 Fig. 3 Fig. 5 Fig. 6 Fig. 7 PMID:16664828

Dye-induced aggregation of single stranded RNA: a mechanistic approach.

PubMed

Biver, Tarita; Ciatto, Carlo; Secco, Fernando; Venturini, Marcella

2006-08-15

The binding of proflavine (D) to single stranded poly(A) (P) was investigated at pH 7.0 and 25 degrees C using T-jump, stopped-flow and spectrophotometric methods. Equilibrium measurements show that an external complex PD(I) and an internal complex PD(II) form upon reaction between P and D and that their concentrations depend on the polymer/dye concentration ratio (C(P)/C(D)). For C(P)/C(D)<2.5, cooperative formation of stacks external to polymer strands prevails (PD(I)). Equilibria and T-jump experiments, performed at I=0.1M and analyzed according to the Schwarz theory for cooperative binding, provide the values of site size (g=1), equilibrium constant for the nucleation step (K( *)=(1.4+/-0.6)x10(3)M(-1)), equilibrium constant for the growth step (K=(1.2+/-0.6)x10(5)M(-1)), cooperativity parameter (q=85) and rate constants for the growth step (k(r)=1.2x10(7)M(-1)s(-1), k(d)=1.1 x 10(2)s(-1)). Stopped-flow experiments, performed at low ionic strength (I=0.01 M), indicate that aggregation of stacked poly(A) strands do occur provided that C(P)/C(D)<2.5.

CPSF30 at the Interface of Alternative Polyadenylation and Cellular Signaling in Plants

PubMed Central

Chakrabarti, Manohar; Hunt, Arthur G.

2015-01-01

Post-transcriptional processing, involving cleavage of precursor messenger RNA (pre mRNA), and further incorporation of poly(A) tail to the 3' end is a key step in the expression of genetic information. Alternative polyadenylation (APA) serves as an important check point for the regulation of gene expression. Recent studies have shown widespread prevalence of APA in diverse systems. A considerable amount of research has been done in characterizing different subunits of so-called Cleavage and Polyadenylation Specificity Factor (CPSF). In plants, CPSF30, an ortholog of the 30 kD subunit of mammalian CPSF is a key polyadenylation factor. CPSF30 in the model plant Arabidopsis thaliana was reported to possess unique biochemical properties. It was also demonstrated that poly(A) site choice in a vast majority of genes in Arabidopsis are CPSF30 dependent, suggesting a pivotal role of this gene in APA and subsequent regulation of gene expression. There are also indications of this gene being involved in oxidative stress and defense responses and in cellular signaling, suggesting a role of CPSF30 in connecting physiological processes and APA. This review will summarize the biochemical features of CPSF30, its role in regulating APA, and possible links with cellular signaling and stress response modules. PMID:26061761

tRNA nuclear export in saccharomyces cerevisiae: in situ hybridization analysis.

PubMed

Sarkar, S; Hopper, A K

1998-11-01

To understand the factors specifically affecting tRNA nuclear export, we adapted in situ hybridization procedures to locate endogenous levels of individual tRNA families in wild-type and mutant yeast cells. Our studies of tRNAs encoded by genes lacking introns show that nucleoporin Nup116p affects both poly(A) RNA and tRNA export, whereas Nup159p affects only poly(A) RNA export. Los1p is similar to exportin-t, which facilitates vertebrate tRNA export. A los1 deletion mutation affects tRNA but not poly(A) RNA export. The data support the notion that Los1p and exportin-t are functional homologues. Because LOS1 is nonessential, tRNA export in vertebrate and yeast cells likely involves factors in addition to exportin-t. Mutation of RNA1, which encodes RanGAP, causes nuclear accumulation of tRNAs and poly(A) RNA. Many yeast mutants, including those with the rna1-1 mutation, affect both pre-tRNA splicing and RNA export. Our studies of the location of intron-containing pre-tRNAs in the rna1-1 mutant rule out the possibility that this results from tRNA export occurring before splicing. Our results also argue against inappropriate subnuclear compartmentalization causing defects in pre-tRNA splicing. Rather, the data support "feedback" of nucleus/cytosol exchange to the pre-tRNA splicing machinery.

tRNA Nuclear Export in Saccharomyces cerevisiae: In Situ Hybridization Analysis

PubMed Central

Sarkar, Srimonti; Hopper, Anita K.

1998-01-01

To understand the factors specifically affecting tRNA nuclear export, we adapted in situ hybridization procedures to locate endogenous levels of individual tRNA families in wild-type and mutant yeast cells. Our studies of tRNAs encoded by genes lacking introns show that nucleoporin Nup116p affects both poly(A) RNA and tRNA export, whereas Nup159p affects only poly(A) RNA export. Los1p is similar to exportin-t, which facilitates vertebrate tRNA export. A los1 deletion mutation affects tRNA but not poly(A) RNA export. The data support the notion that Los1p and exportin-t are functional homologues. Because LOS1 is nonessential, tRNA export in vertebrate and yeast cells likely involves factors in addition to exportin-t. Mutation of RNA1, which encodes RanGAP, causes nuclear accumulation of tRNAs and poly(A) RNA. Many yeast mutants, including those with the rna1-1 mutation, affect both pre-tRNA splicing and RNA export. Our studies of the location of intron-containing pre-tRNAs in the rna1-1 mutant rule out the possibility that this results from tRNA export occurring before splicing. Our results also argue against inappropriate subnuclear compartmentalization causing defects in pre-tRNA splicing. Rather, the data support “feedback” of nucleus/cytosol exchange to the pre-tRNA splicing machinery. PMID:9802895

Highly narrow nanogap-containing Au@Au core-shell SERS nanoparticles: size-dependent Raman enhancement and applications in cancer cell imaging

NASA Astrophysics Data System (ADS)

Hu, Chongya; Shen, Jianlei; Yan, Juan; Zhong, Jian; Qin, Weiwei; Liu, Rui; Aldalbahi, Ali; Zuo, Xiaolei; Song, Shiping; Fan, Chunhai; He, Dannong

2016-01-01

Cellular imaging technologies employing metallic surface-enhanced Raman scattering (SERS) tags have gained much interest toward clinical diagnostics, but they are still suffering from poor controlled distribution of hot spots and reproducibility of SERS signals. Here, we report the fabrication and characterization of high narrow nanogap-containing Au@Au core-shell SERS nanoparticles (GCNPs) for the identification and imaging of proteins overexpressed on the surface of cancer cells. First, plasmonic nanostructures are made of gold nanoparticles (~15 nm) coated with gold shells, between which a highly narrow and uniform nanogap (~1.1 nm) is formed owing to polyA anchored on the Au cores. The well controlled distribution of Raman reporter molecules, such as 4,4'-dipyridyl (44DP) and 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), are readily encoded in the nanogap and can generate strong, reproducible SERS signals. In addition, we have investigated the size-dependent SERS activity of GCNPs and found that with the same laser wavelength, the Raman enhancement discriminated between particle sizes. The maximum Raman enhancement was achieved at a certain threshold of particle size (~76 nm). High narrow nanogap-containing Au@Au core-shell SERS tags (GCTs) were prepared via the functionalization of hyaluronic acid (HA) on GCNPs, which recognized the CD44 receptor, a tumor-associated surface biomarker. And it was shown that GCTs have a good targeting ability to tumour cells and promising prospects for multiplex biomarker detection.Cellular imaging technologies employing metallic surface-enhanced Raman scattering (SERS) tags have gained much interest toward clinical diagnostics, but they are still suffering from poor controlled distribution of hot spots and reproducibility of SERS signals. Here, we report the fabrication and characterization of high narrow nanogap-containing Au@Au core-shell SERS nanoparticles (GCNPs) for the identification and imaging of proteins overexpressed on the surface of cancer cells. First, plasmonic nanostructures are made of gold nanoparticles (~15 nm) coated with gold shells, between which a highly narrow and uniform nanogap (~1.1 nm) is formed owing to polyA anchored on the Au cores. The well controlled distribution of Raman reporter molecules, such as 4,4'-dipyridyl (44DP) and 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), are readily encoded in the nanogap and can generate strong, reproducible SERS signals. In addition, we have investigated the size-dependent SERS activity of GCNPs and found that with the same laser wavelength, the Raman enhancement discriminated between particle sizes. The maximum Raman enhancement was achieved at a certain threshold of particle size (~76 nm). High narrow nanogap-containing Au@Au core-shell SERS tags (GCTs) were prepared via the functionalization of hyaluronic acid (HA) on GCNPs, which recognized the CD44 receptor, a tumor-associated surface biomarker. And it was shown that GCTs have a good targeting ability to tumour cells and promising prospects for multiplex biomarker detection. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr06919j

Comparative Life Cycle Transcriptomics Revises Leishmania mexicana Genome Annotation and Links a Chromosome Duplication with Parasitism of Vertebrates

PubMed Central

Fiebig, Michael; Kelly, Steven; Gluenz, Eva

2015-01-01

Leishmania spp. are protozoan parasites that have two principal life cycle stages: the motile promastigote forms that live in the alimentary tract of the sandfly and the amastigote forms, which are adapted to survive and replicate in the harsh conditions of the phagolysosome of mammalian macrophages. Here, we used Illumina sequencing of poly-A selected RNA to characterise and compare the transcriptomes of L. mexicana promastigotes, axenic amastigotes and intracellular amastigotes. These data allowed the production of the first transcriptome evidence-based annotation of gene models for this species, including genome-wide mapping of trans-splice sites and poly-A addition sites. The revised genome annotation encompassed 9,169 protein-coding genes including 936 novel genes as well as modifications to previously existing gene models. Comparative analysis of gene expression across promastigote and amastigote forms revealed that 3,832 genes are differentially expressed between promastigotes and intracellular amastigotes. A large proportion of genes that were downregulated during differentiation to amastigotes were associated with the function of the motile flagellum. In contrast, those genes that were upregulated included cell surface proteins, transporters, peptidases and many uncharacterized genes, including 293 of the 936 novel genes. Genome-wide distribution analysis of the differentially expressed genes revealed that the tetraploid chromosome 30 is highly enriched for genes that were upregulated in amastigotes, providing the first evidence of a link between this whole chromosome duplication event and adaptation to the vertebrate host in this group. Peptide evidence for 42 proteins encoded by novel transcripts supports the idea of an as yet uncharacterised set of small proteins in Leishmania spp. with possible implications for host-pathogen interactions. PMID:26452044

Transcriptional mapping of the varicella-zoster virus regulatory genes encoding open reading frames 4 and 63.

PubMed Central

Kinchington, P R; Vergnes, J P; Defechereux, P; Piette, J; Turse, S E

1994-01-01

Four of the 68 varicella-zoster virus (VZV) unique open reading frames (ORFs), i.e., ORFs 4, 61, 62, and 63, encode proteins that influence viral transcription and are considered to be positional homologs of herpes simplex virus type 1 (HSV-1) immediate-early (IE) proteins. In order to identify the elements that regulate transcription of VZV ORFs 4 and 63, the encoded mRNAs were mapped in detail. For ORF 4, a major 1.8-kb and a minor 3.0-kb polyadenylated [poly(A)+] RNA were identified, whereas ORF 63-specific probes recognized 1.3- and 1.9-kb poly(A)+ RNAs. Probes specific for sequences adjacent to the ORFs and mapping of the RNA 3' ends indicated that the ORF 4 RNAs were 3' coterminal, whereas the RNAs for ORF 63 represented two different termination sites. S1 nuclease mapping and primer extension analyses indicated a single transcription initiation site for ORF 4 at 38 bp upstream of the ORF start codon. For ORF 63, multiple transcriptional start sites at 87 to 95, 151 to 153, and (tentatively) 238 to 243 bp upstream of the ORF start codon were identified. TATA box motifs at good positional locations were found upstream of all mapped transcription initiation sites. However, no sequences resembling the TAATGARAT motif, which confers IE regulation upon HSV-1 IE genes, were found. The finding of the absence of this motif was supported through analyses of the regulatory sequences of ORFs 4 and 63 in transient transfection assays alongside those of ORFs 61 and 62. Sequences representing the promoters for ORFs 4, 61, and 63 were all stimulated by VZV infection but failed to be stimulated by coexpression with the HSV-1 transactivator Vmw65. In contrast, the promoter for ORF 62, which contains TAATGARAT motifs, was activated by VZV infection and coexpression with Vmw65. These results extend the transcriptional knowledge for VZV and suggest that ORFs 4 and 63 contain regulatory signals different from those of the ORF 62 and HSV-1 IE genes. Images PMID:8189496

Endoplasmic reticulum stress in vasopressin neurons of familial diabetes insipidus model mice: aggregate formation and mRNA poly(A) tail shortening.

PubMed

Arima, Hiroshi; Morishita, Yoshiaki; Hagiwara, Daisuke; Hayashi, Masayuki; Oiso, Yutaka

2014-01-01

The immunoglobulin heavy chain binding protein (BiP) is an endoplasmic reticulum (ER) chaperone, which binds to newly synthesized secretory and transmembrane proteins to facilitate protein folding. BiP mRNA is expressed in the arginine vasopressin (AVP) neurons in the supraoptic nucleus of wild-type mice even in basal conditions, and the expression levels increase in response to dehydration. These data suggest that AVP neurons are subjected to ER stress. Familial neurohypophysial diabetes insipidus (FNDI) is caused by mutations in the gene locus of AVP. The mutant proteins could accumulate in the ER and possibly increase ER stress in the AVP neurons. We bred mice possessing a mutation causing FNDI, which manifested progressive polyuria, as do the patients with FNDI. Electron microscopic analyses demonstrated that aggregates accumulated in the ER of AVP neurons in FNDI mice. Despite polyuria, which could potentially induce dehydration, AVP mRNA expression was decreased in the supraoptic nucleus, and the AVP mRNA poly(A) tail length was shortened in FNDI mice compared with wild-type mice. Incubation of hypothalamic explants of wild-type mice with ER stressors caused shortening of the poly(A) tail length of AVP mRNA, accompanied by decreases in the expression. These data revealed a mechanism by which ER stress decreases poly(A) tail length of AVP mRNA, and this reduces the load of unfolded proteins that form the aggregates in ER of the AVP neurons in FNDI mice.

Characterization of short interspersed elements (SINEs) in a red alga, Porphyra yezoensis.

PubMed

Zhang, Wenbo; Lin, Xiaofei; Peddigari, Suresh; Takechi, Katsuaki; Takano, Hiroyoshi; Takio, Susumu

2007-02-01

Short interspersed element (SINE)-like sequences referred to as PySN1 and PySN2 were identified in a red alga, Porphyra yezoensis. Both elements contained an internal promoter with motifs (A box and B box) recognized by RNA polymerase III, and target site duplications at both ends. Genomic Southern blot analysis revealed that both elements were widely and abundantly distributed on the genome. 3' and 5' RACE suggested that PySN1 was expressed as a chimera transcript with flanking SINE-unrelated sequences and possessed the poly-A tail at the same position near the 3' end of PySN1.

AT-rich sequence elements promote nascent transcript cleavage leading to RNA polymerase II termination

PubMed Central

White, Eleanor; Kamieniarz-Gdula, Kinga; Dye, Michael J.; Proudfoot, Nick J.

2013-01-01

RNA Polymerase II (Pol II) termination is dependent on RNA processing signals as well as specific terminator elements located downstream of the poly(A) site. One of the two major terminator classes described so far is the Co-Transcriptional Cleavage (CoTC) element. We show that homopolymer A/T tracts within the human β-globin CoTC-mediated terminator element play a critical role in Pol II termination. These short A/T tracts, dispersed within seemingly random sequences, are strong terminator elements, and bioinformatics analysis confirms the presence of such sequences in 70% of the putative terminator regions (PTRs) genome-wide. PMID:23258704

Gain and loss of polyadenylation signals during evolution of green algae.

PubMed

Wodniok, Sabina; Simon, Andreas; Glöckner, Gernot; Becker, Burkhard

2007-04-18

The Viridiplantae (green algae and land plants) consist of two monophyletic lineages: the Chlorophyta and the Streptophyta. Most green algae belong to the Chlorophyta, while the Streptophyta include all land plants and a small group of freshwater algae known as Charophyceae. Eukaryotes attach a poly-A tail to the 3' ends of most nuclear-encoded mRNAs. In embryophytes, animals and fungi, the signal for polyadenylation contains an A-rich sequence (often AAUAAA or related sequence) 13 to 30 nucleotides upstream from the cleavage site, which is commonly referred to as the near upstream element (NUE). However, it has been reported that the pentanucleotide UGUAA is used as polyadenylation signal for some genes in volvocalean algae. We set out to investigate polyadenylation signal differences between streptophytes and chlorophytes that may have emerged shortly after the evolutionary split between Streptophyta and Chlorophyta. We therefore analyzed expressed genes (ESTs) from three streptophyte algae, Mesostigma viride, Klebsormidium subtile and Coleochaete scutata, and from two early-branching chlorophytes, Pyramimonas parkeae and Scherffelia dubia. In addition, to extend the database, our analyses included ESTs from six other chlorophytes (Acetabularia acetabulum, Chlamydomonas reinhardtii, Helicosporidium sp. ex Simulium jonesii, Prototheca wickerhamii, Scenedesmus obliquus and Ulva linza) and one streptophyte (Closterium peracerosum). Our results indicate that polyadenylation signals in green algae vary widely. The UGUAA motif is confined to late-branching Chlorophyta. Most streptophyte algae do not have an A-rich sequence motif like that in embryophytes, animals and fungi. We observed polyadenylation signals similar to those of Arabidopsis and other land plants only in Mesostigma. Polyadenylation signals in green algae show considerable variation. A new NUE (UGUAA) was invented in derived chlorophytes and replaced not only the A-rich NUE but the complete poly(A) signal in all chlorophytes investigated except Scherffelia (only NUE replaced) and Pyramimonas (UGUAA completely missing). The UGUAA element is completely absent from streptophytes. However, the structure of the poly(A) signal was often modified in streptophyte algae. In most species investigated, an A-rich NUE is missing; instead, these species seem to rely mainly on U-rich elements.

Regression analysis using dependent Polya trees.

PubMed

Schörgendorfer, Angela; Branscum, Adam J

2013-11-30

Many commonly used models for linear regression analysis force overly simplistic shape and scale constraints on the residual structure of data. We propose a semiparametric Bayesian model for regression analysis that produces data-driven inference by using a new type of dependent Polya tree prior to model arbitrary residual distributions that are allowed to evolve across increasing levels of an ordinal covariate (e.g., time, in repeated measurement studies). By modeling residual distributions at consecutive covariate levels or time points using separate, but dependent Polya tree priors, distributional information is pooled while allowing for broad pliability to accommodate many types of changing residual distributions. We can use the proposed dependent residual structure in a wide range of regression settings, including fixed-effects and mixed-effects linear and nonlinear models for cross-sectional, prospective, and repeated measurement data. A simulation study illustrates the flexibility of our novel semiparametric regression model to accurately capture evolving residual distributions. In an application to immune development data on immunoglobulin G antibodies in children, our new model outperforms several contemporary semiparametric regression models based on a predictive model selection criterion. Copyright © 2013 John Wiley & Sons, Ltd.

Useful Bicistronic Reporter System for Studying Poly(A) Site-Defining cis Elements and Regulation of Alternative Polyadenylation.

PubMed

Deng, Zhongyuan; Zhang, Shen; Gu, Shaohua; Ni, Xinzhi; Zeng, Wenxian; Li, Xianchun

2018-01-17

The link between polyadenylation (pA) and various biological, behavioral, and pathological events of eukaryotes underlines the need to develop in vivo polyadenylation assay methods for characterization of the cis -acting elements, trans -acting factors and environmental stimuli that affect polyadenylation efficiency and/or relative usage of two alternative polyadenylation (APA) sites. The current protein-based CAT or luciferase reporter systems can measure the polyadenylation efficiency of a single pA site or candidate cis element but not the choice of two APA sites. To address this issue, we developed a set of four new bicistronic reporter vectors that harbor either two luciferase or fluorescence protein open reading frames connected with one Internal Ribosome Entry Site (IRES). Transfection of single or dual insertion constructs of these vectors into mammalian cells demonstrated that they could be utilized not only to quantify the strength of a single candidate pA site or cis element, but also to accurately measure the relative usage of two APA sites at both the mRNA (qRT-PCR) and protein levels. This represents the first reporter system that can study polyadenylation efficiency of a single pA site or element and regulation of two APA sites at both the mRNA and protein levels.

Sequences downstream of AAUAAA signals affect pre-mRNA cleavage and polyadenylation in vitro both directly and indirectly.

PubMed Central

Ryner, L C; Takagaki, Y; Manley, J L

1989-01-01

To investigate the role of sequences lying downstream of the conserved AAUAAA hexanucleotide in pre-mRNA cleavage and polyadenylation, deletions or substitutions were constructed in polyadenylation signals from simian virus 40 and adenovirus, and their effects were assayed in both crude and fractionated HeLa cell nuclear extracts. As expected, these sequences influenced the efficiency of both cleavage and polyadenylation as well as the accuracy of the cleavage reaction. Sequences near or upstream of the actual site of poly(A) addition appeared to specify a unique cleavage site, since their deletion resulted, in some cases, in heterogeneous cleavage. Furthermore, the sequences that allowed the simian virus 40 late pre-RNA to be cleaved preferentially by partially purified cleavage activity were also those at the cleavage site itself. Interestingly, sequences downstream of the cleavage site interacted with factors not directly involved in catalyzing cleavage and polyadenylation, since the effects of deletions were substantially diminished when partially purified components were used in assays. In addition, these sequences contained elements that could affect 3'-end formation both positively and negatively. Images PMID:2566911

Identification, cloning, and sequencing of a fragment of Amsacta moorei entomopoxvirus DNA containing the spheroidin gene and three vaccinia virus-related open reading frames.

PubMed Central

Hall, R L; Moyer, R W

1991-01-01

Entomopoxvirus virions are frequently contained within crystalline occlusion bodies, which are composed of primarily a single protein, spheroidin, which is analogous to the polyhedrin protein of baculovirus. The spheroidin gene of Amsacta moorei entomopoxvirus was identified following the microsequencing of polypeptides generated from cyanogen bromide treatment of spheroidin and the subsequent synthesis of oligonucleotide hybridization probes. DNA sequencing of a 6.8-kb region of DNA containing the spheroidin gene showed that the spheroidin protein is derived from a 3.0-kb open reading frame potentially encoding a protein of 115 kDa. Three copies of the heptanucleotide, TTTTTNT, a sequence associated with early gene transcription in the vertebrate poxviruses, and four in-frame translational termination signals were found within 60 bp upstream of the putative spheroidin gene promoter (TAAATG). The spheroidin gene promoter region contains the sequence TAAATG, which is found in many late promoters of the vertebrate poxviruses and which serves as the site of transcriptional initiation, as shown by primer extension. Primer extension experiments also showed that spheroidin gene transcripts contain 5' poly(A) sequences typical of vertebrate poxvirus late transcripts. The 92 bases upstream of the initiating TAAATG are unusually A + T rich and contain only 7 G or C residues. An analysis of open reading frames around the spheroidin gene suggests that the colinear core of "essential genes" typical of the vertebrate poxviruses is absent in A. moorei entomopoxvirus. Images PMID:1942245

The Novel Poly(A) Polymerase Star-PAP is a Signal-Regulated Switch at the 3′-end of mRNAs

PubMed Central

Li, Weimin; Laishram, Rakesh S.; Anderson, Richard A.

2013-01-01

The mRNA 3′-untranslated region (3′-UTR) modulates message stability, transport, intracellular location and translation. We have discovered a novel nuclear poly(A) polymerase termed Star-PAP (nuclear speckle targeted PIPKIα regulated-poly(A) polymerase) that couples with the transcriptional machinery and is regulated by the phosphoinositide lipid messenger phosphatidylinositol-4,5-bisphosphate (PI4,5P2), the central lipid in phosphoinositide signaling. PI4,5P2 is generated primarily by type I phosphatidylinositol phosphate kinases (PIPKI). Phosphoinositides are present in the nucleus including at nuclear speckles compartments separate from known membrane structures. PIPKs regulate cellular functions by interacting with PI4,5P2 effectors where PIPKs generate PI4,5P2 that then modulates the activity of the associated effectors. Nuclear PIPKIα interacts with and regulates Star-PAP, and PI4,5P2 specifically activates Star-PAP in a gene- and signaling-dependent manner. Importantly, other select signaling molecules integrated into the Star-PAP complex seem to regulate Star-PAP activities and processivities toward RNA substrates, and unique sequence elements around the Star-PAP binding sites within the 3′-UTR of target genes contribute to Star-PAP specificity for processing. Therefore, Star-PAP and its regulatory molecules form a signaling nexus at the 3′-end of target mRNAs to control the expression of select group of genes including the ones involved in stress responses. PMID:23306079

The differential expression of alternatively polyadenylated transcripts is a common stress-induced response mechanism that modulates mammalian mRNA expression in a quantitative and qualitative fashion.

PubMed

Hollerer, Ina; Curk, Tomaz; Haase, Bettina; Benes, Vladimir; Hauer, Christian; Neu-Yilik, Gabriele; Bhuvanagiri, Madhuri; Hentze, Matthias W; Kulozik, Andreas E

2016-09-01

Stress adaptation plays a pivotal role in biological processes and requires tight regulation of gene expression. In this study, we explored the effect of cellular stress on mRNA polyadenylation and investigated the implications of regulated polyadenylation site usage on mammalian gene expression. High-confidence polyadenylation site mapping combined with global pre-mRNA and mRNA expression profiling revealed that stress induces an accumulation of genes with differentially expressed polyadenylated mRNA isoforms in human cells. Specifically, stress provokes a global trend in polyadenylation site usage toward decreased utilization of promoter-proximal poly(A) sites in introns or ORFs and increased utilization of promoter-distal polyadenylation sites in intergenic regions. This extensively affects gene expression beyond regulating mRNA abundance by changing mRNA length and by altering the configuration of open reading frames. Our study highlights the impact of post-transcriptional mechanisms on stress-dependent gene regulation and reveals the differential expression of alternatively polyadenylated transcripts as a common stress-induced mechanism in mammalian cells. © 2016 Hollerer et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

Binding of Phenazinium Dye Safranin T to Polyriboadenylic Acid: Spectroscopic and Thermodynamic Study

PubMed Central

Roy, Snigdha; Das, Suman

2014-01-01

Here, we report results from experiments designed to explore the association of the phenazinium dye safranin T (ST, 3,7-diamino-2,8-dimethyl-5-phenylphenazinium chloride) with single and double stranded form of polyriboadenylic acid (hereafter poly-A) using several spectroscopic techniques. We demonstrate that the dye binds to single stranded polyriboadenylic acid (hereafter ss poly-A) with high affinity while it does not interact at all with the double stranded (ds) form of the polynucleotide. Fluorescence and absorption spectral studies reveal the molecular aspects of binding of ST to single stranded form of the polynucleotide. This observation is also supported by the circular dichroism study. Thermodynamic data obtained from temperature dependence of binding constant reveals that association is driven by negative enthalpy change and opposed by negative entropy change. Ferrocyanide quenching studies have shown intercalative binding of ST to ss poly-A. Experiments on viscosity measurements confirm the binding mode of the dye to be intercalative. The effect of [Na+] ion concentration on the binding process suggests the role of electrostatic forces in the complexation. Present studies reveal the utility of the dye in probing nucleic acid structure. PMID:24498422

Binding of phenazinium dye safranin T to polyriboadenylic acid: spectroscopic and thermodynamic study.

PubMed

Pradhan, Ankur Bikash; Haque, Lucy; Roy, Snigdha; Das, Suman

2014-01-01

Here, we report results from experiments designed to explore the association of the phenazinium dye safranin T (ST, 3,7-diamino-2,8-dimethyl-5-phenylphenazinium chloride) with single and double stranded form of polyriboadenylic acid (hereafter poly-A) using several spectroscopic techniques. We demonstrate that the dye binds to single stranded polyriboadenylic acid (hereafter ss poly-A) with high affinity while it does not interact at all with the double stranded (ds) form of the polynucleotide. Fluorescence and absorption spectral studies reveal the molecular aspects of binding of ST to single stranded form of the polynucleotide. This observation is also supported by the circular dichroism study. Thermodynamic data obtained from temperature dependence of binding constant reveals that association is driven by negative enthalpy change and opposed by negative entropy change. Ferrocyanide quenching studies have shown intercalative binding of ST to ss poly-A. Experiments on viscosity measurements confirm the binding mode of the dye to be intercalative. The effect of [Na⁺] ion concentration on the binding process suggests the role of electrostatic forces in the complexation. Present studies reveal the utility of the dye in probing nucleic acid structure.

Rrp6p controls mRNA polyA tail length and its decoration with polyA binding proteins

PubMed Central

Schmid, Manfred; Poulsen, Mathias Bach; Olszewski, Pawel; Pelechano, Vicent; Saguez, Cyril; Gupta, Ishaan; Steinmetz, Lars M.; Moore, Claire; Jensen, Torben Heick

2012-01-01

PolyA (pA) tail binding proteins (PABPs) control mRNA polyadenylation, stability and translation. In a purified system, S. cerevisiae PABPs, Pab1p and Nab2p, are individually sufficient to provide normal pA tail length. However, it is unknown how this occurs in more complex environments. Here we find that the nuclear exosome subunit Rrp6p counteracts the in vitro and in vivo extension of mature pA tails by the non-canonical pA polymerase Trf4p. Moreover, PABP loading onto nascent pA tails is controlled by Rrp6p; while Pab1p is the major PABP, Nab2p only associates in the absence of Rrp6p. This is because Rrp6p can interact with Nab2p and displace it from pA tails, potentially leading to RNA turnover as evidenced for certain pre-mRNAs. We suggest that a nuclear mRNP surveillance step involves targeting of Rrp6p by Nab2p-bound pA-tailed RNPs and that pre-mRNA abundance is regulated at this level. PMID:22683267

2′-O Methylation of Internal Adenosine by Flavivirus NS5 Methyltransferase

PubMed Central

Dong, Hongping; Chang, David C.; Hua, Maggie Ho Chia; Lim, Siew Pheng; Chionh, Yok Hian; Hia, Fabian; Lee, Yie Hou; Kukkaro, Petra; Lok, Shee-Mei; Dedon, Peter C.; Shi, Pei-Yong

2012-01-01

RNA modification plays an important role in modulating host-pathogen interaction. Flavivirus NS5 protein encodes N-7 and 2′-O methyltransferase activities that are required for the formation of 5′ type I cap (m7GpppAm) of viral RNA genome. Here we reported, for the first time, that flavivirus NS5 has a novel internal RNA methylation activity. Recombinant NS5 proteins of West Nile virus and Dengue virus (serotype 4; DENV-4) specifically methylates polyA, but not polyG, polyC, or polyU, indicating that the methylation occurs at adenosine residue. RNAs with internal adenosines substituted with 2′-O-methyladenosines are not active substrates for internal methylation, whereas RNAs with adenosines substituted with N6-methyladenosines can be efficiently methylated, suggesting that the internal methylation occurs at the 2′-OH position of adenosine. Mass spectroscopic analysis further demonstrated that the internal methylation product is 2′-O-methyladenosine. Importantly, genomic RNA purified from DENV virion contains 2′-O-methyladenosine. The 2′-O methylation of internal adenosine does not require specific RNA sequence since recombinant methyltransferase of DENV-4 can efficiently methylate RNAs spanning different regions of viral genome, host ribosomal RNAs, and polyA. Structure-based mutagenesis results indicate that K61-D146-K181-E217 tetrad of DENV-4 methyltransferase forms the active site of internal methylation activity; in addition, distinct residues within the methyl donor (S-adenosyl-L-methionine) pocket, GTP pocket, and RNA-binding site are critical for the internal methylation activity. Functional analysis using flavivirus replicon and genome-length RNAs showed that internal methylation attenuated viral RNA translation and replication. Polymerase assay revealed that internal 2′-O-methyladenosine reduces the efficiency of RNA elongation. Collectively, our results demonstrate that flavivirus NS5 performs 2′-O methylation of internal adenosine of viral RNA in vivo and host ribosomal RNAs in vitro. PMID:22496660

Some Biochemical Properties of an Acido-Thermophilic Archae-Bacterium Sulfolobus Acidocaldarius

NASA Astrophysics Data System (ADS)

Oshima, Tairo; Ohba, Masayuki; Wagaki, Takayoshi

1984-12-01

To elucidate the phylogenic status of archaebacteria, some basic cellular components of an acido-thermophilic archaebacterium,Sulfolobus acidocaldarius, were studied. Poly(A) containing RNA was present in the cells, and performed the role of mRNA in a cell-free extract of reticulocyte or the archaebacteria. Poly(A) containing RNA was also found in other archaebacterial cells. The absence of cap structure was suggested in these RNAs. The cell-free protein synthesis using the archaebacterial extract was inhibited by anisomycin, a specific inhibitor for eukaryotic ribosomes. Two unique membrane-bound ATPases were detected. Based on resistance to H+-ATPase inhibitors, these enzymes seemed not to be F0F1-ATPase.

PolyA_DB 3 catalogs cleavage and polyadenylation sites identified by deep sequencing in multiple genomes

PubMed Central

Wang, Ruijia; Nambiar, Ram; Zheng, Dinghai

2018-01-01

Abstract PolyA_DB is a database cataloging cleavage and polyadenylation sites (PASs) in several genomes. Previous versions were based mainly on expressed sequence tags (ESTs), which had a limited amount and could lead to inaccurate PAS identification due to the presence of internal A-rich sequences in transcripts. Here, we present an updated version of the database based solely on deep sequencing data. First, PASs are mapped by the 3′ region extraction and deep sequencing (3′READS) method, ensuring unequivocal PAS identification. Second, a large volume of data based on diverse biological samples increases PAS coverage by 3.5-fold over the EST-based version and provides PAS usage information. Third, strand-specific RNA-seq data are used to extend annotated 3′ ends of genes to obtain more thorough annotations of alternative polyadenylation (APA) sites. Fourth, conservation information of PAS across mammals sheds light on significance of APA sites. The database (URL: http://www.polya-db.org/v3) currently holds PASs in human, mouse, rat and chicken, and has links to the UCSC genome browser for further visualization and for integration with other genomic data. PMID:29069441

Extreme heterogeneity of polyadenylation sites in mRNAs encoding chloroplast RNA-binding proteins in Nicotiana plumbaginifolia.

PubMed

Klahre, U; Hemmings-Mieszczak, M; Filipowicz, W

1995-06-01

We have previously characterized nuclear cDNA clones encoding two RNA binding proteins, CP-RBP30 and CP-RBP-31, which are targeted to chloroplasts in Nicotiana plumbaginifolia. In this report we describe the analysis of the 3'-untranslated regions (3'-UTRs) in 22 CP-RBP30 and 8 CP-RBP31 clones which reveals that mRNAs encoding both proteins have a very complex polyadenylation pattern. Fourteen distinct poly(A) sites were identified among CP-RBP30 clones and four sites among the CP-RBP31 clones. The authenticity of the sites was confirmed by RNase A/T1 mapping of N. plumbaginifolia RNA. CP-RBP30 provides an extreme example of the heterogeneity known to be a feature of mRNA polyadenylation in higher plants. Using PCR we have demonstrated that CP-RBP genes in N. plumbaginifolia and N. sylvestris, in addition to the previously described introns interrupting the coding region, contain an intron located in the 3' non-coding part of the gene. In the case of the CP-RBP31, we have identified one polyadenylation event occurring in this intron.

Problem solving of student with visual impairment related to mathematical literacy problem

NASA Astrophysics Data System (ADS)

Pratama, A. R.; Saputro, D. R. S.; Riyadi

2018-04-01

The student with visual impairment, total blind category depends on the sense of touch and hearing in obtaining information. In fact, the two senses can receive information less than 20%. Thus, students with visual impairment of the total blind categories in the learning process must have difficulty, including learning mathematics. This study aims to describe the problem-solving process of the student with visual impairment, total blind category on mathematical literacy issues based on Polya phase. This research using test method similar problems mathematical literacy in PISA and in-depth interviews. The subject of this study was a student with visual impairment, total blind category. Based on the result of the research, problem-solving related to mathematical literacy based on Polya phase is quite good. In the phase of understanding the problem, the student read about twice by brushing the text and assisted with information through hearing three times. The student with visual impairment in problem-solving based on the Polya phase, devising a plan by summoning knowledge and experience gained previously. At the phase of carrying out the plan, students with visual impairment implement the plan in accordance with pre-made. In the looking back phase, students with visual impairment need to check the answers three times but have not been able to find a way.

Replacement of the yeast TRP4 3' untranslated region by a hammerhead ribozyme results in a stable and efficiently exported mRNA that lacks a poly(A) tail.

PubMed Central

Düvel, Katrin; Valerius, Oliver; Mangus, David A; Jacobson, Allan; Braus, Gerhard H

2002-01-01

The mRNA poly(A) tail serves different purposes, including the facilitation of nuclear export, mRNA stabilization, efficient translation, and, finally, specific degradation. The posttranscriptional addition of a poly(A) tail depends on sequence motifs in the 3' untranslated region (3' UTR) of the mRNA and a complex trans-acting protein machinery. In this study, we have replaced the 3' UTR of the yeast TRP4 gene with sequences encoding a hammerhead ribozyme that efficiently cleaves itself in vivo. Expression of the TRP4-ribozyme allele resulted in the accumulation of a nonpolyadenylated mRNA. Cells expressing the TRP4-ribozyme mRNA showed a reduced growth rate due to a reduction in Trp4p enzyme activity. The reduction in enzyme activity was not caused by inefficient mRNA export from the nucleus or mRNA destabilization. Rather, analyses of mRNA association with polyribosomes indicate that translation of the ribozyme-containing mRNA is impaired. This translational defect allows sufficient synthesis of Trp4p to support growth of trp4 cells, but is, nevertheless, of such magnitude as to activate the general control network of amino acid biosynthesis. PMID:12003493

Tales around the clock: Poly(A) tails in circadian gene expression.

PubMed

Beta, Rafailia A A; Balatsos, Nikolaos A A

2018-06-17

Circadian rhythms are ubiquitous time-keeping processes in eukaryotes with a period of ~24 hr. Light is perhaps the main environmental cue (zeitgeber) that affects several aspects of physiology and behaviour, such as sleep/wake cycles, orientation of birds and bees, and leaf movements in plants. Temperature can serve as the main zeitgeber in the absence of light cycles, even though it does not lead to rhythmicity through the same mechanism as light. Additional cues include feeding patterns, humidity, and social rhythms. At the molecular level, a master oscillator orchestrates circadian rhythms and organizes molecular clocks located in most cells. The generation of the 24 hr molecular clock is based on transcriptional regulation, as it drives intrinsic rhythmic changes based on interlocked transcription/translation feedback loops that synchronize expression of genes. Thus, processes and factors that determine rhythmic gene expression are important to understand circadian rhythms. Among these, the poly(A) tails of RNAs play key roles in their stability, translational efficiency and degradation. In this article, we summarize current knowledge and discuss perspectives on the role and significance of poly(A) tails and associating factors in the context of the circadian clock. This article is categorized under: RNA Turnover and Surveillance > Regulation of RNA Stability RNA Processing > 3' End Processing. © 2018 Wiley Periodicals, Inc.

Isolation and characterization of cDNA clones for carrot extensin and a proline-rich 33-kDa protein

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chen, J.; Varner, J.E.

1985-07-01

Extensins are hydroxyproline-rich glycoproteins associated with most dicotyledonous plant cell walls. To isolate cDNA clones encoding extensin, the authors started by isolating poly(A) RNA from carrot root tissue, and then translating the RNA in vitro, in the presence of tritiated leucine or proline. A 33-kDa peptide was identified in the translation products as a putative extensin precursor. From a cDNA library constructed with poly(A) RNA from wounded carrots, one cDNA clone (pDC5) was identified that specifically hybridized to poly(A) RNA encoding this 33-kDa peptide. They isolated three cDNA clones (pDC11, pDC12, and pDC16) from another cDNA library using pCD5 asmore » a probe. DNA sequence data, RNA hybridization analysis, and hybrid released in vitro translation indicate that the cDNA clones pDC11 encodes extensin and that cDNA clones pDC12 and pDC16 encode the 33-kDa peptide, which as yet has an unknown identity and function. The assumption that the 33-kDa peptide was an extensin precursor was invalid. RNA hybridization analysis showed that RNA encoded by both clone types is accumulated upon wounding.« less

Structural Features of a Picornavirus Polymerase Involved in the Polyadenylation of Viral RNA

PubMed Central

Kempf, Brian J.; Kelly, Michelle M.; Springer, Courtney L.; Peersen, Olve B.

2013-01-01

Picornaviruses have 3′ polyadenylated RNA genomes, but the mechanisms by which these genomes are polyadenylated during viral replication remain obscure. Based on prior studies, we proposed a model wherein the poliovirus RNA-dependent RNA polymerase (3Dpol) uses a reiterative transcription mechanism while replicating the poly(A) and poly(U) portions of viral RNA templates. To further test this model, we examined whether mutations in 3Dpol influenced the polyadenylation of virion RNA. We identified nine alanine substitution mutations in 3Dpol that resulted in shorter or longer 3′ poly(A) tails in virion RNA. These mutations could disrupt structural features of 3Dpol required for the recruitment of a cellular poly(A) polymerase; however, the structural orientation of these residues suggests a direct role of 3Dpol in the polyadenylation of RNA genomes. Reaction mixtures containing purified 3Dpol and a template RNA with a defined poly(U) sequence provided data consistent with a template-dependent reiterative transcription mechanism for polyadenylation. The phylogenetically conserved structural features of 3Dpol involved in the polyadenylation of virion RNA include a thumb domain alpha helix that is positioned in the minor groove of the double-stranded RNA product and lysine and arginine residues that interact with the phosphates of both the RNA template and product strands. PMID:23468507

Regions of conservation and divergence in the 3' untranslated sequences of genomic RNA from Ross River virus isolates.

PubMed

Faragher, S G; Dalgarno, L

1986-07-20

The 3' untranslated (UT) sequences of the genomic RNAs of five geographic variants of the alphavirus Ross River virus (RRV) were determined and compared with the 3' UT sequence of RRV T48, the prototype strain. Part of the 3' UT region of Getah virus, a close serological relative of RRV, was also sequenced. The RRV 3' UT region varies markedly in length between variants. Large deletions or insertions, sequence rearrangements and single nucleotide substitutions are observed. A sequence tract of 49 to 58 nucleotides, which is repeated as four blocks in the RRV T48 3' UT region, occurs only once in the 3' UT region of one RRV strain (NB5092), indicating that the existence of repeat sequence blocks is not essential for RRV replication. However, the precise sequence of the 3' proximal copy of the repeat block and its position relative to the poly(A) tail were identical in all RRV isolates examined, suggesting that it has an important role in RRV replication. Nucleotide substitutions between RRV variants are distributed non-randomly along the length of the 3' UT region. The sequence of 120 to 130 nucleotides adjacent to the poly(A) tail is strongly conserved. Getah virus RNA contains three repeat sequence blocks in the 3' UT region. These are similar in sequence to those in RRV RNA but differ in their arrangement. Homology between the RRV and Getah 3' UT sequences is greatest in the 3' proximal repeat sequence block that shows three differences in 49 nucleotides. The 3' proximal repeat in Getah RNA occurs at the same position, relative to the poly(A) tail, as in all RRV variants. The RRV and Getah virus 3' UT sequences show extensive homology in the region between the 3' proximal repeat and the poly(A) tail but, apart from the repeat blocks themselves, they show no significant homology elsewhere.

Novel mouse models of oculopharyngeal muscular dystrophy (OPMD) reveal early onset mitochondrial defects and suggest loss of PABPN1 may contribute to pathology.

PubMed

Vest, Katherine E; Phillips, Brittany L; Banerjee, Ayan; Apponi, Luciano H; Dammer, Eric B; Xu, Weiting; Zheng, Dinghai; Yu, Julia; Tian, Bin; Pavlath, Grace K; Corbett, Anita H

2017-09-01

Oculopharyngeal muscular dystrophy (OPMD) is a late onset disease caused by polyalanine expansion in the poly(A) binding protein nuclear 1 (PABPN1). Several mouse models have been generated to study OPMD; however, most of these models have employed transgenic overexpression of alanine-expanded PABPN1. These models do not recapitulate the OPMD patient genotype and PABPN1 overexpression could confound molecular phenotypes. We have developed a knock-in mouse model of OPMD (Pabpn1+/A17) that contains one alanine-expanded Pabpn1 allele under the control of the native promoter and one wild-type Pabpn1 allele. This mouse is the closest available genocopy of OPMD patients. We show that Pabpn1+/A17 mice have a mild myopathic phenotype in adult and aged animals. We examined early molecular and biochemical phenotypes associated with expressing native levels of A17-PABPN1 and detected shorter poly(A) tails, modest changes in poly(A) signal (PAS) usage, and evidence of mitochondrial damage in these mice. Recent studies have suggested that a loss of PABPN1 function could contribute to muscle pathology in OPMD. To investigate a loss of function model of pathology, we generated a heterozygous Pabpn1 knock-out mouse model (Pabpn1+/Δ). Like the Pabpn1+/A17 mice, Pabpn1+/Δ mice have mild histologic defects, shorter poly(A) tails, and evidence of mitochondrial damage. However, the phenotypes detected in Pabpn1+/Δ mice only partially overlap with those detected in Pabpn1+/A17 mice. These results suggest that loss of PABPN1 function could contribute to but may not completely explain the pathology detected in Pabpn1+/A17 mice. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Tenebrio molitor antifreeze protein gene identification and regulation.

PubMed

Qin, Wensheng; Walker, Virginia K

2006-02-15

The yellow mealworm, Tenebrio molitor, is a freeze susceptible, stored product pest. Its winter survival is facilitated by the accumulation of antifreeze proteins (AFPs), encoded by a small gene family. We have now isolated 11 different AFP genomic clones from 3 genomic libraries. All the clones had a single coding sequence, with no evidence of intervening sequences. Three genomic clones were further characterized. All have putative TATA box sequences upstream of the coding regions and multiple potential poly(A) signal sequences downstream of the coding regions. A TmAFP regulatory region, B1037, conferred transcriptional activity when ligated to a luciferase reporter sequence and after transfection into an insect cell line. A 143 bp core promoter including a TATA box sequence was identified. Its promoter activity was increased 4.4 times by inserting an exotic 245 bp intron into the construct, similar to the enhancement of transgenic expression seen in several other systems. The addition of a duplication of the first 120 bp sequence from the 143 bp core promoter decreased promoter activity by half. Although putative hormonal response sequences were identified, none of the five hormones tested enhanced reporter activity. These studies on the mechanisms of AFP transcriptional control are important for the consideration of any transfer of freeze-resistance phenotypes to beneficial hosts.

Fungal and Prokaryotic Activities in the Marine Subsurface Biosphere at Peru Margin and Canterbury Basin Inferred from RNA-Based Analyses and Microscopy

PubMed Central

Pachiadaki, Maria G.; Rédou, Vanessa; Beaudoin, David J.; Burgaud, Gaëtan; Edgcomb, Virginia P.

2016-01-01

The deep sedimentary biosphere, extending 100s of meters below the seafloor harbors unexpected diversity of Bacteria, Archaea, and microbial eukaryotes. Far less is known about microbial eukaryotes in subsurface habitats, albeit several studies have indicated that fungi dominate microbial eukaryotic communities and fungal molecular signatures (of both yeasts and filamentous forms) have been detected in samples as deep as 1740 mbsf. Here, we compare and contrast fungal ribosomal RNA gene signatures and whole community metatranscriptomes present in sediment core samples from 6 and 95 mbsf from Peru Margin site 1229A and from samples from 12 and 345 mbsf from Canterbury Basin site U1352. The metatranscriptome analyses reveal higher relative expression of amino acid and peptide transporters in the less nutrient rich Canterbury Basin sediments compared to the nutrient rich Peru Margin, and higher expression of motility genes in the Peru Margin samples. Higher expression of genes associated with metals transporters and antibiotic resistance and production was detected in Canterbury Basin sediments. A poly-A focused metatranscriptome produced for the Canterbury Basin sample from 345 mbsf provides further evidence for active fungal communities in the subsurface in the form of fungal-associated transcripts for metabolic and cellular processes, cell and membrane functions, and catalytic activities. Fungal communities at comparable depths at the two geographically separated locations appear dominated by distinct taxa. Differences in taxonomic composition and expression of genes associated with particular metabolic activities may be a function of sediment organic content as well as oceanic province. Microscopic analysis of Canterbury Basin sediment samples from 4 and 403 mbsf produced visualizations of septate fungal filaments, branching fungi, conidiogenesis, and spores. These images provide another important line of evidence supporting the occurrence and activity of fungi in the deep subseafloor biosphere. PMID:27375571

Fungal and Prokaryotic Activities in the Marine Subsurface Biosphere at Peru Margin and Canterbury Basin Inferred from RNA-Based Analyses and Microscopy.

PubMed

Pachiadaki, Maria G; Rédou, Vanessa; Beaudoin, David J; Burgaud, Gaëtan; Edgcomb, Virginia P

2016-01-01

The deep sedimentary biosphere, extending 100s of meters below the seafloor harbors unexpected diversity of Bacteria, Archaea, and microbial eukaryotes. Far less is known about microbial eukaryotes in subsurface habitats, albeit several studies have indicated that fungi dominate microbial eukaryotic communities and fungal molecular signatures (of both yeasts and filamentous forms) have been detected in samples as deep as 1740 mbsf. Here, we compare and contrast fungal ribosomal RNA gene signatures and whole community metatranscriptomes present in sediment core samples from 6 and 95 mbsf from Peru Margin site 1229A and from samples from 12 and 345 mbsf from Canterbury Basin site U1352. The metatranscriptome analyses reveal higher relative expression of amino acid and peptide transporters in the less nutrient rich Canterbury Basin sediments compared to the nutrient rich Peru Margin, and higher expression of motility genes in the Peru Margin samples. Higher expression of genes associated with metals transporters and antibiotic resistance and production was detected in Canterbury Basin sediments. A poly-A focused metatranscriptome produced for the Canterbury Basin sample from 345 mbsf provides further evidence for active fungal communities in the subsurface in the form of fungal-associated transcripts for metabolic and cellular processes, cell and membrane functions, and catalytic activities. Fungal communities at comparable depths at the two geographically separated locations appear dominated by distinct taxa. Differences in taxonomic composition and expression of genes associated with particular metabolic activities may be a function of sediment organic content as well as oceanic province. Microscopic analysis of Canterbury Basin sediment samples from 4 and 403 mbsf produced visualizations of septate fungal filaments, branching fungi, conidiogenesis, and spores. These images provide another important line of evidence supporting the occurrence and activity of fungi in the deep subseafloor biosphere.

Influenza Virus Mounts a Two-Pronged Attack on Host RNA Polymerase II Transcription.

PubMed

Bauer, David L V; Tellier, Michael; Martínez-Alonso, Mónica; Nojima, Takayuki; Proudfoot, Nick J; Murphy, Shona; Fodor, Ervin

2018-05-15

Influenza virus intimately associates with host RNA polymerase II (Pol II) and mRNA processing machinery. Here, we use mammalian native elongating transcript sequencing (mNET-seq) to examine Pol II behavior during viral infection. We show that influenza virus executes a two-pronged attack on host transcription. First, viral infection causes decreased Pol II gene occupancy downstream of transcription start sites. Second, virus-induced cellular stress leads to a catastrophic failure of Pol II termination at poly(A) sites, with transcription often continuing for tens of kilobases. Defective Pol II termination occurs independently of the ability of the viral NS1 protein to interfere with host mRNA processing. Instead, this termination defect is a common effect of diverse cellular stresses and underlies the production of previously reported downstream-of-gene transcripts (DoGs). Our work has implications for understanding not only host-virus interactions but also fundamental aspects of mammalian transcription. Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.

Blood-Derived RNA- and microRNA-Hydrolyzing IgG Antibodies in Schizophrenia Patients.

PubMed

Ermakov, E A; Ivanova, S A; Buneva, V N; Nevinsky, G A

2018-05-01

Abzymes with various catalytic activities are the earliest statistically significant markers of existing and developing autoimmune diseases (AIDs). Currently, schizophrenia (SCZD) is not considered to be a typical AID. It was demonstrated recently that antibodies from SCZD patients efficiently hydrolyze DNA and myelin basic protein. Here, we showed for the first time that autoantibodies from 35 SCZD patients efficiently hydrolyze RNA (cCMP > poly(C) > poly(A) > yeast RNA) and analyzed site-specific hydrolysis of microRNAs involved in the regulation of several genes in SCZD (miR-137, miR-9-5p, miR-219-2-3p, and miR-219a-5p). All four microRNAs were cleaved by IgG preparations (n = 21) from SCZD patients in a site-specific manner. The RNase activity of the abzymes correlated with SCZD clinical parameters. The data obtained showed that SCZD patients might display signs of typical autoimmune processes associated with impaired functioning of microRNAs resulting from their hydrolysis by the abzymes.

Comprehensive annotation of Glossina pallidipes salivary gland hypertrophy virus from Ethiopian tsetse flies: a proteogenomics approach

PubMed Central

Kariithi, Henry M.; Cousserans, François; Parker, Nicolas J.; İnce, İkbal Agah; Scully, Erin D.; Boeren, Sjef; Geib, Scott M.; Mekonnen, Solomon; Vlak, Just M.; Parker, Andrew G.; Vreysen, Marc J. B.; Bergoin, Max

2016-01-01

Glossina pallidipes salivary gland hypertrophy virus (GpSGHV; family Hytrosaviridae) can establish asymptomatic and symptomatic infection in its tsetse fly host. Here, we present a comprehensive annotation of the genome of an Ethiopian GpSGHV isolate (GpSGHV-Eth) compared with the reference Ugandan GpSGHV isolate (GpSGHV-Uga; GenBank accession number EF568108). GpSGHV-Eth has higher salivary gland hypertrophy syndrome prevalence than GpSGHV-Uga. We show that the GpSGHV-Eth genome has 190 291 nt, a low G+C content (27.9 %) and encodes 174 putative ORFs. Using proteogenomic and transcriptome mapping, 141 and 86 ORFs were mapped by transcripts and peptides, respectively. Furthermore, of the 174 ORFs, 132 had putative transcriptional signals [TATA-like box and poly(A) signals]. Sixty ORFs had both TATA-like box promoter and poly(A) signals, and mapped by both transcripts and peptides, implying that these ORFs encode functional proteins. Of the 60 ORFs, 10 ORFs are homologues to baculovirus and nudivirus core genes, including three per os infectivity factors and four RNA polymerase subunits (LEF4, 5, 8 and 9). Whereas GpSGHV-Eth and GpSGHV-Uga are 98.1 % similar at the nucleotide level, 37 ORFs in the GpSGHV-Eth genome had nucleotide insertions (n = 17) and deletions (n = 20) compared with their homologues in GpSGHV-Uga. Furthermore, compared with the GpSGHV-Uga genome, 11 and 24 GpSGHV ORFs were deleted and novel, respectively. Further, 13 GpSGHV-Eth ORFs were non-canonical; they had either CTG or TTG start codons instead of ATG. Taken together, these data suggest that GpSGHV-Eth and GpSGHV-Uga represent two different lineages of the same virus. Genetic differences combined with host and environmental factors possibly explain the differential GpSGHV pathogenesis observed in different G. pallidipes colonies. PMID:26801744

Biallelic insertion of a transcriptional terminator via the CRISPR/Cas9 system efficiently silences expression of protein-coding and non-coding RNA genes.

PubMed

Liu, Yangyang; Han, Xiao; Yuan, Junting; Geng, Tuoyu; Chen, Shihao; Hu, Xuming; Cui, Isabelle H; Cui, Hengmi

2017-04-07

The type II bacterial CRISPR/Cas9 system is a simple, convenient, and powerful tool for targeted gene editing. Here, we describe a CRISPR/Cas9-based approach for inserting a poly(A) transcriptional terminator into both alleles of a targeted gene to silence protein-coding and non-protein-coding genes, which often play key roles in gene regulation but are difficult to silence via insertion or deletion of short DNA fragments. The integration of 225 bp of bovine growth hormone poly(A) signals into either the first intron or the first exon or behind the promoter of target genes caused efficient termination of expression of PPP1R12C , NSUN2 (protein-coding genes), and MALAT1 (non-protein-coding gene). Both NeoR and PuroR were used as markers in the selection of clonal cell lines with biallelic integration of a poly(A) signal. Genotyping analysis indicated that the cell lines displayed the desired biallelic silencing after a brief selection period. These combined results indicate that this CRISPR/Cas9-based approach offers an easy, convenient, and efficient novel technique for gene silencing in cell lines, especially for those in which gene integration is difficult because of a low efficiency of homology-directed repair. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Structural and biochemical studies on ATP binding and hydrolysis by the Escherichia coli RNA chaperone Hfq.

PubMed

Hämmerle, Hermann; Beich-Frandsen, Mads; Večerek, Branislav; Rajkowitsch, Lukas; Carugo, Oliviero; Djinović-Carugo, Kristina; Bläsi, Udo

2012-01-01

In Escherichia coli the RNA chaperone Hfq is involved in riboregulation by assisting base-pairing between small regulatory RNAs (sRNAs) and mRNA targets. Several structural and biochemical studies revealed RNA binding sites on either surface of the donut shaped Hfq-hexamer. Whereas sRNAs are believed to contact preferentially the YKH motifs present on the proximal site, poly(A)(15) and ADP were shown to bind to tripartite binding motifs (ARE) circularly positioned on the distal site. Hfq has been reported to bind and to hydrolyze ATP. Here, we present the crystal structure of a C-terminally truncated variant of E. coli Hfq (Hfq(65)) in complex with ATP, showing that it binds to the distal R-sites. In addition, we revisited the reported ATPase activity of full length Hfq purified to homogeneity. At variance with previous reports, no ATPase activity was observed for Hfq. In addition, FRET assays neither indicated an impact of ATP on annealing of two model oligoribonucleotides nor did the presence of ATP induce strand displacement. Moreover, ATP did not lead to destabilization of binary and ternary Hfq-RNA complexes, unless a vast stoichiometric excess of ATP was used. Taken together, these studies strongly suggest that ATP is dispensable for and does not interfere with Hfq-mediated RNA transactions.

Long-read sequencing of nascent RNA reveals coupling among RNA processing events.

PubMed

Herzel, Lydia; Straube, Korinna; Neugebauer, Karla M

2018-06-14

Pre-mRNA splicing is accomplished by the spliceosome, a megadalton complex that assembles de novo on each intron. Because spliceosome assembly and catalysis occur cotranscriptionally, we hypothesized that introns are removed in the order of their transcription in genomes dominated by constitutive splicing. Remarkably little is known about splicing order and the regulatory potential of nascent transcript remodeling by splicing, due to the limitations of existing methods that focus on analysis of mature splicing products (mRNAs) rather than substrates and intermediates. Here, we overcome this obstacle through long-read RNA sequencing of nascent, multi-intron transcripts in the fission yeast Schizosaccharomyces pombe Most multi-intron transcripts were fully spliced, consistent with rapid cotranscriptional splicing. However, an unexpectedly high proportion of transcripts were either fully spliced or fully unspliced, suggesting that splicing of any given intron is dependent on the splicing status of other introns in the transcript. Supporting this, mild inhibition of splicing by a temperature-sensitive mutation in prp2 , the homolog of vertebrate U2AF65, increased the frequency of fully unspliced transcripts. Importantly, fully unspliced transcripts displayed transcriptional read-through at the polyA site and were degraded cotranscriptionally by the nuclear exosome. Finally, we show that cellular mRNA levels were reduced in genes with a high number of unspliced nascent transcripts during caffeine treatment, showing regulatory significance of cotranscriptional splicing. Therefore, overall splicing of individual nascent transcripts, 3' end formation, and mRNA half-life depend on the splicing status of neighboring introns, suggesting crosstalk among spliceosomes and the polyA cleavage machinery during transcription elongation. © 2018 Herzel et al.; Published by Cold Spring Harbor Laboratory Press.

The specificity and flexibility of l1 reverse transcription priming at imperfect T-tracts.

PubMed

Monot, Clément; Kuciak, Monika; Viollet, Sébastien; Mir, Ashfaq Ali; Gabus, Caroline; Darlix, Jean-Luc; Cristofari, Gaël

2013-05-01

L1 retrotransposons have a prominent role in reshaping mammalian genomes. To replicate, the L1 ribonucleoprotein particle (RNP) first uses its endonuclease (EN) to nick the genomic DNA. The newly generated DNA end is subsequently used as a primer to initiate reverse transcription within the L1 RNA poly(A) tail, a process known as target-primed reverse transcription (TPRT). Prior studies demonstrated that most L1 insertions occur into sequences related to the L1 EN consensus sequence (degenerate 5'-TTTT/A-3' sites) and frequently preceded by imperfect T-tracts. However, it is currently unclear whether--and to which degree--the liberated 3'-hydroxyl extremity on the genomic DNA needs to be accessible and complementary to the poly(A) tail of the L1 RNA for efficient priming of reverse transcription. Here, we employed a direct assay for the initiation of L1 reverse transcription to define the molecular rules that guide this process. First, efficient priming is detected with as few as 4 matching nucleotides at the primer 3' end. Second, L1 RNP can tolerate terminal mismatches if they are compensated within the 10 last bases of the primer by an increased number of matching nucleotides. All terminal mismatches are not equally detrimental to DNA extension, a C being extended at higher levels than an A or a G. Third, efficient priming in the context of duplex DNA requires a 3' overhang. This suggests the possible existence of additional DNA processing steps, which generate a single-stranded 3' end to allow L1 reverse transcription. Based on these data we propose that the specificity of L1 reverse transcription initiation contributes, together with the specificity of the initial EN cleavage, to the distribution of new L1 insertions within the human genome.

Single Nucleotide Polymorphisms Can Create Alternative Polyadenylation Signals and Affect Gene Expression through Loss of MicroRNA-Regulation

PubMed Central

Thomas, Laurent F.; Sætrom, Pål

2012-01-01

Alternative polyadenylation (APA) can for example occur when a protein-coding gene has several polyadenylation (polyA) signals in its last exon, resulting in messenger RNAs (mRNAs) with different 3′ untranslated region (UTR) lengths. Different 3′UTR lengths can give different microRNA (miRNA) regulation such that shortened transcripts have increased expression. The APA process is part of human cells' natural regulatory processes, but APA also seems to play an important role in many human diseases. Although altered APA in disease can have many causes, we reasoned that mutations in DNA elements that are important for the polyA process, such as the polyA signal and the downstream GU-rich region, can be one important mechanism. To test this hypothesis, we identified single nucleotide polymorphisms (SNPs) that can create or disrupt APA signals (APA-SNPs). By using a data-integrative approach, we show that APA-SNPs can affect 3′UTR length, miRNA regulation, and mRNA expression—both between homozygote individuals and within heterozygote individuals. Furthermore, we show that a significant fraction of the alleles that cause APA are strongly and positively linked with alleles found by genome-wide studies to be associated with disease. Our results confirm that APA-SNPs can give altered gene regulation and that APA alleles that give shortened transcripts and increased gene expression can be important hereditary causes for disease. PMID:22915998

mRNA interactome capture in mammalian cells.

PubMed

Kastelic, Nicolai; Landthaler, Markus

2017-08-15

Throughout their entire life cycle, mRNAs are associated with RNA-binding proteins (RBPs), forming ribonucleoprotein (RNP) complexes with highly dynamic compositions. Their interplay is one key to control gene regulatory mechanisms from mRNA synthesis to decay. To assay the global scope of RNA-protein interactions, we and others have published a method combining crosslinking with highly stringent oligo(dT) affinity purification to enrich proteins associated with polyadenylated RNA (poly(A)+ RNA). Identification of the poly(A)+ RNA-bound proteome (also: mRNA interactome capture) has by now been applied to a diversity of cell lines and model organisms, uncovering comprehensive repertoires of RBPs and hundreds of novel RBP candidates. In addition to determining the RBP catalog in a given biological system, mRNA interactome capture allows the examination of changes in protein-mRNA interactions in response to internal and external stimuli, altered cellular programs and disease. Copyright © 2017. Published by Elsevier Inc.

The cDNA sequence of mouse Pgp-1 and homology to human CD44 cell surface antigen and proteoglycan core/link proteins.

PubMed

Wolffe, E J; Gause, W C; Pelfrey, C M; Holland, S M; Steinberg, A D; August, J T

1990-01-05

We describe the isolation and sequencing of a cDNA encoding mouse Pgp-1. An oligonucleotide probe corresponding to the NH2-terminal sequence of the purified protein was synthesized by the polymerase chain reaction and used to screen a mouse macrophage lambda gt11 library. A cDNA clone with an insert of 1.2 kilobases was selected and sequenced. In Northern blot analysis, only cells expressing Pgp-1 contained mRNA species that hybridized with this Pgp-1 cDNA. The nucleotide sequence of the cDNA has a single open reading frame that yields a protein-coding sequence of 1076 base pairs followed by a 132-base pair 3'-untranslated sequence that includes a putative polyadenylation signal but no poly(A) tail. The translated sequence comprises a 13-amino acid signal peptide followed by a polypeptide core of 345 residues corresponding to an Mr of 37,800. Portions of the deduced amino acid sequence were identical to those obtained by amino acid sequence analysis from the purified glycoprotein, confirming that the cDNA encodes Pgp-1. The predicted structure of Pgp-1 includes an NH2-terminal extracellular domain (residues 14-265), a transmembrane domain (residues 266-286), and a cytoplasmic tail (residues 287-358). Portions of the mouse Pgp-1 sequence are highly similar to that of the human CD44 cell surface glycoprotein implicated in cell adhesion. The protein also shows sequence similarity to the proteoglycan tandem repeat sequences found in cartilage link protein and cartilage proteoglycan core protein which are thought to be involved in binding to hyaluronic acid.

Structural and Biochemical Studies on ATP Binding and Hydrolysis by the Escherichia coli RNA Chaperone Hfq

PubMed Central

Večerek, Branislav; Rajkowitsch, Lukas; Carugo, Oliviero; Djinović-Carugo, Kristina; Bläsi, Udo

2012-01-01

In Escherichia coli the RNA chaperone Hfq is involved in riboregulation by assisting base-pairing between small regulatory RNAs (sRNAs) and mRNA targets. Several structural and biochemical studies revealed RNA binding sites on either surface of the donut shaped Hfq-hexamer. Whereas sRNAs are believed to contact preferentially the YKH motifs present on the proximal site, poly(A)15 and ADP were shown to bind to tripartite binding motifs (ARE) circularly positioned on the distal site. Hfq has been reported to bind and to hydrolyze ATP. Here, we present the crystal structure of a C-terminally truncated variant of E. coli Hfq (Hfq65) in complex with ATP, showing that it binds to the distal R-sites. In addition, we revisited the reported ATPase activity of full length Hfq purified to homogeneity. At variance with previous reports, no ATPase activity was observed for Hfq. In addition, FRET assays neither indicated an impact of ATP on annealing of two model oligoribonucleotides nor did the presence of ATP induce strand displacement. Moreover, ATP did not lead to destabilization of binary and ternary Hfq-RNA complexes, unless a vast stoichiometric excess of ATP was used. Taken together, these studies strongly suggest that ATP is dispensable for and does not interfere with Hfq-mediated RNA transactions. PMID:23226421

Hepatitis C Virus Core Protein Promotes miR-122 Destabilization by Inhibiting GLD-2

PubMed Central

Kim, Geon-Woo; Lee, Seung-Hoon; Cho, Hee; Kim, Minwoo; Shin, Eui-Cheol; Oh, Jong-Won

2016-01-01

The liver-specific microRNA miR-122, which has essential roles in liver development and metabolism, is a key proviral factor for hepatitis C virus (HCV). Despite its crucial role in the liver and HCV life cycle, little is known about the molecular mechanism of miR-122 expression regulation by HCV infection. Here, we show that the HCV core protein downregulates the abundance of miR-122 by promoting its destabilization via the inhibition of GLD-2, a non-canonical cytoplasmic poly(A) polymerase. The decrease in miR-122 expression resulted in the dysregulation of the known functions of miR-122, including its proviral activity for HCV. By high-throughput sequencing of small RNAs from human liver biopsies, we found that the 22-nucleotide (nt) prototype miR-122 is modified at its 3′ end by 3′-terminal non-templated and templated nucleotide additions. Remarkably, the proportion of miR-122 isomers bearing a single nucleotide tail of any ribonucleotide decreased in liver specimens from patients with HCV. We found that these single-nucleotide-tailed miR-122 isomers display increased miRNA activity and stability over the 22-nt prototype miR-122 and that the 3′-terminal extension is catalyzed by the unique terminal nucleotidyl transferase activity of GLD-2, which is capable of adding any single ribonucleotide without preference of adenylate to the miR-122 3′ end. The HCV core protein specifically inhibited GLD-2, and its interaction with GLD-2 in the cytoplasm was found to be responsible for miR-122 downregulation. Collectively, our results provide new insights into the regulatory role of the HCV core protein in controlling viral RNA abundance and miR-122 functions through miR-122 stability modulation. PMID:27366906

Poly(adenylic acid) complementary DNA real-time polymerase chain reaction in pancreatic ductal juice in patients undergoing pancreaticoduodenectomy.

PubMed

Oliveira-Cunha, Melissa; Byers, Richard J; Siriwardena, Ajith K

2010-03-01

There is a need to develop methods of early diagnosis for pancreatic cancer. Pancreatic juice is easily collected by endoscopic retrograde cholangiopancreatography and may facilitate diagnosis using molecular markers. The aim of this work was to explore the feasibility of measurement of gene expression in RNA isolated from ductal juice. Intraoperative sampling of pancreatic juice was undertaken in 27 patients undergoing pancreaticoduodenectomy for suspected tumor. Total RNA was extracted and used as template for poly(adenylic acid) (poly[A]) polymerase chain reaction (PCR) to generate a globally amplified complementary DNA pool representative of all expressed messenger RNAs. Real-time PCR was performed for trefoil factor 2 (TFF2), carboxypeptidase B1 (CPB1), and kallikrein-related peptidase 3 (KLK3) in a subset of samples; all samples were normalized for 3 reference genes (glyceraldehyde-3-phosphate dehydrogenase [GAPDH], PSMB6, and beta-2-microglobulin [B2M]). The median volume of the pancreatic juice obtained was 1245 microL (range, 50-5000 microL). The RNA integrity number ranged from 1.9 to 10. Reverse transcriptase PCR was positive for pancreas-specific genes (TFF2 and CPB1) and negative for prostatic-specific antigen in all samples. These results demonstrate that RNA analysis of pancreatic juice is feasible using a combination of poly(A) PCR and real-time PCR. In addition, the poly(A) complementary DNA generated can be probed for multiple genes and is indefinitely renewable, thereby representing a molecular block of importance for future research.

Structure of the human gene encoding the protein repair L-isoaspartyl (D-aspartyl) O-methyltransferase.

PubMed

DeVry, C G; Tsai, W; Clarke, S

1996-11-15

The protein L-isoaspartyl/D-aspartyl O-methyltransferase (EC 2.1.1.77) catalyzes the first step in the repair of proteins damaged in the aging process by isomerization or racemization reactions at aspartyl and asparaginyl residues. A single gene has been localized to human chromosome 6 and multiple transcripts arising through alternative splicing have been identified. Restriction enzyme mapping, subcloning, and DNA sequence analysis of three overlapping clones from a human genomic library in bacteriophage P1 indicate that the gene spans approximately 60 kb and is composed of 8 exons interrupted by 7 introns. Analysis of intron/exon splice junctions reveals that all of the donor and acceptor splice sites are in agreement with the mammalian consensus splicing sequence. Determination of transcription initiation sites by primer extension analysis of poly(A)+ mRNA from human brain identifies multiple start sites, with a major site 159 nucleotides upstream from the ATG start codon. Sequence analysis of the 5'-untranslated region demonstrates several potential cis-acting DNA elements including SP1, ETF, AP1, AP2, ARE, XRE, CREB, MED-1, and half-palindromic ERE motifs. The promoter of this methyltransferase gene lacks an identifiable TATA box but is characterized by a CpG island which begins approximately 723 nucleotides upstream of the major transcriptional start site and extends through exon 1 and into the first intron. These features are characteristic of housekeeping genes and are consistent with the wide tissue distribution observed for this methyltransferase activity.

Processed pseudogenes of human endogenous retroviruses generated by LINEs: their integration, stability, and distribution.

PubMed

Pavlícek, Adam; Paces, Jan; Elleder, Daniel; Hejnar, Jirí

2002-03-01

We report here the presence of numerous processed pseudogenes derived from the W family of endogenous retroviruses in the human genome. These pseudogenes are structurally colinear with the retroviral mRNA followed by a poly(A) tail. Our analysis of insertion sites of HERV-W processed pseudogenes shows a strong preference for the insertion motif of long interspersed nuclear element (LINE) retrotransposons. The genomic distribution, stability during evolution, and frequent truncations at the 5' end resemble those of the pseudogenes generated by LINEs. We therefore suggest that HERV-W processed pseudogenes arose by multiple and independent LINE-mediated retrotransposition of retroviral mRNA. These data document that the majority of HERV-W copies are actually nontranscribed promoterless pseudogenes. The current search for HERV-Ws associated with several human diseases should concentrate on a small subset of transcriptionally competent elements.

Novel Method of Estimating Metabolic Rates of Soldiers Engaged in Chemical Biological Defense Training

DTIC Science & Technology

2016-12-01

Simplified example of estimating metabolic rate from core temperature using the SCENARIO thermoregulatory model. 7 4 Edgewood training site, Day 1, core... temperature (TC) and metabolic rate (Ṁ). 10 5 Edgewood training site, Day 2, core temperature (TC) and metabolic rate (Ṁ). 11 6 Hayward...training site, Day 1, core temperature (TC) and metabolic rate (Ṁ). 12 7 Hayward training site, Day 2, core temperature (TC) and metabolic rate (Ṁ). 13

Plant polyadenylation factors: conservation and variety in the polyadenylation complex in plants.

PubMed

Hunt, Arthur G; Xing, Denghui; Li, Qingshun Q

2012-11-20

Polyadenylation, an essential step in eukaryotic gene expression, requires both cis-elements and a plethora of trans-acting polyadenylation factors. The polyadenylation factors are largely conserved across mammals and fungi. The conservation seems also extended to plants based on the analyses of Arabidopsis polyadenylation factors. To extend this observation, we systemically identified the orthologs of yeast and human polyadenylation factors from 10 plant species chosen based on both the availability of their genome sequences and their positions in the evolutionary tree, which render them representatives of different plant lineages. The evolutionary trajectories revealed several interesting features of plant polyadenylation factors. First, the number of genes encoding plant polyadenylation factors was clearly increased from "lower" to "higher" plants. Second, the gene expansion in higher plants was biased to some polyadenylation factors, particularly those involved in RNA binding. Finally, while there are clear commonalities, the differences in the polyadenylation apparatus were obvious across different species, suggesting an ongoing process of evolutionary change. These features lead to a model in which the plant polyadenylation complex consists of a conserved core, which is rather rigid in terms of evolutionary conservation, and a panoply of peripheral subunits, which are less conserved and associated with the core in various combinations, forming a collection of somewhat distinct complex assemblies. The multiple forms of plant polyadenylation complex, together with the diversified polyA signals may explain the intensive alternative polyadenylation (APA) and its regulatory role in biological functions of higher plants.

Efficient Cleavage of Ribosome-Associated Poly(A)-Binding Protein by Enterovirus 3C Protease

PubMed Central

Kuyumcu-Martinez, N. Muge; Joachims, Michelle; Lloyd, Richard E.

2002-01-01

Poliovirus (PV) causes a rapid and drastic inhibition of host cell cap-dependent protein synthesis during infection while preferentially allowing cap-independent translation of its own genomic RNA via an internal ribosome entry site element. Inhibition of cap-dependent translation is partly mediated by cleavage of an essential translation initiation factor, eIF4GI, during PV infection. In addition to cleavage of eIF4GI, cleavage of eIF4GII and poly(A)-binding protein (PABP) has been recently proposed to contribute to complete host translation shutoff; however, the relative importance of eIF4GII and PABP cleavage has not been determined. At times when cap-dependent translation is first blocked during infection, only 25 to 35% of the total cellular PABP is cleaved; therefore, we hypothesized that the pool of PABP associated with polysomes may be preferentially targeted by viral proteases. We have investigated what cleavage products of PABP are produced in vivo and the substrate determinants for cleavage of PABP by 2A protease (2Apro) or 3C protease (3Cpro). Our results show that PABP in ribosome-enriched fractions is preferentially cleaved in vitro and in vivo compared to PABP in other fractions. Furthermore, we have identified four N-terminal PABP cleavage products produced during PV infection and have shown that viral 3C protease generates three of the four cleavage products. Also, 3Cpro is more efficient in cleaving PABP in ribosome-enriched fractions than 2Apro in vitro. In addition, binding of PABP to poly(A) RNA stimulates 3Cpro-mediated cleavage and inhibits 2Apro-mediated cleavage. These results suggest that 3Cpro plays a major role in processing PABP during virus infection and that the interaction of PABP with translation initiation factors, ribosomes, or poly(A) RNA may promote its cleavage by viral 2A and 3C proteases. PMID:11836384

The Specificity and Flexibility of L1 Reverse Transcription Priming at Imperfect T-Tracts

PubMed Central

Viollet, Sébastien; Mir, Ashfaq Ali; Gabus, Caroline; Darlix, Jean-Luc; Cristofari, Gaël

2013-01-01

L1 retrotransposons have a prominent role in reshaping mammalian genomes. To replicate, the L1 ribonucleoprotein particle (RNP) first uses its endonuclease (EN) to nick the genomic DNA. The newly generated DNA end is subsequently used as a primer to initiate reverse transcription within the L1 RNA poly(A) tail, a process known as target-primed reverse transcription (TPRT). Prior studies demonstrated that most L1 insertions occur into sequences related to the L1 EN consensus sequence (degenerate 5′-TTTT/A-3′ sites) and frequently preceded by imperfect T-tracts. However, it is currently unclear whether—and to which degree—the liberated 3′-hydroxyl extremity on the genomic DNA needs to be accessible and complementary to the poly(A) tail of the L1 RNA for efficient priming of reverse transcription. Here, we employed a direct assay for the initiation of L1 reverse transcription to define the molecular rules that guide this process. First, efficient priming is detected with as few as 4 matching nucleotides at the primer 3′ end. Second, L1 RNP can tolerate terminal mismatches if they are compensated within the 10 last bases of the primer by an increased number of matching nucleotides. All terminal mismatches are not equally detrimental to DNA extension, a C being extended at higher levels than an A or a G. Third, efficient priming in the context of duplex DNA requires a 3′ overhang. This suggests the possible existence of additional DNA processing steps, which generate a single-stranded 3′ end to allow L1 reverse transcription. Based on these data we propose that the specificity of L1 reverse transcription initiation contributes, together with the specificity of the initial EN cleavage, to the distribution of new L1 insertions within the human genome. PMID:23675310

ARX polyalanine expansions are highly implicated in familial cases of mental retardation with infantile epilepsy and/or hand dystonia.

PubMed

Cossée, Mireille; Faivre, Laurence; Philippe, Christophe; Hichri, Heifa; de Saint-Martin, Anne; Laugel, Vincent; Bahi-Buisson, Nadia; Lemaitre, Jean-François; Leheup, Bruno; Delobel, Bruno; Demeer, Bénédicte; Poirier, Karine; Biancalana, Valérie; Pinoit, Jean-Michel; Julia, Sophie; Chelly, Jamel; Devys, Didier; Mandel, Jean-Louis

2011-01-01

Mutations in the ARX gene cause both nonsyndromic and several forms of syndromic mental retardation (MR). Two polyalanine (polyA) expansions of ARX are recurrent mutations. The most common one, the c.428_451dup, is associated with a wide spectrum of phenotypes, ranging from the most severe West syndrome to Partington syndrome (MR and hand dystonia), and even nonsyndromic X-linked mental retardation (NS-XLMR). Studies of patients not selected for specific clinical signs showed that the c.428_451dup is relatively frequent in families harboring X-linked MR (7.5%), but less common in familial cases compatible with X-linked NR (1%), and very rare in sporadic cases (0.1%). The c.333_334ins(GCG)7 expansion is less frequent and mainly associated with West syndrome. We screened for both ARX polyA expansions in 98 unrelated patients selected for the presence of NR associated with different types of epilepsy and/or with hand dystonia. We also studied two families with an initial diagnosis of NS-XLMR, one of which was identified as showing linkage to the ARX locus. The c.428_451dup was identified in three patients and the c.333_334ins(GCG)7 in one; all of the patients were from families with two affected brothers. We also found the c.428_451dup in the family linked to ARX, and clinical re-evaluation showed subtle, previously undetected signs. Our study illustrates that ARX polyA expansions are primarily associated with syndromic MR and shows a higher yield (18% in our cohort) when these mutations are screened in familial cases of MR with epilepsy and/or dystonia. Copyright © 2010 Wiley-Liss, Inc.

Gene activity during germination of spores of the fern, Onoclea sensibilis: RNA and protein synthesis and the role of stored mRNA

NASA Technical Reports Server (NTRS)

Raghavan, V.

1991-01-01

Pattern of 3H-uridine incorporation into RNA of spores of Onoclea sensibilis imbibed in complete darkness (non-germinating conditions) and induced to germinate in red light was followed by oligo-dT cellulose chromatography, gel electrophoresis coupled with fluorography and autoradiography. In dark-imbibed spores, RNA synthesis was initiated about 24 h after sowing, with most of the label accumulating in the high mol. wt. poly(A) -RNA fraction. There was no incorporation of the label into poly(A) +RNA until 48 h after sowing. In contrast, photo-induced spores began to synthesize all fractions of RNA within 12 h after sowing and by 24 h, incorporation of 3H-uridine into RNA of irradiated spores was nearly 70-fold higher than that into dark-imbibed spores. Protein synthesis, as monitored by 3H-arginine incorporation into the acid-insoluble fraction and by autoradiography, was initiated in spores within 1-2 h after sowing under both conditions. Autoradiographic experiments also showed that onset of protein synthesis in the cytoplasm of the germinating spore is independent of the transport of newly synthesized nuclear RNA. One-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis of 35S-methionine-labelled proteins revealed a good correspondence between proteins synthesized in a cell-free translation system directed by poly(A) +RNA of dormant spores and those synthesized in vivo by dark-imbibed and photo-induced spores. These results indicate that stored mRNAs of O. sensibilis spores are functionally competent and provide templates for the synthesis of proteins during dark-imbibition and germination.

Complete nucleotide sequence and genome structure of a Japanese isolate of hibiscus latent Fort Pierce virus, a unique tobamovirus that contains an internal poly(A) region in its 3' end.

PubMed

Yoshida, Tetsuya; Kitazawa, Yugo; Komatsu, Ken; Neriya, Yutaro; Ishikawa, Kazuya; Fujita, Naoko; Hashimoto, Masayoshi; Maejima, Kensaku; Yamaji, Yasuyuki; Namba, Shigetou

2014-11-01

In this study, we detected a Japanese isolate of hibiscus latent Fort Pierce virus (HLFPV-J), a member of the genus Tobamovirus, in a hibiscus plant in Japan and determined the complete sequence and organization of its genome. HLFPV-J has four open reading frames (ORFs), each of which shares more than 98 % nucleotide sequence identity with those of other HLFPV isolates. Moreover, HLFPV-J contains a unique internal poly(A) region of variable length, ranging from 44 to 78 nucleotides, in its 3'-untranslated region (UTR), as is the case with hibiscus latent Singapore virus (HLSV), another hibiscus-infecting tobamovirus. The length of the HLFPV-J genome was 6431 nucleotides, including the shortest internal poly(A) region. The sequence identities of ORFs 1, 2, 3 and 4 of HLFPV-J to other tobamoviruses were 46.6-68.7, 49.9-70.8, 31.0-70.8 and 39.4-70.1 %, respectively, at the nucleotide level and 39.8-75.0, 43.6-77.8, 19.2-70.4 and 31.2-74.2 %, respectively, at the amino acid level. The 5'- and 3'-UTRs of HLFPV-J showed 24.3-58.6 and 13.0-79.8 % identity, respectively, to other tobamoviruses. In particular, when compared to other tobamoviruses, each ORF and UTR of HLFPV-J showed the highest sequence identity to those of HLSV. Phylogenetic analysis showed that HLFPV-J, other HLFPV isolates and HLSV constitute a malvaceous-plant-infecting tobamovirus cluster. These results indicate that the genomic structure of HLFPV-J has unique features similar to those of HLSV. To our knowledge, this is the first report of the complete genome sequence of HLFPV.

Expression, crystallization and preliminary crystallographic analysis of RNA-binding protein Hfq (YmaH) from Bacillus subtilis in complex with an RNA aptamer.

PubMed

Baba, Seiki; Someya, Tatsuhiko; Kawai, Gota; Nakamura, Kouji; Kumasaka, Takashi

2010-05-01

The Hfq protein is a hexameric RNA-binding protein which regulates gene expression by binding to RNA under the influence of diverse environmental stresses. Its ring structure binds various types of RNA, including mRNA and sRNA. RNA-bound structures of Hfq from Escherichia coli and Staphylococcus aureus have been revealed to have poly(A) RNA at the distal site and U-rich RNA at the proximal site, respectively. Here, crystals of a complex of the Bacillus subtilis Hfq protein with an A/G-repeat 7-mer RNA (Hfq-RNA) that were prepared using the hanging-drop vapour-diffusion technique are reported. The type 1 Hfq-RNA crystals belonged to space group I422, with unit-cell parameters a = b = 123.70, c = 119.13 A, while the type 2 Hfq-RNA crystals belonged to space group F222, with unit-cell parameters a = 91.92, b = 92.50, c = 114.92 A. Diffraction data were collected to a resolution of 2.20 A from both crystal forms. The hexameric structure of the Hfq protein was clearly shown by self-rotation analysis.

Fourth Graders' Heuristic Problem-Solving Behavior.

ERIC Educational Resources Information Center

Lee, Kil S.

1982-01-01

Eight boys and eight girls from a rural elementary school participated in the investigation. Specific heuristics were adopted from Polya; and the students selected represented two substages of Piaget's concrete operational stage. Five hypotheses were generated, based on observed results and the study's theoretical rationale. (MP)

Nonsense mutations in the alcohol dehydrogenase gene of Drosophila melanogaster correlate with an abnormal 3' end processing of the corresponding pre-mRNA.

PubMed Central

Brogna, S

1999-01-01

From bacteria to mammals, mutations that generate premature termination codons have been shown to result in the reduction in the abundance of the corresponding mRNA. In mammalian cells, more often than not, the reduction happens while the RNA is still associated with the nucleus. Here, it is reported that mutations in the alcohol dehydrogenase gene (Adh) of Drosophila melanogaster that generate premature termination codons lead to reduced levels of cytoplasmic and nuclear mRNA. Unexpectedly, it has been found that the poly(A) tails of Adh mRNAs and pre-mRNAs that carry a premature termination codon are longer than in the wild-type transcript. The more 5' terminal the mutation is, the longer is the poly(A) tail of the transcript. These findings suggest that the integrity of the coding region may be required for accurate mRNA 3' end processing. PMID:10199572

The length of an internal poly(A) tract of hibiscus latent Singapore virus is crucial for its replication.

PubMed

Niu, Shengniao; Cao, Shishu; Huang, Li-Jing; Tan, Kelvin Chee-Leong; Wong, Sek-Man

2015-01-01

Hibiscus latent Singapore virus (HLSV) mutants were constructed to study roles of its internal poly(A) tract (IPAT) in viral replication and coat protein (CP) expression. Shortening of the IPAT resulted in reduced HLSV RNA accumulation and its minimal length required for HLSV CP expression in plants was 24 nt. Disruption of a putative long range RNA-RNA interacting structure between 5' and 3' untranslated regions of HLSV-22A and -24A resulted in reduced viral RNA and undetectable CP accumulation in inoculated leaves. Replacement of the IPAT in HLSV with an upstream pseudoknot domain (UPD) of Tobacco mosaic virus (TMV) or insertion of the UPD to the immediate downstream of a 24 nt IPAT in HLSV resulted in drastically reduced viral RNA replication. Plants infected with a TMV mutant by replacement of the UPD with 43 nt IPAT exhibited milder mosaic symptoms without necrosis. We have proposed a model for HLSV replication. Copyright © 2014 Elsevier Inc. All rights reserved.

Polynucleotide: adenosine glycosidase activity of saporin-L1: effect on DNA, RNA and poly(A).

PubMed Central

Barbieri, L; Valbonesi, P; Gorini, P; Pession, A; Stirpe, F

1996-01-01

The ribosome-inactivating proteins (RIPs) are a family of plant enzymes for which a unique activity has been determined: rRNA N-glycosidase, which removes adenine at a specific universally conserved position (A4324 in the case of rat ribosomes). Here we report that saporin-L1, a RIP from the leaves of Saponaria officinalis, recognizes other substrates, including RNAs from different sources, DNA and poly(A). Saporin-L1 depurinated DNA extensively and released adenine from all adenine-containing polynucleotides tested. Adenine was the only base released from DNA or artificial polynucleotides. The characteristics of the reactions catalysed by saporin-L1 have been determined: optimal pH and temperature, ionic requirements, and the kinetic parameters Km and kcat. The reaction proceeded without cofactors, at low ionic strength, in the absence of Mg2+ and K+. Saporin-L1 had no activity towards various adenine-containing non-polynucleotide compounds (cytokinins, cofactors, nucleotides). This plant protein may now be classified as a polynucleotide: adenosine glycosidase. PMID:8912688

Purification and Properties of Bacteriophage T4-Induced RNA Ligase*

PubMed Central

Silber, Robert; Malathi, V. G.; Hurwitz, Jerard

1972-01-01

An enzyme, purified 300-fold from Escherichia coli infected with bacteriophage T4, catalyzes the conversion of 5′-termini of polyribonucleotides to internal phosphodiester bonds. The reaction requires ATP and Mg++. For every 5′-32P terminus rendered resistant to alkaline phosphatase, an equal amount of AMP and PPi are formed. Various polyribonucleotides are substrates in the reaction; to date, the best substrate is [5′-32P]polyriboadenylate. With the latter substrate, no evidence of intermolecular reaction was obtained. However, the 5′-32P termini of poly(A) rendered resistant to alkaline phosphatase are also resistant to attack by RNase II, polynucleotide phosphorylase, and low concentrations of venom phosphodiesterase. Since the product formed with poly(A) lacks 3′-hydroxyl ends, as measured with these exonucleases, the enzyme appears to convert linear molecules of polyriboadenylate to a circular form by the intramolecular covalent linkage of the 5′-phosphate end to the 3′-hydroxyl terminus. Images PMID:4342972

Statistics of concentrations due to single air pollution sources to be applied in numerical modelling of pollutant dispersion

NASA Astrophysics Data System (ADS)

Tumanov, Sergiu

A test of goodness of fit based on rank statistics was applied to prove the applicability of the Eggenberger-Polya discrete probability law to hourly SO 2-concentrations measured in the vicinity of single sources. With this end in view, the pollutant concentration was considered an integral quantity which may be accepted if one properly chooses the unit of measurement (in this case μg m -3) and if account is taken of the limited accuracy of measurements. The results of the test being satisfactory, even in the range of upper quantiles, the Eggenberger-Polya law was used in association with numerical modelling to estimate statistical parameters, e.g. quantiles, cumulative probabilities of threshold concentrations to be exceeded, and so on, in the grid points of a network covering the area of interest. This only needs accurate estimations of means and variances of the concentration series which can readily be obtained through routine air pollution dispersion modelling.

Impact of extended prostate biopsy including apical anterior region for cancer detection and prediction of surgical margin status for radical prostatectomy.

PubMed

Hashimoto, Kohei; Shinkai, Nobuo; Tanaka, Toshiaki; Masumori, Naoya

2017-06-01

We investigated diagnostic yield of initial biopsy and repeated biopsy including apical cores. We investigated 573 consecutive men with PSA of ≤20 ng/ml who underwent prostate biopsy between 2004 and 2013. The initial 14-core biopsy consisted of the sextant type, lateral sites at the base and middle, lateral apices (la) at anterior horn sites, and apical anterior sites (aa). The repeated 18-core biopsy consisted of the initial 14-core biopsy with four transition zone (TZ) sites at the base (tzb) and middle (tzm). Prostate cancer was diagnosed in 178 (38.9%) of 458 men with the initial 14-core biopsy, and 44 (38.3%) of 115 men with the repeated 18-core biopsy. In the initial biopsy setting, the unique cancer detection rate was high in apical sites (apex, la, and aa: 6.2%, 6.2% and 5.1%, respectively). In the repeated setting, it was high in the TZ site in addition to the apical site (apex, la, aa, tzm, and tzb: 6.8%, 6.8%, 11.4%, 9.1% and 11.4%, respectively). The positive SM rate at the apex was higher in patients whose cancer was detected only in sites other than the sextant region than for those in the sextant region (36.4% vs. 14.8%, P = 0.037). The initial 14-core and the repeated 18-core biopsy scheme including apical anterior cores are feasible for prostate cancer detection. We propose that apical biopsy cores can be used to predict not only the existence of cancer but also surgical margin status at the apex. © The Author 2017. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

A manganese-dependent ribozyme in the 3'-untranslated region of Xenopus Vg1 mRNA.

PubMed

Kolev, Nikolay G; Hartland, Emilia I; Huber, Paul W

2008-10-01

The smallest catalytic RNA identified to date is a manganese-dependent ribozyme that requires only a complex between GAAA and UUU to effect site-specific cleavage. We show here that this ribozyme occurs naturally in the 3'-UTR of Vg1 and beta-actin mRNAs. In accord with earlier studies with model RNAs, cleavage occurs only in the presence of manganese or cadmium ions and proceeds optimally near 30 degrees C and physiological pH. The time course of cleavage in Vg1 mRNA best fits a two-step process in which both steps are first-order. In Vg1 mRNA, the ribozyme is positioned adjacent to a polyadenylation signal, but has no influence on translation of the mRNA in Xenopus oocytes. Putative GAAA ribozyme structures are also near polyadenylation sites in yeast and rat actin mRNAs. Analysis of sequences in the PolyA Cleavage Site and 3'-UTR Database (PACdb) revealed no particular bias in the frequency or distribution of the GAAA motif that would suggest that this ribozyme is currently or was recently used for cleavage to generate processed transcripts. Nonetheless, we speculate that the complementary strands that comprise the ribozyme may account for the origin of sequence elements that direct present-day 3'-end processing of eukaryotic mRNAs.

Alfalfa virus S, a new species in the family Alphaflexiviridae

USDA-ARS?s Scientific Manuscript database

A new species of the family Alphaflexiviridae provisionally named alfalfa virus S (AVS) was discovered in alfalfa samples originating from Sudan. A complete nucleotide sequence of the viral genome consisting of 8,349 nucleotides excluding the 3’ poly(A) tail was determined by high throughput sequenc...

Problem-Solving Test: Expression Cloning of the Erythropoietin Receptor

ERIC Educational Resources Information Center

Szeberenyi, Jozsef

2008-01-01

Terms to be familiar with before you start to solve the test: cytokines, cytokine receptors, cDNA library, cDNA synthesis, poly(A)[superscript +] RNA, primer, template, reverse transcriptase, restriction endonucleases, cohesive ends, expression vector, promoter, Shine-Dalgarno sequence, poly(A) signal, DNA helicase, DNA ligase, topoisomerases,…

The Shoemaker's Knife--An Approach of the Polya Type.

ERIC Educational Resources Information Center

Libeskind, Shlomo; Lott, Johnny W.

1984-01-01

Archimedes' shoemaker's knife problem is interesting in its own right and also allows the demonstration of heuristic teaching ideas and a different method of doing a routine construction. The focus in the article is on the thought processes involved and questions asked when attempting proofs with the problem. (MNS)

Imre Lakatos's Use of Dialogue.

ERIC Educational Resources Information Center

Greig, Judith Maxwell

This paper uses a book, "Proofs and Refutations: The Logic of Mathematical Discovery," as an example of Lakatos's use of dialogue. The book was originally adapted from his dissertation and influenced by Polya and Popper. His discussion of the Euler conjecture is summarized. Three purposes for choosing the dialogue form for the book were…

Genomewide analysis of Drosophila circular RNAs reveals their structural and sequence properties and age-dependent neural accumulation

PubMed Central

Westholm, Jakub O.; Miura, Pedro; Olson, Sara; Shenker, Sol; Joseph, Brian; Sanfilippo, Piero; Celniker, Susan E.; Graveley, Brenton R.; Lai, Eric C.

2014-01-01

Circularization was recently recognized to broadly expand transcriptome complexity. Here, we exploit massive Drosophila total RNA-sequencing data, >5 billion paired-end reads from >100 libraries covering diverse developmental stages, tissues and cultured cells, to rigorously annotate >2500 fruitfly circular RNAs. These mostly derive from back-splicing of protein-coding genes and lack poly(A) tails, and circularization of hundreds of genes is conserved across multiple Drosophila species. We elucidate structural and sequence properties of Drosophila circular RNAs, which exhibit commonalities and distinctions from mammalian circles. Notably, Drosophila circular RNAs harbor >1000 well-conserved canonical miRNA seed matches, especially within coding regions, and coding conserved miRNA sites reside preferentially within circularized exons. Finally, we analyze the developmental and tissue specificity of circular RNAs, and note their preferred derivation from neural genes and enhanced accumulation in neural tissues. Interestingly, circular isoforms increase dramatically relative to linear isoforms during CNS aging, and constitute a novel aging biomarker. PMID:25544350

Sequence specificity for uridylylation of the viral peptide linked to the genome (VPg) of enteroviruses.

PubMed

Schein, Catherine H; Ye, Mengyi; Paul, Aniko V; Oberste, M Steven; Chapman, Nora; van der Heden van Noort, Gerbrand J; Filippov, Dmitri V; Choi, Kyung H

2015-10-01

Enteroviruses (EV) uridylylate a peptide, VPg, as the first step in their replication. VPgpUpU, found free in infected cells, serves as the primer for RNA elongation. The abilities of four polymerases (3D(pol)), from EV-species A-C, to uridylylate VPgs that varied by up to 60% of their residues were compared. Each 3D(pol) was able to uridylylate all five VPgs using polyA RNA as template, while showing specificity for its own genome encoded peptide. All 3D(pol) uridylylated a consensus VPg representing the physical chemical properties of 31 different VPgs. Thus the residues required for uridylylation and the enzymatic mechanism must be similar in diverse EV. As VPg-binding sites differ in co-crystal structures, the reaction is probably done by a second 3D(pol) molecule. The conservation of polymerase residues whose mutation reduces uridylylation but not RNA elongation is compared. Copyright © 2015 Elsevier Inc. All rights reserved.

Genome-wide Analysis of Drosophila Circular RNAs Reveals Their Structural and Sequence Properties and Age-Dependent Neural Accumulation

DOE PAGES

Westholm, Jakub  O.; Miura, Pedro; Olson, Sara; ...

2014-11-26

Circularization was recently recognized to broadly expand transcriptome complexity. Here, we exploit massive Drosophila total RNA-sequencing data, >5 billion paired-end reads from >100 libraries covering diverse developmental stages, tissues, and cultured cells, to rigorously annotate >2,500 fruit fly circular RNAs. These mostly derive from back-splicing of protein-coding genes and lack poly(A) tails, and the circularization of hundreds of genes is conserved across multiple Drosophila species. We elucidate structural and sequence properties of Drosophila circular RNAs, which exhibit commonalities and distinctions from mammalian circles. Notably, Drosophila circular RNAs harbor >1,000 well-conserved canonical miRNA seed matches, especially within coding regions, and codingmore » conserved miRNA sites reside preferentially within circularized exons. Finally, we analyze the developmental and tissue specificity of circular RNAs and note their preferred derivation from neural genes and enhanced accumulation in neural tissues. Interestingly, circular isoforms increase substantially relative to linear isoforms during CNS aging and constitute an aging biomarker.« less

Genome-wide Analysis of Drosophila Circular RNAs Reveals Their Structural and Sequence Properties and Age-Dependent Neural Accumulation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Westholm, Jakub  O.; Miura, Pedro; Olson, Sara

Circularization was recently recognized to broadly expand transcriptome complexity. Here, we exploit massive Drosophila total RNA-sequencing data, >5 billion paired-end reads from >100 libraries covering diverse developmental stages, tissues, and cultured cells, to rigorously annotate >2,500 fruit fly circular RNAs. These mostly derive from back-splicing of protein-coding genes and lack poly(A) tails, and the circularization of hundreds of genes is conserved across multiple Drosophila species. We elucidate structural and sequence properties of Drosophila circular RNAs, which exhibit commonalities and distinctions from mammalian circles. Notably, Drosophila circular RNAs harbor >1,000 well-conserved canonical miRNA seed matches, especially within coding regions, and codingmore » conserved miRNA sites reside preferentially within circularized exons. Finally, we analyze the developmental and tissue specificity of circular RNAs and note their preferred derivation from neural genes and enhanced accumulation in neural tissues. Interestingly, circular isoforms increase substantially relative to linear isoforms during CNS aging and constitute an aging biomarker.« less

The point of no return: The poly(A)-associated elongation checkpoint.

PubMed

Tellier, Michael; Ferrer-Vicens, Ivan; Murphy, Shona

2016-01-01

Cyclin-dependent kinases play critical roles in transcription by RNA polymerase II (pol II) and processing of the transcripts. For example, CDK9 regulates transcription of protein-coding genes, splicing, and 3' end formation of the transcripts. Accordingly, CDK9 inhibitors have a drastic effect on the production of mRNA in human cells. Recent analyses indicate that CDK9 regulates transcription at the early-elongation checkpoint of the vast majority of pol II-transcribed genes. Our recent discovery of an additional CDK9-regulated elongation checkpoint close to poly(A) sites adds a new layer to the control of transcription by this critical cellular kinase. This novel poly(A)-associated checkpoint has the potential to powerfully regulate gene expression just before a functional polyadenylated mRNA is produced: the point of no return. However, many questions remain to be answered before the role of this checkpoint becomes clear. Here we speculate on the possible biological significance of this novel mechanism of gene regulation and the players that may be involved.

The use of mini-samples in palaeomagnetism

NASA Astrophysics Data System (ADS)

Böhnel, Harald; Michalk, Daniel; Nowaczyk, Norbert; Naranjo, Gildardo Gonzalez

2009-10-01

Rock cores of ~25 mm diameter are widely used in palaeomagnetism. Occasionally smaller diameters have been used as well which represents distinct advantages in terms of throughput, weight of equipment and core collections. How their orientation precision compares to 25 mm cores, however, has not been evaluated in detail before. Here we compare the site mean directions and their statistical parameters for 12 lava flows sampled with 25 mm cores (standard samples, typically 8 cores per site) and with 12 mm drill cores (mini-samples, typically 14 cores per site). The site-mean directions for both sample sizes appear to be indistinguishable in most cases. For the mini-samples, site dispersion parameters k on average are slightly lower than for the standard samples reflecting their larger orienting and measurement errors. Applying the Wilcoxon signed-rank test the probability that k or α95 have the same distribution for both sizes is acceptable only at the 17.4 or 66.3 per cent level, respectively. The larger mini-core numbers per site appears to outweigh the lower k values yielding also slightly smaller confidence limits α95. Further, both k and α95 are less variable for mini-samples than for standard size samples. This is interpreted also to result from the larger number of mini-samples per site, which better averages out the detrimental effect of undetected abnormal remanence directions. Sampling of volcanic rocks with mini-samples therefore does not present a disadvantage in terms of the overall obtainable uncertainty of site mean directions. Apart from this, mini-samples do present clear advantages during the field work, as about twice the number of drill cores can be recovered compared to 25 mm cores, and the sampled rock unit is then more widely covered, which reduces the contribution of natural random errors produced, for example, by fractures, cooling joints, and palaeofield inhomogeneities. Mini-samples may be processed faster in the laboratory, which is of particular advantage when carrying out palaeointensity experiments.

Kinetic analysis of bypass of abasic site by the catalytic core of yeast DNA polymerase eta.

PubMed

Yang, Juntang; Wang, Rong; Liu, Binyan; Xue, Qizhen; Zhong, Mengyu; Zeng, Hao; Zhang, Huidong

2015-09-01

Abasic sites (Apurinic/apyrimidinic (AP) sites), produced ∼ 50,000 times/cell/day, are very blocking and miscoding. To better understand miscoding mechanisms of abasic site for yeast DNA polymerase η, pre-steady-state nucleotide incorporation and LC-MS/MS sequence analysis of extension product were studied using pol η(core) (catalytic core, residues 1-513), which can completely eliminate the potential effects of the C-terminal C2H2 motif of pol η on dNTP incorporation. The extension beyond the abasic site was very inefficient. Compared with incorporation of dCTP opposite G, the incorporation efficiencies opposite abasic site were greatly reduced according to the order of dGTP > dATP > dCTP and dTTP. Pol η(core) showed no fast burst phase for any incorporation opposite G or abasic site, suggesting that the catalytic step is not faster than the dissociation of polymerase from DNA. LC-MS/MS sequence analysis of extension products showed that 53% products were dGTP misincorporation, 33% were dATP and 14% were -1 frameshift, indicating that Pol η(core) bypasses abasic site by a combined G-rule, A-rule and -1 frameshift deletions. Compared with full-length pol η, pol η(core) relatively reduced the efficiency of incorporation of dCTP opposite G, increased the efficiencies of dNTP incorporation opposite abasic site and the exclusive incorporation of dGTP opposite abasic site, but inhibited the extension beyond abasic site, and increased the priority in extension of A: abasic site relative to G: abasic site. This study provides further understanding in the mutation mechanism of abasic sites for yeast DNA polymerase η. Copyright © 2015 Elsevier B.V. All rights reserved.

Central Core Laboratory versus Site Interpretation of Coronary CT Angiography: Agreement and Association with Cardiovascular Events in the PROMISE Trial.

PubMed

Lu, Michael T; Meyersohn, Nandini M; Mayrhofer, Thomas; Bittner, Daniel O; Emami, Hamed; Puchner, Stefan B; Foldyna, Borek; Mueller, Martin E; Hearne, Steven; Yang, Clifford; Achenbach, Stephan; Truong, Quynh A; Ghoshhajra, Brian B; Patel, Manesh R; Ferencik, Maros; Douglas, Pamela S; Hoffmann, Udo

2018-04-01

Purpose To assess concordance and relative prognostic utility between central core laboratory and local site interpretation for significant coronary artery disease (CAD) and cardiovascular events. Materials and Methods In the Prospective Multicenter Imaging Study for Evaluation of Chest Pain (PROMISE) trial, readers at 193 North American sites interpreted coronary computed tomographic (CT) angiography as part of the clinical evaluation of stable chest pain. Readers at a central core laboratory also interpreted CT angiography blinded to clinical data, site interpretation, and outcomes. Significant CAD was defined as stenosis greater than or equal to 50%; cardiovascular events were defined as a composite of cardiovascular death or myocardial infarction. Results In 4347 patients (51.8% women; mean age ± standard deviation, 60.4 years ± 8.2), core laboratory and site interpretations were discordant in 16% (683 of 4347), most commonly because of a finding of significant CAD by site but not by core laboratory interpretation (80%, 544 of 683). Overall, core laboratory interpretation resulted in 41% fewer patients being reported as having significant CAD (14%, 595 of 4347 vs 23%, 1000 of 4347; P < .001). Over a median follow-up period of 25 months, 1.3% (57 of 4347) sustained myocardial infarction or cardiovascular death. The C statistic for future myocardial infarction or cardiovascular death was 0.61 (95% confidence interval [CI]: 0.54, 0.68) for the core laboratory and 0.63 (95% CI: 0.56, 0.70) for the sites. Conclusion Compared with interpretation by readers at 193 North American sites, standardized core laboratory interpretation classified 41% fewer patients as having significant CAD. © RSNA, 2017 Online supplemental material is available for this article. Clinical trial registration no. NCT01174550.

How Spherical Are the Archimedean Solids and Their Duals?

ERIC Educational Resources Information Center

Aravind, P. K.

2011-01-01

The Isoperimetric Quotient, or IQ, introduced by G. Polya, characterizes the degree of sphericity of a convex solid. This paper obtains closed form expressions for the surface area and volume of any Archimedean polyhedron in terms of the integers specifying the type and number of regular polygons occurring around each vertex. Similar results are…

Sharing Teaching Ideas: Active Participation in the Classroom through Creative Problem Generation.

ERIC Educational Resources Information Center

Gonzales, Nancy A.; And Others

1996-01-01

Presents an activity to involve students in mathematical communication and creative thinking. The activity is similar to the "pass it along" gossip game in which each person in a chain adds a piece of information. The class analyzes the resulting mathematics problem using George Polya's problem-solving techniques. (MKR)

Complete nucleotide sequence and genome organization of a novel allexivirus from alfalfa (Medicago sativa)

USDA-ARS?s Scientific Manuscript database

A new species of the family Alphaflexiviridae provisionally named Alfalfa virus S (AVS) was diagnosed in alfalfa samples originating from Sudan. A complete nucleotide sequence of the viral genome consisting of 8,349 nucleotides excluding the 3’ poly(A) tail was determined by Illumina NGS technology ...

Statistical properties of alternative national forest inventory area estimators

Treesearch

Francis Roesch; John Coulston; Andrew D. Hill

2012-01-01

The statistical properties of potential estimators of forest area for the USDA Forest Service's Forest Inventory and Analysis (FIA) program are presented and discussed. The current FIA area estimator is compared and contrasted with a weighted mean estimator and an estimator based on the Polya posterior, in the presence of nonresponse. Estimator optimality is...

Problem Solving through an Optimization Problem in Geometry

ERIC Educational Resources Information Center

Poon, Kin Keung; Wong, Hang-Chi

2011-01-01

This article adapts the problem-solving model developed by Polya to investigate and give an innovative approach to discuss and solve an optimization problem in geometry: the Regiomontanus Problem and its application to football. Various mathematical tools, such as calculus, inequality and the properties of circles, are used to explore and reflect…

Complete genome sequence of a novel potyvirus, Callistephus mottle virus identified in Callistephus chinensis

USDA-ARS?s Scientific Manuscript database

The complete genomic sequence of a novel putative member of the genus Potyvirus was detected from Callistephus chinensis (china aster) in South Korea. The genomic RNA consists of 9,859 nucleotides excluding the 3’ poly(A) tail. The Callistephus virus genome, which contains the typical open reading f...

Simple methods for the 3' biotinylation of RNA.

PubMed

Moritz, Bodo; Wahle, Elmar

2014-03-01

Biotinylation of RNA allows its tight coupling to streptavidin and is thus useful for many types of experiments, e.g., pull-downs. Here we describe three simple techniques for biotinylating the 3' ends of RNA molecules generated by chemical or enzymatic synthesis. First, extension with either the Schizosaccharomyces pombe noncanonical poly(A) polymerase Cid1 or Escherichia coli poly(A) polymerase and N6-biotin-ATP is simple, efficient, and generally applicable independently of the 3'-end sequences of the RNA molecule to be labeled. However, depending on the enzyme and the reaction conditions, several or many biotinylated nucleotides are incorporated. Second, conditions are reported under which splint-dependent ligation by T4 DNA ligase can be used to join biotinylated and, presumably, other chemically modified DNA oligonucleotides to RNA 3' ends even if these are heterogeneous as is typical for products of enzymatic synthesis. Third, we describe the use of 29 DNA polymerase for a template-directed fill-in reaction that uses biotin-dUTP and, thanks to the enzyme's proofreading activity, can cope with more extended 3' heterogeneities.

Star-PAP, a poly(A) polymerase, functions as a tumor suppressor in an orthotopic human breast cancer model.

PubMed

Yu, C; Gong, Y; Zhou, H; Wang, M; Kong, L; Liu, J; An, T; Zhu, H; Li, Y

2017-02-02

Star-PAP is a noncanonical poly(A) polymerase and required for the expression of a select set of mRNAs. However, the pathological role of Star-PAP in cancer largely remains unknown. In this study, we observed decreased expression of Star-PAP in breast cancer cell lines and tissues. Ectopic Star-PAP expression inhibited proliferation as well as colony-forming ability of breast cancer cells. In breast cancer patients, high levels of Star-PAP correlated with an improved prognosis. Moreover, by regulating the expression of BIK (BCL2-interacting killer), Star-PAP induced apoptosis of breast cancer cells through the mitochondrial pathway. The growth of breast cancer xenografts in NOD/SCID mice was also inhibited by the doxycycline-induced Star-PAP overexpression. Furthermore, Star-PAP sensitized breast cancer cells to chemotherapy drugs both in vitro and in vivo. In mammary epithelial cells, Star-PAP knockdown partially transformed these cells and induced them to undergo epithelial-mesenchymal transition (EMT). These findings suggested that Star-PAP possesses tumor-suppressing activity and can be a valuable target for developing new cancer therapeutic strategies.

Lineage-specific roles of the cytoplasmic polyadenylation factor CPEB4 in the regulation of melanoma drivers

PubMed Central

Pérez-Guijarro, Eva; Karras, Panagiotis; Cifdaloz, Metehan; Martínez-Herranz, Raúl; Cañón, Estela; Graña, Osvaldo; Horcajada-Reales, Celia; Alonso-Curbelo, Direna; Calvo, Tonantzin G.; Gómez-López, Gonzalo; Bellora, Nicolas; Riveiro-Falkenbach, Erica; Ortiz-Romero, Pablo L.; Rodríguez-Peralto, José L.; Maestre, Lorena; Roncador, Giovanna; de Agustín Asensio, Juan C.; Goding, Colin R.; Eyras, Eduardo; Megías, Diego; Méndez, Raúl; Soengas, María S.

2016-01-01

Nuclear 3'-end-polyadenylation is essential for the transport, stability and translation of virtually all eukaryotic mRNAs. Poly(A) tail extension can also occur in the cytoplasm, but the transcripts involved are incompletely understood, particularly in cancer. Here we identify a lineage-specific requirement of the cytoplasmic polyadenylation binding protein 4 (CPEB4) in malignant melanoma. CPEB4 is upregulated early in melanoma progression, as defined by computational and histological analyses. Melanoma cells are distinct from other tumour cell types in their dependency on CPEB4, not only to prevent mitotic aberrations, but to progress through G1/S cell cycle checkpoints. RNA immunoprecipitation, sequencing of bound transcripts and poly(A) length tests link the melanoma-specific functions of CPEB4 to signalling hubs specifically enriched in this disease. Essential in these CPEB4-controlled networks are the melanoma drivers MITF and RAB7A, a feature validated in clinical biopsies. These results provide new mechanistic links between cytoplasmic polyadenylation and lineage specification in melanoma. PMID:27857118

Expression, crystallization and preliminary crystallographic analysis of RNA-binding protein Hfq (YmaH) from Bacillus subtilis in complex with an RNA aptamer

PubMed Central

Baba, Seiki; Someya, Tatsuhiko; Kawai, Gota; Nakamura, Kouji; Kumasaka, Takashi

2010-01-01

The Hfq protein is a hexameric RNA-binding protein which regulates gene expression by binding to RNA under the influence of diverse environmental stresses. Its ring structure binds various types of RNA, including mRNA and sRNA. RNA-bound structures of Hfq from Escherichia coli and Staphylococcus aureus have been revealed to have poly(A) RNA at the distal site and U-rich RNA at the proximal site, respectively. Here, crystals of a complex of the Bacillus subtilis Hfq protein with an A/G-repeat 7-mer RNA (Hfq–RNA) that were prepared using the hanging-drop vapour-diffusion technique are reported. The type 1 Hfq–RNA crystals belonged to space group I422, with unit-cell parameters a = b = 123.70, c = 119.13 Å, while the type 2 Hfq–RNA crystals belonged to space group F222, with unit-cell parameters a = 91.92, b = 92.50, c = 114.92 Å. Diffraction data were collected to a resolution of 2.20 Å from both crystal forms. The hexameric structure of the Hfq protein was clearly shown by self-rotation analysis. PMID:20445260

GENCODE: the reference human genome annotation for The ENCODE Project.

PubMed

Harrow, Jennifer; Frankish, Adam; Gonzalez, Jose M; Tapanari, Electra; Diekhans, Mark; Kokocinski, Felix; Aken, Bronwen L; Barrell, Daniel; Zadissa, Amonida; Searle, Stephen; Barnes, If; Bignell, Alexandra; Boychenko, Veronika; Hunt, Toby; Kay, Mike; Mukherjee, Gaurab; Rajan, Jeena; Despacio-Reyes, Gloria; Saunders, Gary; Steward, Charles; Harte, Rachel; Lin, Michael; Howald, Cédric; Tanzer, Andrea; Derrien, Thomas; Chrast, Jacqueline; Walters, Nathalie; Balasubramanian, Suganthi; Pei, Baikang; Tress, Michael; Rodriguez, Jose Manuel; Ezkurdia, Iakes; van Baren, Jeltje; Brent, Michael; Haussler, David; Kellis, Manolis; Valencia, Alfonso; Reymond, Alexandre; Gerstein, Mark; Guigó, Roderic; Hubbard, Tim J

2012-09-01

The GENCODE Consortium aims to identify all gene features in the human genome using a combination of computational analysis, manual annotation, and experimental validation. Since the first public release of this annotation data set, few new protein-coding loci have been added, yet the number of alternative splicing transcripts annotated has steadily increased. The GENCODE 7 release contains 20,687 protein-coding and 9640 long noncoding RNA loci and has 33,977 coding transcripts not represented in UCSC genes and RefSeq. It also has the most comprehensive annotation of long noncoding RNA (lncRNA) loci publicly available with the predominant transcript form consisting of two exons. We have examined the completeness of the transcript annotation and found that 35% of transcriptional start sites are supported by CAGE clusters and 62% of protein-coding genes have annotated polyA sites. Over one-third of GENCODE protein-coding genes are supported by peptide hits derived from mass spectrometry spectra submitted to Peptide Atlas. New models derived from the Illumina Body Map 2.0 RNA-seq data identify 3689 new loci not currently in GENCODE, of which 3127 consist of two exon models indicating that they are possibly unannotated long noncoding loci. GENCODE 7 is publicly available from gencodegenes.org and via the Ensembl and UCSC Genome Browsers.

1,4-Benzoquinone reductase from Phanerochaete chrysosporium: cDNA cloning and regulation of expression

DOE Office of Scientific and Technical Information (OSTI.GOV)

Akileswaran, L.; Brock, B.J.; Cereghino, J.L.

1999-02-01

A cDNA clone encoding a quinone reductase (QR) from the white rot basidiomycete Phanerochaete chrysosporium was isolated and sequenced. The cDNA consisted of 1,007 nucleotides and a poly(A) tail and encoded a deduced protein containing 271 amino acids. The experimentally determined eight-amino-acid N-germinal sequence of the purified QR protein from P. chrysosporium matched amino acids 72 to 79 of the predicted translation product of the cDNA. The M{sub r} of the predicted translation product, beginning with Pro-72, was essentially identical to the experimentally determined M{sub r} of one monomer of the QR dimer, and this finding suggested that QR ismore » synthesized as a proenzyme. The results of in vitro transcription-translation experiments suggested that QR is synthesized as a proenzyme with a 71-amino-acid leader sequence. This leader sequence contains two potential KEX2 cleavage sites and numerous potential cleavage sites for dipeptidyl aminopeptidase. The QR activity in cultures of P. chrysosporium increased following the addition of 2-dimethoxybenzoquinone, vanillic acid, or several other aromatic compounds. An immunoblot analysis indicated that induction resulted in an increase in the amount of QR protein, and a Northern blot analysis indicated that this regulation occurs at the level of the qr mRNA.« less

The Oncogenic Fusion Proteins SET-Nup214 and Sequestosome-1 (SQSTM1)-Nup214 Form Dynamic Nuclear Bodies and Differentially Affect Nuclear Protein and Poly(A)+ RNA Export.

PubMed

Port, Sarah A; Mendes, Adélia; Valkova, Christina; Spillner, Christiane; Fahrenkrog, Birthe; Kaether, Christoph; Kehlenbach, Ralph H

2016-10-28

Genetic rearrangements are a hallmark of several forms of leukemia and can lead to oncogenic fusion proteins. One example of an affected chromosomal region is the gene coding for Nup214, a nucleoporin that localizes to the cytoplasmic side of the nuclear pore complex (NPC). We investigated two such fusion proteins, SET-Nup214 and SQSTM1 (sequestosome)-Nup214, both containing C-terminal portions of Nup214. SET-Nup214 nuclear bodies containing the nuclear export receptor CRM1 were observed in the leukemia cell lines LOUCY and MEGAL. Overexpression of SET-Nup214 in HeLa cells leads to the formation of similar nuclear bodies that recruit CRM1, export cargo proteins, and certain nucleoporins and concomitantly affect nuclear protein and poly(A) + RNA export. SQSTM1-Nup214, although mostly cytoplasmic, also forms nuclear bodies and inhibits nuclear protein but not poly(A) + RNA export. The interaction of the fusion proteins with CRM1 is RanGTP-dependent, as shown in co-immunoprecipitation experiments and binding assays. Further analysis revealed that the Nup214 parts mediate the inhibition of nuclear export, whereas the SET or SQSTM1 part determines the localization of the fusion protein and therefore the extent of the effect. SET-Nup214 nuclear bodies are highly mobile structures, which are in equilibrium with the nucleoplasm in interphase and disassemble during mitosis or upon treatment of cells with the CRM1-inhibitor leptomycin B. Strikingly, we found that nucleoporins can be released from nuclear bodies and reintegrated into existing NPC. Our results point to nuclear bodies as a means of preventing the formation of potentially insoluble and harmful protein aggregates that also may serve as storage compartments for nuclear transport factors. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

The Oncogenic Fusion Proteins SET-Nup214 and Sequestosome-1 (SQSTM1)-Nup214 Form Dynamic Nuclear Bodies and Differentially Affect Nuclear Protein and Poly(A)+ RNA Export*

PubMed Central

Port, Sarah A.; Mendes, Adélia; Valkova, Christina; Spillner, Christiane; Fahrenkrog, Birthe; Kaether, Christoph; Kehlenbach, Ralph H.

2016-01-01

Genetic rearrangements are a hallmark of several forms of leukemia and can lead to oncogenic fusion proteins. One example of an affected chromosomal region is the gene coding for Nup214, a nucleoporin that localizes to the cytoplasmic side of the nuclear pore complex (NPC). We investigated two such fusion proteins, SET-Nup214 and SQSTM1 (sequestosome)-Nup214, both containing C-terminal portions of Nup214. SET-Nup214 nuclear bodies containing the nuclear export receptor CRM1 were observed in the leukemia cell lines LOUCY and MEGAL. Overexpression of SET-Nup214 in HeLa cells leads to the formation of similar nuclear bodies that recruit CRM1, export cargo proteins, and certain nucleoporins and concomitantly affect nuclear protein and poly(A)+ RNA export. SQSTM1-Nup214, although mostly cytoplasmic, also forms nuclear bodies and inhibits nuclear protein but not poly(A)+ RNA export. The interaction of the fusion proteins with CRM1 is RanGTP-dependent, as shown in co-immunoprecipitation experiments and binding assays. Further analysis revealed that the Nup214 parts mediate the inhibition of nuclear export, whereas the SET or SQSTM1 part determines the localization of the fusion protein and therefore the extent of the effect. SET-Nup214 nuclear bodies are highly mobile structures, which are in equilibrium with the nucleoplasm in interphase and disassemble during mitosis or upon treatment of cells with the CRM1-inhibitor leptomycin B. Strikingly, we found that nucleoporins can be released from nuclear bodies and reintegrated into existing NPC. Our results point to nuclear bodies as a means of preventing the formation of potentially insoluble and harmful protein aggregates that also may serve as storage compartments for nuclear transport factors. PMID:27613868

Developmental regulation of collagenase-3 mRNA in normal, differentiating osteoblasts through the activator protein-1 and the runt domain binding sites

NASA Technical Reports Server (NTRS)

Winchester, S. K.; Selvamurugan, N.; D'Alonzo, R. C.; Partridge, N. C.

2000-01-01

Collagenase-3 mRNA is initially detectable when osteoblasts cease proliferation, increasing during differentiation and mineralization. We showed that this developmental expression is due to an increase in collagenase-3 gene transcription. Mutation of either the activator protein-1 or the runt domain binding site decreased collagenase-3 promoter activity, demonstrating that these sites are responsible for collagenase-3 gene transcription. The activator protein-1 and runt domain binding sites bind members of the activator protein-1 and core-binding factor family of transcription factors, respectively. We identified core-binding factor a1 binding to the runt domain binding site and JunD in addition to a Fos-related antigen binding to the activator protein-1 site. Overexpression of both c-Fos and c-Jun in osteoblasts or core-binding factor a1 increased collagenase-3 promoter activity. Furthermore, overexpression of c-Fos, c-Jun, and core-binding factor a1 synergistically increased collagenase-3 promoter activity. Mutation of either the activator protein-1 or the runt domain binding site resulted in the inability of c-Fos and c-Jun or core-binding factor a1 to increase collagenase-3 promoter activity, suggesting that there is cooperative interaction between the sites and the proteins. Overexpression of Fra-2 and JunD repressed core-binding factor a1-induced collagenase-3 promoter activity. Our results suggest that members of the activator protein-1 and core-binding factor families, binding to the activator protein-1 and runt domain binding sites are responsible for the developmental regulation of collagenase-3 gene expression in osteoblasts.

Analyzing tree cores to detect petroleum hydrocarbon-contaminated groundwater at a former landfill site in the community of Happy Valley-Goose Bay, eastern Canadian subarctic.

PubMed

Fonkwe, Merline L D; Trapp, Stefan

2016-08-01

This research examines the feasibility of analyzing tree cores to detect benzene, toluene, ethylbenzene, and m, p, o-xylene (BTEX) compounds and methyl tertiary-butyl ether (MTBE) in groundwater in eastern Canada subarctic environments, using a former landfill site in the remote community of Happy Valley-Goose Bay, Labrador. Petroleum hydrocarbon contamination at the landfill site is the result of environmentally unsound pre-1990s disposal of households and industrial solid wastes. Tree cores were taken from trembling aspen, black spruce, and white birch and analyzed by headspace-gas chromatography-mass spectrometry. BTEX compounds were detected in tree cores, corroborating known groundwater contamination. A zone of anomalously high concentrations of total BTEX constituents was identified and recommended for monitoring by groundwater wells. Tree cores collected outside the landfill site at a local control area suggest the migration of contaminants off-site. Tree species exhibit different concentrations of BTEX constituents, indicating selective uptake and accumulation. Toluene in wood exhibited the highest concentrations, which may also be due to endogenous production. Meanwhile, MTBE was not found in the tree cores and is considered to be absent in the groundwater. The results demonstrate that tree-core analysis can be useful for detecting anomalous concentrations of petroleum hydrocarbons, such as BTEX compounds, in subarctic sites with shallow unconfined aquifers and permeable soils. This method can therefore aid in the proper management of contamination during landfill operations and after site closures.

Elaborately designed diblock nanoprobes for simultaneous multicolor detection of microRNAs

NASA Astrophysics Data System (ADS)

Wang, Chenguang; Zhang, Huan; Zeng, Dongdong; Sun, Wenliang; Zhang, Honglu; Aldalbahi, Ali; Wang, Yunsheng; San, Lili; Fan, Chunhai; Zuo, Xiaolei; Mi, Xianqiang

2015-09-01

Simultaneous detection of multiple biomarkers has important prospects in the biomedical field. In this work, we demonstrated a novel strategy for the detection of multiple microRNAs (miRNAs) based on gold nanoparticles (Au NPs) and polyadenine (polyA) mediated nanoscale molecular beacon (MB) probes (denoted p-nanoMBs). Novel fluorescent labeled p-nanoMBs bearing consecutive adenines were designed, of which polyA served as an effective anchoring block binding to the surface of Au NPs, and the appended hairpin block formed an upright conformation that favored the hybridization with targets. Using the co-assembling method and the improved hybridization conformation of the hairpin probes, we achieved high selectivity for specifically distinguishing DNA targets from single-base mismatched DNA targets. We also realized multicolor detection of three different synthetic miRNAs in a wide dynamic range from 0.01 nM to 200 nM with a detection limit of 10 pM. What's more, we even detected miRNAs in a simulated serum environment, which indicated that our method could be used in complex media. Compared with the traditional method, our strategy provides a promising alternative method for the qualitative and quantitative detection of miRNAs.Simultaneous detection of multiple biomarkers has important prospects in the biomedical field. In this work, we demonstrated a novel strategy for the detection of multiple microRNAs (miRNAs) based on gold nanoparticles (Au NPs) and polyadenine (polyA) mediated nanoscale molecular beacon (MB) probes (denoted p-nanoMBs). Novel fluorescent labeled p-nanoMBs bearing consecutive adenines were designed, of which polyA served as an effective anchoring block binding to the surface of Au NPs, and the appended hairpin block formed an upright conformation that favored the hybridization with targets. Using the co-assembling method and the improved hybridization conformation of the hairpin probes, we achieved high selectivity for specifically distinguishing DNA targets from single-base mismatched DNA targets. We also realized multicolor detection of three different synthetic miRNAs in a wide dynamic range from 0.01 nM to 200 nM with a detection limit of 10 pM. What's more, we even detected miRNAs in a simulated serum environment, which indicated that our method could be used in complex media. Compared with the traditional method, our strategy provides a promising alternative method for the qualitative and quantitative detection of miRNAs. Electronic supplementary information (ESI) available: Sequences for oligonucleotides used for this work, dynamic light scattering (DLS) measurements, fluorescent signal intensity with different ratios between p-MBs and A5 oligonucleotides, quantification of the fluorescent p-MB, and UV-Vis spectra for naked AuNPs and the p-nanoMB. See DOI: 10.1039/c5nr04618a

Strong vortex core pinning and Barkhausen-free magnetization response in thin Permalloy disks induced by implantation of 1 × 10{sup 4} Ga{sup +} ions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fani Sani, F., E-mail: fanisani@ualberta.ca, E-mail: mark.freeman@ualberta.ca; Losby, J. E.; Diao, Z.

2014-05-07

Artificial vortex core pinning sites are induced in thin Permalloy disks by point exposure to as few as 10 000 ions from a focused Ga{sup +} beam. These pinning sites yield a first-order change in the magnetization response of the disk. A single site can keep the vortex core pinned over an applied field range comparable to the vortex annihilation field of the unaltered disk. Several widely separated sites can work together to keep the core pinned in one place, while the Barkhausen effect is eliminated from the magnetization curve over a range approaching the saturation moment of the disk.

Leisure Activities for the Development of Creative Intelligence in Mathematical Problem Solving

ERIC Educational Resources Information Center

Castro, Angélica Mercedes Tumbaco; Guerra, Galo Ernestro Cabanilla; Brito, Christian Antonio Pavón; Chávez, Tannia Gabriela Acosta

2018-01-01

The present work studies the influence of leisure activities on the creative intelligence of the students. An experimental pre- and post-test design was carried out with individuals selected in a sampling process. The design identifies the ease of students to place themselves in possible contexts and solve them mathematically through Polya's…

Hermite-Birkhoff interpolation in the nth roots of unity

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cavaretta, A.S. Jr.; Sharma, A.; Varga, R.S.

1980-06-01

Consider, as nodes for polynomial interpolation, the nth roots of unity. For a sufficiently smooth function f(z), we require a polynomial p(z) to interpolate f and certain of its derivatives at each node. It is shown that the so-called Polya conditions, which are necessary for unique interpolation, are in this setting also sufficient.

Pre-Service Class Teacher' Ability in Solving Mathematical Problems and Skills in Solving Daily Problems

ERIC Educational Resources Information Center

Aljaberi, Nahil M.; Gheith, Eman

2016-01-01

This study aims to investigate the ability of pre-service class teacher at University of Petrain solving mathematical problems using Polya's Techniques, their level of problem solving skills in daily-life issues. The study also investigates the correlation between their ability to solve mathematical problems and their level of problem solving…

Investigation of the Effect of Assignment Projects on Mathematical Activity of Graduating Junior High School Students.

ERIC Educational Resources Information Center

Zehavi, Nurit

This study explored student mathematical activity in open problem-solving situations, derived from the work of Polya on problem solving and Skemp on intelligent learning and teaching. Assignment projects with problems for ninth-grade students were developed, whether they elicit the desired cognitive and cogno-affective goals was investigated, and…

The Rhinovirus Subviral A-Particle Exposes 3′-Terminal Sequences of Its Genomic RNA

PubMed Central

Harutyunyan, Shushan; Kowalski, Heinrich

2014-01-01

ABSTRACT Enteroviruses, which represent a large genus within the family Picornaviridae, undergo important conformational modifications during infection of the host cell. Once internalized by receptor-mediated endocytosis, receptor binding and/or the acidic endosomal environment triggers the native virion to expand and convert into the subviral (altered) A-particle. The A-particle is lacking the internal capsid protein VP4 and exposes N-terminal amphipathic sequences of VP1, allowing for its direct interaction with a lipid bilayer. The genomic single-stranded (+)RNA then exits through a hole close to a 2-fold axis of icosahedral symmetry and passes through a pore in the endosomal membrane into the cytosol, leaving behind the empty shell. We demonstrate that in vitro acidification of a prototype of the minor receptor group of common cold viruses, human rhinovirus A2 (HRV-A2), also results in egress of the poly(A) tail of the RNA from the A-particle, along with adjacent nucleotides totaling ∼700 bases. However, even after hours of incubation at pH 5.2, 5′-proximal sequences remain inside the capsid. In contrast, the entire RNA genome is released within minutes of exposure to the acidic endosomal environment in vivo. This finding suggests that the exposed 3′-poly(A) tail facilitates the positioning of the RNA exit site onto the putative channel in the lipid bilayer, thereby preventing the egress of viral RNA into the endosomal lumen, where it may be degraded. IMPORTANCE For host cell infection, a virus transfers its genome from within the protective capsid into the cytosol; this requires modifications of the viral shell. In common cold viruses, exit of the RNA genome is prepared by the acidic environment in endosomes converting the native virion into the subviral A-particle. We demonstrate that acidification in vitro results in RNA exit starting from the 3′-terminal poly(A). However, the process halts as soon as about 700 bases have left the viral shell. Conversely, inside the cell, RNA egress completes in about 2 min. This suggests the existence of cellular uncoating facilitators. PMID:24672023

Evaluation of surficial sediment toxicity and sediment physico-chemical characteristics of representative sites in the Lagoon of Venice (Italy)

NASA Astrophysics Data System (ADS)

Losso, C.; Arizzi Novelli, A.; Picone, M.; Marchetto, D.; Pessa, G.; Molinaroli, E.; Ghetti, P. F.; Volpi Ghirardini, A.

2004-11-01

Toxic hazard in sites with varying types and levels of contamination in the Lagoon of Venice was estimated by means of toxicity bioassays based on the early life-stages of the autochthonous sea urchin Paracentrotus lividus. Elutriate was chosen as the test matrix, due to its ability to highlight potential toxic effects towards sensitive biological components of the water column caused by sediment resuspension phenomena affecting the Lagoon. Surficial sediments (core-top 5 cm deep), directly influenced by resuspension/redeposition processes, and core sediments (core 20 cm deep), recording time-mediated contamination, were sampled in some sites located in the lagoonal area most greatly influenced by anthropogenic activities. Particle size, organic matter and water content were also analysed. In two sites, the results of physical parameters showed that the core-top sediments were coarser than the 20-cm core sediments. Sperm cell toxicity test results showed the negligible acute toxicity of elutriates from all investigated sites. The embryo toxicity test demonstrated a short-term chronic toxicity gradient for elutriates from the 20-cm core sediments, in general agreement both with the expected contamination gradient and with results of the Microtox® solid-phase test. Elutriates of the core-top 5-cm sediments revealed a totally inverted gradient, in comparison with that for the 20-cm core sediments, and the presence of a "hot spot" of contamination in the site chosen as a possible reference. Investigations on ammonia and sulphides as possible confounding factors excluded their contribution to this "hot spot". Integrated physico-chemical and toxicity results on sediments at various depths demonstrated the presence of disturbed sediments in the central basin of the Lagoon of Venice.

Ground-truth collections at the MTI core sites

NASA Astrophysics Data System (ADS)

Garrett, Alfred J.; Kurzeja, Robert J.; Parker, Matthew J.; O'Steen, Byron L.; Pendergast, Malcolm M.; Villa-Aleman, Eliel

2001-08-01

The Savannah River Technology Center (SRTC) selected 13 sites across the continental US and one site in the western Pacific to serve as the primary or core site for collection of ground truth data for validation of MTI science algorithms. Imagery and ground truth data from several of these sites are presented in this paper. These sites are the Comanche Peak, Pilgrim and Turkey Point power plants, Ivanpah playas, Crater Lake, Stennis Space Center and the Tropical Western Pacific ARM site on the island of Nauru. Ground truth data includes water temperatures (bulk and skin), radiometric data, meteorological data and plant operating data. The organizations that manage these sites assist SRTC with its ground truth data collections and also give the MTI project a variety of ground truth measurements that they make for their own purposes. Collectively, the ground truth data from the 14 core sites constitute a comprehensive database for science algorithm validation.

Determining site index accurately in even-aged stands

Treesearch

Gayne G. Erdmann; Ralph M., Jr. Peterson

1992-01-01

Good site index estimates are necessary for intensive forest management. To get tree age used in determining site index, increment cores are commonly used. The diffuse-porous rings of northern hardwoods, though, are difficult to count in cores, so many site index estimates are imprecise. Also, measuring the height of standing trees is more difficult and less accurate...

Analyses of native water, core material, and elutriate samples collected from the Atchafalaya River and Atchafalaya Bay

USGS Publications Warehouse

Demas, Charles R.

1977-01-01

During October and November 1976 the U.S. Geological Survey, in cooperation with the U.S. Army Corps of Engineers, collected native water and core material from 14 sites along the Atchafalya River in Louisiana (from the head of Whiskey Bay Pilot Channel to American Pass) and 5 sites in Atchafalya Bay for evaluation of possible environmental effects of a proposed channel-enlargement project. Core material from all river sites and one bay site was collected to a depth of 50 feet (15 meters). At the remaining bay sites, samples were collected to a depth of less than 6 inches (15 centimeters) using a pipe dredge. Core material and native water were analyzed (separately and as elutriate samples prepared from mixtures) for selected metals, nutrients, organic compounds, and physical characteristics. No interpretation of the data is given. (Woodard-USGS)

Draft Genome Sequence of a Picorna-Like Virus Associated with Gill Tissue in Clinically Normal Brook Trout, Salvelinus fontinalis.

PubMed

Iwanowicz, Luke R; Iwanowicz, Deborah D; Adams, Cynthia R; Galbraith, Heather; Aunins, Aaron; Cornman, Robert S

2017-10-12

Here, we report a draft genome sequence of a picorna-like virus associated with brook trout, Salvelinus fontinalis , gill tissue. The draft genome comprises 8,681 nucleotides, excluding the poly(A) tract, and contains two open reading frames. It is most similar to picorna-like viruses that infect invertebrates.

Draft genome sequence of a picorna-like virus associated with gill tissue in clinically normal brook trout, Salvelinus fontinalis

USGS Publications Warehouse

Iwanowicz, Luke R.; Iwanowicz, Deborah; Adams, Cynthia; Galbraith, Heather S.; Aunins, Aaron W.; Cornman, Robert S.

2017-01-01

Here, we report a draft genome sequence of a picorna-like virus associated with brook trout, Salvelinus fontinalis, gill tissue. The draft genome comprises 8,681 nucleotides, excluding the poly(A) tract, and contains two open reading frames. It is most similar to picorna-like viruses that infect invertebrates.

Star-PAP, a poly(A) polymerase, functions as a tumor suppressor in an orthotopic human breast cancer model

PubMed Central

Yu, C; Gong, Y; Zhou, H; Wang, M; Kong, L; Liu, J; An, T; Zhu, H; Li, Y

2017-01-01

Star-PAP is a noncanonical poly(A) polymerase and required for the expression of a select set of mRNAs. However, the pathological role of Star-PAP in cancer largely remains unknown. In this study, we observed decreased expression of Star-PAP in breast cancer cell lines and tissues. Ectopic Star-PAP expression inhibited proliferation as well as colony-forming ability of breast cancer cells. In breast cancer patients, high levels of Star-PAP correlated with an improved prognosis. Moreover, by regulating the expression of BIK (BCL2-interacting killer), Star-PAP induced apoptosis of breast cancer cells through the mitochondrial pathway. The growth of breast cancer xenografts in NOD/SCID mice was also inhibited by the doxycycline-induced Star-PAP overexpression. Furthermore, Star-PAP sensitized breast cancer cells to chemotherapy drugs both in vitro and in vivo. In mammary epithelial cells, Star-PAP knockdown partially transformed these cells and induced them to undergo epithelial–mesenchymal transition (EMT). These findings suggested that Star-PAP possesses tumor-suppressing activity and can be a valuable target for developing new cancer therapeutic strategies. PMID:28151486

Vector modifications to eliminate transposase expression following piggyBac-mediated transgenesis

PubMed Central

Chakraborty, Syandan; Ji, HaYeun; Chen, Jack; Gersbach, Charles A.; Leong, Kam W.

2014-01-01

Transgene insertion plays an important role in gene therapy and in biological studies. Transposon-based systems that integrate transgenes by transposase-catalyzed “cut-and-paste” mechanism have emerged as an attractive system for transgenesis. Hyperactive piggyBac transposon is particularly promising due to its ability to integrate large transgenes with high efficiency. However, prolonged expression of transposase can become a potential source of genotoxic effects due to uncontrolled transposition of the integrated transgene from one chromosomal locus to another. In this study we propose a vector design to decrease post-transposition expression of transposase and to eliminate the cells that have residual transposase expression. We design a single plasmid construct that combines the transposase and the transpositioning transgene element to share a single polyA sequence for termination. Consequently, the separation of the transposase element from the polyA sequence after transposition leads to its deactivation. We also co-express Herpes Simplex Virus thymidine kinase (HSV-tk) with the transposase. Therefore, cells having residual transposase expression can be eliminated by the administration of ganciclovir. We demonstrate the utility of this combination transposon system by integrating and expressing a model therapeutic gene, human coagulation Factor IX, in HEK293T cells. PMID:25492703

[Study of fusion of bacteriophage f2 double-stranded RNA, poly(A).poly(U), and poly(G).poly(C) in the presence of tetraethylammonium bromide].

PubMed

Permogorov, V I; Tiaglov, B V; Minaev, V E

1980-01-01

The data on the dependence of the melting curve parameters of double-stranded RNA (replicative form of RNA of f2 bacteriophage) poly(A) times poly(U) and poly(G) times poly(C) on the concentration of (C2H5)4NBr were obtained. The RNA melting range width is shown to pass through the minimum value T =2.1+/-0.1degrees at the point of inversion of relative stability of GC and AU pairs that corresponds to 4.0+/-0.1 M concentration of (C2H5)4NBr. Using the melting temperatures of poly(A) times poly(U) and poly(G) times poly(C) the rependence of Tgc-Tau parameter on (C2H5)4NBr concentration was shown. It was concluded from these data that the effect of the double-stranded RNA stacking heterogeneity was negligible in the 0-3 M range of (C2H5)4NBr concentration. Melting curves of RNA were obtained at various values of Tgc-Tau parameter. It was shown that the profile of fine structure of melting curves depends on the value of Tgc-Tau parameter.

Targeted gene deletion of miRNAs in mice by TALEN system.

PubMed

Takada, Shuji; Sato, Tempei; Ito, Yoshiaki; Yamashita, Satoshi; Kato, Tomoko; Kawasumi, Miyuri; Kanai-Azuma, Masami; Igarashi, Arisa; Kato, Tomomi; Tamano, Moe; Asahara, Hiroshi

2013-01-01

Mice are among the most valuable model animal species with an enormous amount of heritage in genetic modification studies. However, targeting genes in mice is sometimes difficult, especially for small genes, such as microRNAs (miRNAs) and targeting genes in repeat sequences. Here we optimized the application of TALEN system for mice and successfully obtained gene targeting technique in mice for intergenic region and series of microRNAs. Microinjection of synthesized RNA of TALEN targeting each gene in one cell stage of embryo was carried out and injected oocytes were transferred into pseudopregnant ICR female mice, producing a high success rate of the targeted deletion of miRNA genes. In our condition, TALEN RNA without poly(A) tail worked better than that of with poly(A) tail. This mutated allele in miRNA was transmitted to the next generation, suggesting the successful germ line transmission of this targeting method. Consistent with our notion of miRNAs maturation mechanism, in homozygous mutant mice of miR-10a, the non- mutated strand of miRNAs expression was completely diminished. This method will lead us to expand and accelerate our genetic research using mice in a high throughput way.

The E3 ubiquitin ligase and RNA-binding protein ZNF598 orchestrates ribosome quality control of premature polyadenylated mRNAs

PubMed Central

Garzia, Aitor; Jafarnejad, Seyed Mehdi; Meyer, Cindy; Chapat, Clément; Gogakos, Tasos; Morozov, Pavel; Amiri, Mehdi; Shapiro, Maayan; Molina, Henrik; Tuschl, Thomas; Sonenberg, Nahum

2017-01-01

Cryptic polyadenylation within coding sequences (CDS) triggers ribosome-associated quality control (RQC), followed by degradation of the aberrant mRNA and polypeptide, ribosome disassembly and recycling. Although ribosomal subunit dissociation and nascent peptide degradation are well-understood, the molecular sensors of aberrant mRNAs and their mechanism of action remain unknown. We studied the Zinc Finger Protein 598 (ZNF598) using PAR-CLIP and revealed that it cross-links to tRNAs, mRNAs and rRNAs, thereby placing the protein on translating ribosomes. Cross-linked reads originating from AAA-decoding tRNALys(UUU) were 10-fold enriched over its cellular abundance, and poly-lysine encoded by poly(AAA) induced RQC in a ZNF598-dependent manner. Encounter with translated polyA segments by ZNF598 triggered ubiquitination of several ribosomal proteins, requiring the E2 ubiquitin ligase UBE2D3 to initiate RQC. Considering that human CDS are devoid of >4 consecutive AAA codons, sensing of prematurely placed polyA tails by a specialized RNA-binding protein is a novel nucleic-acid-based surveillance mechanism of RQC. PMID:28685749

The E3 ubiquitin ligase and RNA-binding protein ZNF598 orchestrates ribosome quality control of premature polyadenylated mRNAs.

PubMed

Garzia, Aitor; Jafarnejad, Seyed Mehdi; Meyer, Cindy; Chapat, Clément; Gogakos, Tasos; Morozov, Pavel; Amiri, Mehdi; Shapiro, Maayan; Molina, Henrik; Tuschl, Thomas; Sonenberg, Nahum

2017-07-07

Cryptic polyadenylation within coding sequences (CDS) triggers ribosome-associated quality control (RQC), followed by degradation of the aberrant mRNA and polypeptide, ribosome disassembly and recycling. Although ribosomal subunit dissociation and nascent peptide degradation are well-understood, the molecular sensors of aberrant mRNAs and their mechanism of action remain unknown. We studied the Zinc Finger Protein 598 (ZNF598) using PAR-CLIP and revealed that it cross-links to tRNAs, mRNAs and rRNAs, thereby placing the protein on translating ribosomes. Cross-linked reads originating from AAA-decoding tRNA Lys (UUU) were 10-fold enriched over its cellular abundance, and poly-lysine encoded by poly(AAA) induced RQC in a ZNF598-dependent manner. Encounter with translated polyA segments by ZNF598 triggered ubiquitination of several ribosomal proteins, requiring the E2 ubiquitin ligase UBE2D3 to initiate RQC. Considering that human CDS are devoid of >4 consecutive AAA codons, sensing of prematurely placed polyA tails by a specialized RNA-binding protein is a novel nucleic-acid-based surveillance mechanism of RQC.

Diatoms and stratigraphically significant silicoflagellates from the Atlantic Margin Coring Project and other Atlantic margin sites

USGS Publications Warehouse

Abbott, W.H.

1980-01-01

In 1976, 19 sites were cored along the U.S. Atlantic Continental Shelf and Slope by the Oceanographic Branch of the U.S. Geological Survey aboard the Glomar Conception. Only 6 sites contained siliceous microfossil assemblages of sufficient quantity and quality for biostratigraphic study. Two of the sites, AMCOR (Atlantic Margin Coring Project) 6002 and AMCOR 6011, contained good Miocene assemblages: a small Pleistocene assemblage occurred at the top of AMCOR 6002. A Late Miocene to Early Pliocene assemblage was encountered in AMCOR 6007B. AMCOR 6013, 6019, and 6021 contained Pleistocene assemblages. In addition to the AMCOR cores, 3 additional Atlantic Margin cores were studied. These were the JOIDES 1 (Caldrill) core, and Atlantic Slope Project (ASP) cores 10 and 22. JOIDES 1 contains a Middle Miocene assemblage similar to AMCOR 6002. ASP 10 contains a Lower Pliocene assemblage and ASP 22 contains a Middle to Late Oligocene and a Pleistocene assemblage. Siliceous assemblages at all sites consisted mainly of shallow shelf species, including brackish and marine benthics and occasionally freshwater forms. Although planktonic forms were present, they were few and most were extant cosmopolitan species. This makes it difficult to correlate the biostratigraphy of the sediments with siliceous microfossil zonations from other oceans. The only biostratigraphic zonations for Atlantic Shelf diatom assemblages are for the Miocene.

Reflections on the history of pre-mRNA processing and highlights of current knowledge: A unified picture

PubMed Central

Darnell, James E.

2013-01-01

Several strong conclusions emerge concerning pre-mRNA processing from both old and newer experiments. The RNAPII complex is involved with pre-mRNA processing through binding of processing proteins to the CTD (carboxyl terminal domain) of the largest RNAPII subunit. These interactions are necessary for efficient processing, but whether factor binding to the CTD and delivery to splicing sites is obligatory or facilitatory is unsettled. Capping, addition of an m7Gppp residue (cap) to the initial transcribed residue of a pre-mRNA, occurs within seconds. Splicing of pre-mRNA by spliceosomes at particular sites is most likely committed during transcription by the binding of initiating processing factors and ∼50% of the time is completed in mammalian cells before completion of the primary transcript. This fact has led to an outpouring in the literature about “cotranscriptional splicing.” However splicing requires several minutes for completion and can take longer. The RNAPII complex moves through very long introns and also through regions dense with alternating exons and introns at an average rate of ∼3 kb per min and is, therefore, not likely detained at each splice site for more than a few seconds, if at all. Cleavage of the primary transcript at the 3′ end and polyadenylation occurs within 30 sec or less at recognized polyA sites, and the majority of newly polyadenylated pre-mRNA molecules are much larger than the average mRNA. Finally, it seems quite likely that the nascent RNA most often remains associated with the chromosomal locus being transcribed until processing is complete, possibly acquiring factors related to the transport of the new mRNA to the cytoplasm. PMID:23440351

Multiple core-hole formation by free-electron laser radiation in molecular nitrogen

NASA Astrophysics Data System (ADS)

Banks, H. I. B.; Little, D. A.; Emmanouilidou, A.

2018-05-01

We investigate the formation of multiple-core-hole states of molecular nitrogen interacting with a free-electron laser pulse. In previous work, we obtained bound and continuum molecular orbitals in the single-center expansion scheme and used these orbitals to calculate photo-ionization and auger decay rates. We extend our formulation to track the proportion of the population that accesses single-site versus two-site double-core-hole (TSDCH) states, before the formation of the final atomic ions. We investigate the pulse parameters that favor the formation of the single-site and TSDCH as well as triple-core-hole states for 525 and 1100 eV photons.

Results of Physical Property Measurements Obtained during the CHIKYU Cruise CK16-01 to Hydrothermal Fields of the Middle Okinawa Trough

NASA Astrophysics Data System (ADS)

Tanikawa, W.; Masaki, Y.; Komori, S.; Torimoto, J.; Makio, M.; Ohta, Y.; Nozaki, T.; Ishibashi, J. I.; Kumagai, H.; Maeda, L.; Hamada, Y.

2016-12-01

The middle Okinawa trough, along the Ryukyu-arc on the margin of the western Pacific, fosters several hydrothermal fields. The cruise CK16-01 of D/V CHIKYU targeted the Iheya-North Field and Noho hydrothermal site. More than ten-days extensive coring was carried out with Logging While Drilling (LWD) and deployment of Kuroko cultivation apparatus between February 29th to March 17th2016. Here we present the results of the physical property measurements obtained using Chikyu's on-board laboratory. Cores were sampled among three sites where the seafloor environments were quite different: the Noho site (C9017), a site between the Natsu and Aki sites of the Iheya-North field (C9021), and the Iheya-North Aki site (C9023). Site C9017 was near the center of the hydrothermal activity, and the obtained core was limited 36 m in length and 30 % in the recovery rate. At 70 mbsf (meters below seafloor), the grain density and bulk density of the sediment reached their maxim (3.7 g/m3 and 2.7 g/cm3, respectively), while thermal conductivity reached its lowest value (0.6 W/m·K). Site C9021 yielded a 54 m core, with a core recovery rate of 50 %. Coarse pumiceous layers were found at 68 mbsf, with a hydrothermally altered layer appearing below 68 mbsf. The mean grain density value was 2.4 g/cm3 and was uniform throughout the core. The mean bulk density value of the pumiceous layers was 1.3 g/cm3, and of the hydrothermally altered layer was 2.1 g/cm3. Site C9023 was close to the active hydrothermal chimneys of the Iheya-North Aki site, and yielded 33 m of core with a core recovery rate of 16 %. Massive sulfide layers were found below 48 mbsf with grain density and bulk density values varying between 2.8-4.7 g/cm3 and 1.5-3.9 g/cm3, respectively. Magnetic susceptibility exhibited a high anomaly in a sedimented anhydrite layer found between 95 and 135 mbsf, and a high porosity and low resistivity zone was found below 150 mbsf. Together, these data from drilling cores and onboard analysis provide the first insights into the physical properties of hydrothermal fields in the Okinawa trough. This work was supported by the Council for Science, Technology and Innovation (CSTI) Cross-ministerial Strategic Innovation Promotion Program (SIP), "Next-generation technology for ocean resources exploration" (Lead agency: JAMSTEC)

Mineralogy of selected sedimentary interbeds at or near the Idaho National Engineering Laboratory, Idaho

USGS Publications Warehouse

Reed, Michael F.; Bartholomay, Roy C.

1994-01-01

The U.S. Geological Survey (USGS) Project Office at the Idaho National Engineering Laboratory (INEL), in cooperation with the U.S. Department of Energy and Idaho State University, analyzed 66 samples from sedimentary interbed cores during a 38-month period beginning in October 1990 to determine bulk and clay mineralogy. These cores had been collected from 19 sites in the Big Lost River Basin, 2 sites in the Birch Creek Basin, and 1 site in the Mud Lake Basin, and were archived at the USGS lithologic core library at the INEL. Mineralogy data indicate that the core samples from the Big Lost River Basin have larger mean and median percentages of quartz, total feldspar, and total clay minerals, but smaller mean and median percentages of calcite than the core samples from the Birch Creek Basin. Core samples from the Mud Lake Basin have abundant quartz, total feldspar, calcite, and total clay minerals.

The point of no return: The poly(A)-associated elongation checkpoint

PubMed Central

Tellier, Michael; Ferrer-Vicens, Ivan; Murphy, Shona

2016-01-01

abstract Cyclin-dependent kinases play critical roles in transcription by RNA polymerase II (pol II) and processing of the transcripts. For example, CDK9 regulates transcription of protein-coding genes, splicing, and 3′ end formation of the transcripts. Accordingly, CDK9 inhibitors have a drastic effect on the production of mRNA in human cells. Recent analyses indicate that CDK9 regulates transcription at the early-elongation checkpoint of the vast majority of pol II-transcribed genes. Our recent discovery of an additional CDK9-regulated elongation checkpoint close to poly(A) sites adds a new layer to the control of transcription by this critical cellular kinase. This novel poly(A)-associated checkpoint has the potential to powerfully regulate gene expression just before a functional polyadenylated mRNA is produced: the point of no return. However, many questions remain to be answered before the role of this checkpoint becomes clear. Here we speculate on the possible biological significance of this novel mechanism of gene regulation and the players that may be involved. PMID:26853452

Analysis of alterative cleavage and polyadenylation by 3′ region extraction and deep sequencing

PubMed Central

Hoque, Mainul; Ji, Zhe; Zheng, Dinghai; Luo, Wenting; Li, Wencheng; You, Bei; Park, Ji Yeon; Yehia, Ghassan; Tian, Bin

2012-01-01

Alternative cleavage and polyadenylation (APA) leads to mRNA isoforms with different coding sequences (CDS) and/or 3′ untranslated regions (3′UTRs). Using 3′ Region Extraction And Deep Sequencing (3′READS), a method which addresses the internal priming and oligo(A) tail issues that commonly plague polyA site (pA) identification, we comprehensively mapped pAs in the mouse genome, thoroughly annotating 3′ ends of genes and revealing over five thousand pAs (~8% of total) flanked by A-rich sequences, which have hitherto been overlooked. About 79% of mRNA genes and 66% of long non-coding RNA (lncRNA) genes have APA; but these two gene types have distinct usage patterns for pAs in introns and upstream exons. Promoter-distal pAs become relatively more abundant during embryonic development and cell differentiation, a trend affecting pAs in both 3′-most exons and upstream regions. Upregulated isoforms generally have stronger pAs, suggesting global modulation of the 3′ end processing activity in development and differentiation. PMID:23241633

Cloning and molecular characterization of scorpion Buthus martensi venom hyaluronidases: a novel full-length and diversiform noncoding isoforms.

PubMed

Xia, Xichao; Liu, Rongzhi; Li, Yi; Xue, Shipeng; Liu, Qingchun; Jiang, Xiao; Zhang, Wenjuan; Ding, Ke

2014-09-01

Hyaluronidase is a common component of scorpion venom and has been considered as "spreading factor" that promotes a fast penetration of the venom in the anaphylactic reaction. In the current study, a novel full-length of hyaluronidase BmHYI and three noncoding isoforms of BmHYII, BmHYIII and BmHYIV were cloned by using a combined strategy based on peptide sequencing and Rapid Amplification of cDNA Ends (RACE). BmHYI has 410 amino acid residues containing the catalytic, positional and five potential N-glycosylation sites. The deduced protein sequence of BmHYI shares significant identity with venom hyaluronidases from bees and snakes. The phylogenetic analysis showed early divergence and independent evolution of BmHYI from other hyaluronidases. An extraordinarily high level of sequence similarity was detected among four sequences. But, BmHYII, BmHYIII and BmHYIV were short of stop-codon in the open reading frame and poly(A) signal in the 3' end. Copyright © 2014 Elsevier B.V. All rights reserved.

Validation of SMAP surface soil moisture products with core validation sites

USDA-ARS?s Scientific Manuscript database

The NASA Soil Moisture Active Passive (SMAP) mission has utilized a set of core validation sites as the primary methodology in assessing the soil moisture retrieval algorithm performance. Those sites provide well-calibrated in situ soil moisture measurements within SMAP product grid pixels for diver...

Water column and bed-sediment core samples collected from Brownlee Reservoir near Oxbow, Oregon, 2012

USGS Publications Warehouse

Fosness, Ryan L.; Naymik, Jesse; Hopkins, Candice B.; DeWild, John F.

2013-01-01

The U.S. Geological Survey, in cooperation with Idaho Power Company, collected water-column and bed-sediment core samples from eight sites in Brownlee Reservoir near Oxbow, Oregon, during May 5–7, 2012. Water-column and bed-sediment core samples were collected at each of the eight sites and analyzed for total mercury and methylmercury. Additional bed-sediment core samples, collected from three of the eight sites, were analyzed for pesticides and other organic compounds, trace metals, and physical characteristics, such as particle size. Total mercury and methylmercury were detected in each of the water column and bed-sediment core samples. Only 17 of the 417 unique pesticide and organic compounds were detected in bed-sediment core samples. Concentrations of most organic wastewater compounds detected in bed sediment were less than the reporting level. Trace metals detected were greater than the reporting level in all the bed-sediment core samples submitted for analysis. The particle size distribution of bed-sediment core samples was predominantly clay mixed with silt.

Speciation and isotopic composition of sedimentary sulfur in the Everglades, Florida, USA

USGS Publications Warehouse

Bates, A.L.; Spiker, E. C.; Holmes, C.W.

1998-01-01

We have studied the sulfur speciation and isotopic composition of two peat cores from Water Conservation Area 2A (WCA 2A) in the Florida Everglades. Core site E1 is affected by agricultural runoff from the Hillsboro Canal which drains the Everglades Agricultural Area; Core site U3 is distant from the canal and relatively unaffected by agricultural runoff. Depth profiles of the total sulfur content of both cores show fairly constant levels (??0.7 wt.%) below about 25-30 cm depth in Core E1 and below 40-45 cm in Core U3. Above these depths, total sulfur increases to as much as 1.52 wt.% in Core E1 and 1.74 wt.% in Core U3, suggesting that more sulfur has entered the sediments and/or that more sulfur is being retained in recent times at both sampling sites. These changes in total sulfur content with depth in Core E1 correlate with changes in total phosphorus that have been observed in other studies at core sites near the Hillsboro Canal. This correlation of total sulfur with phosphorus with depth is not seen in Core U3 located away from the canal, possibly because phosphorus is more effectively retained than sulfur in the organic sediment near the canal. Organic-sulfur (OS) concentrations are at least twice as high as the dusulfide-sulfur (DS) concentrations in the upper parts of both cores suggesting that iron is presently limiting the amount of dusulfide minerals formed in these sediments. The degree of pyritization (DOP) in the upper parts of the cores suggest that sulfide mineralization is limited by the availability of highly reactive iron during the earliest stages of diagenesis. Positive ??34S values for reduced sulfur forms in both cores indicate a relatively restricted sulfate reservoir, consistent with nearly complete reduction of the sulfate available in the sediment at any given time. Differences between the two core appear in the ??34S values for the near-surface sediments. The DS ??34S values in the upper 10.0 cm of sediment are more posotive at site E1, with a mean ??34S value of +12.9???, than at site U3, with a mean ??34S value of +2.9???. These reuslts may indicate that increased rates of organic deposition due to nutrient loading near the canal have increased the rate of sulfate reduction at the E1 site in recent times. Acid-volatile-sulfide (AVS) concentrations are lower than DS and OS concentrations by at least a factor of 10. Increasing ??34S values for AVS with increasing depth in both cores suggests ongoing reduction of a limited porewater sulfate reservoir after deposition. The disulfide and organic-sulfur ??34S values diverge from the ??34S values for AVS with depth, suggesting that most of the transformation of AVS into disulfide minerals or incorporation of sulfur into orgnaic matter occurs in the near-surface sediments. A comparison of organic-sulfur ??34S values in the dominant flora at the U3 site (sawgrass leaves and periphyton) with organic-sulfur ??34S values at the top of the U3 core indicates that there was early incorporation of an isotopically light sulfide species into the orgnaic matter.

Genome-wide RNA-binding analysis of the trypanosome U1 snRNP proteins U1C and U1-70K reveals cis/trans-spliceosomal network

PubMed Central

Preußer, Christian; Rossbach, Oliver; Hung, Lee-Hsueh; Li, Dan; Bindereif, Albrecht

2014-01-01

Trans-splicing in trypanosomes adds a 39-nucleotide mini-exon from the spliced leader (SL) RNA to the 5′ end of each protein-coding sequence. On the other hand, cis-splicing of the few intron-containing genes requires the U1 small nuclear ribonucleoprotein (snRNP) particle. To search for potential new functions of the U1 snRNP in Trypanosoma brucei, we applied genome-wide individual-nucleotide resolution crosslinking-immunoprecipitation (iCLIP), focusing on the U1 snRNP-specific proteins U1C and U1-70K. Surprisingly, U1C and U1-70K interact not only with the U1, but also with U6 and SL RNAs. In addition, mapping of crosslinks to the cis-spliced PAP [poly(A) polymerase] pre-mRNA indicate an active role of these proteins in 5′ splice site recognition. In sum, our results demonstrate that the iCLIP approach provides insight into stable and transient RNA–protein contacts within the spliceosomal network. We propose that the U1 snRNP may represent an evolutionary link between the cis- and trans-splicing machineries, playing a dual role in 5′ splice site recognition on the trans-spliceosomal SL RNP as well as on pre-mRNA cis-introns. PMID:24748659

Construction of plasmid, bacterial expression, purification, and assay of dengue virus type 2 NS5 methyltransferase.

PubMed

Boonyasuppayakorn, Siwaporn; Padmanabhan, Radhakrishnan

2014-01-01

Dengue virus (DENV), a member of mosquito-borne flavivirus, causes self-limiting dengue fever as well as life-threatening dengue hemorrhagic fever and dengue shock syndrome. Its positive sense RNA genome has a cap at the 5'-end and no poly(A) tail at the 3'-end. The viral RNA encodes a single polyprotein, C-prM-E-NS1-NS2A-NS2B-NS3-NS4A-NS4B-NS5. The polyprotein is processed into 3 structural proteins (C, prM, and E) and 7 nonstructural (NS) proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5). NS3 and NS5 are multifunctional enzymes performing various tasks in viral life cycle. The N-terminal domain of NS5 has distinct GTP and S-adenosylmethionine (SAM) binding sites. The role of GTP binding site is implicated in guanylyltransferase (GTase) activity of NS5. The SAM binding site is involved in both N-7 and 2'-O-methyltransferase (MTase) activities involved in formation of type I cap. The C-terminal domain of NS5 catalyzes RNA-dependent RNA polymerase (RdRp) activity involved in RNA synthesis. We describe the construction of the MTase domain of NS5 in an E. coli expression vector, purification of the enzyme, and conditions for enzymatic assays of N7- and 2'O-methyltransferase activities that yield the final type I 5'-capped RNA ((7Me)GpppA2'OMe-RNA).

Draft Genome Sequence of a Picorna-Like Virus Associated with Gill Tissue in Clinically Normal Brook Trout, Salvelinus fontinalis

PubMed Central

2017-01-01

ABSTRACT Here, we report a draft genome sequence of a picorna-like virus associated with brook trout, Salvelinus fontinalis, gill tissue. The draft genome comprises 8,681 nucleotides, excluding the poly(A) tract, and contains two open reading frames. It is most similar to picorna-like viruses that infect invertebrates. PMID:29025930

Education and Thinking: The Role of Knowledge

DTIC Science & Technology

1983-06-21

failing to construct a representation of the problem. Through carefully designed problem " exercises , the program elicits procedures for reasoning and...guiding spirits. Polya -. .,• . . .. ~ -, • ° -. -- , ",r , • , , . . .. • . . , . .-r. Page9 9 recommends that explicit attention be paid to heuristic...reflects, according to Carey, a reorganization of knowledge brought about by school learning and world knowledge: for 4- to 7-year olds, biological

Historical CO2 Records from the Law Dome DE08, DE08-2, and DSS Ice Cores (1006 A.D.-1978 A.D)

DOE Data Explorer

Etheridge, D. M. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Australia; Barnola, J. M. [Laboratoire de Glaciologie et Géophysique de l'Environnement, Saint Martin d'Hères-Cedex, France; Morgan, V. I. [Antarctic CRC and Australian Antarctic Division, Hobart, Tasmania, Australia; Steele, L. P. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Australia; Langenfelds, R. L. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Australia; Francey, R. J. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Australia; Martinez, Monica [Oak Ridge National Laboratory, Oak Ridge, TN (USA)

1998-01-01

The CO2 records presented here are derived from three ice cores obtained at Law Dome, East Antarctica from 1987 to 1993. The Law Dome site satisfies many of the desirable characteristics of an ideal ice core site for atmospheric CO2 reconstructions including negligible melting of the ice sheet surface, low concentrations of impurities, regular stratigraphic layering undisturbed at the surface by wind or at depth by ice flow, and high snow accumulation rate. Further details on the site, drilling, and cores are provided in Etheridge et al. (1996), Etheridge and Wookey (1989), and Morgan et al (1997).

A carbon, nitrogen, and sulfur elemental and isotopic study in dated sediment cores from the Louisiana Shelf

USGS Publications Warehouse

Rosenbauer, R.J.; Swarzenski, P.W.; Kendall, C.; Orem, W.H.; Hostettler, F.D.; Rollog, M.E.

2009-01-01

Three sediment cores were collected off the Mississippi River delta on the Louisiana Shelf at sites that are variably influenced by recurring, summer-time water-column hypoxia and fluvial loadings. The cores, with established chronology, were analyzed for their respective carbon, nitrogen, and sulfur elemental and isotopic composition to examine variable organic matter inputs, and to assess the sediment record for possible evidence of hypoxic events. Sediment from site MRJ03-3, which is located close to the Mississippi Canyon and generally not influenced by summer-time hypoxia, is typical of marine sediment in that it contains mostly marine algae and fine-grained material from the erosion of terrestrial C4 plants. Sediment from site MRJ03-2, located closer to the mouth of the Mississippi River and at the periphery of the hypoxic zone (annual recurrence of summer-time hypoxia >50%), is similar in composition to core MRJ03-3, but exhibits more isotopic and elemental variability down-core, suggesting that this site is more directly influenced by river discharge. Site MRJ03-5 is located in an area of recurring hypoxia (annual recurrence >75%), and is isotopically and elementally distinct from the other two cores. The carbon and nitrogen isotopic composition of this core prior to 1960 is similar to average particulate organic matter from the lower Mississippi River, and approaches the composition of C3 plants. This site likely receives a greater input of local terrestrial organic matter to the sediment. After 1960 and to the present, a gradual shift to higher values of ??13C and ??15N and lower C:N ratios suggests that algal input to these shelf sediments increased as a result of increased productivity and hypoxia. The values of C:S and ??34S reflect site-specific processes that may be influenced by the higher likelihood of recurring seasonal hypoxia. In particular, the temporal variations in the C:S and ??34S down-core are likely caused by changes in the rate of sulfate reduction, and hence the degree of hypoxia in the overlying water column. Based principally on the down-core C:N and C:S ratios and ??13C and ??34S profiles, sites MRJ03-3 and MRJ03-2 generally reflect more marine organic matter inputs, while site MRJ03-5 appears to be more influenced by terrestrial deposition. ?? 2009 Springer-Verlag.

The value of core lab stress echocardiography interpretations: observations from the ISCHEMIA Trial.

PubMed

Kataoka, Akihisa; Scherrer-Crosbie, Marielle; Senior, Roxy; Gosselin, Gilbert; Phaneuf, Denis; Guzman, Gabriela; Perna, Gian; Lara, Alfonso; Kedev, Sasko; Mortara, Andrea; El-Hajjar, Mohammad; Shaw, Leslee J; Reynolds, Harmony R; Picard, Michael H

2015-12-18

Stress echocardiography (SE) is dependent on subjective interpretations. As a prelude to the International Study of Comparative Health Effectiveness with Medical and Invasive Approaches (ISCHEMIA) Trial, potential sites were required to submit two SE, one with moderate or severe left ventricular (LV) myocardial ischemia and one with mild ischemia. We evaluated the concordance of site and core lab interpretations. Eighty-one SE were submitted from 41 international sites. Ischemia was classified by the number of new or worsening segmental LV wall motion abnormalities (WMA): none, mild (1 or 2) or moderate or severe (3 or more) by the sites and the core lab. Core lab classified 6 SE as no ischemia, 35 mild and 40 moderate or greater. There was agreement between the site and core in 66 of 81 total cases (81%, weighted kappa coefficient [K] =0.635). Agreement was similar for SE type - 24 of 30 exercise (80%, K = 0.571) vs. 41 of 49 pharmacologic (84%, K = 0.685). The agreement between poor or fair image quality (27 of 36 cases, 75%, K = 0.492) was not as good as for the good or excellent image quality cases (39 of 45 cases, 87%, K = 0.755). Differences in concordance were noted for degree of ischemia with the majority of discordant interpretations (87%) occurring in patients with no or mild LV myocardial ischemia. While site SE interpretations are largely concordant with core lab interpretations, this appears dependent on image quality and the extent of WMA. Thus core lab interpretations remain important in clinical trials where consistency of interpretation across a range of cases is critical. ClinicalTrials.gov NCT01471522.

Videoteleconferencing for administration of a multisite obstetrics and gynecology core clerkship.

PubMed

Browne, M W; Hines, J F; Satin, A J; Haffner, W H

2000-03-01

Recent changes and trends in health care delivery have required medical schools to use multiple sites to obtain adequate patient exposure for their students. Decentralization of clinical undergraduate medical education may lead to a lack of continuity in curricula, evaluation, and feedback. We describe the use of interactive videoteleconferencing as a tool to link and improve a multi-site undergraduate core clerkship in obstetrics and gynecology. The Uniformed Services University of the Health Sciences, Bethesda, Maryland, currently utilizes five geographically separate sites for its 6-week core clerkship in obstetrics and gynecology. The site coordinators, clerkship director, and administrative personnel from the parent institution meet approximately 3 weeks after the completion of each core clerkship for live, real-time, and interactive broadcast to complete student evaluations, review curricula, and discuss problems with current students and other pertinent educational issues. Videoteleconferencing provides a mechanism to ensure consistency in curriculum and student evaluations and provides administrative support to distant sites. Furthermore, it enables site coordinators to keep the clerkship director abreast of students and clerkship issues.

Perispeckles are major assembly sites for the exon junction core complex

PubMed Central

Daguenet, Elisabeth; Baguet, Aurélie; Degot, Sébastien; Schmidt, Ute; Alpy, Fabien; Wendling, Corinne; Spiegelhalter, Coralie; Kessler, Pascal; Rio, Marie-Christine; Le Hir, Hervé; Bertrand, Edouard; Tomasetto, Catherine

2012-01-01

The exon junction complex (EJC) is loaded onto mRNAs as a consequence of splicing and regulates multiple posttranscriptional events. MLN51, Magoh, Y14, and eIF4A3 form a highly stable EJC core, but where this tetrameric complex is assembled in the cell remains unclear. Here we show that EJC factors are enriched in domains that we term perispeckles and are visible as doughnuts around nuclear speckles. Fluorescence resonance energy transfer analyses and EJC assembly mutants show that perispeckles do not store free subunits, but instead are enriched for assembled cores. At the ultrastructural level, perispeckles are distinct from interchromatin granule clusters that may function as storage sites for splicing factors and intermingle with perichromatin fibrils, where nascent RNAs and active RNA Pol II are present. These results support a model in which perispeckles are major assembly sites for the tetrameric EJC core. This subnuclear territory thus represents an intermediate region important for mRNA maturation, between transcription sites and splicing factor reservoirs and assembly sites. PMID:22419818

In silico evolution of the Drosophila gap gene regulatory sequence under elevated mutational pressure.

PubMed

Chertkova, Aleksandra A; Schiffman, Joshua S; Nuzhdin, Sergey V; Kozlov, Konstantin N; Samsonova, Maria G; Gursky, Vitaly V

2017-02-07

Cis-regulatory sequences are often composed of many low-affinity transcription factor binding sites (TFBSs). Determining the evolutionary and functional importance of regulatory sequence composition is impeded without a detailed knowledge of the genotype-phenotype map. We simulate the evolution of regulatory sequences involved in Drosophila melanogaster embryo segmentation during early development. Natural selection evaluates gene expression dynamics produced by a computational model of the developmental network. We observe a dramatic decrease in the total number of transcription factor binding sites through the course of evolution. Despite a decrease in average sequence binding energies through time, the regulatory sequences tend towards organisations containing increased high affinity transcription factor binding sites. Additionally, the binding energies of separate sequence segments demonstrate ubiquitous mutual correlations through time. Fewer than 10% of initial TFBSs are maintained throughout the entire simulation, deemed 'core' sites. These sites have increased functional importance as assessed under wild-type conditions and their binding energy distributions are highly conserved. Furthermore, TFBSs within close proximity of core sites exhibit increased longevity, reflecting functional regulatory interactions with core sites. In response to elevated mutational pressure, evolution tends to sample regulatory sequence organisations with fewer, albeit on average, stronger functional transcription factor binding sites. These organisations are also shaped by the regulatory interactions among core binding sites with sites in their local vicinity.

The 1500m South Pole Ice Core: Recovering a 40 Ka Environmental Record

NASA Technical Reports Server (NTRS)

Casey, Kimberly Ann; Neumann, Thomas Allen; Fudge, T. J.; Neumann, T. A.; Steig, E. J.; Cavitte, M. G. P.; Blankenship, D. D.

2014-01-01

Supported by the US National Science Foundation, a new 1500 m, approximately 40 ka old ice core will be recovered from South Pole during the 2014/15 and 2015/16 austral summer seasons using the new US Intermediate Depth Drill. The combination of low temperatures, relatively high accumulation rates and low impurity concentrations at South Pole will yield detailed records of ice chemistry and trace atmospheric gases. The South Pole ice core will provide a climate history record of a unique area of the East Antarctic plateau that is partly influenced by weather systems that cross the West Antarctic ice sheet. The ice at South Pole flows at approximately 10m a(exp-1) and the South Pole ice-core site is a significant distance from an ice divide. Therefore, ice recovered at depth originated progressively farther upstream of the coring site. New ground-penetrating radar collected over the drill site location shows no anthropogenic influence over the past approximately 50 years or upper 15 m. Depth-age scale modeling results show consistent and plausible annual-layer thicknesses and accumulation rate histories, indicating that no significant stratigraphic disturbances exist in the upper 1500m near the ice-core drill site.

Contaminant trends in reservoir sediment cores as records of influent stream quality

USGS Publications Warehouse

Van Metre, P.C.; Mahler, B.J.

2004-01-01

When reconstructing water-quality histories from lake and reservoir cores, it is sometimes assumed that the chemical signatures in the cores reflect historical water quality in the influent streams. To investigate this assumption, concentrations of metals, PAHs, and organochlorine compounds in sediment cores were compared to those associated with an influent-stream suspended sediment for three reservoirs in Fort Worth, TX, and two reservoirs in Boston, MA, U.S.A., and interpreted in light of land-use and regulation histories. In evaluating relations between suspended sediments and cores, three levels of preservation were indicated: (1) influent concentrations and historical trends are preserved in cores (metals at all sites; some organic contaminants at some sites); (2) some loss occurs during transport and initial deposition but relative historical trends are preserved in cores (some organic contaminants at some sites); and (3) neither stream concentrations nor relative historical trends are preserved (dieldrin and p,p???-DDT). The degree of preservation of influent concentration histories varied between lakes, particularly for PAHs. The results support the use of sediment cores to infer streamwater-quality histories for many contaminants but indicate that reservoir-bottom sediment samples might underestimate concentrations of organic contaminants in some streams.

When core competence is not enough: functional interplay of the DEAD-box helicase core with ancillary domains and auxiliary factors in RNA binding and unwinding.

PubMed

Rudolph, Markus G; Klostermeier, Dagmar

2015-08-01

DEAD-box helicases catalyze RNA duplex unwinding in an ATP-dependent reaction. Members of the DEAD-box helicase family consist of a common helicase core formed by two RecA-like domains. According to the current mechanistic model for DEAD-box mediated RNA unwinding, binding of RNA and ATP triggers a conformational change of the helicase core, and leads to formation of a compact, closed state. In the closed conformation, the two parts of the active site for ATP hydrolysis and of the RNA binding site, residing on the two RecA domains, become aligned. Closing of the helicase core is coupled to a deformation of the RNA backbone and destabilization of the RNA duplex, allowing for dissociation of one of the strands. The second strand remains bound to the helicase core until ATP hydrolysis and product release lead to re-opening of the core. The concomitant disruption of the RNA binding site causes dissociation of the second strand. The activity of the helicase core can be modulated by interaction partners, and by flanking N- and C-terminal domains. A number of C-terminal flanking regions have been implicated in RNA binding: RNA recognition motifs (RRM) typically mediate sequence-specific RNA binding, whereas positively charged, unstructured regions provide binding sites for structured RNA, without sequence-specificity. Interaction partners modulate RNA binding to the core, or bind to RNA regions emanating from the core. The functional interplay of the helicase core and ancillary domains or interaction partners in RNA binding and unwinding is not entirely understood. This review summarizes our current knowledge on RNA binding to the DEAD-box helicase core and the roles of ancillary domains and interaction partners in RNA binding and unwinding by DEAD-box proteins.

Variability of sea salts in ice and firn cores from Fimbul Ice Shelf, Dronning Maud Land, Antarctica

NASA Astrophysics Data System (ADS)

Paulina Vega, Carmen; Isaksson, Elisabeth; Schlosser, Elisabeth; Divine, Dmitry; Martma, Tõnu; Mulvaney, Robert; Eichler, Anja; Schwikowski-Gigar, Margit

2018-05-01

Major ions were analysed in firn and ice cores located at Fimbul Ice Shelf (FIS), Dronning Maud Land - DML, Antarctica. FIS is the largest ice shelf in the Haakon VII Sea, with an extent of approximately 36 500 km2. Three shallow firn cores (about 20 m deep) were retrieved in different ice rises, Kupol Ciolkovskogo (KC), Kupol Moskovskij (KM), and Blåskimen Island (BI), while a 100 m long core (S100) was drilled near the FIS edge. These sites are distributed over the entire FIS area so that they provide a variety of elevation (50-400 m a.s.l.) and distance (3-42 km) to the sea. Sea-salt species (mainly Na+ and Cl-) generally dominate the precipitation chemistry in the study region. We associate a significant sixfold increase in median sea-salt concentrations, observed in the S100 core after the 1950s, to an enhanced exposure of the S100 site to primary sea-salt aerosol due to a shorter distance from the S100 site to the ice front, and to enhanced sea-salt aerosol production from blowing salty snow over sea ice, most likely related to the calving of Trolltunga occurred during the 1960s. This increase in sea-salt concentrations is synchronous with a shift in non-sea-salt sulfate (nssSO42-) toward negative values, suggesting a possible contribution of fractionated aerosol to the sea-salt load in the S100 core most likely originating from salty snow found on sea ice. In contrast, there is no evidence of a significant contribution of fractionated sea salt to the ice-rises sites, where the signal would be most likely masked by the large inputs of biogenic sulfate estimated for these sites. In summary, these results suggest that the S100 core contains a sea-salt record dominated by the proximity of the site to the ocean, and processes of sea ice formation in the neighbouring waters. In contrast, the ice-rises firn cores register a larger-scale signal of atmospheric flow conditions and a less efficient transport of sea-salt aerosols to these sites. These findings are a contribution to the understanding of the mechanisms behind sea-salt aerosol production, transport and deposition at coastal Antarctic sites, and the improvement of the current Antarctic sea ice reconstructions based on sea-salt chemical proxies obtained from ice cores.

Plausible Reasoning in Tactical Planning.

DTIC Science & Technology

1987-04-01

different contexts and tasks that give rise to mappings, and the set of issues and different resolutions to these issues that have been proposed in the...humans use in reasoning about the wo~ ld ( Polya , 1958; Collins, 1978a). Our work attempts to formalize the plausible inferences that frequently occur in...are interconnected by traces. Each hierarchy represents knowledge about a class of concepts arranged in a tree structure according to some viewpoint

Sage Gene Expression Profiles Characterizing Cure

DTIC Science & Technology

2006-10-01

including suggestions for reducing this burden to Department of Defense, Washington Headquarters Services , Directorate for Information Operations and...all of which have concluded that the effect is largely consistent among high quality studies. If their breast cancer surgery is done in the luteal...at different phases of the estrous cycle. 2. prepare RNA from tumors using dynel poly-A beads 3. prepare double-stranded cDNA 4. check quality of

In vitro labeling strategies for in cellulo fluorescence microscopy of single ribonucleoprotein machines.

PubMed

Custer, Thomas C; Walter, Nils G

2017-07-01

RNA plays a fundamental, ubiquitous role as either substrate or functional component of many large cellular complexes-"molecular machines"-used to maintain and control the readout of genetic information, a functional landscape that we are only beginning to understand. The cellular mechanisms for the spatiotemporal organization of the plethora of RNAs involved in gene expression are particularly poorly understood. Intracellular single-molecule fluorescence microscopy provides a powerful emerging tool for probing the pertinent mechanistic parameters that govern cellular RNA functions, including those of protein coding messenger RNAs (mRNAs). Progress has been hampered, however, by the scarcity of efficient high-yield methods to fluorescently label RNA molecules without the need to drastically increase their molecular weight through artificial appendages that may result in altered behavior. Herein, we employ T7 RNA polymerase to body label an RNA with a cyanine dye, as well as yeast poly(A) polymerase to strategically place multiple 2'-azido-modifications for subsequent fluorophore labeling either between the body and tail or randomly throughout the tail. Using a combination of biochemical and single-molecule fluorescence microscopy approaches, we demonstrate that both yeast poly(A) polymerase labeling strategies result in fully functional mRNA, whereas protein coding is severely diminished in the case of body labeling. © 2016 The Protein Society.

Characterization and chromosomal mapping of the human TFG gene involved in thyroid carcinoma

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mencinger, M.; Panagopoulos, I.; Andreasson, P.

1997-05-01

Homology searches in the Expressed Sequence Tag Database were performed using SPYGQ-rich regions as query sequences to find genes encoding protein regions similar to the N-terminal parts of the sarcoma-associated EWS and FUS proteins. Clone 22911 (T74973), encoding a SPYGQ-rich region in its 5{prime} end, and several other clones that overlapped 22911 were selected. The combined data made it possible to assemble a full-length cDNA sequence. This cDNA sequence is 1677 bp, containing an initiation codon ATG, an open reading frame of 400 amino acids, a poly(A) signal, and a poly(A) tail. We found 100% identity between the 5{prime} partmore » of the consensus sequence and the 598-bp-long sequence named TFG. The TFG sequence is fused to the 3{prime} end of NTRK1, generating the TRK-T3 fusion transcript found in papillary thyroid carcinoma. The cDNA therefore represents the full-length transcript of the TFG gene. TFG was localized to 3q11-q12 by fluorescence in situ hybridization. The 3{prime} and the 5{prime} ends of the TFG cDNA probe hybridized to a 2.2-kb band on Northern blot filters in all tissues examined. 28 refs., 5 figs., 1 tab.« less

RNA binding and replication by the poliovirus RNA polymerase

DOE Office of Scientific and Technical Information (OSTI.GOV)

Oberste, M.S.

1988-01-01

RNA binding and RNA synthesis by the poliovirus RNA-dependent RNA polymerase were studied in vitro using purified polymerase. Templates for binding and RNA synthesis studies were natural RNAs, homopolymeric RNAs, or subgenomic poliovirus-specific RNAs synthesized in vitro from cDNA clones using SP6 or T7 RNA polymerases. The binding of the purified polymerase to poliovirion and other RNAs was studied using a protein-RNA nitrocellulose filter binding assay. A cellular poly(A)-binding protein was found in the viral polymerase preparations, but was easily separated from the polymerase by chromatography on poly(A) Sepharose. The binding of purified polymerase to {sup 32}P-labeled ribohomopolymeric RNAs wasmore » examined, and the order of binding observed was poly(G) >>> poly(U) > poly(C) > poly(A). The K{sub a} for polymerase binding to poliovirion RNA and to a full-length negative strand transcript was about 1 {times} 10{sup 9} M{sup {minus}1}. The polymerase binds to a subgenomic RNAs which contain the 3{prime} end of the genome with a K{sub a} similar to that for virion RNA, but binds less well to 18S rRNA, globin mRNA, and subgenomic RNAs which lack portions of the 3{prime} noncoding region.« less

Musashi Protein-directed Translational Activation of Target mRNAs Is Mediated by the Poly(A) Polymerase, Germ Line Development Defective-2*

PubMed Central

Cragle, Chad; MacNicol, Angus M.

2014-01-01

The mRNA-binding protein, Musashi, has been shown to regulate translation of select mRNAs and to control cellular identity in both stem cells and cancer cells. Within the mammalian cells, Musashi has traditionally been characterized as a repressor of translation. However, we have demonstrated that Musashi is an activator of translation in progesterone-stimulated oocytes of the frog Xenopus laevis, and recent evidence has revealed Musashi's capability to function as an activator of translation in mammalian systems. The molecular mechanism by which Musashi directs activation of target mRNAs has not been elucidated. Here, we report a specific association of Musashi with the noncanonical poly(A) polymerase germ line development defective-2 (GLD2) and map the association domain to 31 amino acids within the C-terminal domain of Musashi. We show that loss of GLD2 interaction through deletion of the binding domain or treatment with antisense oligonucleotides compromises Musashi function. Additionally, we demonstrate that overexpression of both Musashi and GLD2 significantly enhances Musashi function. Finally, we report a similar co-association also occurs between murine Musashi and GLD2 orthologs, suggesting that coupling of Musashi to the polyadenylation apparatus is a conserved mechanism to promote target mRNA translation. PMID:24644291

Characterization of gas hydrate reservoirs by integration of core and log data in the Ulleung Basin, East Sea

USGS Publications Warehouse

Bahk, J.-J.; Kim, G.-Y.; Chun, J.-H.; Kim, J.-H.; Lee, J.Y.; Ryu, B.-J.; Lee, J.-H.; Son, B.-K.; Collett, Timothy S.

2013-01-01

Examinations of core and well-log data from the Second Ulleung Basin Gas Hydrate Drilling Expedition (UBGH2) drill sites suggest that Sites UBGH2-2_2 and UBGH2-6 have relatively good gas hydrate reservoir quality in terms of individual and total cumulative thicknesses of gas-hydrate-bearing sand (HYBS) beds. In both of the sites, core sediments are generally dominated by hemipelagic muds which are intercalated with turbidite sands. The turbidite sands are usually thin-to-medium bedded and mainly consist of well sorted coarse silt to fine sand. Anomalies in infrared core temperatures and porewater chlorinity data and pressure core measurements indicate that “gas hydrate occurrence zones” (GHOZ) are present about 68–155 mbsf at Site UBGH2-2_2 and 110–155 mbsf at Site UBGH2-6. In both the GHOZ, gas hydrates are preferentially associated with many of the turbidite sands as “pore-filling” type hydrates. The HYBS identified in the cores from Site UBGH2-6 are medium-to-thick bedded particularly in the lower part of the GHOZ and well coincident with significant high excursions in all of the resistivity, density, and velocity logs. Gas-hydrate saturations in the HYBS range from 12% to 79% with an average of 52% based on pore-water chlorinity. In contrast, the HYBS from Site UBGH2-2_2 are usually thin-bedded and show poor correlations with both of the resistivity and velocity logs owing to volume averaging effects of the logging tools on the thin HYBS beds. Gas-hydrate saturations in the HYBS range from 15% to 65% with an average of 37% based on pore-water chlorinity. In both of the sites, large fluctuations in biogenic opal contents have significant effects on the sediment physical properties, resulting in limited usage of gamma ray and density logs in discriminating sand reservoirs.

In-situ carbon and nitrogen turnover dynamics and the role of soil functional biodiversity therein; a climate warming simulation study in Alpine ecosystems

NASA Astrophysics Data System (ADS)

Djukic, Ika

2010-05-01

Climate change affects a variety of soil properties and processes. Alpine soils take an extraordinary position in this context because of the vulnerability of mountain regions to climatic changes. We used altitudinal soil translocation to simulate the combined effects of changing climatic conditions and shifting vegetation zones in order to study short- to medium-term soil changes in the Austrian Limestone Alps. We translocated 160 soil cores from an alpine grassland site (1900 m asl) down to a sub-alpine spruce forest (1300 m asl) and a montane beech forest site (900m asl), including reference soil cores at each site to estimate artifacts arising from the method. 15N-labeled maize straw was added (1 kg/m2) to translocated and control soil cores and sampled over a period of 2 years for the analysis of δ13C and δ15N in the bulk soil and extracted phospholipid fatty acids (PLFAs). Additionally, 20 litter bags (at each of the three climatic zones) containing Fagus sylvatica or Pinus nigra litter were inserted into the soil, and decomposition was studied over a two-year period. The basic soil parameters (organic C, total N and pH) were unaffected by translocation within the observation time. Overall, decomposition of Pinus nigra litter was significantly slower compared to Fagus sylvatica, and the decomposition rate of both litter types was inversely related to elevation. The decomposition of the maize straw carbon was significantly faster in the translocated soil cores (sites at 900 and 1300 m asl) than at the original site (1900 m asl). The labelled nitrogen contents in the translocated soil cores showed just marginal differences to the soil cores at the original site. The maize straw application promptly increased the amount of bacterial and fungal PLFAs at all studied sites. Downslope translocated soil cores showed an increase in total microbial biomass and sum of bacteria. The fungal PLFA biomarker 18:2ω6,9 was slightly lower at the new (host) sites compared to the original site. The bacterial to fungal ratio of the translocated soil cores showed a rapid acclimatization to the new (host) soil conditions. Our study demonstrates that rising temperatures in Alpine ecosystems will accelerate decomposition of fresh C pools but also lead to rapid adaptation of the microbial community to the new conditions.

St. Petersburg Coastal and Marine Science Center's Core Archive Portal

USGS Publications Warehouse

Reich, Chris; Streubert, Matt; Dwyer, Brendan; Godbout, Meg; Muslic, Adis; Umberger, Dan

2012-01-01

This Web site contains information on rock cores archived at the U.S. Geological Survey (USGS) St. Petersburg Coastal and Marine Science Center (SPCMSC). Archived cores consist of 3- to 4-inch-diameter coral cores, 1- to 2-inch-diameter rock cores, and a few unlabeled loose coral and rock samples. This document - and specifically the archive Web site portal - is intended to be a 'living' document that will be updated continually as additional cores are collected and archived. This document may also contain future references and links to a catalog of sediment cores. Sediment cores will include vibracores, pushcores, and other loose sediment samples collected for research purposes. This document will: (1) serve as a database for locating core material currently archived at the USGS SPCMSC facility; (2) provide a protocol for entry of new core material into the archive system; and, (3) set the procedures necessary for checking out core material for scientific purposes. Core material may be loaned to other governmental agencies, academia, or non-governmental organizations at the discretion of the USGS SPCMSC curator.

Scale-dependent spatial variability in peatland lead pollution in the southern Pennines, UK.

PubMed

Rothwell, James J; Evans, Martin G; Lindsay, John B; Allott, Timothy E H

2007-01-01

Increasingly, within-site and regional comparisons of peatland lead pollution have been undertaken using the inventory approach. The peatlands of the Peak District, southern Pennines, UK, have received significant atmospheric inputs of lead over the last few hundred years. A multi-core study at three peatland sites in the Peak District demonstrates significant within-site spatial variability in industrial lead pollution. Stochastic simulations reveal that 15 peat cores are required to calculate reliable lead inventories at the within-site and within-region scale for this highly polluted area of the southern Pennines. Within-site variability in lead pollution is dominant at the within-region scale. The study demonstrates that significant errors may be associated with peatland lead inventories at sites where only a single peat core has been used to calculate an inventory. Meaningful comparisons of lead inventories at the regional or global scale can only be made if the within-site variability of lead pollution has been quantified reliably.

The EOS land validation core sites: background information and current status

USGS Publications Warehouse

Morisette, J.; Privette, J.L.; Justice, C.; Olson, D.; Dwyer, John L.; Davis, P.; Starr, D.; Wickland, D.

1999-01-01

The EOS Land Validation Core Sites1 will provide the user community with timely ground, aircraft, and satellite data for EOS science and validation investigations. The sites, currently 24 distributed worldwide, represent a consensus among the instrument teams and validation investigators and represent a range of global biome types (see Figure 1 and Table 1; Privette et al., 1999; Justice et al., 1998). The sites typically have a history of in situ and remote observations and can expect continued monitoring and land cover research activities. In many cases, a Core Site will have a tower equipped with above-canopy instrumentation for nearcontinuous sampling of landscape radiometric, energy and CO2 flux, meteorological variables, and atmospheric aerosol and water vapor data. These will be complemented by intensive field measurement campaigns. The data collected at these sites will provide an important resource for the broader science community. These sites can also provide a foundation for a validation network supported and used by all international space agencies.

Geologic implications of gas hydrates in the offshore of India: results of the National Gas Hydrate Program Expedition 01

USGS Publications Warehouse

Collett, Timothy S.; Boswell, Ray; Cochran, J.R.; Kumar, Pushpendra; Lall, Malcolm; Mazumdar, Aninda; Ramana, Mangipudi Venkata; Ramprasad, Tammisetti; Riedel, Michael; Sain, Kalachand; Sathe, Arun Vasant; Vishwanath, Krishna

2014-01-01

One of the specific objectives of this expedition was to test gas hydrate formation models and constrain model parameters, especially those that account for the formation of concentrated gas hydrate accumulations. The necessary data for characterizing the occurrence of in situ gas hydrate, such as interstitial water chlorinities, core-derived gas chemistry, physical and sedimentological properties, thermal images of the recovered cores, and downhole measured logging data (LWD and/or conventional wireline log data), were obtained from most of the drill sites established during NGHP-01. Almost all of the drill sites yielded evidence for the occurrence of gas hydrate; however, the inferred in situ concentration of gas hydrate varied substantially from site to site. For the most part, the interpretation of downhole logging data, core thermal images, interstitial water analyses, and pressure core images from the sites drilled during NGHP-01 indicate that the occurrence of concentrated gas hydrate is mostly associated with the presence of fractures in the sediments, and in some limited cases, by coarser grained (mostly sand-rich) sediments.

Sedimentology of cores recovered from the Canada Basin of the Arctic Ocean

NASA Astrophysics Data System (ADS)

Edwards, B. D.; Saint-Ange, F.; Pohlman, J.; Higgins, J.; Mosher, D. C.; Lorenson, T. D.; Hart, P.

2011-12-01

Researchers from the United States and Canada are collaborating to understand the tectonic and sedimentary history of the Arctic Ocean between Canada and Alaska. As part of this on-going study, a joint US-Canadian ice breaker expedition operated in parts of the Canada Basin during August 2010. Occasional interruptions of the seismic data acquisition provided the ship time to collect gravity and piston cores at five sites-of-opportunity throughout the basin. High-resolution multibeam bathymetry and chirp sub-bottom profiler data collected immediately prior to coring reveal the fine-scale morphology of each site. Core photographs, X-ray radiographs, and physical property data support the following descriptions. Two piston cores were collected from the Beaufort Sea continental margin in a region of known bottom simulating reflectors (BSRs). Site 1 (2538 m water depth): This core recovered 5.72 m of gas-charged, gray sticky clay and silty-clay from an approximately 1100 m diameter, 130 m high conical mound overlying the crest of a buried anticline. Gas hydrate recovered in the core catcher combined with cracks and voids, methane and other hydrocarbon gasses, pyrite concretions, chemosynthetic clams, carbonate nodules, and soft carbonate masses indicate the likely upward migration of deep-seated fluids. Site 2 (1157 m water depth): This core, positioned 40 km upslope from the gas hydrate core, recovered 3 m of gray sticky silty clay and clayey silt near the base of an erosional scarp. Some voids and fracturing are apparent but carbonate masses and pyrite concretions are absent. Site 3 (3070 m water depth): This core from the top of a seamount discovered in 2009 in the north-central part of the Canada Basin recovered 4.94 m of sediment. More than 3 m of dark brown to yellowish brown, massive interbedded silty clays with sands and matrix-supported gravels (ice rafted debris [IRD]) occur in abrupt contact with underlying reddish yellow to brownish yellow silty clay and gravelly sandy clay interpreted to be altered hydrothermally. Successions of IRD layers create a thinly- to medium-bedded sequence throughout the lower section. Site 4 (3700 m water depth; central Canada Basin): This core recovered 3.4 m of sediment typified by decimeter-thick sequences of stacked graded beds with erosional basal contacts (Bouma sequences) characteristic of turbidite deposition. Site 5 (2081 m water depth; continental slope west of the Canadian archipelago): This core recovered 4.96 m of sediment of which the upper 2 m is silty clay to clayey silt with dispersed sand and granules. This upper section has an irregular, sharp basal contact with an underlying 16-cm-thick clast-supported massive gravel. The gravel has a scoured basal contact and overlies a monotonous gray clayey silt containing dispersed granules of IRD. Sparse and poorly preserved fauna throughout all the cores make age determination difficult; however, given the paucity of sediment cores in the Arctic Ocean, these samples provide vital geophysical groundtruth and sedimentological information about the basin.

CMS Readiness for Multi-Core Workload Scheduling

DOE Office of Scientific and Technical Information (OSTI.GOV)

Perez-Calero Yzquierdo, A.; Balcas, J.; Hernandez, J.

In the present run of the LHC, CMS data reconstruction and simulation algorithms benefit greatly from being executed as multiple threads running on several processor cores. The complexity of the Run 2 events requires parallelization of the code to reduce the memory-per- core footprint constraining serial execution programs, thus optimizing the exploitation of present multi-core processor architectures. The allocation of computing resources for multi-core tasks, however, becomes a complex problem in itself. The CMS workload submission infrastructure employs multi-slot partitionable pilots, built on HTCondor and GlideinWMS native features, to enable scheduling of single and multi-core jobs simultaneously. This provides amore » solution for the scheduling problem in a uniform way across grid sites running a diversity of gateways to compute resources and batch system technologies. This paper presents this strategy and the tools on which it has been implemented. The experience of managing multi-core resources at the Tier-0 and Tier-1 sites during 2015, along with the deployment phase to Tier-2 sites during early 2016 is reported. The process of performance monitoring and optimization to achieve efficient and flexible use of the resources is also described.« less

CMS readiness for multi-core workload scheduling

NASA Astrophysics Data System (ADS)

Perez-Calero Yzquierdo, A.; Balcas, J.; Hernandez, J.; Aftab Khan, F.; Letts, J.; Mason, D.; Verguilov, V.

2017-10-01

In the present run of the LHC, CMS data reconstruction and simulation algorithms benefit greatly from being executed as multiple threads running on several processor cores. The complexity of the Run 2 events requires parallelization of the code to reduce the memory-per- core footprint constraining serial execution programs, thus optimizing the exploitation of present multi-core processor architectures. The allocation of computing resources for multi-core tasks, however, becomes a complex problem in itself. The CMS workload submission infrastructure employs multi-slot partitionable pilots, built on HTCondor and GlideinWMS native features, to enable scheduling of single and multi-core jobs simultaneously. This provides a solution for the scheduling problem in a uniform way across grid sites running a diversity of gateways to compute resources and batch system technologies. This paper presents this strategy and the tools on which it has been implemented. The experience of managing multi-core resources at the Tier-0 and Tier-1 sites during 2015, along with the deployment phase to Tier-2 sites during early 2016 is reported. The process of performance monitoring and optimization to achieve efficient and flexible use of the resources is also described.

Angiographic Validation of the American College of Cardiology Foundation–The Society of Thoracic Surgeons Collaboration on the Comparative Effectiveness of Revascularization Strategies Study

PubMed Central

Chakrabarti, Anjan K.; Grau-Sepulveda, Maria V.; O’Brien, Sean; Abueg, Cassandra; Ponirakis, Angelo; Delong, Elizabeth; Peterson, Eric; Klein, Lloyd W.; Garratt, Kirk N.; Weintraub, William S.; Gibson, C. Michael

2017-01-01

Background The goal of this study was to compare angiographic interpretation of coronary arteriograms by sites in community practice versus those made by a centralized angiographic core laboratory. Methods and Results The study population consisted of 2013 American College of Cardiology–National Cardiovascular Data Registry (ACC–NCDR) records with 2- and 3- vessel coronary disease from 54 sites in 2004 to 2007. The primary analysis compared Registry (NCDR)-defined 2- and 3-vessel disease versus those from an angiographic core laboratory analysis. Vessel-level kappa coefficients suggested moderate agreement between NCDR and core laboratory analysis, ranging from kappa=0.39 (95% confidence intervals, 0.32–0.45) for the left anterior descending artery to kappa=0.59 (95% confidence intervals, 0.55–0.64) for the right coronary artery. Overall, 6.3% (n=127 out of 2013) of those patients identified with multivessel disease at NCDR sites had had 0- or 1-vessel disease by core laboratory reading. There was no directional bias with regard to overcall, that is, 12.3% of cases read as 3-vessel disease by the sites were read as <3-vessel disease by the core laboratory, and 13.9% of core laboratory 3-vessel cases were read as <3-vessel by the sites. For a subset of patients with left main coronary disease, registry overcall was not linked to increased rates of mortality or myocardial infarction. Conclusions There was only modest agreement between angiographic readings in clinical practice and those from an independent core laboratory. Further study will be needed because the implications for patient management are uncertain. PMID:24496239

Angiographic validation of the American College of Cardiology Foundation-the Society of Thoracic Surgeons Collaboration on the Comparative Effectiveness of Revascularization Strategies study.

PubMed

Chakrabarti, Anjan K; Grau-Sepulveda, Maria V; O'Brien, Sean; Abueg, Cassandra; Ponirakis, Angelo; Delong, Elizabeth; Peterson, Eric; Klein, Lloyd W; Garratt, Kirk N; Weintraub, William S; Gibson, C Michael

2014-02-01

The goal of this study was to compare angiographic interpretation of coronary arteriograms by sites in community practice versus those made by a centralized angiographic core laboratory. The study population consisted of 2013 American College of Cardiology-National Cardiovascular Data Registry (ACC-NCDR) records with 2- and 3- vessel coronary disease from 54 sites in 2004 to 2007. The primary analysis compared Registry (NCDR)-defined 2- and 3-vessel disease versus those from an angiographic core laboratory analysis. Vessel-level kappa coefficients suggested moderate agreement between NCDR and core laboratory analysis, ranging from kappa=0.39 (95% confidence intervals, 0.32-0.45) for the left anterior descending artery to kappa=0.59 (95% confidence intervals, 0.55-0.64) for the right coronary artery. Overall, 6.3% (n=127 out of 2013) of those patients identified with multivessel disease at NCDR sites had had 0- or 1-vessel disease by core laboratory reading. There was no directional bias with regard to overcall, that is, 12.3% of cases read as 3-vessel disease by the sites were read as <3-vessel disease by the core laboratory, and 13.9% of core laboratory 3-vessel cases were read as <3-vessel by the sites. For a subset of patients with left main coronary disease, registry overcall was not linked to increased rates of mortality or myocardial infarction. There was only modest agreement between angiographic readings in clinical practice and those from an independent core laboratory. Further study will be needed because the implications for patient management are uncertain.

Site Fidelity in Space Use by Spider Monkeys (Ateles geoffroyi) in the Yucatan Peninsula, Mexico

PubMed Central

Ramos-Fernandez, Gabriel; Smith Aguilar, Sandra E.; Schaffner, Colleen M.; Vick, Laura G.; Aureli, Filippo

2013-01-01

Animal home ranges may vary little in their size and location in the short term but nevertheless show more variability in the long term. We evaluated the degree of site fidelity of two groups of spider monkeys (Ateles geoffroyi) over a 10- and 13-year period, respectively, in the northeastern Yucatan peninsula, Mexico. We used the Local Convex Hull method to estimate yearly home ranges and core areas (defined as the 60% probability contour) for the two groups. Home ranges varied from 7.7 to 49.6 ha and core areas varied from 3.1 to 9.2 ha. We evaluated the degree of site fidelity by quantifying the number of years in which different areas were used as either home ranges or core areas. Large tracts were used only as home ranges and only for a few years, whereas small areas were used as either core area or home range for the duration of the study. The sum of the yearly core areas coincided partially with the yearly home ranges, indicating that home ranges contain areas used intermittently. Home ranges, and especially core areas, contained a higher proportion of mature forest than the larger study site as a whole. Across years and only in one group, the size of core areas was positively correlated with the proportion of adult males in the group, while the size of home ranges was positively correlated with both the proportion of males and the number of tree species included in the diet. Our findings suggest that spider monkey home ranges are the result of a combination of long-term site fidelity and year-to-year use variation to enable exploration of new resources. PMID:23675427

Long-term Glacial History of the West Antarctic Ice Sheet from Cosmogenic Nuclides in a Subglacial Bedrock Core

NASA Astrophysics Data System (ADS)

Spector, P. E.; Stone, J.; Hillebrand, T.; Gombiner, J. H.

2017-12-01

To investigate the response of the West Antarctic Ice Sheet (WAIS) to climatic conditions warmer than present, we are analyzing cosmogenic nuclides in a bedrock core from beneath 150 m of ice at a site near the Pirrit Hills. Our aim is to determine whether the WAIS has thinned in the past, exposing bedrock at this site, and if so, when. This will help to determine the vulnerability of the ice sheet to future warming, and identify climatic thresholds capable of inducing WAIS collapse. We selected a site where the ice-sheet surface lies at 1300 m, approximately halfway from the ice-sheet divide to the grounding line. We expect ice thickness at the site to reflect WAIS dynamics, rather than local meteorology or topography. Ice flow speeds are moderate and ice above the core site is thin enough to remain cold-based, limiting the possibility of subglacial erosion which would compromise the cosmogenic nuclide record. We targeted a subglacial ridge adjacent to an exposed granite nunatak. This lithology provides minerals suitable for analysis of multiple cosmogenic nuclides with different half-lives. Although we aimed to collect two cores from different depths to compare exposure histories, hydrofracture of the basal ice prevented us from reaching the bed at the first drill site. The second hole produced 5.5 m of discontinuous ice core above 8 m of bedrock core. Initial analyses of quartz from the bedrock show low levels of Be-10. Further analyses of Be-10, Al-26, Cl-36 and Ne-21 from the full length of the core will be required to determine whether this is because the surface has never been exposed, or because the cosmogenic nuclide profile has been truncated by glacial erosion. We will present comprehensive cosmogenic nuclide data, and discuss implications for WAIS deglaciation history, at the meeting. Supported by US National Science Foundation awards ANT-1142162 and PLR-1341728.

Closed-loop 15N measurement of N2O and its isotopomers for real-time greenhouse gas tracing

NASA Astrophysics Data System (ADS)

Slaets, Johanna; Mayr, Leopold; Heiling, Maria; Zaman, Mohammad; Resch, Christian; Weltin, Georg; Gruber, Roman; Dercon, Gerd

2016-04-01

Quantifying sources of nitrous oxide is essential to improve understanding of the global N cycle and to develop climate-smart agriculture, as N2O has a global warming potential 300 times higher than CO2. The isotopic signature and the intramolecular distribution (site preference) of 15N are powerful tools to trace N2O, but the application of these methods is limited as conventional methods cannot provide continuous and in situ data. Here we present a method for closed-loop, real time monitoring of the N2O flux, the isotopic signature and the intramolecular distribution of 15N by using off-axis integrated cavity output spectroscopy (ICOS, Los Gatos Research). The developed method was applied to a fertilizer inhibitor experiment, in which N2O emissions were measured on undisturbed soil cores for three weeks. The treatments consisted of enriched urea-N (100 kg urea-N/ha), the same fertilizer combined with the nitrification inhibitor nitrapyrin (375 g/100 kg urea), and control cores. Monitoring the isotopic signature makes it possible to distinguish emissions from soil and fertilizer. Characterization of site preference could additionally provide a tool to identify different microbial processes leading to N2O emissions. Furthermore, the closed-loop approach enables direct measurement on site and does not require removal of CO2 and H2O. Results showed that 75% of total N2O emissions (total=11 346 μg N2O-N/m2) in the fertilized cores originated from fertilizer, while only 55% of total emissions (total=2 450 μg N2ON/m2) stemmed from fertilizer for the cores treated with nitrapyrin. In the controls, N2O derived from soil was only 40% of the size of the corresponding pool from the fertilized cores, pointing towards a priming effect on the microbial community from the fertilizer and demonstrating the bias that could be introduced by relying on non-treated cores to estimate soil emission rates, rather than using the isotopic signature. The site preference increased linearly over time for the cores with fertilizer and those with nitrapyrin, but the increase was stronger for the fertilized cores: during the first 10 days of the experiment, theses cores showed a more negative site preference than the cores with inhibitor, while during the last 10 days, the site preference for the fertilized cores was more positive than that of the inhibitor. This change indicates that the site preference of 15N can be used to distinguish the processes of nitrification and denitrification, the former having been supressed by nitrapyrin in the cores treated with the inhibitor. Low enrichment levels (5% atomic excess in this study) sufficed in order to separate emissions from soil and fertilizer, making the proposed closed-loop approach a cost-effective and practical tool to obtain a continuous, in situ characterization of N2O sources.

Variation of Accumulation Rates Over the Last Eight Centuries on the East Antarctic Plateau Derived from Volcanic Signals in Ice Cores

NASA Technical Reports Server (NTRS)

Anschuetz, H.; Sinisalo, A.; Isaksson, E.; McConnell, J. R.; Hamran, S.-E.; Bisiaux, M. M.; Pasteris, D.; Neumann, T. A.; Winther, J.-G.

2011-01-01

Volcanic signatures in ice-core records provide an excellent means to date the cores and obtain information about accumulation rates. From several ice cores it is thus possible to extract a spatio-temporal accumulation pattern. We show records of electrical conductivity and sulfur from firn cores from the Norwegian-USA scientific traverse during the International Polar Year 2007-2009 (IPY) through East Antarctica. Major volcanic eruptions are identified and used to assess century-scale accumulation changes. The largest changes seem to occur in the most recent decades with accumulation over the period 1963- 2007/08 being up to 25 % different from the long-term record. There is no clear overall trend, some sites show an increase in accumulation over the period 1963 to present while others show a decrease. Almost all of the sites above 3200 m above sea level (asl) suggest a decrease. These sites also show a significantly lower accumulation value than large-scale assessments both for the period 1963 to present and for the long-term mean at the respective drill sites. The spatial accumulation distribution is influenced mainly by elevation and distance to the ocean (continentality), as expected. Ground-penetrating radar data around the drill sites show a spatial variability within 10-20 % over several tens of kilometers, indicating that our drill sites are well representative for the area around them. Our results are important for large-scale assessments of Antarctic mass balance and model validation.

Using Firn Air for Facility Cooling at the WAIS Divide Site

DTIC Science & Technology

2014-09-17

reduce logistics costs at remote field camps where it is critical to maintain proper temperatures to preserve sensitive deep ice cores. We assessed the...feasibility of using firn air for cooling at the West Antarc- tic Ice Sheet (WAIS) Divide ice core drilling site as a means to adequately and...efficiently refrigerate ice cores during storage and processing. We used estimates of mean annual temperature, temperature variations, and firn

Structural correlates of affinity in fetal versus adult endplate nicotinic receptors

NASA Astrophysics Data System (ADS)

Nayak, Tapan Kumar; Chakraborty, Srirupa; Zheng, Wenjun; Auerbach, Anthony

2016-04-01

Adult-type nicotinic acetylcholine receptors (AChRs) mediate signalling at mature neuromuscular junctions and fetal-type AChRs are necessary for proper synapse development. Each AChR has two neurotransmitter binding sites located at the interface of a principal and a complementary subunit. Although all agonist binding sites have the same core of five aromatic amino acids, the fetal site has ~30-fold higher affinity for the neurotransmitter ACh. Here we use molecular dynamics simulations of adult versus fetal homology models to identify complementary-subunit residues near the core that influence affinity, and use single-channel electrophysiology to corroborate the results. Four residues in combination determine adult versus fetal affinity. Simulations suggest that at lower-affinity sites, one of these unsettles the core directly and the others (in loop E) increase backbone flexibility to unlock a key, complementary tryptophan from the core. Swapping only four amino acids is necessary and sufficient to exchange function between adult and fetal AChRs.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Huang, Tingting; Chang, Chin -Yuan; Lohman, Jeremy R.

Comparative analysis of the enediyne biosynthetic gene clusters revealed sets of conserved genes serving as outstanding candidates for the enediyne core. Here we report the crystal structures of SgcJ and its homologue NCS-Orf16, together with gene inactivation and site-directed mutagenesis studies, to gain insight into enediyne core biosynthesis. Gene inactivation in vivo establishes that SgcJ is required for C-1027 production in Streptomyces globisporus. SgcJ and NCS-Orf16 share a common structure with the nuclear transport factor 2-like superfamily of proteins, featuring a putative substrate binding or catalytic active site. Site-directed mutagenesis of the conserved residues lining this site allowed us tomore » propose that SgcJ and its homologues may play a catalytic role in transforming the linear polyene intermediate, along with other enediyne polyketide synthase-associated enzymes, into an enzyme-sequestered enediyne core intermediate. In conclusion, these findings will help formulate hypotheses and design experiments to ascertain the function of SgcJ and its homologues in nine-membered enediyne core biosynthesis.« less

Site 765: Sediment Lithostratigraphy

USGS Publications Warehouse

,

1990-01-01

A 935-m-thick succession of Quaternary through Lower Cretaceous sediments was recovered at Site 765 (Fig. 10). A single core of Quaternary sediment was obtained from Hole 765A; drilling terminated and a new hole was drilled in an attempt to establish the mud line. Quaternary through middle Miocene sediments were cored in Hole 765B down to a depth of 395.6 mbsf. Middle Miocene through Lower Cretaceous sediments were cored in Hole 765C, after washing the interval between 0 and 350.2 mbsf. Exact lithologic correlation of the basal cores from Hole 765B with the upper cores from Hole 765C is not possible because of poor recovery; hence, correlation is based solely on matching sub-bottom depths.

CD/MCD/VTVH-MCD Studies of Escherichia coli Bacterioferritin Support a Binuclear Iron Cofactor Site.

PubMed

Kwak, Yeonju; Schwartz, Jennifer K; Huang, Victor W; Boice, Emily; Kurtz, Donald M; Solomon, Edward I

2015-12-01

Ferritins and bacterioferritins (Bfrs) utilize a binuclear non-heme iron binding site to catalyze oxidation of Fe(II), leading to formation of an iron mineral core within a protein shell. Unlike ferritins, in which the diiron site binds Fe(II) as a substrate, which then autoxidizes and migrates to the mineral core, the diiron site in Bfr has a 2-His/4-carboxylate ligand set that is commonly found in diiron cofactor enzymes. Bfrs could, therefore, utilize the diiron site as a cofactor rather than for substrate iron binding. In this study, we applied circular dichroism (CD), magnetic CD (MCD), and variable-temperature, variable-field MCD (VTVH-MCD) spectroscopies to define the geometric and electronic structures of the biferrous active site in Escherichia coli Bfr. For these studies, we used an engineered M52L variant, which is known to eliminate binding of a heme cofactor but to have very minor effects on either iron oxidation or mineral core formation. We also examined an H46A/D50A/M52L Bfr variant, which additionally disrupts a previously observed mononuclear non-heme iron binding site inside the protein shell. The spectral analyses define a binuclear and an additional mononuclear ferrous site. The biferrous site shows two different five-coordinate centers. After O2 oxidation and re-reduction, only the mononuclear ferrous signal is eliminated. The retention of the biferrous but not the mononuclear ferrous site upon O2 cycling supports a mechanism in which the binuclear site acts as a cofactor for the O2 reaction, while the mononuclear site binds the substrate Fe(II) that, after its oxidation to Fe(III), migrates to the mineral core.

The Heuristic of George Polya and Its Relation to Artificial Intelligence

DTIC Science & Technology

1981-07-01

entitled Mathematical methods in physical science -- odd, in not being the standard freshman fare of physics, chemistry , western civilization and...the questions teachers should use. The first I I Having used a linguistic variation lo convey a matter of substance, a comment on etymology is in...Intelligence to Chemistry : The DEN ORAL project. New York- McGraw-Hill 1980. McDermott, J. Learning to use analogies. Proceedings of the International Joint

Semantics of Procedures: A Cognitive Basis for Maintenance Training Competency

DTIC Science & Technology

1988-04-01

provided by presenting these multiple, interacting views. These 0 conclusions are similar to the views of George Polya , who proposed the use of... exercises . It is also clear that at least some understanding of machine structure and function is required to interpret much of the information in the...check of the problem lists provides the names of all the usual subpects. This accessibility permits the technician to modify the sequence according to

Symposium on Chemical Applications of Topology and Graph Theory, April 18-22, 1983.

DTIC Science & Technology

1983-04-01

illustrated by application to the Lotka - Volterra oscillator. ELECTRICAL NETWORK REPRESENTATION OF n-DIMENSIONAL CHEMICAL MANIFOLDS L. Peusner P.O. Box 380...like molecules and others; the original formulas by Cayley were extended by Polya in a general enumeration theorem, simplified by Otter, and also studied...Gutman, leading to a joint paper which generalized it, using line graphs. Finally, electroneqativity consider- ations tell the strength of a chemical

Naval Research Logistics Quarterly. Volume 27, Number 2.

DTIC Science & Technology

1980-06-01

research area in recent years. Van Zyl [11] provided the first major study on single-product perishable inventories, but the field only began to...Journal of Applied Mathematics 30, 483-500 (1976). [III Van Zyl, G.J.J., "Inventory Control for Perishable Commodities," Unpublished Ph.D. dissertation...Hollatz and B. Bank, "Theorie der Linearen Parametrischen Optimierung," Akademie-Verlag, Berlin (1974). [4] Polya, G. and G. Szego, "Problems and

The cDNA sequence of a neutral horseradish peroxidase.

PubMed

Bartonek-Roxå, E; Eriksson, H; Mattiasson, B

1991-02-16

A cDNA clone encoding a horseradish (Armoracia rusticana) peroxidase has been isolated and characterized. The cDNA contains 1378 nucleotides excluding the poly(A) tail and the deduced protein contains 327 amino acids which includes a 28 amino acid leader sequence. The predicted amino acid sequence is nine amino acids shorter than the major isoenzyme belonging to the horseradish peroxidase C group (HRP-C) and the sequence shows 53.7% identity with this isoenzyme. The described clone encodes nine cysteines of which eight correspond well with the cysteines found in HRP-C. Five potential N-glycosylation sites with the general sequence Asn-X-Thr/Ser are present in the deduced sequence. Compared to the earlier described HRP-C this is three glycosylation sites less. The shorter sequence and fewer N-glycosylation sites give the native isoenzyme a molecular weight of several thousands less than the horseradish peroxidase C isoenzymes. Comparison with the net charge value of HRP-C indicates that the described cDNA clone encodes a peroxidase which has either the same or a slightly less basic pI value, depending on whether the encoded protein is N-terminally blocked or not. This excludes the possibility that HRP-n could belong to either the HRP-A, -D or -E groups. The low sequence identity (53.7%) with HRP-C indicates that the described clone does not belong to the HRP-C isoenzyme group and comparison of the total amino acid composition with the HRP-B group does not place the described clone within this isoenzyme group. Our conclusion is that the described cDNA clone encodes a neutral horseradish peroxidase which belongs to a new, not earlier described, horseradish peroxidase group.

StereoGene: rapid estimation of genome-wide correlation of continuous or interval feature data.

PubMed

Stavrovskaya, Elena D; Niranjan, Tejasvi; Fertig, Elana J; Wheelan, Sarah J; Favorov, Alexander V; Mironov, Andrey A

2017-10-15

Genomics features with similar genome-wide distributions are generally hypothesized to be functionally related, for example, colocalization of histones and transcription start sites indicate chromatin regulation of transcription factor activity. Therefore, statistical algorithms to perform spatial, genome-wide correlation among genomic features are required. Here, we propose a method, StereoGene, that rapidly estimates genome-wide correlation among pairs of genomic features. These features may represent high-throughput data mapped to reference genome or sets of genomic annotations in that reference genome. StereoGene enables correlation of continuous data directly, avoiding the data binarization and subsequent data loss. Correlations are computed among neighboring genomic positions using kernel correlation. Representing the correlation as a function of the genome position, StereoGene outputs the local correlation track as part of the analysis. StereoGene also accounts for confounders such as input DNA by partial correlation. We apply our method to numerous comparisons of ChIP-Seq datasets from the Human Epigenome Atlas and FANTOM CAGE to demonstrate its wide applicability. We observe the changes in the correlation between epigenomic features across developmental trajectories of several tissue types consistent with known biology and find a novel spatial correlation of CAGE clusters with donor splice sites and with poly(A) sites. These analyses provide examples for the broad applicability of StereoGene for regulatory genomics. The StereoGene C ++ source code, program documentation, Galaxy integration scripts and examples are available from the project homepage http://stereogene.bioinf.fbb.msu.ru/. favorov@sensi.org. Supplementary data are available at Bioinformatics online. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com

Isolation and characterization of full-length cDNA clones coding for cholinesterase from fetal human tissues.

PubMed Central

Prody, C A; Zevin-Sonkin, D; Gnatt, A; Goldberg, O; Soreq, H

1987-01-01

To study the primary structure and regulation of human cholinesterases, oligodeoxynucleotide probes were prepared according to a consensus peptide sequence present in the active site of both human serum pseudocholinesterase (BtChoEase; EC 3.1.1.8) and Torpedo electric organ "true" acetylcholinesterase (AcChoEase; EC 3.1.1.7). Using these probes, we isolated several cDNA clones from lambda gt10 libraries of fetal brain and liver origins. These include 2.4-kilobase cDNA clones that code for a polypeptide containing a putative signal peptide and the N-terminal, active site, and C-terminal peptides of human BtChoEase, suggesting that they code either for BtChoEase itself or for a very similar but distinct fetal form of cholinesterase. In RNA blots of poly(A)+ RNA from the cholinesterase-producing fetal brain and liver, these cDNAs hybridized with a single 2.5-kilobase band. Blot hybridization to human genomic DNA revealed that these fetal BtChoEase cDNA clones hybridize with DNA fragments of the total length of 17.5 kilobases, and signal intensities indicated that these sequences are not present in many copies. Both the cDNA-encoded protein and its nucleotide sequence display striking homology to parallel sequences published for Torpedo AcChoEase. These findings demonstrate extensive homologies between the fetal BtChoEase encoded by these clones and other cholinesterases of various forms and species. Images PMID:3035536

Core-Shell Processing of Natural Pigment: Upper Palaeolithic Red Ochre from Lovas, Hungary.

PubMed

Sajó, István E; Kovács, János; Fitzsimmons, Kathryn E; Jáger, Viktor; Lengyel, György; Viola, Bence; Talamo, Sahra; Hublin, Jean-Jacques

2015-01-01

Ochre is the common archaeological term for prehistoric pigments. It is applied to a range of uses, from ritual burials to cave art to medications. While a substantial number of Palaeolithic paint mining pits have been identified across Europe, the link between ochre use and provenance, and their antiquity, has never yet been identified. Here we characterise the mineralogical signature of core-shell processed ochre from the Palaeolithic paint mining pits near Lovas in Hungary, using a novel integration of petrographic and mineralogical techniques. We present the first evidence for core-shell processed, natural pigment that was prepared by prehistoric people from hematitic red ochre. This involved combining the darker red outer shell with the less intensely coloured core to efficiently produce an economical, yet still strongly coloured, paint. We demonstrate the antiquity of the site as having operated between 14-13 kcal BP, during the Epigravettian period. This is based on new radiocarbon dating of bone artefacts associated with the quarry site. The dating results indicate the site to be the oldest known evidence for core-shell pigment processing. We show that the ochre mined at Lovas was exported from the site based on its characteristic signature at other archaeological sites in the region. Our discovery not only provides a methodological framework for future characterisation of ochre pigments, but also provides the earliest known evidence for "value-adding" of products for trade.

Thermal stress in North Western Australian iron ore mining staff.

PubMed

Peiffer, Jeremiah J; Abbiss, Chris R

2013-05-01

Demand for Australian mined iron ore has increased employment within this sector, thus exposing increased numbers of workers to the harsh Australian climate. This study examined the influence of hot (>30°C wet bulb globe temperature) environmental temperatures, consistent with working in North Western Australia, on iron ore mining staff. Core temperature, hydration status, perceived exertion, mood, and fatigue state were measured in 77 participants at three time points (pre-, mid-, and post-shift) during a normal 12-h shift at an open-cut iron ore mining/processing site (n = 31; Site1) and an iron ore processing/shipping site (n = 46; Site2). A significant effect for time was observed for core temperature with greater mean core temperatures measured mid-shift (37.5±0.4°C) and post-shift (37.6±0.3°C) compared with pre-shift values (37.0±0.5°C). All mean core temperature measures were lower than ISO7933 thresholds (38°C) for thermal safety. Mean hydration measures [urine-specific gravity (USG)] were greater at Site1 (1.029±0.006) compared with those at Site2 (1.021±0.007). Furthermore, both pre- and post-shift measures from Site1 and the post-shift measures from Site2 were greater than the threshold for dehydration (USG = 1.020). No differences were observed for mood or perceived exertion over time; however, measures of fatigue state were greater post-shift compared with pre- and mid-shift values for both sites. Our findings indicate that the majority of mine workers in North Western Australia are able to regulate work rate in hot environments to maintain core temperatures below ISO safety guidelines; however, 22% of workers reached or exceeded the safety guidelines, warranting further investigation. Furthermore, hydration practices, especially when off-work, appear inadequate and could endanger health and safety.

A snoRNA modulates mRNA 3′ end processing and regulates the expression of a subset of mRNAs

PubMed Central

Huang, Chunliu; Shi, Junjie; Guo, Yibin; Huang, Weijun; Huang, Shanshan; Ming, Siqi; Wu, Xingui; Zhang, Rui; Ding, Junjun; Zhao, Wei; Jia, Jie; Huang, Xi; Xiang, Andy Peng

2017-01-01

Abstract mRNA 3′ end processing is an essential step in gene expression. It is well established that canonical eukaryotic pre-mRNA 3′ processing is carried out within a macromolecular machinery consisting of dozens of trans-acting proteins. However, it is unknown whether RNAs play any role in this process. Unexpectedly, we found that a subset of small nucleolar RNAs (snoRNAs) are associated with the mammalian mRNA 3′ processing complex. These snoRNAs primarily interact with Fip1, a component of cleavage and polyadenylation specificity factor (CPSF). We have functionally characterized one of these snoRNAs and our results demonstrated that the U/A-rich SNORD50A inhibits mRNA 3′ processing by blocking the Fip1-poly(A) site (PAS) interaction. Consistently, SNORD50A depletion altered the Fip1–RNA interaction landscape and changed the alternative polyadenylation (APA) profiles and/or transcript levels of a subset of genes. Taken together, our data revealed a novel function for snoRNAs and provided the first evidence that non-coding RNAs may play an important role in regulating mRNA 3′ processing. PMID:28911119

Mutagenesis of diploid mammalian genes by gene entrapment

PubMed Central

Lin, Qing; Donahue, Sarah L.; Moore-Jarrett, Tracy; Cao, Shang; Osipovich, Anna B.; Ruley, H. Earl

2006-01-01

The present study describes a genome-wide method for biallelic mutagenesis in mammalian cells. Novel poly(A) gene trap vectors, which contain features for direct cloning vector–cell fusion transcripts and for post-entrapment genome engineering, were used to generate a library of 979 mutant ES cells. The entrapment mutations generally disrupted gene expression and were readily transmitted through the germline, establishing the library as a resource for constructing mutant mice. Cells homozygous for most entrapment loci could be isolated by selecting for enhanced expression of an inserted neomycin-resistance gene that resulted from losses of heterozygosity (LOH). The frequencies of LOH measured at 37 sites in the genome ranged from 1.3 × 10−5 to 1.2 × 10−4 per cell and increased with increasing distance from the centromere, implicating mitotic recombination in the process. The ease and efficiency of obtaining homozygous mutations will (i) facilitate genetic studies of gene function in cultured cells, (ii) permit genome-wide studies of recombination events that result in LOH and mediate a type of chromosomal instability important in carcinogenesis, and (iii) provide new strategies for phenotype-driven mutagenesis screens in mammalian cells. PMID:17062627

Preliminary Physical Stratigraphy and Geophysical Data of the USGS Hope Plantation Core (BE-110), Bertie County, North Carolina

USGS Publications Warehouse

Weems, Robert E.; Seefelt, Ellen L.; Wrege, Beth M.; Self-Trail, Jean M.; Prowell, David C.; Durand, Colleen; Cobbs, Eugene F.; McKinney, Kevin C.

2007-01-01

Introduction In March and April, 2004, the U.S. Geological Survey (USGS), in cooperation with the North Carolina Geological Survey (NCGS) and the Raleigh Water Resources Discipline (WRD), drilled a stratigraphic test hole and well in Bertie County, North Carolina (fig. 1). The Hope Plantation test hole (BE-110-2004) was cored on the property of Hope Plantation near Windsor, North Carolina. The drill site is located on the Republican 7.5 minute quadradrangle at lat 36?01'58'N., long 78?01'09'W. (decimal degrees 36.0329 and 77.0192) (fig. 2). The altitude of the site is 48 ft above mean sea level as determined by Paulin Precise altimeter. This test hole was continuously cored by Eugene F. Cobbs, III and Kevin C. McKinney (USGS) to a total depth of 1094.5 ft. Later, a ground water observation well was installed with a screened interval between 315-329 feet below land surface (fig. 3). Upper Triassic, Lower Cretaceous, Upper Cretaceous, Tertiary, and Quaternary sediments were recovered from the site. The core is stored at the NCGS Coastal Plain core storage facility in Raleigh, North Carolina. In this report, we provide the initial lithostratigraphic summary recorded at the drill site along with site core photographs, data from the geophysical logger, calcareous nannofossil biostratigraphic correlations (Table 1) and initial hydrogeologic interpretations. The lithostratigraphy from this core can be compared to previous investigations of the Elizabethtown corehole, near Elizabethtown, North Carolina in Bladen County (Self-Trail, Wrege, and others, 2004), the Kure Beach corehole, near Wilmington, North Carolina in New Hanover County (Self-Trail, Prowell, and Christopher, 2004), the Esso #1, Esso #2, Mobil #1 and Mobil #2 cores in the Albermarle and Pamlico Sounds (Zarra, 1989), and the Cape Fear River outcrops in Bladen County (Farrell, 1998; Farrell and others, 2001). This core is the third in a series of planned benchmark coreholes that will be used to elucidate the physical stratigraphy, facies, thickness, and hydrogeology of the Tertiary and Cretaceous Coastal Plain sediments of North Carolina.

Field Geometry During the Iceland Basin Event Observed from the North Atlantic Ocean, North Pacific Ocean and the South China Sea

NASA Astrophysics Data System (ADS)

Laj, C.; Kissel, C.; Roberts, A. P.; Hillaire-Marcel, C.; Cortijo, E.

2004-12-01

The Iceland Basin event (IBE), which is named for a record from ODP Site 983 in the North Atlantic Ocean (60.5°N), is a focus of growing interest in our community. This geomagnetic excursion is coeval with the marine oxygen isotope (MIS) stage 7/6 boundary at about 190 ka according to the orbitally tuned SPECMAP time scale. We have detailed new records of this excursion at two new sites from the North Atlantic (core MD99-2247; lat. 59°N, long. 31°W and core MD99-2242; lat. 59°N, long. 47°W), at one site from the North Pacific (ODP Site 884; lat. 51.5°N, long. 168.3°E), and in two others from the South China Sea (ODP Site 1146; lat. 19.5°N, long. 116.3°E and ODP Site 1145; lat. 19.6°N; long. 117.6°E). For all five sites, the event is identified at the MIS 7/6 boundary, which confirms its potential as a precise stratigraphic marker. The average sedimentation rate for this portion of the cores is 7.5, 10 and 15 cm/kyr in North Atlantic, North Pacific and South China sea cores, respectively, which allows high-resolution studies in this time interval. The event is characterized by a marked minimum in the relative paleointensity (sometimes with a double feature) and by large swings in inclination and declination. The VGP latitudes reach 78°S and 45°S for ODP sites 1146 and 1145, respectively, 37°S for ODP Site 884, and 74°S and 41°S for cores MD99-2247 and MD99-2242, respectively. The VGP paths for the four records from the North Atlantic and South China Sea cores are highly similar. For the most detailed of these records, the poles first pass over Africa, then they proceed to Antarctica and return northward over Australia. The paths are less well resolved for ODP Site 1145 and core MD99-2242, for which the poles cross the southern Indian Ocean rather than reaching Antarctica. The VGP paths for the two most detailed records (ODP Site 1146 and MD99-2247) are highly similar and are also similar to the path for ODP Site 983, which also has a high sedimentation rate and is precisely dated. In contrast, but in common with other published records of the IBE, the VGP path for ODP Site 884 loops in the opposite sense and passes from North to South America and then over the Pacific Ocean before returning to high northern latitudes. The striking similarities between multiple records for two observed sets of differing VGP paths for the IBE at locations spanning the northern hemisphere provides evidence for a non-dipolar, but relatively simple, geometry of the transitional field.

Assessment of subsurface chlorinated solvent contamination using tree cores at the front street site and a former dry cleaning facility at the Riverfront Superfund site, New Haven, Missouri, 1999-2003

USGS Publications Warehouse

Schumacher, John G.; Struckhoff, Garrett C.; Burken, Joel G.

2004-01-01

Tree-core sampling has been a reliable and inexpensive tool to quickly assess the presence of shallow (less than about 30 feet deep) tetrachloroethene (PCE) and trichloroethene (TCE) contamination in soils and ground water at the Riverfront Superfund Site. This report presents the results of tree-core sampling that was successfully used to determine the presence and extent of chlorinated solvent contamination at two sites, the Front Street site (operable unit OU1) and the former dry cleaning facility, that are part of the overall Riverfront Superfund Site. Traditional soil and ground-water sampling at these two sites later confirmed the results from the tree-core sampling. Results obtained from the tree-core sampling were used to design and focus subsequent soil and ground-water investigations, resulting in substantial savings in time and site assessment costs. The Front Street site is a small (less than 1-acre) site located on the Missouri River alluvium in downtown New Haven, Missouri, about 500 feet from the south bank of the Missouri River. Tree-core sampling detected the presence of subsurface PCE contamination at the Front Street site and beneath residential property downgradient from the site. Core samples from trees at the site contained PCE concentrations as large as 3,850 mg-h/kg (micrograms in headspace per kilogram of wet core) and TCE concentrations as large as 249 mg-h/kg. Soils at the Front Street site contained PCE concentrations as large as 6,200,000 mg/kg (micrograms per kilogram) and ground-water samples contained PCE concentrations as large as 11,000 mg/L (micrograms per liter). The former dry cleaning facility is located at the base of the upland that forms the south bank of the Missouri River alluvial valley. Tree-core sampling did not indicate the presence of PCE or TCE contamination at the former dry cleaning facility, a finding that was later confirmed by the analyses of soil samples collected from the site. The lateral extent of PCE contamination in trees was in close agreement with the extent of subsurface PCE contamination determined using traditional soil and ground-water sampling methods. Trees growing in soils containing PCE concentrations of 60 to 5,700 mg/kg or larger or overlying ground water containing PCE concentrations from 5 to 11,000 mg/L generally contained detectable concentrations of PCE. The depth to contaminated ground water was about 20 to 25 feet below the land surface. Significant quantitative relations [probability (p) values of less than 0.05 and correlation coefficient (r2) values of 0.88 to 0.90] were found between PCE concentrations in trees and subsurface soils between 4 and 16 feet deep. The relation between PCE concentrations in trees and underlying ground water was less apparent (r2 value of 0.17) and the poor relation is thought to be the result of equilibrium with PCE concentrations in soil and vapor in the unsaturated zone. Based on PCE concentrations detected in trees at the Front Street site and trees growing along contaminated tributaries in other operable units, and from field hydroponic experiments using hybrid poplar cuttings, analysis of tree-core samples appears to be able to detect subsurface PCE contamination in soils at levels of several hundred micrograms per liter or less and PCE concentrations in the range of 8 to 30 mg/L in ground water in direct contact with the roots. Loss of PCE from tree trunks by diffusion resulted in an exponential decrease in PCE concentrations with increasing height above the land surface in most trees. The rate of loss also appeared to be a function of the size and growth characteristics of the tree as some trees exhibited a linear loss with increasing height. Diffusional loss of PCE in small (0.5-inch diameter) trees was observed to occur at a rate more than 10 times larger than in trees 6.5 inches in diameter. Concentrations of PCE also exhibited directional variability around the tree trunks and concentration differe

Incidence of tissue coring during transseptal catheterization when using electrocautery and a standard transseptal needle.

PubMed

Greenstein, Eugene; Passman, Rod; Lin, Albert C; Knight, Bradley P

2012-04-01

The application of radiofrequency electrocautery to a standard, open-ended transseptal needle has been used to facilitate transseptal puncture (TSP). The purpose of this study was to determine the incidence of cardiac tissue coring when this technique is used. A model using excised swine hearts submerged in a saline-filled basin was developed to simulate TSP with electrocautery and a standard transseptal needle. Punctures were performed without the use of electrocautery and by delivering radiofrequency energy to the transseptal needle using a standard electrocautery pen at 3 target sites (fossa ovalis, non-fossa ovalis septum, and aorta). The tissue of the submerged heart was gently tented, and the needle was advanced on delivery of radiofrequency. The devices were retracted, and the needle was flushed in a collection basin. None of the TSPs without cautery caused tissue coring. For TSPs using electrocautery, the frequency of coring was at least 21% for any puncture permutation used in the study and averaged 37% at septal sites (P<0.001 compared with punctures without cautery). Tissue coring occurred in 33 of 96 (35%) punctures through the fossa ovalis and in 38 of 96 (40%) punctures through non-fossa ovalis septum. The frequency of tissue coring at aortic sites was 62 of 96 (65%), which was significantly higher than at the septal sites (P<0.001). In an animal preparation, TSP at the level of the fossa ovalis using electrocautery and a standard open-ended Brockenbrough needle resulted in coring of the septal tissue in 35% of cases (33 of 96 punctures).

The Effect of Ethylene and Propylene Pulses on Respiration, Ripening Advancement, Ethylene-Forming Enzyme, and 1-Aminocyclopropane-1-carboxylic Acid Synthase Activity in Avocado Fruit 12

PubMed Central

Starrett, David A.; Laties, George G.

1991-01-01

When early-season avocado fruit (Persea americana Mill. cv Hass) were treated with ethylene or propylene for 24 hours immediately on picking, the time to the onset of the respiratory climacteric, i.e. the lag period, remained unchanged compared with that in untreated fruit. When fruit were pulsed 24 hours after picking, on the other hand, the lag period was shortened. In both cases, however, a 24 hour ethylene or propylene pulse induced a transient increase in respiration, called the pulse-peak, unaccompanied by ethylene production (IL Eaks [1980] Am Soc Hortic Sci 105: 744-747). The pulse also caused a sharp rise in ethylene-forming enzyme activity in both cases, without any increase in the low level of 1-aminocyclopropane-1-carboxylic acid synthase activity. Thus, the shortening of the lag period by an ethylene pulse is not due to an effect of ethylene on either of the two key enzymes in ethylene biosynthesis. A comparison of two-dimensional polyacrylamide gel electrophoresis polypeptide profiles of in vitro translation products of poly(A+) mRNA from control and ethylene-pulsed fruit showed both up- and down-regulation in response to ethylene pulsing of a number of genes expressed during the ripening syndrome. It is proposed that the pulse-peak or its underlying events reflect an intrinsic element in the ripening process that in late-season or continuously ethylene-treated fruit may be subsumed in the overall climacteric response. A computerized system that allows continuous readout of multiple samples has established that the continued presentation of exogeneous ethylene or propylene to preclimacteric fruit elicits a dual respiration response comprising the merged pulse-peak and climacteric peak in series. The sequential removal of cores from a single fruit has proven an unsatisfactory sampling procedure inasmuch as coring induces wound ethylene, evokes a positive respiration response, and advances ripening. PMID:16668073

RNA-Mediated Gene Duplication and Retroposons: Retrogenes, LINEs, SINEs, and Sequence Specificity

PubMed Central

2013-01-01

A substantial number of “retrogenes” that are derived from the mRNA of various intron-containing genes have been reported. A class of mammalian retroposons, long interspersed element-1 (LINE1, L1), has been shown to be involved in the reverse transcription of retrogenes (or processed pseudogenes) and non-autonomous short interspersed elements (SINEs). The 3′-end sequences of various SINEs originated from a corresponding LINE. As the 3′-untranslated regions of several LINEs are essential for retroposition, these LINEs presumably require “stringent” recognition of the 3′-end sequence of the RNA template. However, the 3′-ends of mammalian L1s do not exhibit any similarity to SINEs, except for the presence of 3′-poly(A) repeats. Since the 3′-poly(A) repeats of L1 and Alu SINE are critical for their retroposition, L1 probably recognizes the poly(A) repeats, thereby mobilizing not only Alu SINE but also cytosolic mRNA. Many flowering plants only harbor L1-clade LINEs and a significant number of SINEs with poly(A) repeats, but no homology to the LINEs. Moreover, processed pseudogenes have also been found in flowering plants. I propose that the ancestral L1-clade LINE in the common ancestor of green plants may have recognized a specific RNA template, with stringent recognition then becoming relaxed during the course of plant evolution. PMID:23984183

3' rapid amplification of cDNA ends (RACE) walking for rapid structural analysis of large transcripts.

PubMed

Ozawa, Tatsuhiko; Kondo, Masato; Isobe, Masaharu

2004-01-01

The 3' rapid amplification of cDNA ends (3' RACE) is widely used to isolate the cDNA of unknown 3' flanking sequences. However, the conventional 3' RACE often fails to amplify cDNA from a large transcript if there is a long distance between the 5' gene-specific primer and poly(A) stretch, since the conventional 3' RACE utilizes 3' oligo-dT-containing primer complementary to the poly(A) tail of mRNA at the first strand cDNA synthesis. To overcome this problem, we have developed an improved 3' RACE method suitable for the isolation of cDNA derived from very large transcripts. By using the oligonucleotide-containing random 9mer together with the GC-rich sequence for the suppression PCR technology at the first strand of cDNA synthesis, we have been able to amplify the cDNA from a very large transcript, such as the microtubule-actin crosslinking factor 1 (MACF1) gene, which codes a transcript of 20 kb in size. When there is no splicing variant, our highly specific amplification allows us to perform the direct sequencing of 3' RACE products without requiring cloning in bacterial hosts. Thus, this stepwise 3' RACE walking will help rapid characterization of the 3' structure of a gene, even when it encodes a very large transcript.

Stimulation of translation by human Unr requires cold shock domains 2 and 4, and correlates with poly(A) binding protein interaction.

PubMed

Ray, Swagat; Anderson, Emma C

2016-03-03

The RNA binding protein Unr, which contains five cold shock domains, has several specific roles in post-transcriptional control of gene expression. It can act as an activator or inhibitor of translation initiation, promote mRNA turnover, or stabilise mRNA. Its role depends on the mRNA and other proteins to which it binds, which includes cytoplasmic poly(A) binding protein 1 (PABP1). Since PABP1 binds to all polyadenylated mRNAs, and is involved in translation initiation by interaction with eukaryotic translation initiation factor 4G (eIF4G), we investigated whether Unr has a general role in translational control. We found that Unr strongly stimulates translation in vitro, and mutation of cold shock domains 2 or 4 inhibited its translation activity. The ability of Unr and its mutants to stimulate translation correlated with its ability to bind RNA, and to interact with PABP1. We found that Unr stimulated the binding of PABP1 to mRNA, and that Unr was required for the stable interaction of PABP1 and eIF4G in cells. siRNA-mediated knockdown of Unr reduced the overall level of cellular translation in cells, as well as that of cap-dependent and IRES-dependent reporters. These data describe a novel role for Unr in regulating cellular gene expression.

Cloning and characterization of a type 3 iodothyronine deiodinase (D3) in the liver of the chondrichtyan Chiloscyllium punctatum.

PubMed

Martínez, Lidia Mayorga; Orozco, Aurea; Villalobos, Patricia; Valverde-R, Carlos

2008-05-01

Thyroid hormone bioactivity is finely regulated at the cellular level by the peripheral iodothyronine deiodinases (D). The study of thyroid function in fish has been restricted mainly to teleosts, whereas the study and characterization of Ds have been overlooked in chondrichthyes. Here we report the cloning and operational characterization of both the native and the recombinant hepatic type 3 iodothyronine deiodinase in the tropical shark Chiloscyllium punctatum. Native and recombinant sD3 show identical catalytic activities: a strong preference for T3-inner-ring deiodination, a requirement for a high concentration of DTT, a sequential reaction mechanism, and resistance to PTU inhibition. The cloned cDNA contains 1298 nucleotides [excluding the poly(A) tail] and encodes a predicted protein of 259 amino acids. The triplet TGA coding for selenocysteine (Sec) is at position 123. The consensus selenocysteine insertion sequence (SECIS) was identified 228 bp upstream of the poly(A) tail and corresponds to form 2. The deduced amino acid sequence was 77% and 72% identical to other D3 cDNAs in fishes and other vertebrates, respectively. As in the case of other piscivore teleost species, shark expresses hepatic D3 through adulthood. This characteristic may be associated with the alimentary strategy in which the protection from an exogenous overload of thyroid hormones could be of physiological importance for thyroidal homeostasis.

Control of storage-protein synthesis during seed development in pea (Pisum sativum L.).

PubMed Central

Gatehouse, J A; Evans, I M; Bown, D; Croy, R R; Boulter, D

1982-01-01

The tissue-specific syntheses of seed storage proteins in the cotyledons of developing pea (Pisum sativum L.) seeds have been demonstrated by estimates of their qualitative and quantitative accumulation by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and rocket immunoelectrophoresis respectively. Vicilin-fraction proteins initially accumulated faster than legumin, but whereas legumin was accumulated throughout development, different components of the vicilin fraction had their predominant periods of synthesis at different stages of development. The translation products in vitro of polysomes isolated from cotyledons at different stages of development reflected the synthesis in vivo of storage-protein polypeptides at corresponding times. The levels of storage-protein mRNA species during development were estimated by 'Northern' hybridization using cloned complementary-DNA probes. This technique showed that the levels of legumin and vicilin (47000-Mr precursors) mRNA species increased and decreased in agreement with estimated rates of synthesis of the respective polypeptides. The relative amounts of these messages, estimated by kinetic hybridization were also consistent. Legumin mRNA was present in leaf poly(A)+ RNA at less than one-thousandth of the level in cotyledon poly(A)+ (polyadenylated) RNA, demonstrating tissue-specific expression. Evidence is presented that storage-protein mRNA species are relatively long-lived, and it is suggested that storage-protein synthesis is regulated primarily at the transcriptional level. Images Fig. 2. Fig. 3. PMID:6897609

Expression of small cytoplasmic transcripts of the rat identifier element in vivo and in cultured cells.

PubMed Central

McKinnon, R D; Danielson, P; Brow, M A; Bloom, F E; Sutcliffe, J G

1987-01-01

We examined the level of expression of small RNA transcripts hybridizing to a rodent repetitive DNA element, the identifier (ID) sequence, in a variety of cell types in vivo and in cultured mammalian cells. A 160-nucleotide (160n) cytoplasmic poly(A)+ RNA (BC1) appeared in late embryonic and early postnatal rat brain development, was enriched in the cerebral cortex, and appeared to be restricted to neural tissue and the anterior pituitary gland. A 110n RNA (BC2) was specifically enriched in brain, especially the postnatal cortex, but was detectable at low levels in peripheral tissues. A third, related 75n poly(A)- RNA (T3) was found in rat brain and at lower levels in peripheral tissues but was very abundant in the testes. The BC RNAs were found in a variety of rat cell lines, and their level of expression was dependent upon cell culture conditions. A rat ID probe detected BC-like RNAs in mouse brain but not liver and detected a 200n RNA in monkey brain but not liver at lower hybridization stringencies. These RNAs were expressed by mouse and primate cell lines. Thus, tissue-specific expression of small ID-sequence-related transcripts is conserved among mammals, but the tight regulation found in vivo is lost by cells in culture. Images PMID:2439903

Highly abundant and stage-specific mRNAs in the obligate pathogen Bremia lactucae.

PubMed

Judelson, H S; Michelmore, R W

1990-01-01

Germinating spores of the obligate pathogen Bremia lactucae (lettuce downy mildew) contain several unusually abundant species of mRNA. Thirty-nine cDNA clones corresponding to prevalent transcripts were isolated from a library synthesized using poly(A)+ RNA from germinating spores; these clones represented only five distinct classes. Each corresponding mRNA accounted for from 0.4 to 9 percent by mass of poly(A)+ RNA from germinating spores and together represented greater than 20 percent of the mRNA. The expression of the corresponding genes, and a gene encoding Hsp70, was analyzed in spores during germination and during growth in planta. The Hsp70 mRNA and mRNA from one abundant cDNA clone (ham34) were expressed constitutively. Two clones (ham9 and ham12) hybridized only to mRNA from spores and germinating spores. Two clones (ham37 and ham27) showed hybridization specific to germinating spores. Quantification of the number of genes homologous to each cDNA clone indicated that four clones corresponded to one or two copies per haploid genome, and one hybridized to an approximately 11-member family of genes. A sequence of the gene corresponding to ham34 was obtained to investigate its function and to identify sequences conferring high levels of gene expression for use in constructing vectors for the transformation of B. lactucae.

The Yucca Mountain Project prototype air-coring test, U12g tunnel, Nevada test site

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ray, J.M.; Newsom, J.C.

1994-12-01

The Prototype Air-Coring Test was conducted at the Nevada Test Site (NTS) G-Tunnel facility to evaluate standard coring techniques, modified slightly for air circulation, for use in testing at a prospective nuclear waste repository at Yucca Mountain, Nevada. Air-coring technology allows sampling of subsurface lithology with minimal perturbation to ambient characteristic such as that required for exploratory holes near aquifers, environmental applications, and site characterization work. Two horizontal holes were cored, one 50 ft long and the other 150 ft long, in densely welded fractured tuff to simulate the difficult drilling conditions anticipated at Yucca Mountain. Drilling data from sevenmore » holes on three other prototype tests in nonwelded tuff were also collected for comparison. The test was used to establish preliminary standards of performance for drilling and dust collection equipment and to assess procedural efficiencies. The Longyear-38 drill achieved 97% recovery for HQ-size core (-2.5 in.), and the Atlas Copco dust collector (DCT-90) captured 1500 lb of fugitive dust in a mine environment with only minor modifications. Average hole production rates were 6-8 ft per 6-h shift in welded tuff and almost 20 ft per shift on deeper holes in nonwelded tuff. Lexan liners were successfully used to encapsulate core samples during the coring process and protect core properties effectively. The Prototype Air-Coring Test demonstrated that horizontal air coring in fractured welded tuff (to at least 150 ft) can be safely accomplished by proper selection, integration, and minor modification of standard drilling equipment, using appropriate procedures and engineering controls. The test also indicated that rig logistics, equipment, and methods need improvement before attempting a large-scale dry drilling program at Yucca Mountain.« less

Heterogeneity of PTEN and ERG expression in prostate cancer on core needle biopsies: implications for cancer risk stratification and biomarker sampling.

PubMed

Shah, Rajal B; Bentley, James; Jeffery, Zach; DeMarzo, Angelo M

2015-05-01

ERG and PTEN biomarkers are increasingly being analyzed on prostate core biopsies (NBXs); ERG as a marker of clonality and number of separately arising tumor foci and PTEN for prognostic information. Yet, in patients with multiple biopsy cores positive for cancer (PCa), there is no standardized approach for interrogation of these biomarkers in terms of the number of positive cores to evaluate. A total of 194 NBX cases containing more than one positive core with cancer were evaluated for ERG overexpression and PTEN loss by immunostaining (immunohistochemistry) of all positive cores. ERG overexpression or PTEN loss in at least one cancer core was present in 111 (57%) and 69 (36%) cases respectively. ERG overexpression was significantly associated with PTEN loss (P < .0001), and PTEN loss was associated with a high Gleason score (P < .0001). Inter- and intra-tumor core staining heterogeneity for ERG overexpression occurred in 42% and 5% cases and for PTEN loss both intra- and inter-tumor core heterogeneity was 68%. PTEN staining was highly discordant between PCa sites regardless of laterality. When the Gleason score was non-uniform across PCa sites, the combination of cores showing the highest Gleason score and largest tumor volume provided the best representation of ERG overexpression (92%) and PTEN loss (98%). When grades were uniform across cancer sites, the highest tumor volume core was generally representative of ERG overexpression (90%) but was less representative for PTEN loss (76%). Our results suggest that knowledge of this heterogeneity is critical for developing optimal yet cost-effective strategies to identify these underlying molecular abnormalities. Copyright © 2015 Elsevier Inc. All rights reserved.

Primary structure of prostaglandin G/H synthase from sheep vesicular gland determined from the complementary DNA sequence.

PubMed Central

DeWitt, D L; Smith, W L

1988-01-01

Prostaglandin G/H synthase (8,11,14-icosatrienoate, hydrogen-donor:oxygen oxidoreductase, EC 1.14.99.1) catalyzes the first step in the formation of prostaglandins and thromboxanes, the conversion of arachidonic acid to prostaglandin endoperoxides G and H. This enzyme is the site of action of nonsteroidal anti-inflammatory drugs. We have isolated a 2.7-kilobase complementary DNA (cDNA) encompassing the entire coding region of prostaglandin G/H synthase from sheep vesicular glands. This cDNA, cloned from a lambda gt 10 library prepared from poly(A)+ RNA of vesicular glands, hybridizes with a single 2.75-kilobase mRNA species. The cDNA clone was selected using oligonucleotide probes modeled from amino acid sequences of tryptic peptides prepared from the purified enzyme. The full-length cDNA encodes a protein of 600 amino acids, including a signal sequence of 24 amino acids. Identification of the cDNA as coding for prostaglandin G/H synthase is based on comparison of amino acid sequences of seven peptides comprising 103 amino acids with the amino acid sequence deduced from the nucleotide sequence of the cDNA. The molecular weight of the unglycosylated enzyme lacking the signal peptide is 65,621. The synthase is a glycoprotein, and there are three potential sites for N-glycosylation, two of them in the amino-terminal half of the molecule. The serine reported to be acetylated by aspirin is at position 530, near the carboxyl terminus. There is no significant similarity between the sequence of the synthase and that of any other protein in amino acid or nucleotide sequence libraries, and a heme binding site(s) is not apparent from the amino acid sequence. The availability of a full-length cDNA clone coding for prostaglandin G/H synthase should facilitate studies of the regulation of expression of this enzyme and the structural features important for catalysis and for interaction with anti-inflammatory drugs. Images PMID:3125548

Crystal structure of SgcJ, an NTF2-like superfamily protein involved in biosynthesis of the nine-membered enediyne antitumor antibiotic C-1027

DOE PAGES

Huang, Tingting; Chang, Chin -Yuan; Lohman, Jeremy R.; ...

2016-10-01

Comparative analysis of the enediyne biosynthetic gene clusters revealed sets of conserved genes serving as outstanding candidates for the enediyne core. Here we report the crystal structures of SgcJ and its homologue NCS-Orf16, together with gene inactivation and site-directed mutagenesis studies, to gain insight into enediyne core biosynthesis. Gene inactivation in vivo establishes that SgcJ is required for C-1027 production in Streptomyces globisporus. SgcJ and NCS-Orf16 share a common structure with the nuclear transport factor 2-like superfamily of proteins, featuring a putative substrate binding or catalytic active site. Site-directed mutagenesis of the conserved residues lining this site allowed us tomore » propose that SgcJ and its homologues may play a catalytic role in transforming the linear polyene intermediate, along with other enediyne polyketide synthase-associated enzymes, into an enzyme-sequestered enediyne core intermediate. In conclusion, these findings will help formulate hypotheses and design experiments to ascertain the function of SgcJ and its homologues in nine-membered enediyne core biosynthesis.« less

Detailed glaciochemical investigations in southern Victoria Land - a proxy climatic record

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mayewski, P.A.

1987-09-01

Local accumulation-basins in the Transantarctic Mountains possess sites suitable for recovering ice-core records that are valuable for studying climate change. These sites are also unique, because they are close to the sites of other ice-core studies and to areas with established terrestrial records. The objective is to study a snowpit in detail and collect ice cores in southern Victoria Land; this work will be similar to the investigations that the authors has conducted in south Greenland and in the Dominion Range near the Beardmore Glacier. The proposed sites are in Convoy, Asgard, and Royal Society ranges. The authors will selectmore » one site at which he will recover two ice cores, each approximately 200 to 300 meters in depth. Samples will be analyzed for major anions (chloride, sulfate, nitrate, fluoride) and cations (sodium, potassium, magnesium, ammonium, silicate), total acidity, conductivity, density, and core stratigraphy with dating provided by cross-calibration of all of the preceding plus total beta-activity, lead-210, oxygen isotopes, and microparticles. This investigation will yield a detailed record of several thousand years of glacial history, climate change, and volcanic activity for southern Victoria Land. This record will be compared to existing terrestrial records to add necessary detail and to other global ice-core records to assess global climatic change. It will also help to document volcanic activity for Mount Erebus as well as other volcanos in the Southern Hemisphere and possibly some in the Northern Hemisphere. With this record, the author will be able to evaluate the influence of volcanic and solar activity on climate as well as add greatly to the understanding of the chemistry of the global atmosphere.« less

Core-Shell Processing of Natural Pigment: Upper Palaeolithic Red Ochre from Lovas, Hungary

PubMed Central

Sajó, István E.; Kovács, János; Fitzsimmons, Kathryn E.; Jáger, Viktor; Lengyel, György; Viola, Bence; Talamo, Sahra; Hublin, Jean-Jacques

2015-01-01

Ochre is the common archaeological term for prehistoric pigments. It is applied to a range of uses, from ritual burials to cave art to medications. While a substantial number of Palaeolithic paint mining pits have been identified across Europe, the link between ochre use and provenance, and their antiquity, has never yet been identified. Here we characterise the mineralogical signature of core-shell processed ochre from the Palaeolithic paint mining pits near Lovas in Hungary, using a novel integration of petrographic and mineralogical techniques. We present the first evidence for core-shell processed, natural pigment that was prepared by prehistoric people from hematitic red ochre. This involved combining the darker red outer shell with the less intensely coloured core to efficiently produce an economical, yet still strongly coloured, paint. We demonstrate the antiquity of the site as having operated between 14–13 kcal BP, during the Epigravettian period. This is based on new radiocarbon dating of bone artefacts associated with the quarry site. The dating results indicate the site to be the oldest known evidence for core-shell pigment processing. We show that the ochre mined at Lovas was exported from the site based on its characteristic signature at other archaeological sites in the region. Our discovery not only provides a methodological framework for future characterisation of ochre pigments, but also provides the earliest known evidence for “value-adding” of products for trade. PMID:26147808

Nuclear factor Y regulates ancient budgerigar hepadnavirus core promoter activity

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shen, Zhongliang; Liu, Yanfeng; Luo, Mengjun

Endogenous viral elements (EVE) in animal genomes are the fossil records of ancient viruses and provide invaluable information on the origin and evolution of extant viruses. Extant hepadnaviruses include avihepadnaviruses of birds and orthohepadnaviruses of mammals. The core promoter (Cp) of hepadnaviruses is vital for viral gene expression and replication. We previously identified in the budgerigar genome two EVEs that contain the full-length genome of an ancient budgerigar hepadnavirus (eBHBV1 and eBHBV2). Here, we found eBHBV1 Cp and eBHBV2 Cp were active in several human and chicken cell lines. A region from nt −85 to −11 in eBHBV1 Cp was critical formore » the promoter activity. Bioinformatic analysis revealed a putative binding site of nuclear factor Y (NF-Y), a ubiquitous transcription factor, at nt −64 to −50 in eBHBV1 Cp. The NF-Y core binding site (ATTGG, nt −58 to −54) was essential for eBHBV1 Cp activity. The same results were obtained with eBHBV2 Cp and duck hepatitis B virus Cp. The subunit A of NF-Y (NF-YA) was recruited via the NF-Y core binding site to eBHBV1 Cp and upregulated the promoter activity. Finally, the NF-Y core binding site is conserved in the Cps of all the extant avihepadnaviruses but not of orthohepadnaviruses. Interestingly, a putative and functionally important NF-Y core binding site is located at nt −21 to −17 in the Cp of human hepatitis B virus. In conclusion, our findings have pinpointed an evolutionary conserved and functionally critical NF-Y binding element in the Cps of avihepadnaviruses. - Highlights: • Endogenous budgerigar hepadnavirus (eBHBV) core promoters (Cps) are active in cells. • NF-Y binding site exists in the Cps of eBHBVs and all the extant avihepadnaviruses. • NF-Y binding and mediated upregulation is critical for eBHBV Cp activity.« less

Methodological issues and preliminary results from a combined sediment fingerprinting and radioisotope dating approach to explore changes in sediment sources with land-use change in the Brantian Catchment, Borneo.

NASA Astrophysics Data System (ADS)

Walsh, Rory; Higton, Sam; Marshall, Jake; Bidin, Kawi; Blake, William; Nainar, Anand

2015-04-01

This paper reports some methodological issues and early results of a project investigating the erosional impacts of land use changes (multiple selective logging and progressive, partial conversion to oil palm) over the last 25-40 years in the 600km2 Brantian river catchment in Sabah, Borneo. A combined sediment fingerprinting and radioisotope dating approach is being applied to sediment cores taken in stream hierarchical fashion across the intermediate catchment scale. Changes in sediment sources and sedimentation rates over time can be captured by changes in the relative importance of geochemical elements with depth in downstream sediment cores, which in turn can be linked to parallel changes in upstream cores by the application of unmixing models and statistical techniques. Radioisotope analysis of the sediment cores allows these changes to be dated and sedimentation rates to be estimated. Work in the neighbouring Segama catchment had successfully demonstrated the potential of such an approach in a rainforest environment (Walsh et al. 2011). The paper first describes steps taken to address methodological issues. The approach relies on taking continuous sediment cores which have aggraded progressively over time and remain relatively undisturbed and uncontaminated. This issue has been tackled (1) through careful core sampling site selection with a focus on lateral bench sites and (2) deployment of techniques such as repeat-measurement erosion bridge transects to assess the contemporary nature of sedimentation to validate (or reject) candidate sites. The issue of sediment storage and uncertainties over lag times has been minimised by focussing on sets of above- and below-confluence sites in the intermediate zone of the catchment, thus minimising sediment transit times between upstream contributing and downstream destination core sites. This focus on the intermediate zone was also driven by difficulties in finding suitable core sites in the mountainous headwaters area due to the prevalence of steep, incised channels without even narrow floodplains. Preliminary results are reported from (1) a field visit to investigate potential sampling sites in July 2014 and (2) initial analysis of a sediment core at a promising lateral bench site. Marked down-profile geochemistry changes of the core indicate a history of phases of high deposition and lateral growth of the channel caused by mobilisation of sediment linked to logging and clearance upstream. Recent channel bed degradation suggests the system has been adjusting a decline in sediment supply with forest recovery since logging in 2005, but a renewed sedimentation phase heralded by > 10 cm deposition at the site in a flood in July 2014 appears to have started linked to partial forest clearance for oil palm. These preliminary results support the ability of a combined fingerprinting and dating approach to reflect the spatial history of land-use change in a catchment undergoing disturbance. Walsh R. P. D. , Bidin K., Blake W.H., Chappell N.A., Clarke M.A., Douglas I., Ghazali R., Sayer A.M., Suhaimi J., Tych W. & Annammala K.V. (2011) Long-term responses of rainforest erosional systems at different spatial scales to selective logging and climatic change. Philosophical Transactions of the Royal Society B, 366, 3340-3353.

Real-Time and Delayed Analysis of Tree and Shrub Cores as Indicators of Subsurface Volatile Organic Compound Contamination, Durham Meadows Superfund Site, Durham, Connecticut, August 29, 2006

USGS Publications Warehouse

Vroblesky, Don A.; Willey, Richard E.; Clifford, Scott; Murphy, James J.

2008-01-01

This study examined volatile organic compound concentrations in cores from trees and shrubs for use as indicators of vadose-zone contamination or potential vapor intrusion by volatile organic compounds into buildings at the Durham Meadows Superfund Site, Durham, Connecticut. The study used both (1) real-time tree- and shrub-core analysis, which involved field heating the core samples for 5 to 10 minutes prior to field analysis, and (2) delayed analysis, which involved allowing the gases in the cores to equilibrate with the headspace gas in the sample vials unheated for 1 to 2 days prior to analysis. General correspondence was found between the two approaches, indicating that preheating and field analysis of vegetation cores is a viable approach to real-time monitoring of subsurface volatile organic compounds. In most cases, volatile organic compounds in cores from trees and shrubs at the Merriam Manufacturing Company property showed a general correspondence to the distribution of volatile organic compounds detected in a soil-gas survey, despite the fact that most of the soil-gas survey data in close proximity to the relevant trees were collected about 3 years prior to the tree-core collection. Most of the trees cored at the Durham Meadows Superfund Site, outside of the Merriam Manufacturing Company property, contained no volatile organic compounds and were in areas where indoor air sampling and soil-gas sampling showed little or no volatile organic compound concentrations. An exception was tree DM11, which contained barely detectable concentrations of trichloroethene near a house where previous investigations found low concentrations of trichloroethene (0.13 to 1.2 parts per billion by volume) in indoor air and 7.7 micrograms per liter of trichloroethene in the ground water. The barely detectable concentration of trichloroethene in tree DM11 and the lack of volatile organic compound detection in nearby tree DM10 (adjacent to the well having 7.7 micrograms of trichloroethene) may be attributable to the relatively large depth to water (17.6 feet), the relatively low soil-vapor trichloroethene concentration, and the large amount of rainfall during and preceding the tree-coring event. The data indicate that real-time and delayed analyses of tree cores are viable approaches to examining subsurface volatile organic compound soil-gas or vadose-zone contamination at the Durham Meadows Superfund Site and other similar sites. Thus, the methods may have application for determining the potential for vapor intrusion into buildings.

CASPASE-9 CARD:CORE DOMAIN INTERACTIONS REQUIRE A PROPERLY-FORMED ACTIVE SITE

PubMed Central

Huber, Kristen L.; Serrano, Banyuhay P.; Hardy, Jeanne A.

2018-01-01

Caspase-9 is a critical factor in the initiation of apoptosis, and as a result is tightly regulated by a number of mechanisms. Caspase-9 contains a Caspase Activation and Recruitment Domain (CARD), which enables caspase-9 to form a tight interaction with the apoptosome, a heptameric activating platform. The caspase-9 CARD has been thought to be principally involved in recruitment to the apoptosome, but its roles outside this interaction have yet to be uncovered. In this work we show that the CARD is involved in physical interactions with the catalytic core of caspase-9 in the absence of the apoptosome; this interaction requires a properly formed caspase-9 active site. The active sites of caspases are composed of four extremely mobile loops. When the active-site loops are not properly ordered, the CARD and core domains of caspase-9 do not interact and behave independently, like loosely tethered beads. When the active-site loop bundle is properly ordered, the CARD domain interacts with the catalytic core, forming a single folding unit. Together these findings provide mechanistic insight into a new level of caspase-9 regulation, prompting speculation that the CARD may also play a role in the recruitment or recognition of substrate. PMID:29500231

Method for detecting core malware sites related to biomedical information systems.

PubMed

Kim, Dohoon; Choi, Donghee; Jin, Jonghyun

2015-01-01

Most advanced persistent threat attacks target web users through malicious code within landing (exploit) or distribution sites. There is an urgent need to block the affected websites. Attacks on biomedical information systems are no exception to this issue. In this paper, we present a method for locating malicious websites that attempt to attack biomedical information systems. Our approach uses malicious code crawling to rearrange websites in the order of their risk index by analyzing the centrality between malware sites and proactively eliminates the root of these sites by finding the core-hub node, thereby reducing unnecessary security policies. In particular, we dynamically estimate the risk index of the affected websites by analyzing various centrality measures and converting them into a single quantified vector. On average, the proactive elimination of core malicious websites results in an average improvement in zero-day attack detection of more than 20%.

Method for Detecting Core Malware Sites Related to Biomedical Information Systems

PubMed Central

Kim, Dohoon; Choi, Donghee; Jin, Jonghyun

2015-01-01

Most advanced persistent threat attacks target web users through malicious code within landing (exploit) or distribution sites. There is an urgent need to block the affected websites. Attacks on biomedical information systems are no exception to this issue. In this paper, we present a method for locating malicious websites that attempt to attack biomedical information systems. Our approach uses malicious code crawling to rearrange websites in the order of their risk index by analyzing the centrality between malware sites and proactively eliminates the root of these sites by finding the core-hub node, thereby reducing unnecessary security policies. In particular, we dynamically estimate the risk index of the affected websites by analyzing various centrality measures and converting them into a single quantified vector. On average, the proactive elimination of core malicious websites results in an average improvement in zero-day attack detection of more than 20%. PMID:25821511

TEMPORAL TRENDS OF AROCLOR 1268 IN THE TAUNTON RIVER ESTUARY: EVIDENCE OF EARLY PRODUCTION, USE AND RELEASE TO THE ENVIRONMENT

EPA Science Inventory

Sediment cores were collected at three sites from the Lower Taunton River in Narragansett Bay, MA, dated and examined for PCBs. At each site, PCBs first appear in the sediment cores at depths corresponding to 1929 based on a combination of radiometric and anthropogenic dating pr...

PolyA Single Strand DNA Translocation Through an Alpha-Hemolysin Pore Stem

NASA Technical Reports Server (NTRS)

OKeeffe, James; Cozmuta, Ioana; Stolc, Viktor

2003-01-01

A new model for the polymer-pore interaction energy is introduced, based on an atomic-scale description of coulombic polymer-pore interaction. The enhanced drift velocity, experimentally observed for short polymers, is successfully accounted for, using this interaction energy model. For R/R(sub 0)>4 (R(sub 0)=7 angstroms) the translocation velocity approaches the free space drift velocity v(sub 0). This motivates the need to appropriately derivatize artificial nanopores, where R>R(sub 0).

Aerospace Medicine and Biology. A Continuing Bibliography with Indexes

DTIC Science & Technology

1987-12-01

GLYCERALDEHYDE AS A SOURCE OF SIMULATION ABOUT THE ORIGIN OF LIFE ENERGY AND MONOMERS FOR PREBIOTIC CONDENSATION URSULA NIESERT (Freiburg, Universitaet, Freiburg...primitive prebiotic replication system. The adsorption of Poly-C, Poly-U, Poly-A, Poly-G, and 5’-AMP, 5’-GMP, 5’-CMP and 5’-UMP onto gypsum was studied. It...Cambridge University Press. 1986. 378 p. For individual titles see A87-49035 to A87-49049. Topics discussed include prebiotic systems and evolutionary

Bradley Fighting Vehicle Gunnery: An Analysis of Engagement Strategies for the M242 25-mm Automatic Gun

DTIC Science & Technology

1993-03-01

source for this estimate of eight rounds per BMP target. According to analyst Donna Quirido, AMSAA does not provide or support any such estimate (30...engagement or in the case of. the Bradley, stabilization inaccuracies. According to Helgert: These errors give rise to aim-wander, a term that derives from...the same area. (6:14_5) The resulting approximation to the truncated normal integral has a maximum relative error of 0.0075. Using Polya -Williams, an

Historic CH4 Records from Antarctic and Greenland Ice Cores, Antarctic Firn Data, and Archived Air Samples from Cape Grim, Tasmania

DOE Data Explorer

Etheridge, D. M. [Division of Atmospheric Research, CSIRO, Aspendale, Victoria, Australia; Steele, L. P. [Division of Atmospheric Research, CSIRO, Aspendale, Victoria, Australia; Francey, R. J. [Division of Atmospheric Research, CSIRO, Aspendale, Victoria, Australia; Langenfelds, R. L. [Division of Atmospheric Research, CSIRO, Aspendale, Victoria, Australia

2002-01-01

The Antarctic CH4 records presented here are derived from three ice cores obtained at Law Dome, East Antarctica (66°44'S, 112°50'E, 1390 meters above mean sea level). Law Dome has many qualities of an ideal ice core site for the reconstruction of past concentrations of atmospheric gases; these qualities include: negligible melting of the ice sheet surface, low concentrations of impurities, regular stratigraphic layering undisturbed by wind stress at the surface or differential ice flow at depth, and a high snow accumulation rate. Further details on the site, drilling, and cores are provided by Etheridge et al. (1998), Etheridge et al. (1996), Etheridge and Wookey (1989), and Morgan et al. (1997). The two Greenland ice cores are from the Summit region (72°34' N, 37°37' W, 3200 meters above mean sea level). Lower snow accumulation rate there results in lower air-age resolution, and measurements presented here cover only the pre-industrial period (until 1885). More details about these measurements are presented in Etheridge et al. (1998). Additionally, this site contains firn data from Core DE08-2, and archived air samples from Cape Grim, Tasmania, for comparison.

Ground water chlorinated ethenes in tree trunks: Case studies, influence of recharge, and potential degradation mechanism

USGS Publications Warehouse

Vroblesky, D.A.; Clinton, B.D.; Vose, J.M.; Casey, C.C.; Harvey, G.J.; Bradley, P.M.

2004-01-01

Trichloroethene (TCE) was detected in cores of trees growing above TCE-contaminated ground at three sites: the Carswell Golf Course in Texas, Air Force Plant PJKS in Colorado, and Naval Weapons Station Charleston in South Carolina. This was true even when the depth to water was 7.9 m or when the contaminated aquifer was confined beneath ???3 m of clay. Additional ground water contaminants detected in the tree cores were cis-1,2-dichloroethene at two sites and tetrachloroethene at one site. Thus, tree coring can be a rapid and effective means of locating shallow subsurface chlorinated ethenes and possibly identifying zones of active TCE dechlorination. Tree cores collected over time were useful in identifying the onset of ground water contamination. Several factors affecting chlorinated ethene concentrations in tree cores were identified in this investigation. The factors include ground water chlorinated ethene concentrations and depth to ground water contamination. In addition, differing TCE concentrations around the trunk of some trees appear to be related to the roots deriving water from differing areas. Opportunistic uptake of infiltrating rainfall can dilute prerain TCE concentrations in the trunk. TCE concentrations in core headspace may differ among some tree species. In some trees, infestation of bacteria in decaying heartwood may provide a TCE dechlorination mechanism within the trunk.

Test of tree core sampling for screening of toxic elements in soils from a Norwegian site.

PubMed

Algreen, Mette; Rein, Arno; Legind, Charlotte N; Amundsen, Carl Einar; Karlson, Ulrich Gosewinkel; Trapp, Stefan

2012-04-01

Tree core samples have been used to delineate organic subsurface plumes. In 2009 and 2010, samples were taken at trees growing on a former dump site in Norway and analyzed for arsenic (As), cadmium (Cd), chromium (Cr), copper (Cu), nickel (Ni), and zinc (Zn). Concentrations in wood were in averages (dw) 30 mg/kg for Zn, 2 mg/kg for Cu, and < 1 mg/kg for Cd, Cr, As and Ni. The concentrations in wood samples from the polluted test site were compared to those derived from a reference site. For all except one case, mean concentrations from the test site were higher than those from the reference site, but the difference was small and not always significant. Differences between tree species were usually higher than differences between reference and test site. Furthermore, all these elements occur naturally, and Cu, Ni, and Zn are essential minerals. Thus, all trees will have a natural background of these elements, and the occurrence alone does not indicate soil pollution. For the interpretation of the results, a comparison to wood samples from an unpolluted reference site with same species and similar soil conditions is required. This makes the tree core screening method less reliable for heavy metals than, e.g., for chlorinated solvents.

High-resolution gamma ray attenuation density measurements on mining exploration drill cores, including cut cores

NASA Astrophysics Data System (ADS)

Ross, P.-S.; Bourke, A.

2017-01-01

Physical property measurements are increasingly important in mining exploration. For density determinations on rocks, one method applicable on exploration drill cores relies on gamma ray attenuation. This non-destructive method is ideal because each measurement takes only 10 s, making it suitable for high-resolution logging. However calibration has been problematic. In this paper we present new empirical, site-specific correction equations for whole NQ and BQ cores. The corrections force back the gamma densities to the "true" values established by the immersion method. For the NQ core caliber, the density range extends to high values (massive pyrite, 5 g/cm3) and the correction is thought to be very robust. We also present additional empirical correction factors for cut cores which take into account the missing material. These "cut core correction factors", which are not site-specific, were established by making gamma density measurements on truncated aluminum cylinders of various residual thicknesses. Finally we show two examples of application for the Abitibi Greenstone Belt in Canada. The gamma ray attenuation measurement system is part of a multi-sensor core logger which also determines magnetic susceptibility, geochemistry and mineralogy on rock cores, and performs line-scan imaging.

a Mineralogical Analysis of Hspdp Core Samples from the Northern Awash Pliocene Hadar Formation, Ethiopia: the tale of AN East African Paleolake

NASA Astrophysics Data System (ADS)

Davis, D. M.; Deocampo, D.; Rabideaux, N. M.; Campisano, C. J.

2017-12-01

The Northern Awash Valley is located in the southwestern portion of the Afar Depression in Ethiopia. During the 2014 field season, two core sites were drilled as part of the Hominin Sites and Paleolakes Drilling Project (HSPDP), recovering a total of 600 m of sediment from both localities (NAO and NAW). Mineralogical analyses of the bulk sediments and clays from the Hadar Formation have helped to begin constructing a more complete picture of the paleoenvironmental conditions of the Northern Awash during the Pliocene. This work is an attempt to begin to answer the questions about salinity/alkalinity of Hadar Paleolake as well as its sulfur content. The two sites, NAW and NAO, are about 3 kilometers apart and presumably part of the same paleolake basin. The data has shown that this area was much more humid during the Pliocene than it is today and that most of the minerals in the paleolake basin are detrital, save for calcite, Mg-calcite, gypsum, pyrite, and a few zeolites. 060 analyses of the clays in the cores show that the lake was a freshwater lake three million years ago, up until its eventual evaporation. Interestingly, the lack of trioctahedral clays is an indication that the lake water did not become highly saline and alkaline during its evaporation. An interesting contrast between the two cores shows that NAO contains gypsum, but so far, no pyrite. NAW, on the other hand, contains both pyrite and gypsum. The pyrite in NAW is mostly at the bottom of the core, whereas, the gypsum is intermittent throughout the core and much more sparse toward the bottom. This line of evidence suggests that the two sites within the lake may have experienced different redox conditions. It may be that the NAO core was sampled in a shallower part of the lake whereas the NAW core sampled a deeper section.

Dechlorane plus, a chlorinated flame retardant, in the Great Lakes.

PubMed

Hoh, Eunha; Zhu, Lingyan; Hites, Ronald A

2006-02-15

A highly chlorinated flame retardant, Dechlorane Plus (DP), was detected and identified in ambient air, fish, and sediment samples from the Great Lakes region. The identity of this compound was confirmed by comparing its gas chromatographic retention times and mass spectra with those of authentic material. This compound exists as two gas chromatographically separable stereoisomers (syn and anti), the structures of which were characterized by one- and two-dimensional proton nuclear magnetic resonance. DP was detected in most air samples, even at remote sites. The atmospheric DP concentrations were higher at the eastern Great Lakes sites (Sturgeon Point, NY, and Cleveland, OH) than those at the western Great Lakes sites (Eagle Harbor, MI, Chicago, IL, and Sleeping Bear Dunes, MI). Atthe Sturgeon Point site, DP concentrations once reached 490 pg/m3. DP atmospheric concentrations were comparable to those of BDE-209 at the eastern Great Lakes sites. DP was also found in sediment cores from Lakes Michigan and Erie. The peak DP concentrations were comparable to BDE-209 concentrations in the sediment core from Lake Erie butwere about 30 times lower than BDE-209 concentrations in the core from Lake Michigan. In the sediment cores, the DP concentrations peaked around 1975-1980, and the surficial concentrations were 10-80% of peak concentrations. Higher DP concentrations in air samples from Sturgeon Point, NY, and in the sediment core from Lake Erie suggest that DP's manufacturing facility in Niagara Falls, NY, may be a source. DP was also detected in archived fish (walleye) from Lake Erie, suggesting that this compound is, at least partially, bioavailable.

Late Quaternary lake-level changes constrained by radiocarbon and stable isotope studies on sediment cores from Lake Titicaca, South America

NASA Astrophysics Data System (ADS)

Rowe, Harold D.; Guilderson, Thomas P.; Dunbar, Robert B.; Southon, John R.; Seltzer, Geoffrey O.; Mucciarone, David A.; Fritz, Sherilyn C.; Baker, Paul A.

2003-09-01

We present and compare AMS- 14C geochronologies for sediment cores recovered from Lake Titicaca, South America. Radiocarbon dates from three core sites constrain the timing of late Quaternary paleoenvironmental changes in the Central Andes and highlight the site-specific factors that limit the radiocarbon geochronometer. With the exception of mid-Holocene sediments, all cores are generally devoid of macrophyte fragments, thus bulk organic fractions are used to build core chronologies. Comparisons of radiocarbon results for chemically defined fractions (bulk decalcified, humate, humin) suggest that ages derived from all fractions are generally coherent in the post-13,500 yr BP time interval. In the pre-13,500 yr BP time interval, ages derived from humate extracts are significantly younger (300-7000 years) than ages from paired humin residues. Gross age incoherencies between paired humate and humin sub-fractions in pre-13,500 yr BP sediments from all core sites probably reflect the net downward migration of humates. Ages derived from bulk decalcified fractions at our shallow water (90 m) and deep water (230 m) core sites consistently fall between ages derived from humate and humin sub-fractions in the pre-13,500 yr BP interval, reflecting that the bulk decalcified fraction is predominantly a mixture of humate and humin sub-fractions. Bulk decalcified ages from the pre-13,500 yr BP interval at our intermediate depth core site (150 m) are consistently older than humate (youngest) and humin sub-fractions. This uniform, reproducible pattern can be explained by the mobilization of a relatively older organic sub-fraction during and after the re-acidification step following the alkaline treatment of the bulk sediment. The inferred existence of this 'alkali-mobile, acid-soluble' sub-fraction implies a different depositional/post-depositional history that is potentially associated with a difference in source material. While internally consistent geochronologies can be developed for the Lake Titicaca sequence using different organic fractions, mobile organic sub-fractions and fractions containing mobile sub-fractions should generally be avoided in geochronology studies. Consequently, we believe humin and/or bulk decalcified ages provide the most consistent chronologies for the post-13,500 yr BP interval, and humin ages provide the most representative ages for sedimentation prior to 13,500 yr BP interval. Using the age model derived from the deep water core site and a previously published isotope-based lake-level reconstruction, we present a qualitative record of lake level in the context of several ice-core records from the western hemisphere. We find the latest Pleistocene lake-level response to changing insolation began during or just prior to the Bølling/Allerød period. Using the isotope-based lake-level reconstruction, we also find the 85-m drop in lake level that occurred during the mid-Holocene was synchronous with an increase in the variability of ice-core δ18O from a nearby icecap, but was not reflected in any of the polar ice-core records recovered from the interior of Antarctica and Greenland.

Chemical gradients in sediment cores from an EPA reference site off the Farallon Islands - Assessing chemical indicators of dredged material disposal in the deep sea

USGS Publications Warehouse

Bothner, Michael H.; Gill, P.W.; Boothman, W.S.; Taylor, B.B.; Karl, Herman A.

1998-01-01

Heavy metal and organic contaminants have been determined in undisturbed sediment cores from the US Environmental Protection Agency reference site for dredged material on the continental slope off San Francisco. As expected, the concentrations are significantly lower than toxic effects guidelines, but concentrations of PCBs, PAHs, Hg, Pb, and Clostridium perfringens (a bacterium spore found in sewage) were nearly two or more times greater in the surface sediments than in intervals deeper in the cores. These observations indicate the usefulness of measuring concentration gradients in sediments at the San Francisco deep ocean disposal site (SF-DODS) where a thin (0.5 cm thick) layer of dredged material has been observed beyond the boundary. This thin layer has not been chemically characterized by the common practice of homogenizing over the top 10 cm. An estimated 300 million cubic yards of dredged material from San Francisco Bay are expected to be discharged at the SF-DODS site during the next 50 years. Detailed depth analysis of sediment cores would add significant new information about the fate and effects of dredged material in the deep sea.

Functional organization of the Sm core in the crystal structure of human U1 snRNP.

PubMed

Weber, Gert; Trowitzsch, Simon; Kastner, Berthold; Lührmann, Reinhard; Wahl, Markus C

2010-12-15

U1 small nuclear ribonucleoprotein (snRNP) recognizes the 5'-splice site early during spliceosome assembly. It represents a prototype spliceosomal subunit containing a paradigmatic Sm core RNP. The crystal structure of human U1 snRNP obtained from natively purified material by in situ limited proteolysis at 4.4 Å resolution reveals how the seven Sm proteins, each recognize one nucleotide of the Sm site RNA using their Sm1 and Sm2 motifs. Proteins D1 and D2 guide the snRNA into and out of the Sm ring, and proteins F and E mediate a direct interaction between the Sm site termini. Terminal extensions of proteins D1, D2 and B/B', and extended internal loops in D2 and B/B' support a four-way RNA junction and a 3'-terminal stem-loop on opposite sides of the Sm core RNP, respectively. On a higher organizational level, the core RNP presents multiple attachment sites for the U1-specific 70K protein. The intricate, multi-layered interplay of proteins and RNA rationalizes the hierarchical assembly of U snRNPs in vitro and in vivo.

The Persistence of Mode 1 Technology in the Korean Late Paleolithic

PubMed Central

Lee, Hyeong Woo

2013-01-01

Ssangjungri (SJ), an open-air site with several Paleolithic horizons, was recently discovered in South Korea. Most of the identified artifacts are simple core and flake tools that indicate an expedient knapping strategy. Bifacially worked core tools, which might be considered non-classic bifaces, also have been found. The prolific horizons at the site were dated by accelerator mass spectrometry (AMS) to about 30 kya. Another newly discovered Paleolithic open-air site, Jeungsan (JS), shows a homogeneous lithic pattern during this period. The dominated artifact types and usage of raw materials are similar in character to those from SJ, although JS yielded a larger number of simple core and flake tools with non-classic bifaces. Chronometric analysis by AMS and optically stimulated luminescence (OSL) indicate that the prime stratigraphic levels at JS also date to approximately 30 kya, and the numerous conjoining pieces indicate that the layers were not seriously affected by post-depositional processes. Thus, it can be confirmed that simple core and flake tools were produced at temporally and culturally independent sites until after 30 kya, supporting the hypothesis of a wide and persistent use of simple technology into the Late Pleistocene. PMID:23724113

Electrochemically Controlled Reconstitution of Immobilized Ferritins for Bioelectronic Applications

NASA Technical Reports Server (NTRS)

Kim, Jae-Woo; Choi, Sang H.; Lillehei, Peter T.; Chu, Sang-Hong; King, Glen C.; Watt, Gerald D.

2007-01-01

Site-specific reconstituted nanoparticles were fabricated via electrochemically-controlled biomineralization through the immobilization of biomolecules. The work reported herein includes the immobilization of ferritin with various surface modifications, the electrochemical biomineralization of ferritins with different inorganic cores, and the electrocatalytic reduction of oxygen on the reconstituted Pt-cored ferritins. Protein immobilization on the substrate is achieved by anchoring ferritins with dithiobis-N-succinimidyl propionate (DTSP). A reconstitution process of site-specific electrochemical biomineralization with a protein cage loads ferritins with different core materials. The ferritin acts as a nano-scale template, a biocompatible cage, and a separator between the nanoparticles. This first demonstration of electrochemically controlled site-specific reconstitution of biomolecules provides a new tool for biomineralization and opens the way to produce the bio-templated nanoparticles by electrochemical control. The nanosized platinum-cored ferritins on gold displayed good catalytic activity for the electrochemical reduction of oxygen, which is applicable to biofuel cell applications. This results in a smaller catalyst loading on the electrodes for fuel cells or other bioelectronic devices.

NASA's Next Generation Space Geodesy Network

NASA Technical Reports Server (NTRS)

Desai, S. D.; Gross, R. S.; Hilliard, L.; Lemoine, F. G.; Long, J. L.; Ma, C.; McGarry, J. F.; Merkowitz, S. M.; Murphy, D.; Noll, C. E.;

Chlorine-36 and cesium-137 in ice-core samples from mid-latitude glacial sites in the Northern Hemisphere

USGS Publications Warehouse

Green, J.R.; Cecil, L.D.; Synal, H.-A.; Kreutz, K.J.; Wake, C.P.; Naftz, D.L.; Frape, S.K.

2000-01-01

Chlorine-36 (36Cl) concentrations, 36Cl/Cl ratios, and 36Cl fluxes in ice-core samples collected from the Upper Fremont Glacier (UFG) in the Wind River Mountain Range, Wyoming, United States and the Nangpai Gosum Glacier (NGG) in the Himalayan Mountains, Nepal, were determined and compared with published results from the Dye-3 ice-core drilling site on the Greenland Ice Sheet. Cesium-137 (137Cs) concentrations in the NGG also were determined. The background fluxes for 36Cl for each glacial site were similar: (1.6??0.3)??10-2 atoms/cm2 s for the UFG samples, (0.7??0.1)??10-2 atoms/cm2 s for the NGG samples, and (0.4??0.1)??10-2 atoms/cm2 s for the Dye-3 samples. The 36Cl fluxes in ice that was deposited as snow during peak atmospheric nuclear weapon test (1957-1958) were (33??1)??10-2 atoms/cm2 s for the UFG site, (291??3)??10-2 atoms/cm2 s for the NGG site, and (124??5)??10-2 atoms/ cm2 s for the Dye-3 site. A weapon test period 137Cs concentration of 0.79??0.05 Bq/kg in the NGG ice core also was detected in the same section of ice that contained the largest 36Cl concentration. ?? 2000 Elsevier Science B.V. All rights reserved.

A Composite Depth Scale for Sediments from Crevice Lake, Montana

USGS Publications Warehouse

Rosenbaum, J.G.; Skipp, G.; Honke, J.; Chapman, C.

2010-01-01

As part of a study to derive records of past environmental change from lake sediments in the western United States, a set of cores was collected from Crevice Lake, Montana, in late February and early March 2001. Crevice Lake (latitude 45.000N, longitude 110.578W, elevation 1,713 meters) lies adjacent to the Yellowstone River at the north edge of Yellowstone National Park. The lake is more than 31 meters deep and has a surface area of 7.76 hectares. The combination of small surface area and significant depth promote anoxic bottom-water conditions that preserve annual laminations (varves) in the sediment. Three types of cores were collected through the ice. The uppermost sediments were obtained in freeze cores that preserved the sediment water interface. Two sites were cored with a 5-centimeter diameter corer. Five cores were taken with a 2-meter-long percussion piston corer. The percussion core uses a plastic core liner with an inside diameter of 9 centimeters. Coring was done at two sites. Because of the relatively large diameter of the percussion cores, samples from these cores were used for a variety of analyses including pollen, charcoal, diatoms, stable isotopes, organic and inorganic carbon, elemental analyses, and magnetic properties.

A conserved apomixis-specific polymorphism is correlated with exclusive exonuclease expression in premeiotic ovules of apomictic boechera species.

PubMed

Corral, José M; Vogel, Heiko; Aliyu, Olawale M; Hensel, Götz; Thiel, Thomas; Kumlehn, Jochen; Sharbel, Timothy F

2013-12-01

Apomixis (asexual seed production) is characterized by meiotically unreduced egg cell production (apomeiosis) followed by its parthenogenetic development into offspring that are genetic clones of the mother plant. Fertilization (i.e. pseudogamy) of the central cell is important for the production of a functional endosperm with a balanced 2:1 maternal:paternal genome ratio. Here, we present the APOLLO (for apomixis-linked locus) gene, an Aspartate Glutamate Aspartate Aspartate histidine exonuclease whose transcripts are down-regulated in sexual ovules entering meiosis while being up-regulated in apomeiotic ovules at the same stage of development in plants of the genus Boechera. APOLLO has both "apoalleles," which are characterized by a set of linked apomixis-specific polymorphisms, and "sexalleles." All apomictic Boechera spp. accessions proved to be heterozygous for the APOLLO gene (having at least one apoallele and one sexallele), while all sexual genotypes were homozygous for sexalleles. Apoalleles contained a 20-nucleotide polymorphism present in the 5' untranslated region that contains specific transcription factor-binding sites for ARABIDOPSIS THALIANA HOMEOBOX PROTEIN5, LIM1 (for LINEAGE ABNORMAL11, INSULIN1, MECHANOSENSORY PROTEIN3), SORLIP1AT (for SEQUENCES OVERREPRESENTED IN LIGHT-INDUCED PROMOTERS IN ARABIDOPSIS THALIANA1), SORLIP2AT, and POLYA SIGNAL1. In the same region, sexalleles contain transcription factor-binding sites for DNA BINDING WITH ONE FINGER2, DNA BINDING WITH ONE FINGER3, and PROLAMIN BOX-BINDING FACTOR. Our results suggest that the expression of a single deregulated allele could induce the cascade of events leading to asexual female gamete formation in an apomictic plant.

Validation of SMAP Surface Soil Moisture Products with Core Validation Sites

NASA Technical Reports Server (NTRS)

Colliander, A.; Jackson, T. J.; Bindlish, R.; Chan, S.; Das, N.; Kim, S. B.; Cosh, M. H.; Dunbar, R. S.; Dang, L.; Pashaian, L.;

caCORE: a common infrastructure for cancer informatics.

PubMed

Covitz, Peter A; Hartel, Frank; Schaefer, Carl; De Coronado, Sherri; Fragoso, Gilberto; Sahni, Himanso; Gustafson, Scott; Buetow, Kenneth H

2003-12-12

Sites with substantive bioinformatics operations are challenged to build data processing and delivery infrastructure that provides reliable access and enables data integration. Locally generated data must be processed and stored such that relationships to external data sources can be presented. Consistency and comparability across data sets requires annotation with controlled vocabularies and, further, metadata standards for data representation. Programmatic access to the processed data should be supported to ensure the maximum possible value is extracted. Confronted with these challenges at the National Cancer Institute Center for Bioinformatics, we decided to develop a robust infrastructure for data management and integration that supports advanced biomedical applications. We have developed an interconnected set of software and services called caCORE. Enterprise Vocabulary Services (EVS) provide controlled vocabulary, dictionary and thesaurus services. The Cancer Data Standards Repository (caDSR) provides a metadata registry for common data elements. Cancer Bioinformatics Infrastructure Objects (caBIO) implements an object-oriented model of the biomedical domain and provides Java, Simple Object Access Protocol and HTTP-XML application programming interfaces. caCORE has been used to develop scientific applications that bring together data from distinct genomic and clinical science sources. caCORE downloads and web interfaces can be accessed from links on the caCORE web site (http://ncicb.nci.nih.gov/core). caBIO software is distributed under an open source license that permits unrestricted academic and commercial use. Vocabulary and metadata content in the EVS and caDSR, respectively, is similarly unrestricted, and is available through web applications and FTP downloads. http://ncicb.nci.nih.gov/core/publications contains links to the caBIO 1.0 class diagram and the caCORE 1.0 Technical Guide, which provide detailed information on the present caCORE architecture, data sources and APIs. Updated information appears on a regular basis on the caCORE web site (http://ncicb.nci.nih.gov/core).

Microhabitat Conditions in Wyoming's Sage-Grouse Core Areas: Effects on Nest Site Selection and Success.

PubMed

Dinkins, Jonathan B; Smith, Kurt T; Beck, Jeffrey L; Kirol, Christopher P; Pratt, Aaron C; Conover, Michael R

2016-01-01

The purpose of our study was to identify microhabitat characteristics of greater sage-grouse (Centrocercus urophasianus) nest site selection and survival to determine the quality of sage-grouse habitat in 5 regions of central and southwest Wyoming associated with Wyoming's Core Area Policy. Wyoming's Core Area Policy was enacted in 2008 to reduce human disturbance near the greatest densities of sage-grouse. Our analyses aimed to assess sage-grouse nest selection and success at multiple micro-spatial scales. We obtained microhabitat data from 928 sage-grouse nest locations and 819 random microhabitat locations from 2008-2014. Nest success was estimated from 924 nests with survival data. Sage-grouse selected nests with greater sagebrush cover and height, visual obstruction, and number of small gaps between shrubs (gap size ≥0.5 m and <1.0 m), while selecting for less bare ground and rock. With the exception of more small gaps between shrubs, we did not find any differences in availability of these microhabitat characteristics between locations within and outside of Core Areas. In addition, we found little supporting evidence that sage-grouse were selecting different nest sites in Core Areas relative to areas outside of Core. The Kaplan-Meier nest success estimate for a 27-day incubation period was 42.0% (95% CI: 38.4-45.9%). Risk of nest failure was negatively associated with greater rock and more medium-sized gaps between shrubs (gap size ≥2.0 m and <3.0 m). Within our study areas, Wyoming's Core Areas did not have differing microhabitat quality compared to outside of Core Areas. The close proximity of our locations within and outside of Core Areas likely explained our lack of finding differences in microhabitat quality among locations within these landscapes. However, the Core Area Policy is most likely to conserve high quality habitat at larger spatial scales, which over decades may have cascading effects on microhabitat quality available between areas within and outside of Core Areas.

Analyses of water, bank material, bottom material, and elutriate samples collected near Belzoni, Mississippi (upper Yazoo projects)

USGS Publications Warehouse

Brightbill, David B.; Treadway, Joseph B.

1980-01-01

Four core-material-sampling sites and one bottom-material site were chosen by the U.S. Army Corps of Engineers to represent areas of proposed dredging activity along a 24.9-mile reach of the upper Yazoo River. Five receiving-water sites also were selected to represent the water that will contact the proposed dredged material. Chemical and physical analyses were performed upon core or bottom material and native-water (receiving-water) samples from these sites as well as upon elutriate samples of the mixture of sediment and receiving water. The results of these analyses are presented without interpertation. (USGS)

Third-generation site characterization: Cryogenic core collection, nuclear magnetic resonance, and electrical resistivity

NASA Astrophysics Data System (ADS)

Kiaalhosseini, Saeed

In modern contaminant hydrology, management of contaminated sites requires a holistic characterization of subsurface conditions. Delineation of contaminant distribution in all phases (i.e., aqueous, non-aqueous liquid, sorbed, and gas), as well as associated biogeochemical processes in a complex heterogeneous subsurface, is central to selecting effective remedies. Arguably, a factor contributing to the lack of success of managing contaminated sites effectively has been the limitations of site characterization methods that rely on monitoring wells and grab sediment samples. The overarching objective of this research is to advance a set of third-generation (3G) site characterization methods to overcome shortcomings of current site characterization techniques. 3G methods include 1) cryogenic core collection (C3) from unconsolidated geological subsurface to improve recovery of sediments and preserving key attributes, 2) high-throughput analysis (HTA) of frozen core in the laboratory to provide high-resolution, depth discrete data of subsurface conditions and processes, 3) resolution of non-aqueous phase liquid (NAPL) distribution within the porous media using a nuclear magnetic resonance (NMR) method, and 4) application of a complex resistivity method to track NAPL depletion in shallow geological formation over time. A series of controlled experiments were conducted to develop the C 3 tools and methods. The critical aspects of C3 are downhole circulation of liquid nitrogen via a cooling system, the strategic use of thermal insulation to focus cooling into the core, and the use of back pressure to optimize cooling. The C3 methods were applied at two contaminated sites: 1) F.E. Warren (FEW) Air Force Base near Cheyenne, WY and 2) a former refinery in the western U.S. The results indicated that the rate of core collection using the C3 methods is on the order of 30 foot/day. The C3 methods also improve core recovery and limits potential biases associated with flowing sands. HTA of frozen core was employed at the former refinery and FEW. Porosity and fluid saturations (i.e., aqueous, non-aqueous liquid, and gas) from the former refinery indicate that given in situ freezing, the results are not biased by drainage of pore fluids from the core during sample collection. At FEW, a comparison between the results of HTA of the frozen core collected in 2014 and the results of site characterization using unfrozen core, (second-generation (2G) methods) at the same locations (performed in 2010) indicate consistently higher contaminant concentrations using C 3. Many factors contribute to the higher quantification of contaminant concentrations using C3. The most significant factor is the preservation of the sediment attributes, in particular, pore fluids and volatile organic compounds (VOCs) in comparison to the unfrozen conventional sediment core. The NMR study was performed on laboratory-fabricated sediment core to resolve NAPL distribution within the porous media qualitatively and quantitatively. The fabricated core consisted of Colorado silica sand saturated with deionized water and trichloroethylene (TCE). The cores were scanned with a BRUKER small-animal scanner (2.3 Tesla, 100 MHz) at 20 °C and while the core was frozen at -25 °C. The acquired images indicated that freezing the water within the core suppressed the NMR signals of water-bound hydrogen. The hydrogen associated with TCE was still detectable since the TCE was in its liquid state (melting point of TCE is -73 °C). Therefore, qualitative detection of TCE within the sediment core was performed via the NMR scanning by freezing the water. A one-dimensional NMR scanning method was used for quantification of TCE mass distribution within the frozen core. However, the results indicated inconsistency in estimating the total TCE mass within the porous media. Downhole NMR logging was performed at the former refinery in the western U.S. to detect NAPL and to discriminate NAPL from water in the formation. The results indicated that detection of NMR signals to discriminate NAPL from water is compromised by the noise stemming from the active facilities and/or power lines passing over the site. A laboratory experiment was performed to evaluate the electrical response of unconsolidated porous media through time (30 days) while NAPL was being depleted. Sand columns (Colorado silica sand) contaminated with methyl tert-butyl ether (MTBE, a light non-aqueous phase liquid (LNAPL)) were studied. A multilevel electrode system was used to measure electrical resistivity of impacted sand by imposing alternative current. The trend of reduction in resistivity through the depth of columns over time followed depletion of LNAPL by volatilization. Finally, a field experiment was performed at the former refinery in the western U.S. to track natural losses of LNAPL over time. Multilevel systems consisting of water samplers, thermocouples, and electrodes were installed at a clean zone (background zone) and an LNAPL-impacted zone. In situ measurements of complex resistivity and temperature were taken and water sampling was performed for each depth (from 3 to 14 feet below the ground surface at one-foot spacing) within almost a year. At both locations, the results indicated decreases in apparent resistivity below the water table over time. This trend was supported by the geochemistry of the pore fluids. Overall, results indicate that application of the electrical resistivity method to track LNAPL depletion at field sites is difficult due to multiple conflicting factors affecting the geoelectrical response of LNAPL-impacted zones over time.

Grain Size Biasing of 230Th-derived Focusing Factors in the Panama Basin

NASA Astrophysics Data System (ADS)

Loveley, M. R.; Marcantonio, F.; Lyle, M. W.; Ibrahim, R.; Wang, J. K.; Hertzberg, J. E.

2014-12-01

In this study, we attempt to understand how differing grain size classes in Panama Basin sediments may create biasing of 230Th as a constant-flux proxy. Greater amounts of 230Th are contained in fine grained particles, which, if fractionated from coarser grained counterparts may lead to biasing of 230Th-derived mass accumulation rates (MARs) and sediment focusing factors. We examined sediments that span the past 25 kyr from four new sediment cores retrieved from two different localities close to the ridges that bound the Panama Basin. Each locality contained paired sites that were seismically interpreted to have undergone sediment redistribution, i.e., thick focused sites versus thin winnowed sites. Two sediment cores were retrieved from the northern part of the Panama basin, Cocos Ridge, (MV1014-01-"4JC", 5° 44.7'N 85° 45.5' W, 1730 m depth; MV1014-01-"8JC", 6° 14.0'N 86° 2.6' W, 1993 m depth), and two were retrieved from the southern part of the basin, Carnegie Ridge, (MV1014-02-"11JC", 0° 41.6'S 85° 20.0' W, 2452 m depth; MV1014-02-"17JC" 0° 10.8'S 85° 52.0' W, 2846 m depth). Cores 4JC and 11JC lie closer to the ridge tops of the Cocos and Carnegie Ridges, respectively, and have thin sediment drapes, while deeper cores, 8JC (Cocos) and 17JC (Carnegie), have thicker sediment drapes and lie downslope from the ridge top cores. Age-model-derived sand MARs, which likely represent the vertical rain of particles that cannot be transported by bottom currents, are similar at each of the paired sites in Holocene and glacial time slices. However, 230Th-normalized MARs are about 50% lower, on average at each of the paired sites during the same time slices. Both Holocene and glacial samples from "thin" cores (4,11JC) contain, surprisingly, significant amounts (up to 50%) of the 230Th within the coarse grained (>63 μm) fraction which makes up 40-70% of the bulk samples analyzed. On the contrary, Holocene and glacial samples from "thick" cores, (8,17JC), contain the greatest amounts of 230Th (up to 49%) in the finest grain-sized fraction (<4μm), which makes up 26-40% of the bulk samples analyzed. Although, redistribution of sediment has taken place, our analysis indicates that 230Th-derived focusing factors are being overestimated at thick sites and underestimated at thin sites.

RNA Modulates the Interaction between Influenza A Virus NS1 and Human PABP1.

PubMed

Arias-Mireles, Bryan H; de Rozieres, Cyrus M; Ly, Kevin; Joseph, Simpson

2018-05-25

Nonstructural protein 1 (NS1) is a multifunctional protein involved in preventing host-interferon response in influenza A virus (IAV). Previous studies have indicated that NS1 also stimulates the translation of viral mRNA by binding to conserved sequences in the viral 5'-UTR. Additionally, NS1 binds to poly(A) binding protein 1 (PABP1) and eukaryotic initiation factor 4G (eIF4G). The interaction of NS1 with the viral 5'-UTR, PABP1, and eIF4G has been suggested to specifically enhance the translation of viral mRNAs. In contrast, we report that NS1 does not directly bind to sequences in the viral 5'-UTR, indicating that NS1 is not responsible for providing the specificity to stimulate viral mRNA translation. We also monitored the interaction of NS1 with PABP1 using a new, quantitative FRET assay. Our data show that NS1 binds to PABP1 with high affinity; however, the binding of double-stranded RNA (dsRNA) to NS1 weakens the binding of NS1 to PABP1. Correspondingly, the binding of PABP1 to NS1 weakens the binding of NS1 to double-stranded RNA (dsRNA). In contrast, the affinity of PABP1 for binding to poly(A) RNA is not significantly changed by NS1. We propose that the modulation of NS1·PABP1 interaction by dsRNA may be important for the viral cycle.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ogami, Koichi; Cho, Rihe; Hoshino, Shin-ichi, E-mail: hoshino@phar.nagoya-cu.ac.jp

Highlights: ► So far, only an enzymatically inactive isoform of PAPD7 was reported. ► The novel isoform: PAPD7 l shows robust nucleotidyl transferase activity. ► The newly identified amino terminal region is required for the activity. ► PAPD7 l localizes to the nucleoplasm. ► The N terminal region identified is also required for the nuclear localization. - Abstract: Non-canonical poly(A) polymerases (ncPAPs) catalyze the addition of poly(A) tail to the 3′ end of RNA to play pivotal roles in the regulation of gene expression and also in quality control. Here we identified a novel isoform of the 7th member ofmore » ncPAPs: PAPD7 (PAPD7 l), which contains 230 extra amino acids at the amino terminus of the previously identified PAPD7 (PAPD7 s). In sharp contrast to the inactive PAPD7 s, PAPD7 l showed robust nucleotidyl transferase activity when tethered to an RNA. A region required for the activity was localized to 187–219 aa, and this region was also required for the nuclear retention of PAPD7 l. Western blot analysis revealed that 94 kDa band (corresponding to PAPD7 l) but not 62 kDa band (corresponding to PAPD7 s) detected by PAPD7 antibody was specifically depleted by treatment with PAPD7 siRNA in both HeLa and U2OS cells. These results suggest that PAPD7 l is the major and active isoform of PAPD7 expressed in cells.« less

Human SUV3 helicase regulates growth rate of the HeLa cells and can localize in the nucleoli.

PubMed

Szewczyk, Maciej; Fedoryszak-Kuśka, Natalia; Tkaczuk, Katarzyna; Dobrucki, Jurek; Waligórska, Agnieszka; Stępień, Piotr P

2017-01-01

The human SUV3 helicase (SUV3, hSUV3, SUPV3L1) is a DNA/RNA unwinding enzyme belonging to the class of DexH-box helicases. It localizes predominantly in the mitochondria, where it forms an RNA-degrading complex called mitochondrial degradosome with exonuclease PNP (polynucleotide phosphorylase). Association of this complex with the polyA polymerase can modulate mitochondrial polyA tails. Silencing of the SUV3 gene was shown to inhibit the cell cycle and to induce apoptosis in human cell lines. However, since small amounts of the SUV3 helicase were found in the cell nuclei, it was not clear whether the observed phenotypes of SUV3 depletion were of mitochondrial or nuclear origin. In order to answer this question we have designed gene constructs able to inhibit the SUV3 activity exclusively in the cell nuclei. The results indicate that the observed growth rate impairment upon SUV3 depletion is due to its nuclear function(s). Unexpectedly, overexpression of the nuclear-targeted wild-type copies of the SUV3 gene resulted in a higher growth rate. In addition, we demonstrate that the SUV3 helicase can be found in the HeLa cell nucleoli, but it is not detectable in the DNA-repair foci. Our results indicate that the nucleolar-associated human SUV3 protein is an important factor in regulation of the cell cycle.

What is an Oceanic Core Complex?

NASA Astrophysics Data System (ADS)

Schroeder, T.; Cheadle, M. J.

2007-12-01

The Mid-Atlantic Ridge (MAR) 75km north and south of the 15-20 Fracture Zone (FZ) has produced upper oceanic lithosphere composed dominantly of mantle peridotite with gabbro intrusions. In the absence of diapirism, mantle peridotite can only be exposed on the seafloor by extensional faulting, thus the sea floor geology and bathymetry provide widespread evidence for extensive low-angle faulting. However, only 3% of the seafloor in this region has the domal morphology characteristic of features that have been termed oceanic core complexes; suggesting that other processes, in addition to low-angle faulting, are responsible for the generation of domal core complexes. Most low-angle faults near the 15-20 FZ form gently dipping (10-15°), 10-15km-wide dip slopes on the flanks of 2000m relief bathymetric ridges that are up to 15-40km long (parallel to the MAR). Core recovered from ODP Leg 209 drill holes in these ridges is dominantly peridotite with small (<50m thick) gabbro intrusions. The peridotite is cut by a very high density of brittle faults dipping at both steep and gentle angles. Several holes also contain long-lived shear zones/faults in their upper reaches in which strain was localized at granulite facies, indicated by mylonitic olivine and cpx, and remained active during cooling to sub-greenschist grade, indicated by cross-cutting of progressively lower-grade syn-deformation mineral assemblages. These observations suggest that seafloor spreading is largely accommodated here by slip on low-angle faults, and that these faults are correctly termed detachment faults. Holes drilled into a domal oceanic core complex north of the 15-20 FZ during Leg 209 (ODP Site 1275) recovered dominantly gabbro and not mantle peridotite. This hole is cut by significantly fewer brittle and ductile faults than the peridotite drilled at the non-core-complex detachment fault sites. The detachment fault in the upper reaches (50m) of Site 1275 was localized at temperatures near feldspar's ductile-to-brittle transition, indicated by cataclasis with minor crystal plastic flow in plagioclase, and a lack of pervasive pure-ductile deformation. Amphibole-plagioclase thermometry in the fault yields equilibrium temperatures from 600-650°C, compared to equilibrium temperatures of 750-850°C for the gabbro outside the fault. The presence of talc- chlorite schists and cataclasites cutting the higher-temperature deformation textures indicate fault activity down- temperature from amphibolite through greenschist facies. This core-complex-bounding fault contrasts with the fault that bounds the Atlantis Bank Core Complex on the Southwest Indian Ridge (SWIR). There, the fault is 100m thick and strain was initially localized at granulite grade (>800°C) (Mehl & Hirth, 2007); significantly hotter than the Site 1275 fault. Therefore, the formation of core-complex morphology does not seem to depend on the initial faulting conditions. Both oceanic core complexes that have been drilled besides Site 1275, Atlantis Massif at 30°N (IODP Hole 1309D) on the MAR and Atlantis Bank on the SWIR (ODP Hole 735B), are also comprised dominantly of gabbro. This suggests that magma supply may be an essential requirement for core complex formation and raises the question whether all domal oceanic core complexes are cored by gabbro? We also ask whether the term 'oceanic core complex' should be restricted to these domal features and not applied to detachment-bound, non- domal, peridotite-cored ridges; or if these should be considered two sub-classes of oceanic core complexes.

CORE_TF: a user-friendly interface to identify evolutionary conserved transcription factor binding sites in sets of co-regulated genes

PubMed Central

Hestand, Matthew S; van Galen, Michiel; Villerius, Michel P; van Ommen, Gert-Jan B; den Dunnen, Johan T; 't Hoen, Peter AC

2008-01-01

Background The identification of transcription factor binding sites is difficult since they are only a small number of nucleotides in size, resulting in large numbers of false positives and false negatives in current approaches. Computational methods to reduce false positives are to look for over-representation of transcription factor binding sites in a set of similarly regulated promoters or to look for conservation in orthologous promoter alignments. Results We have developed a novel tool, "CORE_TF" (Conserved and Over-REpresented Transcription Factor binding sites) that identifies common transcription factor binding sites in promoters of co-regulated genes. To improve upon existing binding site predictions, the tool searches for position weight matrices from the TRANSFACR database that are over-represented in an experimental set compared to a random set of promoters and identifies cross-species conservation of the predicted transcription factor binding sites. The algorithm has been evaluated with expression and chromatin-immunoprecipitation on microarray data. We also implement and demonstrate the importance of matching the random set of promoters to the experimental promoters by GC content, which is a unique feature of our tool. Conclusion The program CORE_TF is accessible in a user friendly web interface at . It provides a table of over-represented transcription factor binding sites in the users input genes' promoters and a graphical view of evolutionary conserved transcription factor binding sites. In our test data sets it successfully predicts target transcription factors and their binding sites. PMID:19036135

Data report: Permeabilities of eastern equatorial Pacific and Peru margin sediments

USGS Publications Warehouse

Gamage, Kusali; Bekins, Barbara A.; Screaton, Elizabeth; Jørgensen, Bo B.; D'Hondt, Steven L.; Miller, D. Jay

2006-01-01

Constant-flow permeability tests were conducted on core samples from Ocean Drilling Program Leg 201 from the eastern equatorial Pacific and the Peru margin. Eighteen whole-round core samples from Sites 1225, 1226, 1227, 1230, and 1231 were tested for vertical permeabilities. Sites 1225, 1226, and 1231 represent sediments of the open ocean, whereas Sites 1227 and 1230 represent sediments of the ocean margin. Measured vertical permeabilities vary from ~8 x 10–19 m2 to ~1 x 10–16 m2 for a porosity range of 45%–90%.

Intercalation of proflavine and a platinum derivative of proflavine into double-helical Poly(A).

PubMed

Ciatto, C; D'Amico, M L; Natile, G; Secco, F; Venturini, M

1999-11-01

The equilibria and kinetics of the interactions of proflavine (PR) and its platinum-containing derivative [PtCl(tmen)(2)HNC(13)H(7)(NHCH(2)CH(2))(2)](+) (PRPt) with double-stranded poly(A) have been investigated by spectrophotometry and Joule temperature-jump relaxation at ionic strength 0.1 M, 25 degrees C, and pH 5.2. Spectrophotometric measurements indicate that base-dye interactions are prevailing. T-jump experiments with polarized light showed that effects due to field-induced alignment could be neglected. Both of the investigated systems display two relaxation effects. The kinetic features of the reaction are discussed in terms of a two-step series mechanism in which a precursor complex DS(I) is formed in the fast step, which is then converted to a final complex in the slow step. The rate constants of the fast step are k(1) = (2.5 +/- 0.4) x 10(6) M(-1) s(-1), k(-1) = (2.4 +/- 0.1) x 10(3) s(-1) for poly(A)-PR and k(1) = (2.3 +/- 0.1) x 10(6) M(-1) s(-1), k(-1) = (1.6 +/- 0.2) x 10(3) s(-1) for poly(A)-PRPt. The rate constants for the slow step are k(2) = (4.5 +/- 0.5) x 10(2) s(-1), k(-2) = (1.7 +/- 0.1) x 10(2) s(-1) for poly(A)-PR and k(2) = 9.7 +/- 1.2 s(-1), k(-2) = 10.6 +/- 0.2 s(-1) for poly(A)-PRPt. Spectrophotometric measurements yield for the equilibrium constants and site size the values K = (4.5 +/- 0.1) x 10(3) M(-1), n = 1.3 +/- 0.5 for poly(A)-PR and K = (2.9 +/- 0.1) x 10(3) M(-1), n = 2.3 +/- 0.6 for poly(A)-PRPt. The values of k(1) are similar and lower than expected for diffusion-limited reactions. The values of k(-1) are similar as well. It is suggested that the formation of DS(I) involves only the proflavine residues in both systems. In contrast, the values of k(2) and k(-2) in poly(A)-PRPt are much lower than in poly(A)-PR. The results suggest that in the complex DS(II) of poly(A)-PRPt both proflavine and platinum residues are intercalated. In addition, a very slow process was detected and ascribed to the covalent binding of Pt(II) to the adenine.

Intercalation of proflavine and a platinum derivative of proflavine into double-helical Poly(A)

PubMed Central

Ciatto, C; D'Amico, ML; Natile, G; Secco, F; Venturini, M

1999-01-01

The equilibria and kinetics of the interactions of proflavine (PR) and its platinum-containing derivative [PtCl(tmen)(2)HNC(13)H(7)(NHCH(2)CH(2))(2)](+) (PRPt) with double-stranded poly(A) have been investigated by spectrophotometry and Joule temperature-jump relaxation at ionic strength 0.1 M, 25 degrees C, and pH 5.2. Spectrophotometric measurements indicate that base-dye interactions are prevailing. T-jump experiments with polarized light showed that effects due to field-induced alignment could be neglected. Both of the investigated systems display two relaxation effects. The kinetic features of the reaction are discussed in terms of a two-step series mechanism in which a precursor complex DS(I) is formed in the fast step, which is then converted to a final complex in the slow step. The rate constants of the fast step are k(1) = (2.5 +/- 0.4) x 10(6) M(-1) s(-1), k(-1) = (2.4 +/- 0.1) x 10(3) s(-1) for poly(A)-PR and k(1) = (2.3 +/- 0.1) x 10(6) M(-1) s(-1), k(-1) = (1.6 +/- 0.2) x 10(3) s(-1) for poly(A)-PRPt. The rate constants for the slow step are k(2) = (4.5 +/- 0.5) x 10(2) s(-1), k(-2) = (1.7 +/- 0.1) x 10(2) s(-1) for poly(A)-PR and k(2) = 9.7 +/- 1.2 s(-1), k(-2) = 10.6 +/- 0.2 s(-1) for poly(A)-PRPt. Spectrophotometric measurements yield for the equilibrium constants and site size the values K = (4.5 +/- 0.1) x 10(3) M(-1), n = 1.3 +/- 0.5 for poly(A)-PR and K = (2.9 +/- 0.1) x 10(3) M(-1), n = 2.3 +/- 0.6 for poly(A)-PRPt. The values of k(1) are similar and lower than expected for diffusion-limited reactions. The values of k(-1) are similar as well. It is suggested that the formation of DS(I) involves only the proflavine residues in both systems. In contrast, the values of k(2) and k(-2) in poly(A)-PRPt are much lower than in poly(A)-PR. The results suggest that in the complex DS(II) of poly(A)-PRPt both proflavine and platinum residues are intercalated. In addition, a very slow process was detected and ascribed to the covalent binding of Pt(II) to the adenine. PMID:10545371

INNOVATIVE TECHNOLOGY EVALUATION REPORT ...

EPA Pesticide Factsheets

The Split Core Sampler for Submerged Sediments (Split Core Sampler) designed and fabricated by Arts Manufacturing & Supply, Inc., was demonstrated under the U.S. Environmental Protection Agency (EPA) Superfund Innovative Technology Evaluation Program in April and May 1999 at sites in EPA Regions 1 and 5, respectively. In addition to assessing ease of sampler operation, key objectives of the demonstration included evaluating the samplers ability to (1) consistently collect a given volume of sediment, (2) consistently collect sediment in a given depth interval, (3) collect samples with consistent characteristics from a homogenous layer of sediment, and (4) collect samples under a variety of site conditions. This report describes the demonstration results for the Split Core Sampler and two conventional samplers (the Hand Corer and Vibrocorer) used as reference samplers. During the demonstration, the Split Core Sampler performed as well as or better than the reference samplers. Based on visual observations, both the Split Core Sampler and reference samplers collected partially compressed samples of consolidated and unconsolidated sediments from the sediment surface downward; sample representativeness may be questionable because of core shortening and core compression. Sediment stratification was preserved for both consolidated and unconsolidated sediment samples collected by the Split Core Sampler and reference samplers. No sampler was able to collect samples

A high resolution record of chlorine-36 nuclear-weapons-tests fallout from Central Asia

USGS Publications Warehouse

Green, J.R.; Cecil, L.D.; Synal, H.-A.; Santos, J.; Kreutz, K.J.; Wake, C.P.

2004-01-01

The Inilchek Glacier, located in the Tien Shan Mountains, central Asia, is unique among mid-latitude glaciers because of its relatively large average annual accumulation. In July 2000, two ice cores of 162 and 167 meters (m) in length were collected from the Inilchek Glacier for (chlorine-36) 36Cl analysis a part of a collaborative international effort to study the environmental changes archived in mid-latitude glaciers worldwide. The average annual precipitation at the collection site was calculated to be 1.6 m. In contrast, the reported average annual accumulations at the high-latitude Dye-3 glacial site, Greenland, the mid-latitude Guliya Ice Cap, China, and the mid-latitude Upper Fremont Glacier, Wyoming, USA, were 0.52, 0.16 and 0.76 m, respectively. The resolution of the 36Cl record in one of the Inilchek ice cores was from 2 to 10 times higher than the resolution of the records at these other sites and could provide an opportunity for detailed study of environmental changes that have occurred over the past 150 years. Despite the differences in accumulation among these various glacial sites, the 36Cl profile and peak concentrations for the Inilchek ice core were remarkably similar in shape and magnitude to those for ice cores from these other sites. The 36Cl peak concentration from 1958, the year during the mid-1900s nuclear-weapons-tests period when 36Cl fallout was largest, was preserved in the Inilchek core at a depth of 90.56 m below the surface of the glacier (74.14-m-depth water equivalent) at a concentration of 7.7 ?? 105 atoms of 36Cl/gram (g) of ice. Peak 36Cl concentrations from Dye-3, Guliya and the Upper Fremont glacial sites were 7.1 ?? 105, 5.4 ?? 105 and 0.7 ?? 105 atoms of 36Cl/g of ice, respectively. Measurements of 36Cl preserved in ice cores improve estimates of historical worldwide atmospheric deposition of this isotope and allow the sources of 36Cl in ground water to be better identified. ?? 2004 Published by Elsevier B.V.

Hominin Sites and Paleolakes Drilling Project. Chew Bahir, southern Ethiopia: How to get from three tonnes of sediment core to > 500 ka of continuous climate history?

NASA Astrophysics Data System (ADS)

Foerster, Verena; Asrat, Asfawossen; Cohen, Andrew S.; Gromig, Raphael; Günter, Christina; Junginger, Annett; Lamb, Henry F.; Schaebitz, Frank; Trauth, Martin H.

2016-04-01

In search of the environmental context of the evolution and dispersal of Homo sapiens and our close relatives within and beyond the African continent, the ICDP-funded Hominin Sites and Paleolakes Drilling Project (HSPDP) has recently cored five fluvio-lacustrine archives of climate change in East Africa. The sediment cores collected in Ethiopia and Kenya are expected to provide valuable insights into East African environmental variability during the last ~3.5 Ma. The tectonically-bound Chew Bahir basin in the southern Ethiopian rift is one of the five sites within HSPDP, located in close proximity to the Lower Omo River valley, the site of the oldest known fossils of anatomically modern humans. In late 2014, the two cores (279 and 266 m long respectively, HSPDP-CHB14-2A and 2B) were recovered, summing up to nearly three tonnes of mostly calcareous clays and silts. Deciphering an environmental record from multiple records, from the source region of modern humans could eventually allow us to reconstruct the pronounced variations of moisture availability during the transition into Middle Stone Age, and its implications for the origin and dispersal of Homo sapiens. Here we present the first results of our analysis of the Chew Bahir cores. Following the HSPDP protocols, the two parallel Chew Bahir sediment cores have been merged into one single, 280 m long and nearly continuous (>90%) composite core on the basis of a high resolution MSCL data set (e.g., magnetic susceptibility, gamma ray density, color intensity transects, core photographs). Based on the obvious cyclicities in the MSCL, correlated with orbital cycles, the time interval covered by our sediment archive of climate change is inferred to span the last 500-600 kyrs. Combining our first results from the long cores with the results from the accomplished pre-study of short cores taken in 2009/10 along a NW-SE transect across the basin (Foerster et al., 2012, Trauth et al., 2015), we have developed a hypothesis linking climate forcing and paleoenvironmental signal formation processes in the basin (e.g. the inverse correlation of the K flux with precipitation in the catchment), providing an important prerequisite for understanding the environmental record contained in the long sediment cores. The good recovery and anticipated high time resolution of the cores will give us a continuous record of environmental fluctuations on decadal to orbital timescales, which will allow us to test current hypotheses of the influence of climate on human evolution and dispersal.

Estimates of in situ gas hydrate concentration from resistivity monitoring of gas hydrate bearing sediments during temperature equilibration

USGS Publications Warehouse

Riedel, M.; Long, P.E.; Collett, T.S.

2006-01-01

As part of Ocean Drilling Program Leg 204 at southern Hydrate Ridge off Oregon we have monitored changes in sediment electrical resistivity during controlled gas hydrate dissociation experiments. Two cores were used, each filled with gas hydrate bearing sediments (predominantly mud/silty mud). One core was from Site 1249 (1249F-9H3), 42.1 m below seafloor (mbsf) and the other from Site 1248 (1248C-4X1), 28.8 mbsf. At Site 1247, a third experiment was conducted on a core without gas hydrate (1247B-2H1, 3.6 mbsf). First, the cores were imaged using an infra-red (IR) camera upon recovery to map the gas hydrate occurrence through dissociation cooling. Over a period of several hours, successive runs on the multi-sensor track (includes sensors for P-wave velocity, resistivity, magnetic susceptibility and gamma-ray density) were carried out complemented by X-ray imaging on core 1249F-9H3. After complete equilibration to room temperature (17-18??C) and complete gas hydrate dissociation, the final measurement of electrical resistivity was used to calculate pore-water resistivity and salinities. The calculated pore-water freshening after dissociation is equivalent to a gas hydrate concentration in situ of 35-70% along core 1249F-9H3 and 20-35% for core 1248C-4X1 assuming seawater salinity of in situ pore fluid. Detailed analysis of the IR scan, X-ray images and split-core photographs showed the hydrate mainly occurred disseminated throughout the core. Additionally, in core 1249F-9H3, a single hydrate filled vein, approximately 10 cm long and dipping at about 65??, was identified. Analyses of the logging-while-drilling (LWD) resistivity data revealed a structural dip of 40-80?? in the interval between 40 and 44 mbsf. We further analyzed all resistivity data measured on the recovered core during Leg 204. Generally poor data quality due to gas cracks allowed analyses to be carried out only at selected intervals at Sites 1244, 1245, 1246, 1247, 1248, 1249, and 1252. With a few exceptions, data from these intervals yield low to no gas hydrate concentration, which corresponds to estimates from downhole resistivity logs. However, since the gas cracking may be the result of gas hydrate dissociation, this is a biased sampling. Cores that had contained some gas hydrate may have been excluded. ?? 2005 Elsevier B.V. All rights reserved.

Generation of RNA in abiotic conditions.

NASA Astrophysics Data System (ADS)

di Mauro, Ernesto

Generation of RNA in abiotic conditions. Ernesto Di Mauro Dipartimento di Genetica Bi-ologia Molecolare, Universit` "Sapienza" Roma, Italy. a At least four conditions must be satisfied for the spontaneous generation of (pre)-genetic poly-mers: 1) availability of precursors that are activated enough to spontaneously polymerize. Preliminary studies showed that (a) nucleic bases and acyclonucleosides can be synthesized from formamide H2NCOH by simply heating with prebiotically available mineral catalysts [last reviewed in (1)], and that b) nucleic bases can be phosphorylated in every possible posi-tion [2'; 3'; 5'; cyclic 2',3'; cyclic 3',5' (2)]. The higher stability of the cyclic forms allows their accumulation. 2) A polymerization mechanism. A reaction showing the formation of RNA polymers starting from prebiotically plausible precursors (3',5' cyclic GMP and 3', 5'cyclic AMP) was recently reported (3). Polymerization in these conditions is thermodynamically up-hill and an equilibrium is attained that limits the maximum length of the polymer produced to about 40 nucleotides for polyG and 100 nucleotides for polyA. 3) Ligation of the synthesized oligomers. If this type of reaction could occur according to a terminal-joining mechanism and could generate canonical 3',5' phosphodiester bonds, exponential growth would be obtained of the generated oligomers. This type of reaction has been reported (4) , limited to homogeneous polyA sequences and leading to the production of polyA dimers and tetramers. What is still missing are: 4) mechanisms that provide the proof of principle for the generation of sequence complexity. We will show evidence for two mechanisms providing this proof of principle for simple complementary sequences. Namely: abiotic sequence complementary-driven terminal ligation and sequence-complementary terminal growth. In conclusion: all the steps leading to the generation of RNA in abiotic conditions are satisfied. (1) R Saladino, C Crestini, F. Ciciriello, S. Pino, G. Costanzo, E. Di Mauro. From formamide to RNA: the roles of formamide and water in the evolution of chemical information. Research In Microbiology, Special Issue on The Origin of life and Microbiology (2009) 160:441-448. (2) Costanzo, G., Saladino, R., Crestini, C., Ciciriello, F., and Di Mauro, E. Nucleoside phos-phorylation by phosphate minerals. J. Biol. Chem. (2007) 282: 16729-16735. (3) Samanta Pino, Fabiana Ciciriello, Giovanna Costanzo and E. Di Mauro, Nonenzymatic RNA Ligation in Water J. Biol. Chem. (2008), 283: 36494-36503 (4) Costanzo, G., Pino, S., Ciciriello, F., and Di Mauro, E. Generation of long RNA chains in water. J Biol Chem. (2009) 284:33206-33216.

Data from a thick unsaturated zone underlying Oro Grande and Sheep Creek washes in the western part of the Mojave Desert, near Victorville, San Bernardino County, California

USGS Publications Warehouse

Izbicki, John A.; Clark, Dennis A.; Pimental, Maria I.; Land, Michael; Radyk, John C.; Michel, Robert L.

2000-01-01

This report presents data on the physical properties of unsaturated alluvial deposits and on the chemical and isotopic composition of soil water and soil gas collected at 12 monitoring sites in the western part of the Mojave Desert, near Victorville, California. Sites were installed using the ODEX air-hammer method. Seven sites were located in the active channels of Oro Grande and Sheep Creek Washes. The remaining five sites were located away from the active washes. Most sites were drilled to a depth of about 100 feet below land surface; two sites were drilled to the water table almost 650 feet below land surface. Drilling procedures, lithologic and geophysical data, and site construction and instrumentation are described. Core material was analyzed for water content, bulk density, water potential, particle size, and water retention. The chemical composition of leachate from almost 1,000 subsamples of cores and cuttings was determined. Water extracted from selected subsamples of cores was analyzed for tritium and the stable isotopes of oxygen and hydrogen. Water from suction-cup lysimeters and soil-gas samples also were analyzed for chemical and isotopic composition. In addition, data on the chemical and isotopic composition of bulk precipitation from five sites and on ground water from two water-table wells are reported.

Site characterization design and techniques used at the Southern Shipbuilding Corporation site

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mueller, J.P.; Geraghty, C.A.; Moore, G.W.

1995-12-31

The Southern Shipbuilding Corporation (SSC) site is an inactive barge/ship manufacturing and repair facility situated on approximately 54 acres in Slidell, St. Tammany Parish, Louisiana. Two unlined surface impoundments (North and South impoundments) are situated on the northwest portion of the site and are surrounded on three sides by Bayou Bonfouca. These impoundments are the sources of carcinogenic polynuclear aromatic hydrocarbon (CPAH) contamination at the site. Inadequate containment has resulted in the release of impoundment wastes into the bayou. To evaluate potential response alternatives for the site, an Engineering Evaluation/Cost Analysis (EE/CA) field investigation was conducted from July through Octobermore » 1994. A two phase sampling approach was used in combination with innovative and traditional sampling techniques, field screening technologies, and exploitation of the visual characteristics of the waste to determine the extent of waste migration with limited off-site laboratory confirmation. A skid-mounted mobile drilling unit, secured to a specialized sampling platform designed for multiple applications, was used for collection of sediment cores from the bayou as well as tarry sludge cores from the impoundments. Field screening of core samples was accomplished on site using an organic vapor analyzer and a total petroleum hydrocarbon (TPH) field analyzer. Pollutants of concern include metals, cyanide, dioxin, and organic compounds. This paper presents details on the sampling design and characterization techniques used to accomplish the EE/CA field investigation.« less

Biologically Derived Nanoparticle Arrays via a Site-Specific Reconstitution of Ferritin and their Electrochemistry

NASA Technical Reports Server (NTRS)

Kim, Jae-Woo; Choi, Sang H.; Lillehei, Peter T.; King, Glen C.; Elliott, James R.; Chu, Sang-Hyon; Park, Yeonjoon; Watt, Gerald D.

2004-01-01

Nanoparticle arrays biologically derived from an electrochemically-controlled site-specific biomineralization were fabricated on a gold substrate through the immobilization process of biomolecules. The work reported herein includes the immobilization of ferritin with various surface modifications, the electrochemical biomineralization of ferritins with different inorganic cores, the fabrication of self-assembled arrays with the immobilized ferritin, and the electrochemical characterization of various core materials. Protein immobilization on the substrate is achieved by anchoring ferritins with dithiobis-N-succinimidyl propionate (DTSP). A reconstitution process of electrochemical site-specific biomineralization with a protein cage loads ferritins with different core materials such as Pt, Co, Mn, and Ni. The ferritin acts as a nano-scale template, a biocompatible cage, and a separator between the nanoparticles. The nano-sized metalcored ferritins on a gold substrate displayed a good electrochemical activity for the electron transport and storage, which is suitable for bioelectronics applications such as biofuel cell, bionanobattery, biosensors, etc. Keywords: Ferritin, immobilization, site-specific reconstitution, biomineralization, and bioelectronics

Variations in the depths of sulfate-methane transition zone (SMTZ) in UBGH2-6 drilling site in Ulleung Basin, East sea of Korea

NASA Astrophysics Data System (ADS)

Kim, Y.; Chun, J. H.; Bahk, J. J.; Ryu, B. J.; Um, I. K.

2016-12-01

The second Ulleung Basin Gas hydrate Drilling Expedition (UBGH2) was conducted in the Ulleung Basin, East Sea of Korea in 2010. Gas hydrates were observed in depth interval from 140 mbsf (meter below seafloor) to 160 mbsf in core sediment taken from UBGH2-6 drilling site, located in the north-western part of the basin (2,164 m water depth). To characterize the geochemical process for UBGH2-6 core sediments, pore fluid samples and headspace gas samples were extracted from core sediments and analyzed SO42- and CH4 concentrations. Based on SO42- and CH4 concentrations, sulfate-methane transition zone (SMTZ), where SO42- is depleted to zero and CH4 starts to increase was defined at a depth of approximately 6.50 mbsf in 2010. And in order to identify the variations in the depths of SMTZ at UBGH2-6 drilling site since 2010 (UBGH2), whole-round piston cores were collected from UBGH2-6 drilling site from 2013 to 2015. We analyzed SO42- and CH4 concentrations and identified the SMTZ for the last 3 years. The depths of SMTZ for the cores obtained from 2013, 2014 and 2015 are approximately 3.50 mbsf, 5.00 mbsf, and 5.00 mbsf respectively. The analysis results indicate that the SMTZ in 2013, 2014, and 2015 are shallower than the SMTZ of 2010.

Microarray Meta-Analysis of RNA-Binding Protein Functions in Alternative Polyadenylation

PubMed Central

Hu, Wenchao; Liu, Yuting; Yan, Jun

2014-01-01

Alternative polyadenylation (APA) is a post-transcriptional mechanism to generate diverse mRNA transcripts with different 3′UTRs from the same gene. In this study, we systematically searched for the APA events with differential expression in public mouse microarray data. Hundreds of genes with over-represented differential APA events and the corresponding experiments were identified. We further revealed that global APA differential expression occurred prevalently in tissues such as brain comparing to peripheral tissues, and biological processes such as development, differentiation and immune responses. Interestingly, we also observed widespread differential APA events in RNA-binding protein (RBP) genes such as Rbm3, Eif4e2 and Elavl1. Given the fact that RBPs are considered as the main regulators of differential APA expression, we constructed a co-expression network between APAs and RBPs using the microarray data. Further incorporation of CLIP-seq data of selected RBPs showed that Nova2 represses and Mbnl1 promotes the polyadenylation of closest poly(A) sites respectively. Altogether, our study is the first microarray meta-analysis in a mammal on the regulation of APA by RBPs that integrated massive mRNA expression data under a wide-range of biological conditions. Finally, we present our results as a comprehensive resource in an online website for the research community. PMID:24622240

Molecular cloning of an inducible serine esterase gene from human cytotoxic lymphocytes.

PubMed Central

Trapani, J A; Klein, J L; White, P C; Dupont, B

1988-01-01

A cDNA clone encoding a human serine esterase gene was isolated from a library constructed from poly(A)+ RNA of allogeneically stimulated, interleukin 2-expanded peripheral blood mononuclear cells. The clone, designated HSE26.1, represents a full-length copy of a 0.9-kilobase mRNA present in human cytotoxic cells but absent from a wide variety of noncytotoxic cell lines. Clone HSE26.1 contains an 892-base-pair sequence, including a single 741-base-pair open reading frame encoding a putative 247-residue polypeptide. The first 20 amino acids of the polypeptide form a leader sequence. The mature protein is predicted to have an unglycosylated Mr of approximately equal to 26,000 and contains a single potential site for N-linked glycosylation. The nucleotide and predicted amino acid sequences of clone HSE26.1 are homologous with all murine and human serine esterases cloned thus far but are most similar to mouse granzyme B (70% nucleotide and 68% amino acid identity). HSE26.1 protein is expressed weakly in unstimulated peripheral blood mononuclear cells but is strongly induced within 6-hr incubation in medium containing phytohemagglutinin. The data suggest that the protein encoded by HSE26.1 plays a role in cell-mediated cytotoxicity. Images PMID:3261871

Hfq variant with altered RNA binding functions

PubMed Central

Ziolkowska, Katarzyna; Derreumaux, Philippe; Folichon, Marc; Pellegrini, Olivier; Régnier, Philippe; Boni, Irina V.; Hajnsdorf, Eliane

2006-01-01

The interaction between Hfq and RNA is central to multiple regulatory processes. Using site-directed mutagenesis, we have found a missense mutation in Hfq (V43R) which strongly affects2 the RNA binding capacity of the Hfq protein and its ability to stimulate poly(A) tail elongation by poly(A)-polymerase in vitro. In vivo, overexpression of this Hfq variant fails to stimulate rpoS–lacZ expression and does not restore a normal growth rate in hfq null mutant. Cells in which the wild-type gene has been replaced by the hfqV43R allele exhibit a phenotype intermediate between those of the wild-type and of the hfq minus or null strains. This missense mutation derepresses Hfq synthesis. However, not all Hfq functions are affected by this mutation. For example, HfqV43R represses OppA synthesis as strongly as the wild-type protein. The dominant negative effect of the V43R mutation over the wild-type allele suggests that hexamers containing variant and genuine subunits are presumably not functional. Finally, molecular dynamics studies indicate that the V43R substitution mainly changes the position of the K56 and Y55 side chains involved in the Hfq–RNA interaction but has probably no effect on the folding and the oligomerization of the protein. PMID:16449205

The complete genome sequence of a south Indian isolate of Rice tungro spherical virus reveals evidence of genetic recombination between distinct isolates.

PubMed

Sailaja, B; Anjum, Najreen; Patil, Yogesh K; Agarwal, Surekha; Malathi, P; Krishnaveni, D; Balachandran, S M; Viraktamath, B C; Mangrauthia, Satendra K

2013-12-01

In this study, complete genome of a south Indian isolate of Rice tungro spherical virus (RTSV) from Andhra Pradesh (AP) was sequenced, and the predicted amino acid sequence was analysed. The RTSV RNA genome consists of 12,171 nt without the poly(A) tail, encoding a putative typical polyprotein of 3,470 amino acids. Furthermore, cleavage sites and sequence motifs of the polyprotein were predicted. Multiple alignment with other RTSV isolates showed a nucleotide sequence identity of 95% to east Indian isolates and 90% to Philippines isolates. A phylogenetic tree based on complete genome sequence showed that Indian isolates clustered together, while Vt6 and PhilA isolates of Philippines formed two separate clusters. Twelve recombination events were detected in RNA genome of RTSV using the Recombination Detection Program version 3. Recombination analysis suggested significant role of 5' end and central region of genome in virus evolution. Further, AP and Odisha isolates appeared as important RTSV isolates involved in diversification of this virus in India through recombination phenomenon. The new addition of complete genome of first south Indian isolate provided an opportunity to establish the molecular evolution of RTSV through recombination analysis and phylogenetic relationship.

Collection of LAI and FPAR Data Over The Terra Core Sites

NASA Technical Reports Server (NTRS)

Myneni, Ranga B.; Knjazihhin, J.; Tian, Y.; Wang, Y.

2001-01-01

The objective of our effort was to collect and archive data on LAI (leaf area index) and FPAR (Fraction of Photosynthetically active Radiation absorbed by vegetation) at the EOS Core validation sites as well as to validate and evaluate global fields of LAI and FPAR derived from atmospherically corrected MODIS (Moderate Resolution Imaging Spectrometer) surface reflectance data by comparing these fields with the EOS Core validation data set. The above has been accomplished by: (a) the participation in selected field campaigns within the EOS Validation Program; (b) the processing of the collected data so that suitable comparison between field measurements and the MODIS LAI/FPAR fields can be made; (c) the comparison of the MODAS LAI/FRAM fields with the EOS Terra Core validation data set.

Chew Bahir: A Key Site within the Hominin Sites and Paleolakes Drilling Project, towards a Half Million-Year Climate Record from Southern Ethiopia

NASA Astrophysics Data System (ADS)

Schaebitz, F.; Asrat, A.; Lamb, H. F.; Trauth, M. H.; Foerster, V. E.; Junginger, A.; Raub, T. D.; Gromig, R.; Viehberg, F. A.; Roberts, H. M.; Cohen, A.

2015-12-01

Chew Bahir, a saline mudflat today, is one of the five sites in East Africa, drilled within the framework of HSPDP (Hominin Site and Paleolakes Drilling Project). It is also one of the key sites of the Collaborative Research Centre (CRC-806) "Our way to Europe" aiming at the reconstruction of environmental conditions in the source region of modern man (H. sapiens). It is suggested that a changing environment could have triggered the mobility and dispersal of modern man. The oldest known fossils of anatomical modern humans (~195 ka BP) were found in the Omo basin, not more than 90km westwards of our drill site. The deposits in the tectonic basin of Chew Bahir in southern Ethiopia were cored in Nov. 2014 in two boreholes down to 280 m and 260 m below surface respectively. The overlapping long cores (drilled ~20 m apart from each other), were opened, scanned, described and sampled in low resolution in April 2015. The recovered sediments mostly contain green-greyish to light coloured and brown to reddish clays and silty clays, interbedded with some laminated mica-rich sand layers and occurrences of carbonate concretions and nodules, which decrease upcore. Here we will present a first set of results on the composite core, comprising mainly lithology and magnetic susceptibility (MS). Based on known sedimentation rates from pre-studies performed on short cores across the basin, we anticipate the deep drilled cores to cover at least 500 ka BP. Moreover, new insights into the role of post-depositional alteration, especially of clay minerals and zeolites, will be presented as a contribution to an improved understanding of formation processes. The results support the identification of wet and dry climate periods in the past. Those pronounced variations of moisture availability, are thought to have influenced the evolution and mobility of Homo sapiens sapiens.

Initial Results from the Deep Drilling of Lake Junin, Perú

NASA Astrophysics Data System (ADS)

Rodbell, D. T.; Abbott, M. B.; Weidhaas, N.; Hatfield, R. G.; Woods, A.; Hillman, A. L.; Tapia, P. M.; Chen, C. Y.; McGee, D.; Stoner, J. S.

2016-12-01

Lake Junín (11.0°S, 76.2°W, 4085 masl) is an intermontane, high-elevation lake in the inner-tropics of the Southern Hemisphere that spans 300 km2. With a maximum water depth of 12m, Lake Junin is dammed at its northern and southern ends by alluvial fans that emanate from glacial valleys in both cordillera. These fans can be traced to moraines that are >250 ka, indicating that the lake is at least this old. During the maximum extent of late Cenozoic glaciation, glaciers reached the lake edge but at no time over the last 1 million years, or more, has Lake Junín been overridden by ice. Lake Junín is thus one of the few lakes in the tropical Andes that predates the maximum extent of glaciation and is in a geomorphic position to record the waxing and waning of alpine glaciers in nearby cordillera. Sediment cores obtained between 1980 and 1996 reveal that sediment deposited during the last glacial cycle ( 30-16 ka) is dominated by glacial flour whereas sediment deposited during the last 16 ka consists predominantly of authigenic calcite (marl) with ostracod carapaces punctuated with intervals of gyttja and peat. In July and August of 2015, piston cores were obtained from three sites in Lake Junin. Multiple overlapping cores from the deepest water site (Site 1) extend to 100 m below lake floor (mblf), and those from two shallow water, paleoglacier-proximal sites (Sites 2 and 3) extend 23 and 51 mblf, respectively. Samples acquired at 8-cm resolution from Site 1 were analyzed for total organic carbon (TOC) and total inorganic carbon [as Ca(Mg)CO3; TIC] by coulometry. Total carbon (TC) was analyzed by combusting 10 mg samples at 1000°C and quantifying the resultant CO2 by coulometry whereas TIC was analyzed by reacting 10 mg samples in 6N H3PO4 and quantifying the resultant CO2 by coulometry; TOC was determined from TOC=TC-TIC. Over the last glacial postglacial cycle (last 30 ka), mean CaCO3 and TOC concentrations in Site 1 cores are higher ( 33% and 7.4%, respectively) than those in shallow water settings ( 9.5% and 4%). Similarly, mean magnetic susceptibility (MS) is lower in Site 1 cores (6.9 SI) than in the most paleoglacier-proximal shallow water site (Site 2, 9.4 SI). Site 1 records 7 glacial and interglacial cycles whereas shallow water locations appear to be dominated by sediment deposited during the last glacial-interglacial cycle.

The Chesapeake Bay Impact Crater: An Educational Investigation for Students into the Planetary Impact Process and its Environmental Consequences

NASA Technical Reports Server (NTRS)

Levine, Arlene S.

2008-01-01

Planetary impact craters are a common surface feature of many planetary bodies, including the Earth, the Moon, Mars, Mercury, Venus, and Jupiter s moons, Ganymede and Callisto. The NASA Langley Research Center in Hampton, VA, is located about 5 km inside the outer rim of the Chesapeake Bay Impact Crater. The Chesapeake Bay Impact Crater, with a diameter of 85 km is the sixth largest impact crater on our planet. The U.S. Geological Survey (USGS), in collaboration with the NASA Langley Research Center, the Virginia Department of Environmental Quality (VDEQ), the Hampton Roads Planning District Commission (HRPDC), and the Department of Geology of the College of William and Mary (WM) drilled into and through the crater at the NASA Langley Research Center and obtained a continuous core to a depth of 2075.9 ft (632.73 meters) from the Chesapeake Bay Impact Crater. At the NASA Langley location, the granite basement depth was at 2046 ft (623.87 meters). This collaborative drilling activity provided a unique educational opportunity and ongoing educational partnership between USGS, NASA Langley and the other collaborators. NASA Langley has a decade-long, ongoing educational partnership with the Colonial Coast Council of the Girl Scouts. The core drilling and on site analysis and cataloguing of the core segments provided a unique opportunity for the Girl Scouts to learn how geologists work in the field, their tools for scientific investigation and evaluation, how they perform geological analyses of the cores in an on-site tent and learn about the formation of impact craters and the impact of impacting bodies on the sub-surface, the surface, the oceans and atmosphere of the target body. This was accomplished with a two-part activity. Girl Scout day camps and local Girl Scout troops were invited to Langley Research Center Conference Center, where more than 300 Girl Scouts, their leaders and adult personnel were given briefings by scientists and educators from the USGS, NASA, VDEQ, HRPDC and WM on the principles of geology, the formation of impact craters, the consequences of the impacting body on the atmosphere, ocean, surface and sub-surface, the geological, chemical and biological analyses of the core and the cataloguing and storage of the core segments, etc. After the briefings, the Girl Scouts visited the drilling site where they inspected the core drilling rig, examined the core samples and discussed the drilling procedures, cores and interpretation of the cores with scientists and educators from the organizations conducting the core drilling. Demonstrations at the drilling site included demonstrations of impacting objects hitting multi-colored layered mud targets at different angles of entry. The multi-colored layers of mud were instructive in mapping out the distribution of impact-ejected material around the impact crater. The presentation will include a series of photographs of the Girl Scout participating in activities at the Chesapeake Bay Impact Crater drill site, including retrieving cores from the drilling rig, inspecting the core samples and participating in the impact-crater formation demonstrations.

98. View of IBM digital computer model 7090 magnet core ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

98. View of IBM digital computer model 7090 magnet core installation. ITT Artic Services, Inc., Official photograph BMEWS Site II, Clear, AK, by unknown photographer, 17 September 1965. BMEWS, clear as negative no. A-6606. - Clear Air Force Station, Ballistic Missile Early Warning System Site II, One mile west of mile marker 293.5 on Parks Highway, 5 miles southwest of Anderson, Anderson, Denali Borough, AK

Functional Analysis of Chk2-Kiaa0170 Interaction

DTIC Science & Technology

2006-09-01

terminal repeat; NEO, neomycin resistance gene; pA, poly-A; PGK, phosphoglycerate kinase-1; BTK , Bru- ton’s tyrosine kinase; SA and SD, splice acceptor...Briefly, MEFs were lysed in buffer I (50 mM HEPES [pH 7.5], 150 mM NaCl, 1 mM EDTA, 0.05% NP40, and protease and phosphatase inhibitors ) for 5 min on...0.5% DOC, 0.1% SDS, and protease and phosphatase inhibitors ) on ice for 20 min. The extracts were centrifuged at 14,000 rpm for 20 min at 4ºC. The

Detection of a divergent variant of grapevine virus F by next-generation sequencing.

PubMed

Molenaar, Nicholas; Burger, Johan T; Maree, Hans J

2015-08-01

The complete genome sequence of a South African isolate of grapevine virus F (GVF) is presented. It was first detected by metagenomic next-generation sequencing of field samples and validated through direct Sanger sequencing. The genome sequence of GVF isolate V5 consists of 7539 nucleotides and contains a poly(A) tail. It has a typical vitivirus genome arrangement that comprises five open reading frames (ORFs), which share only 88.96 % nucleotide sequence identity with the existing complete GVF genome sequence (JX105428).

Novel dicistrovirus from bat guano.

PubMed

Reuter, Gábor; Pankovics, Péter; Gyöngyi, Zoltán; Delwart, Eric; Boros, Akos

2014-12-01

A novel dicistrovirus (strain NB-1/2011/HUN, KJ802403) genome was detected from guano collected from an insectivorous bat (species Pipistrellus pipistrellus) in Hungary, using viral metagenomics. The complete genome of NB-1 is 9136 nt in length, excluding the poly(A) tail. NB-1 has a genome organization typical of a dicistrovirus with multiple 3B(VPg) and a cripavirus-like intergenic region (IGR)-IRES. NB-1 shares only 41 % average amino acid sequence identity with capsid proteins of Himetobi P virus, indicating a potential novel species in the genus Cripavirus, family Dicistroviridae.

Tree Ring Chronology Indexes and Reconstructions of Precipitation in Central Iowa, USA (1984) (NDP-002)

DOE Data Explorer

Blasing, T. J. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Duvick, D. N. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Building Technologies Research and Integration Center (BTRIC)

2012-01-01

Tree core samples (4 mm in diameter) were extracted from the trunks of white oak (Quercus alba) at three sites in central Iowa (Duvick Back Woods, Ledges State Park, and Pammel). At least 60 trees were sampled at each site, and at least two cores were taken from each tree. The growth rings of each core were dated by calendar year and measured; the measurements were then transformed into dimensionless ring-width indices and correlated with annual precipitation. Data were collected for the years 1680 through 1979. Each tree ring was characterized by the site, year, tree-ring-width index, number of core samples, decade year, and the annual reconstructed precipitation estimate. These data have more than 50% of their variance in common with the known annual statewide average precipitation for Iowa and serve as useful indicators of the precipitation and drought history of the region for the past 300 years. The data are in two files: tree-ring-chronology data (8 kB) and the annual reconstructed precipitation data for central Iowa (2 kB).

Linear extension rates and fluctuations of trace metals in Porites sp. from around Peninsular Malaysia

NASA Astrophysics Data System (ADS)

Amir, Liyana; Mohamed, Che Abd Rahim

2018-04-01

Coral cores were collected from P. Payar, Port Dickson, P. Redang and P. Tioman. The length of cores represented data spanning from year 2009 - 2015. Satellite sea surface temperatures from year 2009 - 2015 were obtained from the Reynolds and Smith dataset. Sr/Ca concentrations were measured from the coral powder taken at 1mm intervals along the vertical growth axis. Sea Surface Temperature (SST) was significantly higher during year 2010 in all four locations and linear extension was observed to have declined in year 2010 compared to year 2009 in cores from both sites. This decline coincides with the higher SST observed in year 2010 as a result of the El Niño event. Correlation analysis showed that Sr/Ca ratios in cores from all sites have a significant inverse relationship with SST. Analysis of the trace metals such as Pb, Ba, Cr and Cu produced results that were within the reported range in coral skeleton. Concentrations were significantly higher in Port Dickson and the lowest in P. Redang. These findings could be due to differences in terrestrial input at respective reef sites.

Analyses of water, core material, and elutriate samples collected near Buras, Louisiana (New Orleans to Venice, Louisiana, Hurricane Protection Project)

USGS Publications Warehouse

Leone, Harold A.

1977-01-01

Eight core-material-sampling sites were chosen by the U.S. Army Corps of Engineers as possible borrow areas for fill material to be used in levee contruction near Buras, La. Eleven receiving-water sites also were selected to represent the water that will contact the porposed levees. Analyses of selected nutrients, metals, pesticides, and other organic constitutents were performed upon these bed-material and native-water samples as well as upon elutriate samples of specific core material-receiving water systems. The results of these analyses are presented without interpretation. (Woodard-USGS)

Agreement between core laboratory and study investigators for imaging scores in a thrombectomy trial.

PubMed

Fahed, Robert; Ben Maacha, Malek; Ducroux, Célina; Khoury, Naim; Blanc, Raphaël; Piotin, Michel; Lapergue, Bertrand

2018-05-14

We aimed to assess the agreement between study investigators and the core laboratory (core lab) of a thrombectomy trial for imaging scores. The Alberta Stroke Program Early CT Score (ASPECTS), the European Collaborative Acute Stroke Study (ECASS) hemorrhagic transformation (HT) classification, and the Thrombolysis In Cerebral Infarction (TICI) scores as recorded by study investigators were compared with the core lab scores in order to assess interrater agreement, using Cohen's unweighted and weighted kappa statistics. There were frequent discrepancies between study sites and core lab for all the scores. Agreement for ASPECTS and ECASS HT classification was less than substantial, with disagreement occurring in more than one-third of cases. Agreement was higher on MRI-based scores than on CT, and was improved after dichotomization on both CT and MRI. Agreement for TICI scores was moderate (with disagreement occurring in more than 25% of patients), and went above the substantial level (less than 10% disagreement) after dichotomization (TICI 0/1/2a vs 2b/3). Discrepancies between scores assessed by the imaging core lab and those reported by study sites occurred in a significant proportion of patients. Disagreement in the assessment of ASPECTS and day 1 HT scores was more frequent on CT than on MRI. The agreement for the dichotomized TICI score (the trial's primary outcome) was substantial, with less than 10% of disagreement between study sites and core lab. NCT02523261, Post-results. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

Upward extension of the Nankai accretionary prism megasplay fault into slope basin strata. Insights from drilling at IODP Expedition 338 Site C0022

NASA Astrophysics Data System (ADS)

Fabbri, O.; Oohashi, K.; Kanagawa, K.; Yamaguchi, A.

2013-12-01

Megasplay faults have been recognized on seismic reflection profiles across several convergent margins in the world. Understanding the behavior of these faults during large to very large inter-plate earthquakes is a major challenge in assessing strong-motion and tsunami hazards at or near subduction zones. One of the goals of the IODP NanTroSEIZE project is to drill across and to obtain data from the megasplay fault crossing the Nankai accretionary prism off Kii peninsula (Kumano transect), SW Japan. This fault is considered to have been activated during the 1944 Tonankai earthquake (Baba et al., 2006 ; Moore et al., 2007). Drilling and coring during IODP Expedition 316 (Expedition 316 Scientists, 2009) showed that the megasplay fault at 300 mbsf at Site C0004 consists in a 60 m thick package of fractured and brecciated rocks. Combined analysis of 3D reflection data in the vicinity of Site C0004 and core data from sites C0004 and C0008 (Strasser et al., 2009 ; Kimura et al., 2011) suggest that the lower boundary of the megasplay fault ceased activity at about 1.55 Ma while its upper boundary has remained active since about 1.95 Ma and probably 1.24 Ma. In order to determine whether the megasplay fault upper boundary crosscuts slope sediments or is sealed by them, drilling at IODP Site C0022 was carried out during Expedition 338. Two 420 m deep holes were drilled: C0022A (LWD) and C0022B (coring). At Hole C0022A, LWD resistivity images show that the 85-105.5 mbsf interval is fractured and extends above and below a ca. 1 m thick interval characterized by a low resistivity value at 100-101 mbsf. Structures observed in cores from Hole C0022B confirm LWD data. While gently dipping elsewhere, bedding in the 73-146 mbsf interval is steep, commonly exceeding 30°. This bedding dip increase may be a consequence of fault activity (folding ?). Though the low-resistivity interval at 100-101 mbsf could not be sampled at Hole C0022B (no recovery between 95.5 and 99.5 mbsf), cores immediately from above this interval show three ca. 2 cm thick zones of claystone characterized by a marked planar fabric bearing faint striations raking at about 90°. Preliminary biostratigraphic dating in Hole C0022B indicate age reversals at 80.5, 137.5 and 145.5 mbsf, suggesting reverse offset bringing older strata over younger strata. Drilling at IODP Site C0022 confirms that a branch of the megasplay fault previously cored at Expedition 316 Site C0004 extends upwards and southeastwards. The core zone of this branch lies at about 100 mbsf and is about 1 m thick. The presence of weakly foliated claystone suggests aseismic motion immediately above the core zone. The lack of samples from the core zone prevents to determine whether motion was aseismic or not.

Reconstructing the Quaternary evolution of the world's most active silicic volcanic system: insights from an ˜1.65 Ma deep ocean tephra record sourced from Taupo Volcanic Zone, New Zealand

NASA Astrophysics Data System (ADS)

Allan, Aidan S. R.; Baker, Joel A.; Carter, Lionel; Wysoczanksi, Richard J.

2008-12-01

The Taupo Volcanic Zone (TVZ), central North Island, New Zealand, is the most frequently active Quaternary rhyolitic system in the world. Silicic tephras recovered from Ocean Drilling Programme Site 1123 (41°47.16'S, 171°29.94'W; 3290 m water depth) in the southwest Pacific Ocean provide a well-dated record of explosive TVZ volcanism since ˜1.65 Ma. We present major, minor and trace element data for 70 Quaternary tephra layers from Site 1123 determined by electron probe microanalysis (1314 analyses) and laser ablation inductively coupled plasma mass spectrometry (654 analyses). Trace element data allow for the discrimination of different tephras with similar major element chemistries and the establishment of isochronous tie-lines between three sediment cores (1123A, 1123B and 1123C) recovered from Site 1123. These tephra tie-lines are used to evaluate the stratigraphy and orbitally tuned stable isotope age model of the Site 1123 composite record. Trace element fingerprinting of tephras identifies ˜4.5 m and ˜7.9 m thick sections of repeated sediments in 1123A (49.0-53.5 mbsf [metres below seafloor]) and 1123C (48.1-56.0 mbsf), respectively. These previously unrecognised repeated sections have resulted in significant errors in the Site 1123 composite stratigraphy and age model for the interval 1.15-1.38 Ma and can explain the poor correspondence between δ18O profiles for Site 1123 and Site 849 (equatorial Pacific) during this interval. The revised composite stratigraphy for Site 1123 shows that the 70 tephra layers, when correlated between cores, correspond to ˜37-38 individual eruptive events (tephras), 7 of which can be correlated to onshore TVZ deposits. The frequency of large-volume TVZ-derived silicic eruptions, as recorded by the deposition of tephras at Site 1123, has not been uniform through time. Rather it has been typified by short periods (25-50 ka) of intense activity bracketed by longer periods (100-130 ka) of quiescence. The most active period (at least 1 event per 7 ka) occurred between ˜1.53 and 1.66 Ma, corresponding to the first ˜130 ka of TVZ rhyolitic magmatism. Since 1.2 Ma, ˜80% of tephras preserved at Site 1123 and the more proximal Site 1124 were erupted and deposited during glacial periods. This feature may reflect either enhanced atmospheric transport of volcanic ash to these sites (up to 1000 km from source) during glacial conditions or, more speculatively, that these events are triggered by changes in crustal stress accumulation associated with large amplitude sea-level changes. Only 8 of the ˜37-38 Site 1123 tephra units (˜20%) can be found in all three cores, and 22 tephra units (˜60%) are only present in one of the three cores. Whether a tephra is preserved in all three cores does not have any direct relationship to eruptive volume. Instead it is postulated that tephra preservation at Site 1123 is 'patchy' and influenced by the vigorous nature of their deposition to the deep ocean floor as vertical density currents. At this site, at least 5 cores would need to have been drilled within a proximity of 10's to 100's of metres of each other to yield a >99% chance of recovering all the silicic tephras deposited on the ocean surface above it in the past 1.65 Ma.

Field Performance of Three-Phase Amorphous Metal Core Distribution Transformers at Pearl Harbor, Hawaii

DTIC Science & Technology

1990-08-01

transformer core, such as loose or fractured core laminations . A sound level meter with an A- weighting frequency network was used for the...loaded on flatbed trucks as shown in Figure 2 and permanently installed at various sites throughout the Pearl Harbor complex. Figure 3 shows the final

Controlled Synthesis of Pd/Pt Core Shell Nanoparticles Using Area-selective Atomic Layer Deposition

PubMed Central

Cao, Kun; Zhu, Qianqian; Shan, Bin; Chen, Rong

2015-01-01

We report an atomic scale controllable synthesis of Pd/Pt core shell nanoparticles (NPs) via area-selective atomic layer deposition (ALD) on a modified surface. The method involves utilizing octadecyltrichlorosilane (ODTS) self-assembled monolayers (SAMs) to modify the surface. Take the usage of pinholes on SAMs as active sites for the initial core nucleation, and subsequent selective deposition of the second metal as the shell layer. Since new nucleation sites can be effectively blocked by surface ODTS SAMs in the second deposition stage, we demonstrate the successful growth of Pd/Pt and Pt/Pd NPs with uniform core shell structures and narrow size distribution. The size, shell thickness and composition of the NPs can be controlled precisely by varying the ALD cycles. Such core shell structures can be realized by using regular ALD recipes without special adjustment. This SAMs assisted area-selective ALD method of core shell structure fabrication greatly expands the applicability of ALD in fabricating novel structures and can be readily applied to the growth of NPs with other compositions. PMID:25683469

Large-scale identification of core-fucosylated glycopeptide sites in pancreatic cancer serum using mass spectrometry.

PubMed

Tan, Zhijing; Yin, Haidi; Nie, Song; Lin, Zhenxin; Zhu, Jianhui; Ruffin, Mack T; Anderson, Michelle A; Simeone, Diane M; Lubman, David M

2015-04-03

Glycosylation has significant effects on protein function and cell metastasis, which are important in cancer progression. It is of great interest to identify site-specific glycosylation in search of potential cancer biomarkers. However, the abundance of glycopeptides is low compared to that of nonglycopeptides after trypsin digestion of serum samples, and the mass spectrometric signals of glycopeptides are often masked by coeluting nonglycopeptides due to low ionization efficiency. Selective enrichment of glycopeptides from complex serum samples is essential for mass spectrometry (MS)-based analysis. Herein, a strategy has been optimized using LCA enrichment to improve the identification of core-fucosylation (CF) sites in serum of pancreatic cancer patients. The optimized strategy was then applied to analyze CF glycopeptide sites in 13 sets of serum samples from pancreatic cancer, chronic pancreatitis, healthy controls, and a standard reference. In total, 630 core-fucosylation sites were identified from 322 CF proteins in pancreatic cancer patient serum using an Orbitrap Elite mass spectrometer. Further data analysis revealed that 8 CF peptides exhibited a significant difference between pancreatic cancer and other controls, which may be potential diagnostic biomarkers for pancreatic cancer.

Seismic-reflection data on the eastern U.S. continental shelf acquired by M. V. L'OLONNOIS as part of the Atlantic Margin Coring Project (AMCOR) of the U.S. Geological Survey, July-September 1976

USGS Publications Warehouse

Robb, James M.

1980-01-01

In 1976 the U.S. Geological Survey undertook a program to sample the eastern United States Shelf for stratigraphic information by drilling a set of core holes. Results of this Atlantic Margin Coring Program (AMCOR) have been reported by Hathaway and others. Sites were chosen from seismic-reflection data and were reviewed by a safety panel to minimize the risk of penetrating any hydrocarbon accumulation which might lead to environmental contamination.The M-V-L'OLONNOIS, the service ship for the drilling operation, was fitted with seismic-reflection profiling equipment (listed below), to run seismic-reflection profiles before drilling began on each hole. This provided additional assurance that no closed structures would be penetrated and allowed minor adjustment with the site selection. A total of 491 km of high-resolution seismic profiles was collected on 22 sites.Equipment used (specifics for each site noted on records): Bolt Air Guns 1-40 cubic inch chambers EPC Recorder Teledyne Minisparker (last two sites) Navigation used two Internav 101 Loran-C receivers.

Historical contributions of phosphorus from natural and agricultural sources and implications for stream water quality, Cheney Reservoir watershed, south-central Kansas

USGS Publications Warehouse

Pope, Larry M.; Milligan, Chad R.; Mau, David Phillip

2002-01-01

An examination of soil cores collected from 43 nonagricultural coring sites in the Cheney Reservoir watershed of south-central Kansas was conducted by the U.S. Geological Survey in September 1999. The cores were collected as part of an ongoing cooperative study with the city of Wichita, Kansas. The 43 sites (mostly cemeteries) were thought to have total phosphorus concentrations in the soil that are representative of natural conditions (unaffected by human activity). The purpose of this report is to present the analysis and evaluation of these soil cores, to quantify the phosphorus contributions to Cheney Reservoir from natural and agricultural sources, and to provide estimates of stream-water-quality response to natural concentrations of total phosphorus in the soil. Analysis of soil cores from the 43 sites produced natural concentrations of total phosphorus that ranged from 74 to 539 milligrams per kilogram with a median concentration of 245 milligrams per kilogram in 2-inch soil cores and from 50 to 409 milligrams per kilogram with a median concentration of 166 milligrams per kilogram in 8-inch soil cores. Natural concentrations of total phosphorus in soil were statistically larger in samples from coring sites in the eastern half of the watershed than in samples from coring sites in the western half of the watershed. This result partly explains a previously determined west-to-east increase in total phosphorus yields in streams of the Cheney Reservoir watershed. A comparison of total phosphorus concentrations in soil under natural conditions to the historical mean total phosphorus concentration in agriculturally enriched bottom sediment in Cheney Reservoir indicated that agricultural activities within the watershed have increased total phosphorus concentrations in watershed soil that is transported in streams to about 2.9 times natural concentrations. Retention efficiencies for phosphorus and sediment historically transported to Cheney Reservoir were calculated at 92 and 99 percent, respectively. Most of the phosphorus was retained in bottom sediment. Sediment accumulation in Cheney Reservoir was less than reservoir design-life specifications on the basis of the age of the reservoir. Estimates of mean total phosphorus concentrations for selected streams in the Cheney Reservoir watershed under natural concentrations of total phosphorus in soil and a historic set of watershed conditions indicate that water from two of the five streamflow sampling sites would not meet the total phosphorus water-quality goal of 0.10 milligram per liter established by the Cheney Reservoir Watershed Task Force Committee. These results imply that the water-quality goal for total phosphorus in some streams of the watershed may not be met simply by reducing the amount of phosphorus applied. Instead, meeting the goal could involve a combination of approaches-for example, reducing the agricultural distribution of phosphorus and implementing changes in watershed activities to mitigate phosphorus movement to surface water.

Microhabitat Conditions in Wyoming’s Sage-Grouse Core Areas: Effects on Nest Site Selection and Success

PubMed Central

Beck, Jeffrey L.; Kirol, Christopher P.; Pratt, Aaron C.; Conover, Michael R.

2016-01-01

The purpose of our study was to identify microhabitat characteristics of greater sage-grouse (Centrocercus urophasianus) nest site selection and survival to determine the quality of sage-grouse habitat in 5 regions of central and southwest Wyoming associated with Wyoming’s Core Area Policy. Wyoming’s Core Area Policy was enacted in 2008 to reduce human disturbance near the greatest densities of sage-grouse. Our analyses aimed to assess sage-grouse nest selection and success at multiple micro-spatial scales. We obtained microhabitat data from 928 sage-grouse nest locations and 819 random microhabitat locations from 2008–2014. Nest success was estimated from 924 nests with survival data. Sage-grouse selected nests with greater sagebrush cover and height, visual obstruction, and number of small gaps between shrubs (gap size ≥0.5 m and <1.0 m), while selecting for less bare ground and rock. With the exception of more small gaps between shrubs, we did not find any differences in availability of these microhabitat characteristics between locations within and outside of Core Areas. In addition, we found little supporting evidence that sage-grouse were selecting different nest sites in Core Areas relative to areas outside of Core. The Kaplan-Meier nest success estimate for a 27-day incubation period was 42.0% (95% CI: 38.4–45.9%). Risk of nest failure was negatively associated with greater rock and more medium-sized gaps between shrubs (gap size ≥2.0 m and <3.0 m). Within our study areas, Wyoming’s Core Areas did not have differing microhabitat quality compared to outside of Core Areas. The close proximity of our locations within and outside of Core Areas likely explained our lack of finding differences in microhabitat quality among locations within these landscapes. However, the Core Area Policy is most likely to conserve high quality habitat at larger spatial scales, which over decades may have cascading effects on microhabitat quality available between areas within and outside of Core Areas. PMID:27002531

48 CFR 301.607-71 - FAC-P/PM levels and requirements.

Code of Federal Regulations, 2011 CFR

2011-10-01

.... (2) The FAI describes the following three sets of general core competencies on its Web site: (3..., systems engineering, test and evaluation, contracting, and business. (6) Specific core competencies also...

40 CFR 35.6305 - Obtaining supplies.

Code of Federal Regulations, 2011 CFR

2011-07-01

....6325 through 35.6340, and 35.6350. Supplies obtained with Core Program funds must be for non-site-specific purposes. All purchases of supplies under the Core Program must comply with the requirements in...

40 CFR 35.6305 - Obtaining supplies.

Code of Federal Regulations, 2010 CFR

2010-07-01

....6325 through 35.6340, and 35.6350. Supplies obtained with Core Program funds must be for non-site-specific purposes. All purchases of supplies under the Core Program must comply with the requirements in...

Contaminated Sediment Core Profiling

EPA Science Inventory

Evaluating the environmental risk of sites containing contaminated sediments often poses major challenges due in part to the absence of detailed information available for a given location. Sediment core profiling is often utilized during preliminary environmental investigations ...

The Hominin Sites and Paleolakes Drilling Project: inferring the environmental context of human evolution from eastern African rift lake deposits

NASA Astrophysics Data System (ADS)

Cohen, A.; Campisano, C.; Arrowsmith, R.; Asrat, A.; Behrensmeyer, A. K.; Deino, A.; Feibel, C.; Hill, A.; Johnson, R.; Kingston, J.; Lamb, H.; Lowenstein, T.; Noren, A.; Olago, D.; Owen, R. B.; Potts, R.; Reed, K.; Renaut, R.; Schäbitz, F.; Tiercelin, J.-J.; Trauth, M. H.; Wynn, J.; Ivory, S.; Brady, K.; O'Grady, R.; Rodysill, J.; Githiri, J.; Russell, J.; Foerster, V.; Dommain, R.; Rucina, S.; Deocampo, D.; Russell, J.; Billingsley, A.; Beck, C.; Dorenbeck, G.; Dullo, L.; Feary, D.; Garello, D.; Gromig, R.; Johnson, T.; Junginger, A.; Karanja, M.; Kimburi, E.; Mbuthia, A.; McCartney, T.; McNulty, E.; Muiruri, V.; Nambiro, E.; Negash, E. W.; Njagi, D.; Wilson, J. N.; Rabideaux, N.; Raub, T.; Sier, M. J.; Smith, P.; Urban, J.; Warren, M.; Yadeta, M.; Yost, C.; Zinaye, B.

2016-02-01

The role that climate and environmental history may have played in influencing human evolution has been the focus of considerable interest and controversy among paleoanthropologists for decades. Prior attempts to understand the environmental history side of this equation have centered around the study of outcrop sediments and fossils adjacent to where fossil hominins (ancestors or close relatives of modern humans) are found, or from the study of deep sea drill cores. However, outcrop sediments are often highly weathered and thus are unsuitable for some types of paleoclimatic records, and deep sea core records come from long distances away from the actual fossil and stone tool remains. The Hominin Sites and Paleolakes Drilling Project (HSPDP) was developed to address these issues. The project has focused its efforts on the eastern African Rift Valley, where much of the evidence for early hominins has been recovered. We have collected about 2 km of sediment drill core from six basins in Kenya and Ethiopia, in lake deposits immediately adjacent to important fossil hominin and archaeological sites. Collectively these cores cover in time many of the key transitions and critical intervals in human evolutionary history over the last 4 Ma, such as the earliest stone tools, the origin of our own genus Homo, and the earliest anatomically modern Homo sapiens. Here we document the initial field, physical property, and core description results of the 2012-2014 HSPDP coring campaign.

Search for continuous paleoclimatic record in Holocene lacustrine sediments from Lake District, Chile (40°S)

NASA Astrophysics Data System (ADS)

Bertrand, S.; Fagel, N.

2003-04-01

Our aim is to reconstruct a continuous Holocene climatic evolution related to ENSO variability in southern Chile. We focus on the sedimentary infilling of two glacial lakes from the Lake District Area (38-40°S). The preliminar sedimentological analysis must allow to define the key-site and the best palaeoclimatic proxies. This area, at the foothill of the Cordillera de Los Andes, has been affected by an intense Quaternary volcanic activity and by several historical earthquakes (e.g., Valdivia 1960). After preliminary seismic investigation, four cores were recovered in two lakes selected at the two ends of a N-S transect. (1) Icalma lake (12 km2, 38°S) is located in the Cordillera de Los Andes at an elevation of 1150m and results of the infilling of a glacial umbilic. The two 8m sediment cores consist of an alternation of laminated silts and volcanic layers. The sedimentary record is strongly disturbed by numerous seismic or volcanic events. The cumulated volcanogenic-derived material represents up to 50 % of the core length. In particular, the cores record at -4.50m a pumice layer widespread in the watershed and dated at 2900 yr BP. One core contains pluricentrimetric layers of wood accumulation. They could be due to earthquake impact on the vegetational cover in the watershed. An ubiquitous 6cm-thick slump described at -40 cm may be related with the 1960 Valdivia earthquake. (2) On Puyehue lake (164 km2, 40°S, elevation 185 m), two cores (7 and 11m) have been collected in both underflow and interflow sites. The interflow site (PUII) shows a very well laminated sediment, with only small disturbances due to volcanic and seismic activities. This core will be the key-site for the paleoclimatic study. The second core (PUI) is very rich in organic matter. The sediment is strongly destratified by numerous gas bubbles (methane). These characteristics are due to the dense vegetational cover in the watershed and to the core location near the delta of the main river. This preliminary study emphasizes the influence of geodynamic conditions on sedimentology of lacustrine deposits in a geodynamically active region. The contribution of the punctual tectonic and volcanic-derived layers to sediment thickness must be taken into account before calculation of the age-depth model.

Temporal shifts in reef lagoon sediment composition, Discovery Bay, Jamaica

NASA Astrophysics Data System (ADS)

Perry, Christopher T.; Taylor, Kevin G.; Machent, Philip G.

2006-03-01

Discovery Bay, north Jamaica, forms a large (1.5 km wide), deep (up to 56 m) embayment that acts as a sink for reef-derived and lagoonal carbonate sediments. Since the mid-1960s, the bay has also provided a sink for inputs of bauxite sediment that are spilled during loading at a boat terminal constructed within Discovery Bay. Bauxite has accumulated across much of the southern section of the bay with surficial sediments presently composed of up to 35 weight% non-carbonate. Cores recovered from sites on the western side of the bay provide a stratigraphic record of this history of bauxite contamination across water depths from 5 to 25 m. The bauxite-influenced upper sediment horizons are clearly visible in each core from the distinctive red-brown colouration of the sediment. These sediments are composed of approximately 10% non-carbonate (bauxite) and have Fe contents of around 2-3000 μg/g (up to 7000 μg/g). The thickness of this upper bauxite-contaminated sequence increases down transect (approximately 18 cm in the shallowest core, to around 47 cm in the deepest core), and in each core overlies a sequence of 'clean' lagoon carbonates. These typically are poorly sorted carbonate sands with variable amounts of coral rubble. Down-core data on CaCO 3 and Fe content provide a chemical record of decreasing sediment contamination with depth, with the lower 'clean' carbonates composed of only around 2% non-carbonate and <700 μg/g Fe. Down-core sediment-constituent data also indicate significant changes in sediment production at the shallowest sites. At depths of 5 and 10 m, sediment assemblages have shifted from diverse assemblages of coral, mollusc, Amphiroa and Halimeda in the clean lagoon sands, to assemblages dominated by Halimeda and Amphiroa within the surficial sediments. At the deeper sites, no major down-core shifts in sediment constituents occur. These sites thus record a rather complex history of changes in sediment composition and chemistry. Clear shifts in chemistry and stratigraphy occur in all the cores and reflect progressive bauxite contamination in the near-surface horizons. These inputs, however, do not appear to have directly affected carbonate production, since the major constituent changes appear to be a response to more regional declines in coral community and reef status.

Petrophysical characterization of first ever drilled core samples from an active CO2 storage site, the German Ketzin Pilot Site - Comparison with long term experiments

NASA Astrophysics Data System (ADS)

Zemke, Kornelia; Liebscher, Axel

2014-05-01

Petrophysical properties like porosity and permeability are key parameters for a safe long-term storage of CO2 but also for the injection operation itself. These parameters may change during and/or after the CO2 injection due to geochemical reactions in the reservoir system that are triggered by the injected CO2. Here we present petrophysical data of first ever drilled cores from a newly drilled well at the active CO2 storage site - the Ketzin pilot site in the Federal State of Brandenburg, Germany. By comparison with pre-injection baseline data from core samples recovered prior to injection, the new samples provide the unique opportunity to evaluate the impact of CO2 on pore size related properties of reservoir and cap rocks at a real injection site under in-situ reservoir conditions. After injection of 61 000 tons CO2, an additional well was drilled and new rock cores were recovered. In total 100 core samples from the reservoir and the overlaying caprock were investigated by NMR relaxation. Permeability of 20 core samples was estimated by nitrogen and porosity by helium pycnometry. The determined data are comparable between pre-injection and post-injection core samples. The lower part of the reservoir sandstone is unaffected by the injected CO2. The upper part of the reservoir sandstone shows consistently slightly lower NMR porosity and permeability values in the post-injection samples when compared to the pre-injection data. This upper sandstone part is above the fluid level and CO2 present as a free gas phase and a possible residual gas saturation of the cores distorted the NMR results. The potash-containing drilling fluid can also influence these results: NMR investigation of twin samples from inner and outer parts of the cores show a reduced fraction of larger pores for the outer core samples together with lower porosities and T2 times. The drill mud penetration depth can be controlled by the added fluorescent tracer. Due to the heterogeneous character of the Stuttgart Formation it is difficult to estimate definite CO2 induced changes from petrophysical measurements. The observed changes are only minor. Several batch experiments on Ketzin samples drilled prior injection confirm the results from investigation of the in-situ rock cores. Core samples of the pre-injection wells were exposed to CO2 and brine in autoclaves over various time periods. Samples were characterized prior to and after the experiments by NMR and Mercury Injection Porosimetry (MIP). The results are consistent with the logging data and show only minor change. Unfortunately, also in these experiments observed mineralogical and petrophysical changes were within the natural heterogeneity of the Ketzin reservoir and precluded unequivocal conclusions. However, given the only minor differences between post-injection well and pre-injection well, it is reasonable to assume that the potential dissolution-precipitation processes appear to have no severe consequences on reservoir and cap rock integrity or on the injection behaviour. This is also in line with the continuously recorded injection operation parameter. These do not point to any changes in reservoir injectivity.|

Cretaceous paleoceanography of the western North Atlantic Ocean

USGS Publications Warehouse

Arthur, Michael A.; Dean, Walter E.

1986-01-01

In this paper we summarize available information on the Cretaceous lithostratigraphy and paleoceanography of the western North Atlantic. The data and some of our interpretations draw in large part on papers published in the Deep Sea Drilling Project (DSDP) volumes. We have attempted to cite relevant references when possible, but space limitations make it difficult to give proper credit to all sources; we apologize for any omissions.Organic carbon (Corg) and carbonate (CaCO3) analyses were tabulated for each site from papers in the DSDP Initial Report volumes and other published works (e.g., Summerhayes,1981). Corg, CaCO3, and non-CaCO3 mass accumulation rates (MARS) were calculated using core by core averages of component percentages for the more continuously cored sites; core averages for wet bulk density and porosity (from DSDP data files); biostratigraphies of de Graciansky and others (1982), Roth and Bowdler (1981), and Cool (1982); and the time scales of the Decade of North American Geology (Palmer, 1983; Kent and Gradstein, this volume) or Harland and others (1982; see Plate 1).Backtracked paleodepths for western North Atlantic DSDP Sites from Tucholke and Vogt (1979) with the revised stratigraphy of de Graciansky and others (1982) were used in plotting Corg and CaCO3 in Figures 2, 3, 4 and 5 (see also Thierstein, 1979).Backtracking curves of seafloor paleodepth versus age (Sclater and others, 1977; Tucholke and Vogt, 1979) for selected western North Atlantic DSDP sites. Average CaCO3 concentrations per core are shown by code number

Report of the Peer Review Panel on the early site suitability evaluation of the Potential Repository Site at Yucca Mountain, Nevada; Yucca Mountain Site Characterization Project

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

1992-01-01

The US Department of Energy (DOE) Yucca mountain Site Characterization Project Office (YMPO) assigned Science Applications International Corporation (SAIC), the Technical and Management Support Services (T&MSS) contractor to the YmPo, the task of conducting an Early Site Suitability Evaluation (ESSE) of the Yucca mountain site as a potential site for a high-level radioactive waste repository. First, the assignment called for the development of a method to evaluate a single site against the DOE General Guidelines for Recommendation of Sites for Nuclear Waste Repositories, 10 CFR Part 960. Then, using this method, an evaluation team, the ESSE Core Team, of seniormore » YMP scientists, engineers, and technical experts, evaluated new information obtained about the site since publication of the final Environmental Assessment (DOE, 1986) to determine if new suitability/unsuitability findings could be recommended. Finally, the Core Team identified further information and analyses needed to make final determinations for each of the guidelines. As part of the task, an independent peer review of the ESSE report has been conducted. Expertise was solicited that covered the entire spectrum of siting guidelines in 10 CFR Part 960 in order to provide a complete, in-depth critical review of the data evaluated and cited in the ESSE report, the methods used to evaluate the data, and the conclusions and recommendations offered by the report. Fourteen nationally recognized technical experts (Table 2) served on the Peer Review Panel. The comments from the Panel and the responses prepared by the ESSE Core Team, documented on formal Comment Response Forms, constitute the body of this document.« less

Sampling for Soil Carbon Stock Assessment in Rocky Agricultural Soils

NASA Technical Reports Server (NTRS)

Beem-Miller, Jeffrey P.; Kong, Angela Y. Y.; Ogle, Stephen; Wolfe, David

2016-01-01

Coring methods commonly employed in soil organic C (SOC) stock assessment may not accurately capture soil rock fragment (RF) content or soil bulk density (rho (sub b)) in rocky agricultural soils, potentially biasing SOC stock estimates. Quantitative pits are considered less biased than coring methods but are invasive and often cost-prohibitive. We compared fixed-depth and mass-based estimates of SOC stocks (0.3-meters depth) for hammer, hydraulic push, and rotary coring methods relative to quantitative pits at four agricultural sites ranging in RF content from less than 0.01 to 0.24 cubic meters per cubic meter. Sampling costs were also compared. Coring methods significantly underestimated RF content at all rocky sites, but significant differences (p is less than 0.05) in SOC stocks between pits and corers were only found with the hammer method using the fixed-depth approach at the less than 0.01 cubic meters per cubic meter RF site (pit, 5.80 kilograms C per square meter; hammer, 4.74 kilograms C per square meter) and at the 0.14 cubic meters per cubic meter RF site (pit, 8.81 kilograms C per square meter; hammer, 6.71 kilograms C per square meter). The hammer corer also underestimated rho (sub b) at all sites as did the hydraulic push corer at the 0.21 cubic meters per cubic meter RF site. No significant differences in mass-based SOC stock estimates were observed between pits and corers. Our results indicate that (i) calculating SOC stocks on a mass basis can overcome biases in RF and rho (sub b) estimates introduced by sampling equipment and (ii) a quantitative pit is the optimal sampling method for establishing reference soil masses, followed by rotary and then hydraulic push corers.

Did the onset of high amplitude glacio-eustatic cycles trigger mass-transport processes on the Northwest Shelf of Australia? Insights from IODP expedition 356

NASA Astrophysics Data System (ADS)

Gallagher, S. J.; McCaffrey, J.; Wallace, M. W.; Keep, M.; Fulthorpe, C.; Bogus, K.; McHugh, C.

2017-12-01

Mass-transport processes on continental margins may have catastrophic consequences, causing tsunamis, major rock falls and avalanches and can destroy offshore hydrocarbon installations. Mass-transport deposits (MTD's) with volumes 17 to >162 km3 are common along the northwest margin of Australia. One of the largest is the Gorgon slide which is offshore from Barrow Island with a minimum volume of 250 km3. Age estimates for slides on the Northwest Shelf are variable and range from Miocene to Recent (Gorgon MTD), late Pliocene to Recent (Thebe/Bonaventure MTD's) and Pleistocene to Recent. This age uncertainty is related to a lack of cored sections through these slides and relies on pre-existing ages and correlations from poorly dated sections (usually industry well sections with minimal samples in the upper 500 m) distal from the MTD's. Therefore, the age, origin and history of these MTD's is not well known. A recent International Ocean Discovery Program Expedition (IODP Expedition 356) to the region obtained a series of continuous cores from the upper 600m to 1.1 km of the Northern Carnarvon and Roebuck Basins. Four sites were cored adjacent to hydrocarbon wells; West Tryal Rocks-2 (Site U1461), Fisher-1 (Site U1462), Picard-1 (Site U1463) and Minilya-1 (Site U1464). Site U1461 yielded 100% core recovery through the Gorgon Slide. Preliminary data from this section suggests that it is relatively young (<1 Ma) with ongoing activity from 0.5 Ma continuing to today. We suggest neotectonism combined with the onset of high amplitude glacio-eustatic cycles may have been triggering factors for this slide.

Variations in soil detachment rates after wildfire as a function of soil depth, flow properties, and root properties

USGS Publications Warehouse

Moody, John A.; Nyman, Peter

2013-01-01

Wildfire affects hillslope erosion through increased surface runoff and increased sediment availability, both of which contribute to large post-fire erosion events. Relations between soil detachment rate, soil depth, flow and root properties, and fire impacts are poorly understood and not represented explicitly in commonly used post-fire erosion models. Detachment rates were measured on intact soil cores using a modified tilting flume. The cores were mounted flush with the flume-bed and a measurement was made on the surface of the core. The core was extruded upward, cut off, and another measurement was repeated at a different depth below the original surface of the core. Intact cores were collected from one site burned by the 2010 Fourmile Canyon (FMC) fire in Colorado and from one site burned by the 2010 Pozo fire in California. Each site contained contrasting vegetation and soil types. Additional soil samples were collected alongside the intact cores and were analyzed in the laboratory for soil properties (organic matter, bulk density, particle-size distribution) and for root properties (root density and root-length density). Particle-size distribution and root properties were different between sites, but sites were similar in terms of bulk density and organic matter. Soil detachment rates had similar relations with non-uniform shear stress and non-uniform unit stream power. Detachment rates within single sampling units displayed a relatively weak and inconsistent relation to flow variables. When averaged across all clusters, the detachment rate displayed a linear relation to shear stress, but variability in soil properties meant that the shear stress accounted for only a small proportion of the overall variability in detachment rates (R2 = 0.23; R2 is the coefficient of determination). Detachment rate was related to root-length density in some clusters (R2 values up to 0.91) and unrelated in others (R2 values 2 value improved and the range of exponents became narrower by applying a multivariate regression model where boundary shear stress and root-length density were included as explanatory variables. This suggests that an erodibility parameter which incorporates the effects of both flow and root properties on detachment could improve the representation of sediment availability after wildfire.

Analysis of core-periphery organization in protein contact networks reveals groups of structurally and functionally critical residues.

PubMed

Isaac, Arnold Emerson; Sinha, Sitabhra

2015-10-01

The representation of proteins as networks of interacting amino acids, referred to as protein contact networks (PCN), and their subsequent analyses using graph theoretic tools, can provide novel insights into the key functional roles of specific groups of residues. We have characterized the networks corresponding to the native states of 66 proteins (belonging to different families) in terms of their core-periphery organization. The resulting hierarchical classification of the amino acid constituents of a protein arranges the residues into successive layers - having higher core order - with increasing connection density, ranging from a sparsely linked periphery to a densely intra-connected core (distinct from the earlier concept of protein core defined in terms of the three-dimensional geometry of the native state, which has least solvent accessibility). Our results show that residues in the inner cores are more conserved than those at the periphery. Underlining the functional importance of the network core, we see that the receptor sites for known ligand molecules of most proteins occur in the innermost core. Furthermore, the association of residues with structural pockets and cavities in binding or active sites increases with the core order. From mutation sensitivity analysis, we show that the probability of deleterious or intolerant mutations also increases with the core order. We also show that stabilization centre residues are in the innermost cores, suggesting that the network core is critically important in maintaining the structural stability of the protein. A publicly available Web resource for performing core-periphery analysis of any protein whose native state is known has been made available by us at http://www.imsc.res.in/ ~sitabhra/proteinKcore/index.html.

Occurrence of and trends in selected sediment-associated contaminants in Caddo Lake, East Texas, 1940-2002

USGS Publications Warehouse

Wilson, Jennifer T.

2003-01-01

Bottom-sediment cores were collected from four sites in Caddo Lake in East Texas during May 2002 for analyses of radionuclides (for age dating), organochlorine pesticides, polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), and major and trace elements, and to describe the occurrence and trends of these sediment-associated contaminants. The Goose Prairie Creek and Harrison Bayou sites receive drainage from an area that includes parts of the now-closed Longhorn Army Ammunitions Plant. The mid-lake site is relatively close to dense oil and gas operations in the lake. The Carter Lake site receives minimal discharge from developed areas. Sediment age (deposition) dates represented in the cores ranged from 1940 to 2002. The only organochlorine compounds detected in all core samples were the DDT degradation products DDE or DDD, and PCB Aroclors 1242, 1254, and 1260 were detected only at the Goose Prairie Creek site. One or more of the DDE concentrations at all sites exceeded a consensus-based threshold effect concentration (on benthic biota), but none exceeded a consensus-based probable effect concentration. The Goose Prairie Creek site had significant downward trends in concentrations of organochlorine compounds, except for no trend in DDE concentrations. The Ammunitions Plant is a possible historical source of the few organochlorine compounds detected at the Goose Prairie Creek and Harrison Bayou sites. PAH concentrations at all sites were below respective threshold effect concentrations. Highest PAH concentrations at all four sites were of C2- alkylated naphthalenes. Nearly all statistically significant PAH trends in the cores were downward. On the basis of PAH source-indicator ratios, the majority of PAH compounds appear to have originated from uncombusted sources such as leaks or spills from oil and gas operations or vehicles (automobiles, boats, aircraft) in the Caddo Lake area. Concentrations of several of the eight trace elements with threshold effect concentrations and probable effect concentrations (among 26 analyzed) were above the respective threshold effect concentrations, but all, except one lead concentration at the Goose Prairie Creek site (deposited about 1961), were below respective probable effect concentrations. Among trace element concentrations at the four sites, lead and mercury were consistently relatively high at the Goose Prairie Creek site. Again the Ammunitions Plant, because of its proximity and history of industrial activities, is the suspected primary source. Statistically significant trends in trace element concentrations were mixed, but more were downward than upward. Computations to indicate the dominant source (atmospheric fallout or drainage area) of mercury to the Caddo Lake sediment core sites (except Carter Lake) indicate that about one-third of the mercury at the Goose Prairie Creek site might result from drainage area sources. No drainage area sources were indicated for the Harrison Bayou and mid-lake sites. Arsenic, cadmium, and zinc concentrations were highest at the Carter Lake site. No relation between the relatively higher trace element concentrations and any potential source of contamination in the Carter Lake drainage area (for example, oil and gas operations, a road, a boat ramp) is indicated.

Effects of urbanization and long-term rainfall on the occurrence of organic compounds and trace elements in reservoir sediment cores, streambed sediment, and fish tissue from the Santa Ana River basin, California, 1998

USGS Publications Warehouse

Burton, Carmen A.

2002-01-01

Organcochlorine compounds, semivolatile-organic compounds (SVOC), and trace elements were analyzed in reservoir sediment cores, streambed sediment, and fish tissue in the Santa Ana River Basin as part of the U.S. Geological Survey's National Water-Quality Assessment Program. Three reservoirs were sampled in areas that have different degrees of urbanization. Streambed sediment and fish tissue collected at 12 sites were divided into two groups, urban and nonurban. More organochlorine compounds were detected in reservoir sediment cores, streambed sediment and fish tissue, and at higher concentrations at urban sites than at nonurban sites. At all sites, except West Street Basin, concentrations of organochlorine compounds were lower than the probable-effect concentration (PEC). At the highly urbanized West Street Basin, chlordane and p,p'-DDE exceeded the PEC throughout the historical record. The less stringent threshold-effect concentration (TEC) was exceeded for six compounds at eight sites. Most of the organochlorine compounds detected in streambed sediment and fish tissue were at urban sites on the Santa Ana River as opposed to its tributaries, suggesting accumulation and persistence in the river. More SVOCs were detected in reservoir sediment cores and streambed sediment, and at higher concentrations, at urban sites than at nonurban sites. At all the sites, except West Street Basin, concentrations of SVOCs were lower than the PEC. At West Street Basin, chrysene, pyrene, and total polycyclic-aromatic hydrocarbons exceeded the PEC throughout the historical record. The TEC was exceeded for 10 compounds at 3 sites. Most of the SVOCs were detected in streambed sediment at urban sites on tributaries to the Santa Ana River rather than the mainstem itself. The less frequent occurrence and lower concentrations in the Santa Ana River suggest that SVOCs are less persistent than organochlorine compounds, possibly as a result of volatization, gradation, or dilution. Most trace-element detections in reservoir sediment cores and streambed sediment were at urban sites, and the concentrations were generally higher than at nonurban sites. Lead and zinc exceeded their PECs at West Street Basin throughout the historical record; copper exceeded its PEC at Canyon Lake, an area of urban growth. The TEC was exceeded for 10 compounds at 11 sites. Frequency of detection and concentration did not differ between tributary and Santa Ana River sites, which may be attributed to the fact that trace elements occur naturally. Four trace elements (arsenic, copper, mercury, and selenium) had higher concentrations in fish tissue at nonurban sites than at urban sites. Concentrations decreased over time for organochlorine compounds at all three reservoirs, probably a result of the discontinued use of many of the compounds. Decreasing trends in SVOCs and trace elements were observed at West Street Basin, but increasing trends were observed at Canyon Lake. Concentrations of organochlorine compounds, SVOCs, and trace elements were higher during periods of above average rainfall at both West Street Basin and Canyon Lake.

Paleolimnology of Lake Tanganyika, East Africa, over the past 100 k yr

USGS Publications Warehouse

Scholz, C.A.; King, J.W.; Ellis, G.S.; Swart, Peter K.; Stager, J.C.; Colman, Steven M.

2003-01-01

New sediment core data from a unique slow-sedimentation rate site in Lake Tanganyika contain a much longer and continuous record of limnological response to climate change than have been previously observed in equatorial regions of central Africa. The new core site was first located through an extensive seismic reflection survey over the Kavala Island Ridge (KIR), a sedimented basement high that separates the Kigoma and Kalemie Basins in Lake Tanganyika. Proxy analyses of paleoclimate response carried out on core T97-52V include paleomagnetic and index properties, TOC and isotopic analyses of organic carbon, and diatom and biogenic silica analyses. A robust age model based on 11 radiocarbon (AMS) dates indicates a linear, continuous sedimentation rate nearly an order of magnitude slower here compared to other core sites around the lake. This age model indicates continuous sedimentation over the past 79 k yr, and a basal age in excess of 100 k yr. The results of the proxy analyses for the past ??? 20 k yr are comparable to previous studies focused on that interval in Lake Tanganyika, and show that the lake was about 350 m lower than present at the Last Glacial Maximum (LGM). Repetitive peaks in TOC and corresponding drops in ??13C over the past 79 k yr indicate periods of high productivity and mixing above the T97-52V core site, probably due to cooler and perhaps windier conditions. From ??? 80 through ??? 58 k yr the ??13C values are relatively negative (-26 to -28???) suggesting predominance of algal contributions to bottom sediments at this site during this time. Following this interval there is a shift to higher values of ??13C, indicating a possible shift to C-4 pathway-dominated grassland-type vegetation in the catchment, and indicating cooler, dryer conditions from ??? 55 k yr through the LGM. Two seismic sequence boundaries are observed at shallow stratigraphic levels in the seismic reflection data, and the upper boundary correlates to a major discontinuity near the base of T97-52V. We interpret these discontinuities to reflect major, prolonged drops in lake level below the core site (393 m), with the lower boundary correlating to marine oxygen isotope Stage 6. This suggests that the previous glacial period was considerably cooler and more arid in the equatorial tropics than was the last glacial period.

Increasing Stability and Activity of Core-Shell Catalysts by Preferential Segregation of Oxide on Edges and Vertexes: Oxygen Reduction on Ti-Au@Pt/C

DOE PAGES

Hu, J.; Wu, L.; Kuttiyiel, K.; ...

2016-06-30

We describe a new class of core-shell nanoparticle catalysts having edges and vertexes covered by refractory metal oxide that preferentially segregates onto these catalyst sites. The monolayer shell is deposited on the oxidefree core atoms. The oxide on edges and vertexes induces high catalyst’s stability and activity. The catalyst and synthesis are exemplified by fabrication of Au nanoparticles doped by Ti atoms that segregate as oxide onto low–coordination sites of edges and vertexes. Pt monolayer shell deposited on Au sites has the mass and specific activities for the oxygen reduction reaction about 13 and 5 times higher than those ofmore » commercial Pt/C catalysts. The durability tests show no activity loss after 10000 potential cycles from 0.6 to 1.0V. The superior activity and durability of the Ti-Au@Pt catalyst originate from protective Ti oxide located at the most dissolution-prone edge and vertex sites, and Au-supported active and stable Pt shell.« less

CKI isoforms α and ε regulate Star–PAP target messages by controlling Star–PAP poly(A) polymerase activity and phosphoinositide stimulation

PubMed Central

Laishram, Rakesh S.; Barlow, Christy A.; Anderson, Richard A.

2011-01-01

Star–PAP is a non-canonical, nuclear poly(A) polymerase (PAP) that is regulated by the lipid signaling molecule phosphatidylinositol 4,5 bisphosphate (PI4,5P2), and is required for the expression of a select set of mRNAs. It was previously reported that a PI4,5P2 sensitive CKI isoform, CKIα associates with and phosphorylates Star–PAP in its catalytic domain. Here, we show that the oxidative stress-induced by tBHQ treatment stimulates the CKI mediated phosphorylation of Star–PAP, which is critical for both its polyadenylation activity and stimulation by PI4,5P2. CKI activity was required for the expression and efficient 3′-end processing of its target mRNAs in vivo as well as the polyadenylation activity of Star–PAP in vitro. Specific CKI activity inhibitors (IC261 and CKI7) block in vivo Star–PAP activity, but the knockdown of CKIα did not equivalently inhibit the expression of Star–PAP targets. We show that in addition to CKIα, Star–PAP associates with another CKI isoform, CKIε in the Star–PAP complex that phosphorylates Star–PAP and complements the loss of CKIα. Knockdown of both CKI isoforms (α and ε) resulted in the loss of expression and the 3′-end processing of Star–PAP targets similar to the CKI activity inhibitors. Our results demonstrate that CKI isoforms α and ε modulate Star–PAP activity and regulates Star–PAP target messages. PMID:21729869

The developmental transcriptome of the mosquito Aedes aegypti, an invasive species and major arbovirus vector.

PubMed

Akbari, Omar S; Antoshechkin, Igor; Amrhein, Henry; Williams, Brian; Diloreto, Race; Sandler, Jeremy; Hay, Bruce A

2013-09-04

Mosquitoes are vectors of a number of important human and animal diseases. The development of novel vector control strategies requires a thorough understanding of mosquito biology. To facilitate this, we used RNA-seq to identify novel genes and provide the first high-resolution view of the transcriptome throughout development and in response to blood feeding in a mosquito vector of human disease, Aedes aegypti, the primary vector for Dengue and yellow fever. We characterized mRNA expression at 34 distinct time points throughout Aedes development, including adult somatic and germline tissues, by using polyA+ RNA-seq. We identify a total of 14,238 novel new transcribed regions corresponding to 12,597 new loci, as well as many novel transcript isoforms of previously annotated genes. Altogether these results increase the annotated fraction of the transcribed genome into long polyA+ RNAs by more than twofold. We also identified a number of patterns of shared gene expression, as well as genes and/or exons expressed sex-specifically or sex-differentially. Expression profiles of small RNAs in ovaries, early embryos, testes, and adult male and female somatic tissues also were determined, resulting in the identification of 38 new Aedes-specific miRNAs, and ~291,000 small RNA new transcribed regions, many of which are likely to be endogenous small-interfering RNAs and Piwi-interacting RNAs. Genes of potential interest for transgene-based vector control strategies also are highlighted. Our data have been incorporated into a user-friendly genome browser located at www.Aedes.caltech.edu, with relevant links to Vectorbase (www.vectorbase.org).

Enzymes Involved in Post-transcriptional RNA Metabolism in Gram-negative bacteria

PubMed Central

Mohanty, Bijoy K.

2018-01-01

Gene expression in Gram-negative bacteria is regulated at many levels, including transcription initiation, RNA processing, RNA/RNA interactions, mRNA decay, and translational controls involving enzymes that alter translational efficiency. In this chapter we discuss the various enzymes that control transcription, translation and RNA stability through RNA processing and degradation. RNA processing is essential to generate functional RNAs, while degradation helps control the steady-state level of each individual transcript. For example, all the pre-tRNAs are transcribed with extra nucleotides at both their 5′ and 3′ termini, which are subsequently processed to produce mature tRNAs that can be aminoacylated. Similarly, rRNAs that are transcribed as part of a 30S polycistronic transcript, are matured to individual 16S, 23S and 5S rRNAs. Decay of mRNAs plays a key role in gene regulation through controlling the steady-state level of each transcript, which is essential for maintaining appropriate protein levels. In addition, degradation of both translated and non-translated RNAs recycles nucleotides to facilitate new RNA synthesis. To carry out all these reactions Gram-negative bacteria employ a large number of endonucleases, exonucleases, RNA helicases, and poly(A) polymerase as well as proteins that regulate the catalytic activity of particular ribonucleases. Under certain stress conditions an additional group of specialized endonucleases facilitate the cell’s ability to adapt and survive. Many of the enzymes, such as RNase E, RNase III, polynucleotide phosphorylase, RNase R, and poly(A) polymerase I participate in multiple RNA processing and decay pathways. PMID:29676246

[Interaction of trivaline with single-stranded polyribonucleotides].

PubMed

Strel'tsov, S A; Lysov, Iu P; Semenov, T E; Vengerov, Iu Iu; Khorlin, A A; Surovaia, A N; Gurskiĭ, G V

1991-01-01

Binding of tripeptide H-Val3-(NH)2-Dns (TVP) to polyribonucleotides was studied by fluorescence methods, circular and flow linear dichroism, equilibrium dialysis and electron microscopy. It was found that TVP binds to poly(U) in monomer, dimer and tetramer forms with binding constants of about 10(3), 40, 18.10(4) M, respectively. The cooperativity parameter for peptide dimer binding is 2000. The peptide forms tetramer complexes with poly(A), poly(C), poly(G) also. The formation of a complex between the peptide tetramer and nucleic acid is accompanied by a significant increase in the fluorescence intensity. The cooperative binding of TVP dimers to poly(U), poly(A), poly(C) is accompanied by a dramatic decrease in the flexibility of polynucleotide chains. However, it has a small effect (if any) on the flexibility of the poly(G) chain. The observed similarity of thermodynamic, optical and hydrodynamic++ properties of TVP complexes with single-stranded and double-stranded nucleic acids may reflect a similarity in the geometries of peptide complexes with nucleic acids. Electron microscopy studies show that peptide binding to poly(U) and dsDNA leads to compactization of the nucleic acids caused by interaction between the peptide tetramers bound to a nucleic acid. At the first stage of the compactization process the well-organized rod-like particles are formed, each consisting of one or more single-stranded polynucleotide fibers. Increasing the peptide concentration stimulates a side-by-side association and folding of the rods with the formation of macromolecular "leech-like" structures with the thickness of 20-50 nm.

Separation and parallel sequencing of the genomes and transcriptomes of single cells using G&T-seq.

PubMed

Macaulay, Iain C; Teng, Mabel J; Haerty, Wilfried; Kumar, Parveen; Ponting, Chris P; Voet, Thierry

2016-11-01

Parallel sequencing of a single cell's genome and transcriptome provides a powerful tool for dissecting genetic variation and its relationship with gene expression. Here we present a detailed protocol for G&T-seq, a method for separation and parallel sequencing of genomic DNA and full-length polyA(+) mRNA from single cells. We provide step-by-step instructions for the isolation and lysis of single cells; the physical separation of polyA(+) mRNA from genomic DNA using a modified oligo-dT bead capture and the respective whole-transcriptome and whole-genome amplifications; and library preparation and sequence analyses of these amplification products. The method allows the detection of thousands of transcripts in parallel with the genetic variants captured by the DNA-seq data from the same single cell. G&T-seq differs from other currently available methods for parallel DNA and RNA sequencing from single cells, as it involves physical separation of the DNA and RNA and does not require bespoke microfluidics platforms. The process can be implemented manually or through automation. When performed manually, paired genome and transcriptome sequencing libraries from eight single cells can be produced in ∼3 d by researchers experienced in molecular laboratory work. For users with experience in the programming and operation of liquid-handling robots, paired DNA and RNA libraries from 96 single cells can be produced in the same time frame. Sequence analysis and integration of single-cell G&T-seq DNA and RNA data requires a high level of bioinformatics expertise and familiarity with a wide range of informatics tools.

Alkaline Earth Core Level Photoemission Spectroscopy of High-Temperature Superconductors

NASA Technical Reports Server (NTRS)

Vasquez, R.

1993-01-01

This paper examines photoemission measurements of the alkaline Earth core levels of high-temperature superconductors and related materials, models that seek to explain the large negative shifts observed relative to the corresponding alkaline Earth metals, and the effect of lattice site disorder on the core level spectra and the presence or absence of intrinsic surface peaks.

A Study on Benthic Foraminifera Assemblages in the Upper Slope off Southwest Taiwan

NASA Astrophysics Data System (ADS)

Yeh, Jen-Chu; Lin, Andrew T.; Chien, Chih-Wei

2016-04-01

This study attempts to establish the spatial distribution of benthic foraminifera in the upper accretionary wedge off SW Taiwan. A few box cores (each core up to 49 cm thick) are retrieved onboard R/V Ocean Researcher I during 1092 cruise in 2014 at water depths ranging from 1,135 to 1,586 m lying in between the Good Weather Ridge and the Yuan-An Ridge. Analyses on grain size reveal that the sediment size ranges from clay to silt for all sites with the exception of YT1 site, where a small percentage of fine sand (< 20%) is found to distribute evenly in a 32 cm-thick box core. Core images from X-radiographs show some layers of foraminifera ooze and rare traces of bioturbation. Age of sedimentation is obtained by using 210Pb dating method. The 210Pb concentration profile decays exponentially down core, indicating sedimentation from suspension. The measured sedimentation rate ranges from 0.47 to 2.4 mm/yr. Site YT1 has the lowest sedimentation rate (around 0.47 mm/yr), leading to high abundance of individual benthic foraminiferal species. Living foraminiferal individuals were distinguished from dead assemblages by Rose Bengal staining method during the cruise. Our results show that the dominant living species of all studied cores is Chilostomella oolina, with subsidiary occurrences of Bulimina aculeata, Bolivinita quadrilateral, and Lenticulina spp. etc. Cluster analysis suggests that the forams have similar spatial distribution pattern at all studied sites, indicating uniform and stable hemipelagic sedimentation. Analyses of dead assemblages reveal a remarkable decrease in the abundance of Bulimina and Uvigerina for the last 100 years at YT-2 site, with increasing abundance of Chilostomella. This indicates that the water masses may have turned from suboxic to dysoxic conditions since c. 100 year ago. This is the first study to report the living benthic foraminifera distribution in water depths up to c. 1,600 m off SW Taiwan, providing a basis for future studies. Keywords: benthic foraminifera, upper slope, Taiwan

Evidence of Superstoichiometric H/d Lenr Active Sites and High-Temperature Superconductivity in a Hydrogen-Cycled Pd/PdO

NASA Astrophysics Data System (ADS)

Lipson, A. G.; Castano, C. H.; Miley, G. H.; Lyakhov, B. F.; Tsivadze, A. Yu.; Mitin, A. V.

Electron transport and magnetic properties have been studied in a 12.5 μm thick Pd foil with a thermally grown oxide and a low-residual concentration of hydrogen. This foil was deformed by cycling across the Pd hydride miscibility gap and the residual hydrogen was trapped at dislocation cores. Anomalies of both resistance and magnetic susceptibility have been observed below 70 K, indicating the appearance of excess conductivity and a diamagnetic response that we interpret in terms of filamentary superconductivity. These anomalies are attributed to a condensed hydrogen-rich phase at dislocation cores. The role of deuterium rich dislocation cores as LENR active sites is discussed.

A Conserved Apomixis-Specific Polymorphism Is Correlated with Exclusive Exonuclease Expression in Premeiotic Ovules of Apomictic Boechera Species1[W][OPEN

PubMed Central

Corral, José M.; Vogel, Heiko; Aliyu, Olawale M.; Hensel, Götz; Thiel, Thomas; Kumlehn, Jochen; Sharbel, Timothy F.

2013-01-01

Apomixis (asexual seed production) is characterized by meiotically unreduced egg cell production (apomeiosis) followed by its parthenogenetic development into offspring that are genetic clones of the mother plant. Fertilization (i.e. pseudogamy) of the central cell is important for the production of a functional endosperm with a balanced 2:1 maternal:paternal genome ratio. Here, we present the APOLLO (for apomixis-linked locus) gene, an Aspartate Glutamate Aspartate Aspartate histidine exonuclease whose transcripts are down-regulated in sexual ovules entering meiosis while being up-regulated in apomeiotic ovules at the same stage of development in plants of the genus Boechera. APOLLO has both “apoalleles,” which are characterized by a set of linked apomixis-specific polymorphisms, and “sexalleles.” All apomictic Boechera spp. accessions proved to be heterozygous for the APOLLO gene (having at least one apoallele and one sexallele), while all sexual genotypes were homozygous for sexalleles. Apoalleles contained a 20-nucleotide polymorphism present in the 5′ untranslated region that contains specific transcription factor-binding sites for ARABIDOPSIS THALIANA HOMEOBOX PROTEIN5, LIM1 (for LINEAGE ABNORMAL11, INSULIN1, MECHANOSENSORY PROTEIN3), SORLIP1AT (for SEQUENCES OVERREPRESENTED IN LIGHT-INDUCED PROMOTERS IN ARABIDOPSIS THALIANA1), SORLIP2AT, and POLYA SIGNAL1. In the same region, sexalleles contain transcription factor-binding sites for DNA BINDING WITH ONE FINGER2, DNA BINDING WITH ONE FINGER3, and PROLAMIN BOX-BINDING FACTOR. Our results suggest that the expression of a single deregulated allele could induce the cascade of events leading to asexual female gamete formation in an apomictic plant. PMID:24163323

CARBON MONOXIDE FLUXES OF DIFFERENT SOIL LAYERS IN UPLAND CANADIAN BOREAL FORESTS

EPA Science Inventory

Dark or low-light carbon monoxide fluxes at upland Canadian boreal forest sites were measured on-site with static chambers and with a laboratory incubation technique using cores from different depths at the same sites. Three different upland black spruce sites, burned in 1987,199...

Evolutionary acquisition and loss of saxitoxin biosynthesis in dinoflagellates: the second "core" gene, sxtG.

PubMed

Orr, Russell J S; Stüken, Anke; Murray, Shauna A; Jakobsen, Kjetill S

2013-04-01

Saxitoxin and its derivatives are potent neurotoxins produced by several cyanobacteria and dinoflagellate species. SxtA is the initial enzyme in the biosynthesis of saxitoxin. The dinoflagellate full mRNA and partial genomic sequences have previously been characterized, and it appears that sxtA originated in dinoflagellates through a horizontal gene transfer from a bacterium. So far, little is known about the remaining genes involved in this pathway in dinoflagellates. Here we characterize sxtG, an amidinotransferase enzyme gene that putatively encodes the second step in saxitoxin biosynthesis. In this study, the entire sxtG transcripts from Alexandrium fundyense CCMP1719 and Alexandrium minutum CCMP113 were amplified and sequenced. The transcripts contained typical dinoflagellate spliced leader sequences and eukaryotic poly(A) tails. In addition, partial sxtG transcript fragments were amplified from four additional Alexandrium species and Gymnodinium catenatum. The phylogenetic inference of dinoflagellate sxtG, congruent with sxtA, revealed a bacterial origin. However, it is not known if sxtG was acquired independently of sxtA. Amplification and sequencing of the corresponding genomic sxtG region revealed noncanonical introns. These introns show a high interspecies and low intraspecies variance, suggesting multiple independent acquisitions and losses. Unlike sxtA, sxtG was also amplified from Alexandrium species not known to synthesize saxitoxin. However, amplification was not observed for 22 non-saxitoxin-producing dinoflagellate species other than those of the genus Alexandrium or G. catenatum. This result strengthens our hypothesis that saxitoxin synthesis has been secondarily lost in conjunction with sxtA for some descendant species.

RNA is an integral component of chromatin that contributes to its structural organization.

PubMed

Rodríguez-Campos, Antonio; Azorín, Fernando

2007-11-14

Chromatin structure is influenced by multiples factors, such as pH, temperature, nature and concentration of counterions, post-translational modifications of histones and binding of structural non-histone proteins. RNA is also known to contribute to the regulation of chromatin structure as chromatin-induced gene silencing was shown to depend on the RNAi machinery in S. pombe, plants and Drosophila. Moreover, both in Drosophila and mammals, dosage compensation requires the contribution of specific non-coding RNAs. However, whether RNA itself plays a direct structural role in chromatin is not known. Here, we report results that indicate a general structural role for RNA in eukaryotic chromatin. RNA is found associated to purified chromatin prepared from chicken liver, or cultured Drosophila S2 cells, and treatment with RNase A alters the structural properties of chromatin. Our results indicate that chromatin-associated RNAs, which account for 2%-5% of total chromatin-associated nucleic acids, are polyA(-) and show a size similar to that of the DNA contained in the corresponding chromatin fragments. Chromatin-associated RNA(s) are not likely to correspond to nascent transcripts as they are also found bound to chromatin when cells are treated with alpha-amanitin. After treatment with RNase A, chromatin fragments of molecular weight >3.000 bp of DNA showed reduced sedimentation through sucrose gradients and increased sensitivity to micrococcal nuclease digestion. This structural transition, which is observed both at euchromatic and heterochromatic regions, proceeds without loss of histone H1 or any significant change in core-histone composition and integrity.

Density of Emerald Ash Borer (Coleoptera: Buprestidae) Adults and Larvae at Three Stages of the Invasion Wave.

PubMed

Burr, Stephen J; McCullough, Deborah G; Poland, Therese M

2018-02-08

Emerald ash borer (EAB) (Agrilus planipennis Fairmaire) (Coleoptera: Buprestidae), an invasive phloem-feeding buprestid, has killed hundreds of millions of ash (Fraxinus spp.) trees in the United States and two Canadian provinces. We evaluated EAB persistence in post-invasion sites and compared EAB adult captures and larval densities in 24 forested sites across an east-west gradient in southern Michigan representing the Core (post-invasion), Crest (high EAB populations), and Cusp (recently infested areas) of the EAB invasion wave. Condition of green ash (Fraxinus pennsylvanica Marsh) trees were recorded in fixed radius plots and linear transects in each site. Ash mortality was highest in Core sites in the southeast, moderate in Crest sites in central southern Michigan, and low in Cusp sites in the southwest. Traps and trap trees in Crest sites accounted for 75 and 60% of all EAB beetles captured in 2010 and 2011, respectively. Populations of EAB were present in all Core sites and traps in these sites captured 13% of all beetles each year. Beetle captures and larval densities at Cusp sites roughly doubled between 2010 and 2011, reflecting the increasing EAB populations. Sticky bands on girdled trees captured the highest density of EAB beetles per m2 of area, while baited double-decker traps had the highest detection rates and captured the most beetles. Larval densities were higher on girdled ash than on similar ungirdled trees and small planted trees. Woodpecker predation and a native larval parasitoid were present in all three invasion regions but had minor effects on ash survival and EAB densities. © The Author(s) 2018. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Correlation of reaction sites during the chlorine extraction by hydrogen atom from Cl /Si(100)-2×1

NASA Astrophysics Data System (ADS)

Hsieh, Ming-Feng; Chung, Jen-Yang; Lin, Deng-Sung; Tsay, Shiow-Fon

2007-07-01

The Cl abstraction by gas-phase H atoms from a Cl-terminated Si(100) surface was investigated by scanning tunneling microscopy (STM), high-resolution core level photoemission spectroscopy, and computer simulation. The core level measurements indicate that some additional reactions occur besides the removal of Cl. The STM images show that the Cl-extracted sites disperse randomly in the initial phase of the reaction, but form small clusters as more Cl is removed, indicating a correlation between Cl-extracted sites. These results suggest that the hot-atom process may occur during the atom-adatom collision.

Primary Care-Mental Health Integration in the Veterans Affairs Health System: Program Characteristics and Performance.

PubMed

Cornwell, Brittany L; Brockmann, Laurie M; Lasky, Elaine C; Mach, Jennifer; McCarthy, John F

2018-06-01

The Veterans Health Administration (VHA) has achieved substantial national implementation of primary care-mental health integration (PC-MHI) services. However, little is known regarding program characteristics, variation in characteristics across settings, or associations between program fidelity and performance. This study identified core elements of PC-MHI services and evaluated their associations with program characteristics and performance. A principal-components analysis (PCA) of reports from 349 sites identified factors associated with PC-MHI fidelity. Analyses assessed the correlation among factors and between each factor and facility type (medical center or community-based outpatient clinic), primary care population size, and performance indicators (receipt of PC-MHI services, same-day access to mental health and primary care services, and extended duration of services). PCA identified seven factors: core implementation, care management (CM) assessments and supervision, CM supervision receipt, colocated collaborative care (CCC) by prescribing providers, CCC by behavioral health providers, participation in patient aligned care teams (PACTs) for special populations, and treatment of complex mental health conditions. Sites serving larger populations had greater core implementation scores. Medical centers and sites serving larger populations had greater scores for CCC by prescribing providers, CM assessments and supervision, and participation in PACTs. Greater core implementation scores were associated with greater same-day access. Sites with greater scores for CM assessments and supervision had lower scores for treatment of complex conditions. Outpatient clinics and sites serving smaller populations experienced challenges in integrated care implementation. To enhance same-day access, VHA should continue to prioritize PC-MHI implementation. Providing brief, problem-focused care may enhance CM implementation.

Mineralization and nitrification patterns at eight northeastern USA forested research sites

USGS Publications Warehouse

Ross, D.S.; Lawrence, G.B.; Fredriksen, G.

2004-01-01

Nitrogen transformation rates in eight northeastern US research sites were measured in soil samples taken in the early season of 2000 and the late season of 2001. Net mineralization and nitrification rates were determined on Oa or A horizon samples by two different sampling methods - intact cores and repeated measurements on composite samples taken from around the cores. Net rates in the composite samples (n=30) showed three different temporal patterns: high net nitrification with minimal NH4+ accumulation, high net nitrification and high NH4+ accumulation, and minimal net nitrification and moderate NH4+ accumulation. The 4-week net rates in intact cores were about half that of the rates from the composite samples but were well related (R2 > 0.70). Composite samples from sites that exhibited high net nitrification were incubated with acetylene and net nitrification was completely stopped, suggesting an autotrophic pathway. Gross mineralization and nitrification (2000 only) rates were estimated using the isotope dilution technique. Gross rates of nitrification and consumption in intact cores were relatively low. Gross rates of mineralization and net rates of nitrification were both related to the soil C/N ratio, with higher rates generally occurring in sites containing Acer saccharum as a dominant or co-dominant species. The comparison of methods suggests that all provide a similar hierarchy of potential rates but that the degree of net nitrification is strongly influenced by the degree of sample disturbance. Differences between sites appear to be related to an interaction of soil (C/N) and vegetation (A. saccharum contribution) characteristics. ?? 2003 Elsevier B.V. All rights reserved.

Columnar cell change with atypia (flat epithelial atypia) on breast core biopsy-outcomes following open excision.

PubMed

Biggar, Magdalena A; Kerr, Kris M; Erzetich, Lisa M; Bennett, Ian C

2012-01-01

Columnar cell change with atypia (CCCA) is a relatively recently recognized pathologic breast entity considered to be a risk factor for subsequent development of breast carcinoma. The aim of this study was to investigate the significance of finding CCCA on breast core biopsy, by establishing the frequency of other breast pathology on subsequently performed surgical excision specimens. All cases with CCCA as the most advanced lesion on core biopsy were reviewed. After excision, another advanced proliferative lesion was identified in 17 (33%) patients, including three patients (6%) with in situ or invasive carcinoma. An additional five patients (10%) were concurrently diagnosed with primary breast carcinoma at other sites. These findings indicate that when CCCA is found on core biopsy, open surgical biopsy of the relevant area should be performed and that workup of both breasts should be undertaken to exclude coexistent breast carcinoma at alternative sites. © 2012 Wiley Periodicals, Inc.

Current status of SPINNORs designs

DOE Office of Scientific and Technical Information (OSTI.GOV)

Su'ud, Zaki

2010-06-22

This study discuss about the SPINNOR (Small Power Reactor, Indonesia, No On-site Refuelling) and the VSPINNOR (Very Small Power Reactor, Indonesia, No On-site Refuelling) which are small lead-bismuth cooled nuclear power reactors with fast neutron spectrum that could be operated for more than 10 or 15 years without on-site refuelling. They are based on the concept of a long-life core reactor developed in Indonesia since early 1990 in collaboration with the Research Laboratory for Nuclear Reactors of the Tokyo Institute of Technology (RLNR TITech). The reactor cores are designed to have near zero (less then one effective delayed neutron fraction)more » burn-up reactivity swing during the whole course of their operation to avoid a possibility of prompt criticality accident. The basic concept is that central region of the reactor core is filled with fertile (blanket) material. During the reactor operation fissile material accumulates in this central region, which helps to compensate fissile material loss in the peripheral core region and also contributes to negative coolant loss reactivity effect. A concept of high fuel volume fraction in the core is applied to achieve smaller size of a critical reactor. In this paper we consider to add Np-237 to the fuel to enhance non proliferation characteristics of the systems. The effect of Np-237 amount variation is discussed.« less

Results of chemical and isotopic analyses of sediment and water from alluvium of the Canadian River near a closed municipal landfill, Norman, Oklahoma

USGS Publications Warehouse

Breit, George N.; Tuttle, Michele L.W.; Cozzarelli, Isabelle M.; Christenson, Scott C.; Jaeschke, Jeanne B.; Fey, David L.; Berry, Cyrus J.

2005-01-01

Results of physical and chemical analyses of sediment and water collected near a closed municipal landfill at Norman, Oklahoma are presented in this report. Sediment analyses are from 40 samples obtained by freeze-shoe coring at 5 sites, and 14 shallow (depth <1.3 m) sediment samples. The sediment was analyzed to determine grain size, the abundance of extractable iron species and the abundances and isotopic compositions of forms of sulfur. Water samples included pore water from the freeze-shoe core, ground water, and surface water. Pore water from 23 intervals of the core was collected and analyzed for major and trace dissolved species. Thirteen ground-water samples obtained from wells within a few meters of the freeze-shoe core sites and one from the landfill were analyzed for major and trace elements as well as the sulfur and oxygen isotope composition of dissolved sulfate. Samples of surface water were collected at 10 sites along the Canadian River from New Mexico to central Oklahoma. These river-water samples were analyzed for major elements, trace elements, and the isotopic composition of dissolved sulfate.

Methane Transmission and Oxidation throughout the Soil Column from Three Central Florida Sites

NASA Astrophysics Data System (ADS)

Bond-Lamberty, B. P.; Fansler, S.; Becker, K. E.; Hinkle, C. R.; Bailey, V. L.

2015-12-01

When methane (CH4) is generated in anoxic soil sites, it may be subsequently re-oxidized to carbon dioxide (CO2). Understanding the controls on, and magnitudes of, these processes is necessary to accurately represent greenhouse gas production and emission from soils. We used a laboratory incubation to examine the influence of variable conditions on methane transmission and oxidation, and identify critical reaction zones throughout the soil column. Sandy soils were sampled from three different sites at Disney Wilderness Preserve (DWP), Florida, USA: a depression marsh characterized by significant surface organic matter accumulation, a dry pine flatwood site with water intrusion and organic horizon at depth (200+ cm); and an intermediate-drainage site. Contiguous, 30-cm long cores were sampled from N=4 random boreholes at each site, from the surface to the water table (varying from 90 to 240 cm). In the lab, each core was monitored for 50 hours to quantify baseline (pretreatment) gas fluxes before injection with 6 ml CH4 (an amount commensurate with previous field collar measurements) at the base of each core. We then monitored CH4 and CO2 evolution for 100 hours after injection, calculating per-gas and total C evolution. Methane emissions spiked ~10 hours after injection for all cores, peaking at 0.001 μmol/g soil/hr, ~30x larger than pre-injection flux rates. On a C basis, CO2 emissions were orders of magnitude larger, and rose significantly after injection, with elevated rates generally sustained throughout the incubation. Cores from the depression marsh and shallower depths had significantly higher fluxes of both gases. We estimate that 99.1% of the original CH4 injection was oxidized to CO2. These findings suggest either that the methane measured in the field at DWP originates from within a few centimeters of the surface, or that it is produced in much larger quantities deeper in the profile before most is subsequently oxidized. This highlights the need for better understanding and modeling the multiple processes that result in soil-atmosphere CO2 and CH4 fluxes.

The Pleistocene Eastern Equatorial Pacific: Insights from a New Carnegie Platform Stratigraphic Record

NASA Astrophysics Data System (ADS)

Gwizd, S.; Lea, D. W.

2016-12-01

Renewed interest in a classic Eastern Equatorial Pacific paleoceanographic site at 3° 35.85' S, 83° 57.79' W, previous site of cores V19-29 and TR163-31, prompted a re-coring in 2009 using the recently developed CDH giant piston coring system on cruise KNR195-5. Giant piston core CDH-36 (3225 m depth, 42.61 m length) nearly triples the length of previous cores at this site. When spliced together with companion multicore MC-34A (0.36 m length), these two cores generate continuous stratigraphy throughout most of the middle Pleistocene, and include recognized stratigraphic tie points ash layer "L" (Ninkovich and Shackleton, 1975) and the extinction of pink G. ruber. A new age model utilizing Bayesian analysis of 17 N. dutertrei radiocarbon dates in MC-34A and the top 3.4 m of CDH-36, and alignment of a new CDH-36 δ18O record with the LR04 benthic stack (Lisiecki and Raymo, 2005), demonstrates that this new "Carnegie Platform" (CP) record extends from 0 to 720 ka (MIS 18), tripling the timescale of previous studies, with an average sedimentation rate of 7 cm/kyr. The CP C. wuellerstorfi δ18O and δ13C records reveal strong consistencies in timing and extent of glacial and interglacial episodes with previously studied regional records. Coarse fraction percentage (%CF) ([coarse fraction/bulk dry sample] * 100) is also evaluated throughout the CP core in order to qualitatively assess dissolution cycles. The CP %CF dataset primarily records Pleistocene dissolution cycles, yet exhibits variability representative of potential local bathymetric and hydrographic effects. The timing of %CF cyclicity is consistent with processes which affect basin-wide calcium carbonate dissolution cycles, including changes in terrestrial carbon input to the oceans and changes in water mass ventilation (Shackleton, 1977; Toggweiler et al., 2006; Sexton and Barker, 2012). Establishing the stratigraphy of the CP record provides the first step towards a more thorough and extended analysis of this important Eastern Equatorial Pacific site.

Pre-cometary ice composition from hot core chemistry.

PubMed

Tornow, Carmen; Kührt, Ekkehard; Motschmann, Uwe

2005-10-01

Pre-cometary ice located around star-forming regions contains molecules that are pre-biotic compounds or pre-biotic precursors. Molecular line surveys of hot cores provide information on the composition of the ice since it sublimates near these sites. We have combined a hydrostatic hot core model with a complex network of chemical reactions to calculate the time-dependent abundances of molecules, ions, and radicals. The model considers the interaction between the ice and gas phase. It is applied to the Orion hot core where high-mass star formation occurs, and to the solar-mass binary protostar system IRAS 16293-2422. Our calculations show that at the end of the hot core phase both star-forming sites produce the same prebiotic CN-bearing molecules. However, in the Orion hot core these molecules are formed in larger abundances. A comparison of the calculated values with the abundances derived from the observed line data requires a chemically unprocessed molecular cloud as the initial state of hot core evolution. Thus, it appears that these objects are formed at a much younger cloud stage than previously thought. This implies that the ice phase of the young clouds does not contain CN-bearing molecules in large abundances before the hot core has been formed. The pre-biotic molecules synthesized in hot cores cause a chemical enrichment in the gas phase and in the pre-cometary ice. This enrichment is thought to be an important extraterrestrial aspect of the formation of life on Earth and elsewhere.

Deep Radiostratigraphy of the East Antarctic Plateau: Connecting the Dome C and Vostok Ice Core Sites

NASA Technical Reports Server (NTRS)

Cavitte, Marie G. P.; Blankenship, Donald D.; Young, Duncan A.; Schroeder, Dustin M.; Parrenin, Frederic; Lemeur, Emmanuel; Macgregor, Joseph A.; Siegert, Martin J.

2016-01-01

Several airborne radar-sounding surveys are used to trace internal reflections around the European Project for Ice Coring in Antarctica Dome C and Vostok ice core sites. Thirteen reflections, spanning the last two glacial cycles, are traced within 200 km of Dome C, a promising region for million-year-old ice, using the University of Texas Institute for Geophysics High-Capacity Radar Sounder. This provides a dated stratigraphy to 2318 m depth at Dome C. Reflection age uncertainties are calculated from the radar range precision and signal-to-noise ratio of the internal reflections. The radar stratigraphy matches well with the Multichannel Coherent Radar Depth Sounder (MCoRDS) radar stratigraphy obtained independently. We show that radar sounding enables the extension of ice core ages through the ice sheet with an additional radar-related age uncertainty of approximately 1/3-1/2 that of the ice cores. Reflections are extended along the Byrd-Totten Glacier divide, using University of Texas/Technical University of Denmark and MCoRDS surveys. However, core-to-core connection is impeded by pervasive aeolian terranes, and Lake Vostok's influence on reflection geometry. Poor radar connection of the two ice cores is attributed to these effects and suboptimal survey design in affected areas. We demonstrate that, while ice sheet internal radar reflections are generally isochronal and can be mapped over large distances, careful survey planning is necessary to extend ice core chronologies to distant regions of the East Antarctic ice sheet.

Initial results from geophysical surveys and shallow coring of the Northeast Greenland Ice Stream (NEGIS)

NASA Astrophysics Data System (ADS)

Vallelonga, P.; Christianson, K.; Alley, R. B.; Anandakrishnan, S.; Christian, J. E. M.; Dahl-Jensen, D.; Gkinis, V.; Holme, C.; Jacobel, R. W.; Karlsson, N. B.; Keisling, B. A.; Kipfstuhl, S.; Kjær, H. A.; Kristensen, M. E. L.; Muto, A.; Peters, L. E.; Popp, T.; Riverman, K. L.; Svensson, A. M.; Tibuleac, C.; Vinther, B. M.; Weng, Y.; Winstrup, M.

2014-07-01

The Northeast Greenland Ice Stream (NEGIS) is the sole interior Greenlandic ice stream. Fast flow initiates near the summit dome, and the ice stream terminates approximately 1000 km downstream in three large outlet glaciers that calve into the Greenland Sea. To better understand this important system, in the summer of 2012 we drilled a 67 m firn core and conducted ground-based radio-echo sounding (RES) and active-source seismic surveys at a site approximately 150 km downstream from the onset of streaming flow (NEGIS firn core, 75°37.61' N, 35°56.49' W). The site is representative of the upper part of the ice stream, while also being in a crevasse-free area for safe surface operations. Annual cycles were observed for insoluble dust, sodium and ammonium concentrations and for electrolytic conductivity, allowing a seasonally resolved chronology covering the past 400 yr. Annual layer thicknesses averaged 0.11 m ice equivalent (i.e.) for the period 1607-2011, although accumulation varied between 0.08 and 0.14 m i.e., likely due to flow-related changes in surface topography. Tracing of RES layers from the NGRIP (North Greenland Ice Core Project) ice core site shows that the ice at NEGIS preserves a climatic record of at least the past 51 kyr. We demonstrate that deep ice core drilling in this location can provide a reliable Holocene and late-glacial climate record, as well as helping to constrain the past dynamics and ice-lithosphere interactions of the Greenland Ice Sheet.

A Sketch of the Taiwan Zebrafish Core Facility.

PubMed

You, May-Su; Jiang, Yun-Jin; Yuh, Chiou-Hwa; Wang, Chien-Ming; Tang, Chih-Hao; Chuang, Yung-Jen; Lin, Bo-Hung; Wu, Jen-Leih; Hwang, Sheng-Ping L

2016-07-01

In the past three decades, the number of zebrafish laboratories has significantly increased in Taiwan. The Taiwan Zebrafish Core Facility (TZCF), a government-funded core facility, was launched to serve this growing community. The Core Facility was built on two sites, one located at the National Health Research Institutes (NHRI, called Taiwan Zebrafish Core Facility at NHRI or TZeNH) and the other is located at the Academia Sinica (Taiwan Zebrafish Core Facility at AS a.k.a. TZCAS). The total surface area of the TZCF is about 180 m(2) encompassing 2880 fish tanks. Each site has a separate quarantine room and centralized water recirculating systems, monitoring key water parameters. To prevent diseases, three main strategies have been implemented: (1) imported fish must be quarantined; (2) only bleached embryos are introduced into the main facilities; and (3) working practices were implemented to minimize pathogen transfer between stocks and facilities. Currently, there is no health program in place; however, a fourth measure for the health program, specific regular pathogen tests, is being planned. In March 2015, the TZCF at NHRI has been AAALAC accredited. It is our goal to ensure that we provide "disease-free" fish and embryos to the Taiwanese research community.

Geologic logs of geotechnical cores from the subsurface Sacramento-San Joaquin Delta, California

USGS Publications Warehouse

Maier, Katherine L.; Ponti, Daniel J.; Tinsley, John C.; Gatti, Emma; Pagenkopp, Mark

2014-01-01

This report presents and summarizes descriptive geologic logs of geotechnical cores collected from 2009–12 in the Sacramento–San Joaquin Delta, California, by the California Department of Water Resources. Graphic logs are presented for 1,785.7 ft of retained cores from 56 borehole sites throughout the Sacramento-San Joaquin Delta. Most core sections are from a depth of ~100–200 feet. Cores primarily contain mud, silt, and sand lithologies. Tephra (volcanic ash and pumice), paleosols, and gravels are also documented in some core sections. Geologic observations contained in the core logs in this report provide stratigraphic context for subsequent sampling and data for future chronostratigraphic subsurface correlations.

Effects of oil from the 2010 Macondo well blowout on marsh foraminifera of Mississippi and Louisiana, USA.

PubMed

Brunner, Charlotte A; Yeager, Kevin M; Hatch, Rachel; Simpson, Sondra; Keim, Joseph; Briggs, Kevin B; Louchouarn, Patrick

2013-08-20

Foraminifera responded to both heavy and light oiling of marshes relative to unoiled control sites by changes to both standing stock and depth of habitation (DOH) in sediment following the 2010 Macondo well blowout. Push cores were taken from the middle marsh at sites classified as unoiled, lightly oiled, and heavily oiled based on concentrations of total polycyclic aromatic hydrocarbons ([TPAH]). Cores were sliced and stained with rose Bengal to detect live specimens of foraminifera. Short-term, sediment-mixing depths were determined using the penetration depths of excess (234)Th, and sedimentary organic carbon and carbonate were measured to distinguish depositional environments. Marsh foraminifera reacted to the highest oil concentration (5,000-18,000 ng/g of TPAH) by reducing standing stock and shortening the DOH compared with the control sites. At a second, less heavily oiled site, foraminifera responded with a shallower DOH, but with a boom in standing stock. Deformed, dead foraminifera occurred in all heavily oiled cores-but not elsewhere. Live foraminifera responded with a population boom at lightly oiled sites with [TPAH] near 1,100 ng/g. Changes in standing stock and DOH with [TPAH] suggest disturbance to the marsh food web, apparently due to oil pollution, and support the use of foraminifera as sentinel species.

Subsurface geology of a potential waste emplacement site, Salt Valley Anticline, Grand County, Utah

USGS Publications Warehouse

Hite, R.J.

1977-01-01

The Salt Valley anticline, which is located about 32 km northeast of Moab, Utah, is perhaps one of the most favorable waste emplacement sites in the Paradox basin. The site, which includes about 7.8 km 2, is highly accessible and is adjacent to a railroad. The anticline is one of a series of northwest-trending salt anticlines lying along the northeast edge of the Paradox basin. These anticlines are cored by evaporites of the Paradox Member of the Hermosa Formation of Middle Pennsylvanian age. The central core of the Salt Valley anticline forms a ridgelike mass of evaporites that has an estimated amplitude of 3,600 m. The evaporite core consists of about 87 percent halite rock, which includes some potash deposits; the remainder is black shale, silty dolomite, and anhydrite. The latter three lithologies are referred to as 'marker beds.' Using geophysical logs from drill holes on the anticline, it is possible to demonstrate that the marker beds are complexly folded and faulted. Available data concerning the geothermal gradient and heatflow at the site indicate that heat from emplaced wastes should be rapidly dissipated. Potentially exploitable resources of potash and petroleum are present at Salt Valley. Development of these resources may conflict with use of the site for waste emplacement.

Synoptic controls on precipitation pathways and snow delivery to high-accumulation ice core sites in the Ross Sea region, Antarctica

NASA Astrophysics Data System (ADS)

Sinclair, K. E.; Bertler, N. A. N.; Trompetter, W. J.

2010-11-01

Dominant storm tracks to two ice core sites on the western margin of the Ross Sea, Antarctica (Skinner Saddle (SKS) and Evans Piedmont Glacier), are investigated to establish key synoptic controls on snow accumulation. This is critical in terms of understanding the seasonality, source regions, and transport pathways of precipitation delivered to these sites. In situ snow depth and meteorological observations are used to identify major accumulation events in 2007-2008, which differ considerably between sites in terms of their magnitude and seasonal distribution. While snowfall at Evans Piedmont Glacier occurs almost exclusively during summer and spring, Skinner Saddle receives precipitation year round with a lull during the months of April and May. Cluster analysis of daily back trajectories reveals that the highest-accumulation days at both sites result from fast-moving air masses, associated with synoptic-scale low-pressure systems. There is evidence that short-duration pulses of snowfall at SKS also originate from mesocyclone development over the Ross Ice Shelf and local moisture sources. Changes in the frequency and seasonal distribution of these mechanisms of precipitation delivery will have a marked impact on annual accumulation over time and will therefore need careful consideration during the interpretation of stable isotope and geochemical records from these ice cores.

Paleomagnetic and rock magnetic study of the IODP Site U1332 sediments - relative paleointensity during Eocene and Oligocene

NASA Astrophysics Data System (ADS)

Yamamoto, Y.; Acton, G.; Channell, J. E.; Palmer, E. C.; Richter, C.; Yamazaki, T.

2011-12-01

Integrated Ocean Drilling Program (IODP) Expeditions 320 and 321 recovered sediment cores from equatorial Pacific. Cores were taken at eight Sites (U1331-U1338) and onboard measurements showed that those from Sites U1331, U1332, U1333 and U1334 covered Eocene and/or Oligocene (Expedition 320/321 Scientists, 2010). Although many efforts have been made to reveal relative geomagnetic paleointensity variations in geologic time, those prior to ca. 3 m.y. have been not yet reported except a few studies (e.g. ca. 23-34 Ma, Tauxe and Hartl, 1997). This study concentrates on paleomagnetic and rock magnetic measurements on the Site U1332 sediment core. The measurements include stepwise alternating field demagnetization of the natural remanent magnetization (NRM), the anhysteretic remanent magnetization (ARM) and the isothermal remanent magnetization (IRM). The magnetostrartigraphy constructed from the NRM data show that the sedimentary section extends from the early Oligocene to middle Eocene (23.030-41.358 Ma). Intensity variation of ARM and IRM is within about a factor of six throughout the core. Magnetic grain size proxy, ARM/IRM, differ between Eocene (about 0.11) and Oligocene (about 0.14). These suggest that relative paleointensity (RPI) estimation is basically possible if we divide the core into Eocene and Oligocene periods. RPI estimates have been done by using ARM and IRM as normalizers for NRM. RPIs by ARM and IRM generally show consistent variations. However, several experimental results imply that RPI by IRM may be more preferable. We will compare the U1332 RPI record with the U1331, U1333 and U1334 RPI records.

Summer temperatures inferred from varved lacustrine sediment at Iceberg Lake in southcentral Alaska

NASA Astrophysics Data System (ADS)

Diedrich, K.; Loso, M. G.

2010-12-01

Iceberg Lake, a glacier-dammed lake in southcentral Alaska, has been previously shown to record over 1,500 years of continuous laminated lacustrine sediment deposition. Because previous work was based on examination of subaerial outcrops exposed by stream incision in the bed of the jökulhlaup-drained lake, the length of the record was limited by the extent of the outcrops. In August of 2010, we returned to core the remote lake; our goal was recovery of the complete sedimentary record in the lake, extending perhaps back to the onset of late Holocene glaciation—around 3-5 ka in this region. We used a Vibarcorer system to recover sediment cores from two locations, one near the site of previous work and another at the distal end of the lake. The longest cores recovered were 5.2 meters and 6.2 meters at the proximal and distal sites, respectively. Based on the average lamination thickness established previously at the proximal site (4.7 mm), these cores should each represent over 1000 years of sediment accumulation, and likely much longer at the distal site, where laminations are expected to be thinner. Having established previously that the lake’s laminations are annual varves and that they are positively correlated with summer (melt-season) temperatures, our analysis is focused on documenting a long time-series of annual sediment accumulation and summer-layer particle size. Both measurements will be used to interpret the history of summer temperatures. The cores may also provide sedimentary evidence of the timing of advances/retreats of nearby glaciers, including the Tana Glacier and Bagley Icefield, helping to clarify the poorly-constrained timing of neoglaciation in Southern Alaska. The paleoclimate record produced at Iceberg Lake will be included in the Arctic System Science 8ka project

Thin optical display panel

DOEpatents

Veligdan, James Thomas

1997-01-01

An optical display includes a plurality of optical waveguides each including a cladding bound core for guiding internal display light between first and second opposite ends by total internal reflection. The waveguides are stacked together to define a collective display thickness. Each of the cores includes a heterogeneous portion defining a light scattering site disposed longitudinally between the first and second ends. Adjacent ones of the sites are longitudinally offset from each other for forming a longitudinal internal image display over the display thickness upon scattering of internal display light thereagainst for generating a display image. In a preferred embodiment, the waveguides and scattering sites are transparent for transmitting therethrough an external image in superposition with the display image formed by scattering the internal light off the scattering sites for defining a heads up display.

200 Years of Pb deposition throughout the Czech Republic: Patterns and sources

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vile, M.A.; Wieder, R.K.; Novak, M.

2000-01-01

Historical rates of Pb deposition were determined over the past 150--200 years for eight sites throughout the Czech Republic using {sup 210}Pb-dated, Sphagnum-derived peat cores. Maximum historical Pb deposition was greater at sites in the northern and western parts of the Czech Republic than at sites in the southern part of the Czech Republic. Lead deposition patterns generally reflect increasing industrialization over the past 100--200 years, especially in the post-World War II era. For seven of the eight sites, maximum Pb deposition occurred between 1965 and 1992, corresponding to a period of peak production and burning of lignite coal. Amore » decrease in Pb deposition rates since 1975--1980 was evident in seven of the sites. The most recent Pb deposition rates (1992), estimated from the uppermost peat core sections, averaged 32, 11, and 7 mg m{sup {minus}2} yr{sup {minus}1} for the northern, western, and southern sites, respectively, are higher than current Pb deposition in the eastern United States of 4 mg m{sup {minus}2} yr{sup {minus}1}. Lead deposition rates prior to Czech industrialization, estimated from the deepest dateable peat core sections, averaged 8, 5, and 1 mg m{sup {minus}2} yr{sup {minus}1} for the northern, western, and southern sites, respectively. Using acid-insoluble ash concentrations in peat and peat magnetic susceptibility determinations, the authors were able to identify past periods of elevated Pb deposition related to local mining of Pb-containing ore deposits at three of the sites and periods of elevated Pb deposition from fossil fuel combustion at five of the sites. Without stable Pb isotopic determinations, the importance of leaded gasoline-derived Pb could not be determined.« less

Landscape archaeological research and 3D modelling of the Neolithic site of Barcin Höyük, Northwest Turkey

NASA Astrophysics Data System (ADS)

Groenhuijzen, Mark; Kluiving, Sjoerd; Gerritsen, Fokke

2013-04-01

Barcin Höyük is a dwelling mound in the Yenişehir valley in Northwest Turkey. It is found to be one of the oldest farming communities in the region, with an archaeological record stretching from the Neolithic up to the Roman period, with some finds dating to the Byzantine period. An earlier geoarchaeological study was performed in 2009, revealing interesting deposits from a marsh or lake, and two possible small rivers or streams. The current study forms a continuation of the previous research, aimed at improving our understanding of the interrelationship of the site and the landscape, especially around the early neolithisation process. The following research questions have been investigated: is it possible to bring more detail into the knowledge of the landscape around the site through denser and more detailed coring, can a manner of time-control on the sedimentation be found, and is a 3D-model a suitable tool for storing and analysing data to improve our understanding of the landscape and the site? Data was gathered from hand auger corings placed in the vicinity of the excavation site. Soil samples were systematically gathered from these corings, of which a selection was subjected to laboratory analyses. The methods used here are grain size analysis, thermogravimetric analysis and end-member analysis. Spatial analytical tools, such as ArcMap and ArcScene were used to store and analyse all data, more specifically in order to construct the 3D-subsurface model. The deepest sedimentary unit encountered in the corings can be ascribed to a lacustrine environment, inferring that a lake might have been present at the site location prior to the first Neolithic habitation of Barcin Höyük. Two subsequent layers of gravel and coarse sand are found within the lacustrine unit and can be correlated around the site. In the 3D-subsurface model constructed for the site, these layers show a distinct elevation with a relief of almost 2.5 metres. These results can be interpreted as a presence of a large lake prior to the site, with the coarse layers representing beach deposits at the edge of the lake. The first habitation phase occurred on a natural elevation in the landscape left after a lake retreat, represented by the elevation in the coarse layers. A large amount of archaeological material was discovered in the corings, providing an opportunity for linking sedimentation with the archaeological record. The presence of a natural elevation near a lake edge might be a reason for the first habitation phase to begin. The archaeological material found in the corings proved useful in relative dating some of the layers and provides an indication of the sedimentation speed. The 3D-model of the subsurface is successful in providing more insight into the horizontal and vertical distribution of the deposits around the site of Barcin Höyük, and is also useful for combining and comparing data from this research with other sources.

Vapor-phase exchange of perchloroethene between soil and plants

USGS Publications Warehouse

Struckhoff, G.C.; Burken, J.G.; Schumacher, J.G.

2005-01-01

Tree core concentrations of tetrachloroethylene (perchloroethene, PCE) at the Riverfront Superfund Site in New Haven, MO, were found to mimic the profile of soil phase concentrations. The observed soil-tree core relationship was stronger than that of groundwater PCE to tree core concentrations at the same site. Earlier research has shown a direct, linear relationship between tree core and groundwater concentrations of chlorinated solvents and other organics. Laboratory-scale experiments were performed to elucidate this phenomenon, including determining partitioning coefficients of PCE between plant tissues and air and between plant tissues and water, measured to be 8.1 and 49 L/kg, respectively. The direct relationship of soil to tree core PCE concentrations was hypothesized to be caused by diffusion between tree roots and the soil vapor phase in the subsurface. The central findings of this research are discovering the importance of subsurface vapor-phase transfer for VOCs and uncovering a direct relationship between soil vapor-phase chlorinated solvents and uptake rates that impact contaminant translocation from the subsurface and transfer into the atmosphere. ?? 2005 American Chemical Society.

Interim Safe Storage of Plutonium Production Reactors at the US DOE Hanford Site - 13438

DOE Office of Scientific and Technical Information (OSTI.GOV)

Schilperoort, Daryl L.; Faulk, Darrin

2013-07-01

Nine plutonium production reactors located on DOE's Hanford Site are being placed into an Interim Safe Storage (ISS) period that extends to 2068. The Environmental Impact Statement (EIS) for ISS [1] was completed in 1993 and proposed a 75-year storage period that began when the EIS was finalized. Remote electronic monitoring of the temperature and water level alarms inside the safe storage enclosure (SSE) with visual inspection inside the SSE every 5 years are the only planned operational activities during this ISS period. At the end of the ISS period, the reactor cores will be removed intact and buried inmore » a landfill on the Hanford Site. The ISS period allows for radioactive decay of isotopes, primarily Co-60 and Cs-137, to reduce the dose exposure during disposal of the reactor cores. Six of the nine reactors have been placed into ISS by having an SSE constructed around the reactor core. (authors)« less

Geologic implications of gas hydrates in the offshore of India: Krishna-Godavari Basin, Mahanadi Basin, Andaman Sea, Kerala-Konkan Basin

USGS Publications Warehouse

Kumar, Pushpendra; Collett, Timothy S.; Boswell, Ray; Cochran, James R.; Lall, Malcolm; Mazumdar, Aninda; Ramana, Mangipudi Venkata; Ramprasad, Tammisetti; Riedel, Michael; Sain, Kalachand; Sathe, Arun Vasant; Vishwanath, Krishna; Yadav, U.S.

2014-01-01

NGHP-01 yielded evidence of gas hydrate from downhole log and core data obtained from all the sites in the Krishna–Godavari Basin, the Mahanadi Basin, and in the Andaman Sea. The site drilled in the Kerala–Konkan Basin during NGHP-01 did not yield any evidence of gas hydrate. Most of the downhole log-inferred gas hydrate and core-recovered gas hydrate were characterized as either fracture-filling in clay-dominated sediments or as pore-filling or grain-displacement particles disseminated in both fine- and coarse-grained sediments. Geochemical analyses of gases obtained from sediment cores recovered during NGHP-01 indicated that the gas in most all of the hydrates in the offshore of India is derived from microbial sources; only one site in the Andaman Sea exhibited limited evidence of a thermogenic gas source. The gas hydrate petroleum system concept has been used to effectively characterize the geologic controls on the occurrence of gas hydrates in the offshore of India.

Accumulation of p21 proteins at DNA damage sites independent of p53 and core NHEJ factors following irradiation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Koike, Manabu, E-mail: m_koike@nirs.go.jp; Yutoku, Yasutomo; Graduate School of Science, Chiba University, Chiba 263-8522

2011-08-19

Highlights: {yields} p21 accumulated rapidly at laser-irradiated sites via its C-terminal region. {yields} p21 colocalized with the DSB marker {gamma}-H2AX and the DSB sensor Ku80. {yields} Accumulation of p21 is dependent on PCNA, but not p53 and the NHEJ core factors. {yields} Accumulation activity of p21 was conserved among human and animal cells. {yields} p21 is a useful tool as a detection marker of DNA damaged sites. -- Abstract: The cyclin-dependent kinase (CDK) inhibitor p21 plays key roles in p53-dependent DNA-damage responses, i.e., cell cycle checkpoints, senescence, or apoptosis. p21 might also play a role in DNA repair. p21 focimore » arise at heavy-ion-irradiated DNA-double-strand break (DSB) sites, which are mainly repaired by nonhomologous DNA-end-joining (NHEJ). However, no mechanisms of p21 accumulation at double-strand break (DSB) sites have been clarified in detail. Recent works indicate that Ku70 and Ku80 are essential for the accumulation of other NHEJ core factors, e.g., DNA-PKcs, XRCC4 and XLF, and other DNA damage response factors, e.g., BRCA1. Here, we show that p21 foci arise at laser-irradiated sites in cells from various tissues from various species. The accumulation of EGFP-p21 was detected in not only normal cells, but also transformed or cancer cells. Our results also showed that EGFP-p21 accumulated rapidly at irradiated sites, and colocalized with the DSB marker {gamma}-H2AX and with the DSB sensor protein Ku80. On the other hand, the accumulation occurred in Ku70-, Ku80-, or DNA-PKcs-deficient cell lines and in human papillomavirus 18-positive cells, whereas the p21 mutant without the PCNA-binding region (EGFP-p21(1-146)) failed to accumulate at the irradiated sites. These findings suggest that the accumulation of p21, but not functional p53 and the NHEJ core factors, is dependent on PCNA. These findings also suggest that the accumulation activity of p21 at DNA damaged sites is conserved among human and animal cells, and p21 is a useful tool as a detection marker of DNA damaged sites.« less

Initial Geochemistry Data of the Lake Ohrid (Macedonia, Albania) DEEP -Site Sediment Record: The ICDP Scopsco Drilling Project

NASA Astrophysics Data System (ADS)

Francke, A.; Wagner, B.; Sulpizio, R.; Zanchetta, G.; Leicher, N.; Gromig, R.; Krastel, S.; Lindhorst, K.; Wilke, T.

2014-12-01

Ancient lakes, with sediment records spanning >1 million years, are very rare. The UNESCO World Heritage site of Lake Ohrid on the Balkans is thought to be the oldest lake in Europe. With 212 endemic species described to date, it is also a hotspot of evolution. In order to unravel the geological and evolutionary history of the lake, an international group of scientists, conducted a deep drilling campaign in spring 2013 under the umbrella of the ICDP SCOPSCO project (Scientific Collaboration on Past Speciation Conditions in Lake Ohrid). Overall, about 2,100 m of sediments were recovered from four drill sites. At the main drill site (DEEP-site) in central parts of the lake where seismic data indicated a maximum sediment fill of ca. 700 m, a total of more than 1,500 m of sediments were recovered until a penetration depth of 569 m. Currently, core opening, core description, XRF and MSCL scanning, sub-sampling (16 cm resolution), and inorganic and organic geochemical as well as sedimentological analyses of the sediment cores from the DEEP site are in progress at the University of Cologne. Previous studies at Lake Ohrid have shown that interglacial periods are characterized by high TIC and TOC contents, likely associated with high contents of calcite and organic matter in the sediments. In contrast, during glacial periods negligible TIC and low TOC contents correspond to high K counts indicating enhanced supply of clastic material. Similar patterns can be observed in the biogeochemical analyses of the subsamples and in the XRF data of the DEEP site record. Following these variations on a glacial-interglacial time scale, TIC and TOC data obtained from the subsamples and from core catcher samples indicate that the DEEP site sequence provides a 1.2 million year old continuous record of environmental and climatological variability in the Balkan Region. The age control can be further improved by first findings of macroscopic tephra horizons. Peaks in K, Sr, Zr, and magnetic susceptibility might indicate the occurrence of additional cryptotephra layers in the sediment sequence.

Climatic records of the last and penultimate deglaciations in the South Atlantic and South Indian Ocean

NASA Astrophysics Data System (ADS)

Michel, Elisabeth; Waelbroeck, Claire; Govin, Aline; Skinner, Luke; Vàzquez Riveiros, Natalia; Dewilde, Fabien; Isguder, Gulay; Rebaubier, Hélène

2013-04-01

Surface and deep-water records of Termination I and II in two twin South Atlantic deep-sea cores (44°09' S, 14°14' W, 3770 m depth) and one South Indian core (46°29' S, 88°01' E, 3420 m depth) are presented. Sea surface temperature has been reconstructed based on planktonic foraminifera census counts in all cases, as well as Mg/Ca of G. bulloides and N. pachyderma s. over the last deglaciation. The uncertainty on reconstructed SST using different statistical methods and different faunal databases is assessed. Over the last deglaciation, combined 14C dating and correlation of the SST record with the air temperature signal recorded in Antarctic ice cores allowed us to correct for variable surface reservoir ages in the South Atlantic core (Skinner et al., 2010). Preliminary dating of the South Indian core over the last termination has been done by correlation of its magnetic signal with those of a neighboring 14C dated core (Smart et al., 2010). We have refined the later age scale using the Atlantic core age scale as reference. Benthic isotopic signals in the South Atlantic and South Indian cores over the last deglaciation exhibit the same amplitude and timing. Our results thus indicate that bottom waters at the South Indian site remained isolated from better ventilated deep waters of northern origin until ~15 ka (Waelbroeck et al., 2011). Over Termination II, the two cores have been dated by correlation of their SST records with the air temperature signal recorded in EDC versus the EDC3 age scale (Govin et al., 2009; 2012). A careful examination of the various sources of uncertainty on the derived dating has been performed. Benthic and planktonic isotopic signals reveal analogies but also differences with respect to the last termination. SST was significantly warmer during the Last Interglacial than during the Holocene in both sites. South Atlantic deep waters were also significantly better ventilated during the Last Interglacial than during the Holocene, whereas bottom water ventilation was similar during these two interglacials at the South Indian site.

Genome Therapy of Myotonic Dystrophy Type 1 iPS Cells for Development of Autologous Stem Cell Therapy.

PubMed

Gao, Yuanzheng; Guo, Xiuming; Santostefano, Katherine; Wang, Yanlin; Reid, Tammy; Zeng, Desmond; Terada, Naohiro; Ashizawa, Tetsuo; Xia, Guangbin

2016-08-01

Myotonic dystrophy type 1 (DM1) is caused by expanded Cytosine-Thymine-Guanine (CTG) repeats in the 3'-untranslated region (3' UTR) of the Dystrophia myotonica protein kinase (DMPK) gene, for which there is no effective therapy. The objective of this study is to develop genome therapy in human DM1 induced pluripotent stem (iPS) cells to eliminate mutant transcripts and reverse the phenotypes for developing autologous stem cell therapy. The general approach involves targeted insertion of polyA signals (PASs) upstream of DMPK CTG repeats, which will lead to premature termination of transcription and elimination of toxic mutant transcripts. Insertion of PASs was mediated by homologous recombination triggered by site-specific transcription activator-like effector nuclease (TALEN)-induced double-strand break. We found genome-treated DM1 iPS cells continue to maintain pluripotency. The insertion of PASs led to elimination of mutant transcripts and complete disappearance of nuclear RNA foci and reversal of aberrant splicing in linear-differentiated neural stem cells, cardiomyocytes, and teratoma tissues. In conclusion, genome therapy by insertion of PASs upstream of the expanded DMPK CTG repeats prevented the production of toxic mutant transcripts and reversal of phenotypes in DM1 iPS cells and their progeny. These genetically-treated iPS cells will have broad clinical application in developing autologous stem cell therapy for DM1.

Interplay between Alternative Splicing and Alternative Polyadenylation Defines the Expression Outcome of the Plant Unique OXIDATIVE TOLERANT-6 Gene.

PubMed

Li, Qingshun Q; Liu, Zhaoyang; Lu, Wenjia; Liu, Man

2017-05-17

Pre-mRNA alternative splicing and alternative polyadenylation have been implicated to play important roles during eukaryotic gene expression. However, much remains unknown regarding the regulatory mechanisms and the interactions of these two processes in plants. Here we focus on an Arabidopsis gene OXT6 (Oxidative Tolerant-6) that has been demonstrated to encode two proteins through alternative splicing and alternative polyadenylation. Specifically, alternative polyadenylation at Intron-2 of OXT6 produces a transcript coding for AtCPSF30, an Arabidopsis ortholog of 30 kDa subunit of the Cleavage and Polyadenylation Specificity Factor. On the other hand, alternative splicing of Intron-2 generates a longer transcript encoding a protein named AtC30Y, a polypeptide including most part of AtCPSF30 and a YT521B domain. To investigate the expression outcome of OXT6 in plants, a set of mutations were constructed to alter the splicing and polyadenylation patterns of OXT6. Analysis of transgenic plants bearing these mutations by quantitative RT-PCR revealed a competition relationship between these two processes. Moreover, when both splice sites and poly(A) signals were mutated, polyadenylation became the preferred mode of OXT6 processing. These results demonstrate the interplay between alternative splicing and alternative polyadenylation, and it is their concerted actions that define a gene's expression outcome.

Highly Efficient Targeted Mutagenesis in Mice Using TALENs

PubMed Central

Panda, Sudeepta Kumar; Wefers, Benedikt; Ortiz, Oskar; Floss, Thomas; Schmid, Bettina; Haass, Christian; Wurst, Wolfgang; Kühn, Ralf

2013-01-01

Targeted mouse mutants are instrumental for the analysis of gene function in health and disease. We recently provided proof-of-principle for the fast-track mutagenesis of the mouse genome, using transcription activator-like effector nucleases (TALENs) in one-cell embryos. Here we report a routine procedure for the efficient production of disease-related knockin and knockout mutants, using improved TALEN mRNAs that include a plasmid-coded poly(A) tail (TALEN-95A), circumventing the problematic in vitro polyadenylation step. To knock out the C9orf72 gene as a model of frontotemporal lobar degeneration, TALEN-95A mutagenesis induced sequence deletions in 41% of pups derived from microinjected embryos. Using TALENs together with mutagenic oligodeoxynucleotides, we introduced amyotrophic lateral sclerosis patient-derived missense mutations in the fused in sarcoma (Fus) gene at a rate of 6.8%. For the simple identification of TALEN-induced mutants and their progeny we validate high-resolution melt analysis (HRMA) of PCR products as a sensitive and universal genotyping tool. Furthermore, HRMA of off-target sites in mutant founder mice revealed no evidence for undesired TALEN-mediated processing of related genomic sequences. The combination of TALEN-95A mRNAs for enhanced mutagenesis and of HRMA for simplified genotyping enables the accelerated, routine production of new mouse models for the study of genetic disease mechanisms. PMID:23979585

Nuclear poly(A)-binding protein aggregates misplace a pre-mRNA outside of SC35 speckle causing its abnormal splicing

PubMed Central

Klein, Pierre; Oloko, Martine; Roth, Fanny; Montel, Valérie; Malerba, Alberto; Jarmin, Susan; Gidaro, Teresa; Popplewell, Linda; Perie, Sophie; Lacau St Guily, Jean; de la Grange, Pierre; Antoniou, Michael N.; Dickson, George; Butler-Browne, Gillian; Bastide, Bruno; Mouly, Vincent; Trollet, Capucine

2016-01-01

A short abnormal polyalanine expansion in the polyadenylate-binding protein nuclear-1 (PABPN1) protein causes oculopharyngeal muscular dystrophy (OPMD). Mutated PABPN1 proteins accumulate as insoluble intranuclear aggregates in muscles of OPMD patients. While the roles of PABPN1 in nuclear polyadenylation and regulation of alternative poly(A) site choice have been established, the molecular mechanisms which trigger pathological defects in OPMD and the role of aggregates remain to be determined. Using exon array, for the first time we have identified several splicing defects in OPMD. In particular, we have demonstrated a defect in the splicing regulation of the muscle-specific Troponin T3 (TNNT3) mutually exclusive exons 16 and 17 in OPMD samples compared to controls. This splicing defect is directly linked to the SC35 (SRSF2) splicing factor and to the presence of nuclear aggregates. As reported here, PABPN1 aggregates are able to trap TNNT3 pre-mRNA, driving it outside nuclear speckles, leading to an altered SC35-mediated splicing. This results in a decreased calcium sensitivity of muscle fibers, which could in turn plays a role in muscle pathology. We thus report a novel mechanism of alternative splicing deregulation that may play a role in various other diseases with nuclear inclusions or foci containing an RNA binding protein. PMID:27507886

Substitution of histidine-137 by glutamine abolishes the catalytic activity of the ribosome-inactivating protein alpha-sarcin.

PubMed Central

Lacadena, J; Mancheño, J M; Martinez-Ruiz, A; Martínez del Pozo, A; Gasset, M; Oñaderra, M; Gavilanes, J G

1995-01-01

The alpha-sarcin cytotoxin is an extracellular fungal protein that inhibits protein biosynthesis by specifically cleaving one phosphodiester bond of the 28 S rRNA. The His137 residue of alpha-sarcin is suggested to be involved in the catalytic activity of this protein, based on the observed sequence similarity with some fungal ribonucleases. Replacement of this residue by Gln (H137Q mutant variant of alpha-sarcin) abolishes the ribonuclease activity of the protein. This has been demonstrated for an homogeneous preparation of the H137Q alpha-sarcin by measuring its effect against both intact rabbit ribosomes and the homopolymer poly(A). The conformation of H137Q alpha-sarcin is highly similar to that of the wild-type protein, which has been analysed by CD and fluorescence spectroscopy. Both H137Q and wild-type alpha-sarcin exhibit identical CD spectra in the peptide-bond region, indicating that no changes at the level of the secondary structure are produced upon mutation. Only minor differences are observed in both near-UV CD and fluorescence emission spectra in comparison to those of the wild-type protein. Moreover, H137Q alpha-sarcin interacts with phospholipid vesicles, promoting the same effects as the native cytotoxin. Therefore, we propose that His137 is part of the ribonucleolytic active site of the cytotoxin alpha-sarcin. Images Figure 4 PMID:7626023

PABPN1-Dependent mRNA Processing Induces Muscle Wasting

PubMed Central

Raz, Yotam; van Putten, Maaike; Paniagua-Soriano, Guillem; Krom, Yvonne D.; Florea, Bogdan I.; Raz, Vered

2016-01-01

Poly(A) Binding Protein Nuclear 1 (PABPN1) is a multifunctional regulator of mRNA processing, and its expression levels specifically decline in aging muscles. An expansion mutation in PABPN1 is the genetic cause of oculopharyngeal muscle dystrophy (OPMD), a late onset and rare myopathy. Moreover, reduced PABPN1 expression correlates with symptom manifestation in OPMD. PABPN1 regulates alternative polyadenylation site (PAS) utilization. However, the impact of PAS utilization on cell and tissue function is poorly understood. We hypothesized that altered PABPN1 expression levels is an underlying cause of muscle wasting. To test this, we stably down-regulated PABPN1 in mouse tibialis anterior (TA) muscles by localized injection of adeno-associated viruses expressing shRNA to PABPN1 (shPab). We found that a mild reduction in PABPN1 levels causes muscle pathology including myofiber atrophy, thickening of extracellular matrix and myofiber-type transition. Moreover, reduced PABPN1 levels caused a consistent decline in distal PAS utilization in the 3’-UTR of a subset of OPMD-dysregulated genes. This alternative PAS utilization led to up-regulation of Atrogin-1, a key muscle atrophy regulator, but down regulation of proteasomal genes. Additionally reduced PABPN1 levels caused a reduction in proteasomal activity, and transition in MyHC isotope expression pattern in myofibers. We suggest that PABPN1-mediated alternative PAS utilization plays a central role in aging-associated muscle wasting. PMID:27152426

Autoselection of cytoplasmic yeast virus like elements encoding toxin/antitoxin systems involves a nuclear barrier for immunity gene expression.

PubMed

Kast, Alene; Voges, Raphael; Schroth, Michael; Schaffrath, Raffael; Klassen, Roland; Meinhardt, Friedhelm

2015-05-01

Cytoplasmic virus like elements (VLEs) from Kluyveromyces lactis (Kl), Pichia acaciae (Pa) and Debaryomyces robertsiae (Dr) are extremely A/T-rich (>75%) and encode toxic anticodon nucleases (ACNases) along with specific immunity proteins. Here we show that nuclear, not cytoplasmic expression of either immunity gene (PaORF4, KlORF3 or DrORF5) results in transcript fragmentation and is insufficient to establish immunity to the cognate ACNase. Since rapid amplification of 3' ends (RACE) as well as linker ligation of immunity transcripts expressed in the nucleus revealed polyadenylation to occur along with fragmentation, ORF-internal poly(A) site cleavage due to the high A/T content is likely to prevent functional expression of the immunity genes. Consistently, lowering the A/T content of PaORF4 to 55% and KlORF3 to 46% by gene synthesis entirely prevented transcript cleavage and permitted functional nuclear expression leading to full immunity against the respective ACNase toxin. Consistent with a specific adaptation of the immunity proteins to the cognate ACNases, cross-immunity to non-cognate ACNases is neither conferred by PaOrf4 nor KlOrf3. Thus, the high A/T content of cytoplasmic VLEs minimizes the potential of functional nuclear recruitment of VLE encoded genes, in particular those involved in autoselection of the VLEs via a toxin/antitoxin principle.

The X-ray Crystal Structures of Human {alpha}-Phosphomannomutase 1 Reveal the Structural Basis of Congenital Disorder of Glycosylation Type 1a

DOE Office of Scientific and Technical Information (OSTI.GOV)

Silvaggi,N.; Zhang, C.; Lu, Z.

2006-01-01

Carbohydrate-deficient glycoprotein syndrome type 1a (CDG-1a) is a congenital disease characterized by severe defects in nervous system development. It is caused by mutations in alpha -phosphomannomutase (of which there are two isozymes, {alpha}-PMM1 and {alpha}-PPM2). Here we report the X-ray crystal structures of human {alpha}-PMM1 in the open conformation, with and without the bound substrate, {alpha}-D-mannose 1-phosphate. {alpha}-PMM1, like most Haloalkanoic Acid Dehalogenase Superfamily (HADSF) members, consists of two domains, the cap and core, which open to bind substrate and then close to provide a solvent exclusive environment for catalysis. The substrate phosphate group is observed at a positively chargedmore » site of the cap domain, rather than at the core domain phosphoryl-transfer site defined by the D19 nucleophile and Mg{sup 2+} cofactor. This suggests that substrate binds first to the cap and then is swept into the active site upon cap closure. The orientation of the acid/base residue D21 suggests that {alpha}-PMM uses a different method of protecting the aspartylphosphate from hydrolysis than the HADSF member {beta}-phosphoglucomutase. It is hypothesized that the electrostatic repulsion of positive charges at the interface of the cap and core domains stabilizes {alpha}-PMM1 in the open conformation, and that the negatively charged substrate binds to the cap, thereby facilitating its closure over the core domain. The two isozymes {alpha}-PMM1 and {alpha}-PMM2 are shown to have a conserved active-site structure and to display similar kinetic properties. Analysis of the known mutation sites in the context of the structures reveals the genotype-phenotype relationship underlying CDG-1a.« less

Documenting the location of systematic transrectal ultrasound-guided prostate biopsies: correlation with multi-parametric MRI.

PubMed

Turkbey, Baris; Xu, Sheng; Kruecker, Jochen; Locklin, Julia; Pang, Yuxi; Shah, Vijay; Bernardo, Marcelino; Baccala, Angelo; Rastinehad, Ardeshir; Benjamin, Compton; Merino, Maria J; Wood, Bradford J; Choyke, Peter L; Pinto, Peter A

2011-03-29

During transrectal ultrasound (TRUS)-guided prostate biopsies, the actual location of the biopsy site is rarely documented. Here, we demonstrate the capability of TRUS-magnetic resonance imaging (MRI) image fusion to document the biopsy site and correlate biopsy results with multi-parametric MRI findings. Fifty consecutive patients (median age 61 years) with a median prostate-specific antigen (PSA) level of 5.8 ng/ml underwent 12-core TRUS-guided biopsy of the prostate. Pre-procedural T2-weighted magnetic resonance images were fused to TRUS. A disposable needle guide with miniature tracking sensors was attached to the TRUS probe to enable fusion with MRI. Real-time TRUS images during biopsy and the corresponding tracking information were recorded. Each biopsy site was superimposed onto the MRI. Each biopsy site was classified as positive or negative for cancer based on the results of each MRI sequence. Sensitivity, specificity, and receiver operating curve (ROC) area under the curve (AUC) values were calculated for multi-parametric MRI. Gleason scores for each multi-parametric MRI pattern were also evaluated. Six hundred and 5 systemic biopsy cores were analyzed in 50 patients, of whom 20 patients had 56 positive cores. MRI identified 34 of 56 positive cores. Overall, sensitivity, specificity, and ROC area values for multi-parametric MRI were 0.607, 0.727, 0.667, respectively. TRUS-MRI fusion after biopsy can be used to document the location of each biopsy site, which can then be correlated with MRI findings. Based on correlation with tracked biopsies, T2-weighted MRI and apparent diffusion coefficient maps derived from diffusion-weighted MRI are the most sensitive sequences, whereas the addition of delayed contrast enhancement MRI and three-dimensional magnetic resonance spectroscopy demonstrated higher specificity consistent with results obtained using radical prostatectomy specimens.

10 CFR 960.3-1-4-2 - Site nomination for characterization.

Code of Federal Regulations, 2012 CFR

2012-01-01

... testing of core samples for the evaluation of geochemical and engineering rock properties, and chemical... industrial activities; and extrapolations of regional data to estimate site-specific characteristics and...

10 CFR 960.3-1-4-2 - Site nomination for characterization.

Code of Federal Regulations, 2013 CFR

2013-01-01

... testing of core samples for the evaluation of geochemical and engineering rock properties, and chemical... industrial activities; and extrapolations of regional data to estimate site-specific characteristics and...

10 CFR 960.3-1-4-2 - Site nomination for characterization.

Code of Federal Regulations, 2011 CFR

2011-01-01

... testing of core samples for the evaluation of geochemical and engineering rock properties, and chemical... industrial activities; and extrapolations of regional data to estimate site-specific characteristics and...

10 CFR 960.3-1-4-2 - Site nomination for characterization.

Code of Federal Regulations, 2014 CFR

2014-01-01

... testing of core samples for the evaluation of geochemical and engineering rock properties, and chemical... industrial activities; and extrapolations of regional data to estimate site-specific characteristics and...

Localized Deformation Beginning more than 15 km Beneath the Mid-Atlantic Ridge, 14 to 16 N

NASA Astrophysics Data System (ADS)

Kelemen, P.

2003-12-01

ODP Leg 209 drilled 19 holes at 8 sites along the Mid-Atlantic Ridge from 14° 43 to 15° N. All sites were surveyed by submersible, and chosen to be < 200 m from peridotite or dunite exposed on the seafloor; outcrops of gabbroic rock were also close to some sites. One of our primary goals was to constrain the mechanism of mantle upwelling, corner flow and exhumation of shallow mantle rocks. Drilling at Sites 1268, 1270-72, 1274 and 1275 penetrated 1075 meters, and recovered 354 m of core. At Sites 1268 and 1270-72 we recovered 25% gabbroic rocks and 75% residual mantle peridotite. Core from Site 1274 was mainly residual peridotite, while core from Site 1275 was mainly gabbroic. Most of the residual peridotites have nearly undeformed, protogranular textures. Orthopyroxenes are interstitial to olivine or even poikilitic. Rare, isolated clinopyroxene grains are also interstitial. Skeletal spinel grains have mm-scale extensions in three dimensions, with no discernable shape fabric. These textures are clearly different from porphyroclastic textures typical in ophiolites and fracture zone dredges. As described elsewhere at this meeting, impregnated peridotites contain olivine, 2 pyroxenes, plagioclase and spinel, and equilibrated at 0.54 GPa (+/-0.14 GPa, 2σ ) and 1220° C (+/-16° C, 2σ ) [Kinzler & Grove, JGR 92]. Melts entered the thermal boundary layer beneath the Mid-Atlantic Ridge at about 20 km [e.g., Sleep, JGR 75; Reid & Jackson, MGR 82; Grove et al JGR 92; Cannat JGR 96; Michael & Chase CMP 97; Braun et al., EPSL 00], and began to crystallize within impregnated peridotites and as discrete plutons intruding peridotite. Gabbroic rocks and peridotites from most sites underwent large tectonic rotations since aquiring remanent magnetization. At some sites, rotations may have exceeded 60° around near-horizontal axes parallel to the Mid-Atlantic Ridge. Such large rotations are unlikely to have been accomodated along a single fault, and instead blocks were progressively rotated along a series of fault systems. Sites 1270 (25% gabbroic) and 1275 (75% gabbroic) were drilled into large, low angle fault surfaces previously identified as oceanic core complexes. Deformation at Site 1270 is similar to most other Sites, while core from Site 1275 is the most weakly deformed. However, high pressure igneous assemblages indicate that some Site 1275 rocks were exhumed from depths of 20 km or more, as were residual peridotites at the other Sites. At all sites except 1269, 1273 and 1275, we recovered high temperature mylonitic shear zones (mainly with impregnated peridotite mineralogy, but also mylonitic gabbros), and intervals of low temperature fault gouge. Shear zones and faults are not all parallel; numerous, cross-cutting planes of localized deformation formed at > 1000° C to < 100° C, from > 15 km depth to near the seafloor. These accommodated nearly all subsolidus deformation during corner flow and exhumation of residual peridotites (plus high pressure igneous rocks), in keeping with the inference that the thermal boundary layer in this region extends to at least 20 km. Penetrative, viscous deformation of blocks between shear zones and faults was minor. If this is a general process at slow-spreading ridges, then one would predict that shallow mantle anisotropy in the Atlantic would be less pronounced than in the Pacific, consistent with recent seismic data [FAIM Experiment, Gaherty, Collins et al, this session].

Benthic foraminiferal faunal and geochemical proxies as tracers for paleoenvironmental and paleoceanographic changes in the western Mediterranean over the last 24 ka

NASA Astrophysics Data System (ADS)

Pérez-Asensio, José N.; Cacho, Isabel; Frigola, Jaime; Pena, Leopoldo D.; Sierro, Francisco J.; Asioli, Alessandra; Kuhlmann, Jannis; Huhn, Katrin

2017-04-01

Paleoenvironmental and paleoceanographic changes in the western Mediterranean are reconstructed for the last 24 ka using a combination of benthic foraminiferal assemblages and geochemical proxies measured on benthic foraminiferal shells (Mg/Ca-deep water temperatures and stable isotopes). The studied materials are sediment cores HER-GC-UB06 and MD95-2043recovered at 946 m and 1841 m, respectively, from the Alboran Sea. At present, both core sites are bathed by the Western Mediterranean Deep Water (WMDW), although UB06 core is close to the boundary with the overlying Levantine Intermediate Water (LIW). Therefore, past variability of both water masses can potentially be recorded by the benthic foraminiferal proxies from the studied sites. Benthic foraminiferal assemblages and geochemical data show fluctuations in bottom-water ventilation, organic matter accumulation and deep-water temperatures related to WMDW and LIW circulation. During the glacial interval, an alternation of events showing better ventilation (higher abundance of Cibicides pachyderma) with lower temperatures and events of warmer deep water temperatures with poorer ventilation (Nonionella iridea assemblage, lower abundance of C. pachyderma) are observed. This variability might reflect stronger WMDW formation during the Last Glacial Maximum (LGM) and Heinrich Stadial 1. During the Bølling-Allerød and Younger Dryas (YD) periods, cold temperatures and the lowest oxygenation rates are recorded coinciding with the highest abundance of deep infaunal taxa on both UB06 and MD95-2043 cores. This interval was coetaneous to the deposition of an Organic Rich Layer in the Alboran Sea. However, a re-ventilation trend started at the end of the YD in the shallower site (UB06 core) whereas low-oxygen conditions prevailed until the end of the early Holocene in the deep site (MD95-2043 core). During the early Holocene a significant deep water temperature increase occurred at the shallower site suggesting the replacement of WMDW by warmer water mass, likely LIW. In the middle Holocene, highly variable bottom-water oxygenation and temperatures are observed showing warmer deep waters with less oxygen content (higher deep and intermediate infaunal abundances). The late Holocene (last 4 ka) was characterized by slightly cooler deep water temperatures and enhanced oxygen levels supporting that WMDW became dominant at the shallower site. These observations reveal that Mediterranean thermohaline system has been highly variable during the studied period supporting its high sensitivity to changing climate conditions. These results open a new insight into the Mediterranean sensitivity to Holocene climate variability.

Deciphering Equatorial Pacific Deep Sea Sediment Transport Regimes by Core-Log-Seismic Integration

NASA Astrophysics Data System (ADS)

Ortiz, E.; Tominaga, M.; Marcantonio, F.

2017-12-01

Investigating deep-sea sediment transportation and deposition regimes is a key to accurately understand implications from geological information recorded by pelagic sediments, e.g. climate signals. However, except for physical oceanographic particle trap experiments, geochemical analyses of in situsediments, and theoretical modeling of the relation between the bottom currents and sediment particle flux, it has remained a challenging task to document the movement of deep sea sediments, that takes place over time. We utilized high-resolution, multichannel reflection seismic data from the eastern equatorial Pacific region with drilling and logging results from two Integrated Ocean Drilling Program (IODP) sites, the Pacific Equatorial Age Transect (PEAT) 7 (Site U1337) and 8 (Site U1338), to characterize sediment transportation regimes on 18-24 Ma oceanic crust. Site U1337, constructed by a series of distinct abyssal hills and abyssal basins; Site U1338, located 570 km SE from Site U1337 site and constructed by a series of ridges, seamounts, and abyssal hills. These sites are of particular interest due to their proximity to the equatorial productivity zone, areas with high sedimentation rates and preservation of carbonate-bearing sediment that provide invaluable insights on equatorial Pacific ecosystems and carbon cycle. We integrate downhole geophysical logging data as well as geochemistry and physical properties measurements on recovered cores from IODP Sites U1337 and U1338 to comprehensively examine the mobility of deep-sea sediments and sediment diagenesis over times in a quasi-3D manner. We also examine 1100 km of high resolution underway seismic surveys from site survey lines in between PEAT 7 and 8 in order to investigate changes in sediment transportation between both sites. Integrating detailed seismic interpretations, high resolution core data, and 230Th flux measurements we aim to create a detailed chronological sedimentation and sediment diagenesis history of this area.

A Chronostratigraphic and Environment Magnetic Study of Drill Cores Collected in the Mozambique Channel on Cruise MD13

NASA Astrophysics Data System (ADS)

Ray, D. J.; DeBone, K. N.; Smesny, J.; Chadinha, C.; Mitchell, B.; Acton, G. D.; Kulhanek, D. K.

2017-12-01

The South Atlantic Petroleum Company donated drill cores from 18 sites cored in October 2013 in the Mozambique Channel off Madagascar by the RV Marion Dufresne. Up to 30.11 m of sediment was recovered from cores collected at two localities, referred to as Juan de Nova ( 16.5°S, 42.9°E) and Belo Profound ( 19.5°S, 42.2°E), in water depths of 2,157-2,795 m. The cores are composed mostly of mixtures of fairly homogenous biogenic ooze and clay that is highly bioturbated, with turbidites occurring relatively commonly (about every meter or two) in most of the cores, although rarely in others (e.g., Core MD13-3506). The core color varies between light tan, olive gray, light brown, and light reddish brown. The darker colors probably reflect higher proportion of terrigenous input and the lighter intervals more biogenic input. We collected rock magnetic and paleomagnetic data along the cores and conducted initial calcareous nannofossil biostratigraphic analyses in order to provide stratigraphic and chronologic constraints and to investigate paleoenvironment changes. Records of magnetic susceptibility are used to correlate stratigraphy between sites and then are correlated to well-dated global records of climate change, such as the marine oxygen isotope record. The correlation to oxygen isotope records, which is constrained by biostratigraphic and paleomagnetic observations, helps refine the chronostratigraphy and allows us to interpret the variations in lithology and magnetic properties in the context of global climate change.

Contaminant Gradients in Trees: Directional Tree Coring Reveals Boundaries of Soil and Soil-Gas Contamination with Potential Applications in Vapor Intrusion Assessment.

PubMed

Wilson, Jordan L; Samaranayake, V A; Limmer, Matthew A; Schumacher, John G; Burken, Joel G

2017-12-19

Contaminated sites pose ecological and human-health risks through exposure to contaminated soil and groundwater. Whereas we can readily locate, monitor, and track contaminants in groundwater, it is harder to perform these tasks in the vadose zone. In this study, tree-core samples were collected at a Superfund site to determine if the sample-collection location around a particular tree could reveal the subsurface location, or direction, of soil and soil-gas contaminant plumes. Contaminant-centroid vectors were calculated from tree-core data to reveal contaminant distributions in directional tree samples at a higher resolution, and vectors were correlated with soil-gas characterization collected using conventional methods. Results clearly demonstrated that directional tree coring around tree trunks can indicate gradients in soil and soil-gas contaminant plumes, and the strength of the correlations were directly proportionate to the magnitude of tree-core concentration gradients (spearman's coefficient of -0.61 and -0.55 in soil and tree-core gradients, respectively). Linear regression indicates agreement between the concentration-centroid vectors is significantly affected by in planta and soil concentration gradients and when concentration centroids in soil are closer to trees. Given the existing link between soil-gas and vapor intrusion, this study also indicates that directional tree coring might be applicable in vapor intrusion assessment.

Contaminant gradients in trees: Directional tree coring reveals boundaries of soil and soil-gas contamination with potential applications in vapor intrusion assessment

USGS Publications Warehouse

Wilson, Jordan L.; Samaranayake, V.A.; Limmer, Matthew A.; Schumacher, John G.; Burken, Joel G.

2017-01-01

Contaminated sites pose ecological and human-health risks through exposure to contaminated soil and groundwater. Whereas we can readily locate, monitor, and track contaminants in groundwater, it is harder to perform these tasks in the vadose zone. In this study, tree-core samples were collected at a Superfund site to determine if the sample-collection location around a particular tree could reveal the subsurface location, or direction, of soil and soil-gas contaminant plumes. Contaminant-centroid vectors were calculated from tree-core data to reveal contaminant distributions in directional tree samples at a higher resolution, and vectors were correlated with soil-gas characterization collected using conventional methods. Results clearly demonstrated that directional tree coring around tree trunks can indicate gradients in soil and soil-gas contaminant plumes, and the strength of the correlations were directly proportionate to the magnitude of tree-core concentration gradients (spearman’s coefficient of -0.61 and -0.55 in soil and tree-core gradients, respectively). Linear regression indicates agreement between the concentration-centroid vectors is significantly affected by in-planta and soil concentration gradients and when concentration centroids in soil are closer to trees. Given the existing link between soil-gas and vapor intrusion, this study also indicates that directional tree coring might be applicable in vapor intrusion assessment.

Changes in Black Carbon Deposition to Antarctica from Two Ice Core Records, A.D. 1850-2000

NASA Technical Reports Server (NTRS)

Bisiaux, Marion M.; Edward, Ross; McConnell, Joseph R.; Curran, Mark A. J.; VanOmmen, Tas D.; Smith, Andrew M.; Neumann, Thomas A.; Pasteris, Daniel R.; Penner, Joyce E.; Taylor, Kendrick

2012-01-01

Continuous flow analysis was based on a steady sample flow and in-line detection of BC and other chemical substances as described in McConnell et al. (2007). In the cold room, previously cut one meter ice core sticks of 3x3cm, are melted continuously on a heated melter head specifically designed to eliminate contamination from the atmosphere or by the external parts of the ice. The melted ice from the most inner part of the ice stick is continuously pumped by a peristaltic pump and carried to a clean lab by Teflon lines. The recorded signal is continuous, integrating a sample volume of about 0.05 mL, for which the temporal resolution depends on the speed of melting, ice density and snow accumulation rate at the ice core drilling site. For annual accumulation derived from the WAIS and Law Dome ice cores, we assumed 3.1 cm water equivalent uncertainty in each year's accumulation from short scale spatial variability (glaciological noise) which was determined from several measurements of annual accumulation in multiple parallel ice cores notably from the WAIS Divide ice core site (Banta et al., 2008) and from South Pole site (McConnell et al., 1997; McConnell et al., 2000). Refractory black carbon (rBC) concentrations were determined using the same method as in (Bisiaux et al., 2011) and adapted to continuous flow measurements as described by (McConnell et al., 2007). The technique uses a single particle intracavity laser induced incandescence photometer (SP2, Droplet Measurement Technologies, Boulder, Colorado) coupled to an ultrasonic nebulizer/desolvation (CETAC UT5000) Flow Injection Analysis (FIA). All analyses, sample preparation etc, were performed in a class 100 cleanroom using anti contamination "clean techniques". The samples were not acidified.

Natural gas geochemistry of sediments drilled on the 2005 Gulf of Mexico JIP cruise

USGS Publications Warehouse

Lorenson, T.D.; Claypool, G.E.; Dougherty, J.A.

2008-01-01

In April and May 2005, cores were acquired and sub-sampled for gases in lease blocks Atwater Valley 13 and 14 and Keathley Canyon 151 during deep subseafloor drilling conducted as part of the JIP study of gas hydrates in the northern Gulf of Mexico. Sample types included sediment headspace gas, free gas derived from sediment gas exsolution, and gas exsolution from controlled degassing of pressurized cores. The gases measured both onboard and in shore-based labs were nitrogen, oxygen, hydrogen sulfide, carbon dioxide, and the hydrocarbons methane through hexane. The presence of seafloor mounds, seismic anomalies, a shallow sulfate-methane interface, and similar gas compositions and isotopic compositions near the seafloor and at depth suggest an upward flux of methane at both sites. Sediment gases at the Atwater Valley sites, where seafloor mounds and adjacent sediments were cored, strongly suggest a microbial source of methane, with very little thermogenic gas input. Sediment gas from all cores contained from about 96 to 99.9% methane, with the balance composed primarily of carbon dioxide. Methane to ethane ratios were greater than 1000, and often over 10,000. Gases from cores at Keathley Canyon were similar to those at Atwater Valley, however, deeper cores from Keathley Canyon contained more ethane, propane, and butane suggesting mixing with minor concentrations thermogenic gas. The isotopic composition of methane, ethane, and carbon dioxide were measured, and ??13C values range from -84.3 to -71.5???, -65.2 to -46.8???, and -23.5 to -3.0???, respectively, all consistent with microbial gas sources, early diagenesis of organic matter and perhaps biodegradation of petroleum. The presence of deep microbial gas at these sites here and elsewhere highlights a potentially significant, predominantly microbial gas source in the northern Gulf of Mexico.

Hepatitis B Virus Core Gene Mutations Which Block Nucleocapsid Envelopment

PubMed Central

Koschel, Matthias; Oed, Daniela; Gerelsaikhan, Tudevdagwa; Thomssen, Reiner; Bruss, Volker

2000-01-01

Recently we generated a panel of hepatitis B virus core gene mutants carrying single insertions or deletions which allowed efficient expression of the core protein in bacteria and self-assembly of capsids. Eleven of these mutations were introduced into a eukaryotic core gene expression vector and characterized by trans complementation of a core-negative HBV genome in cotransfected human hepatoma HuH7 cells. Surprisingly, four mutants (two insertions [EFGA downstream of A11 and LDTASALYR downstream of R39] and two deletions [Y38-R39-E40 and L42]) produced no detectable capsids. The other seven mutants supported capsid formation and pregenome packaging/viral minus- and plus-strand-DNA synthesis but to different levels. Four of these seven mutants (two insertions [GA downstream of A11 and EHCSP downstream of P50] and two deletions [S44 and A80]) allowed virion morphogenesis and secretion. The mutant carrying a deletion of A80 at the tip of the spike protruding from the capsid was hepatitis B virus core antigen negative but wild type with respect to virion formation, indicating that this site might not be crucial for capsid-surface protein interactions during morphogenesis. The other three nucleocapsid-forming mutants (one insertion [LS downstream of S141] and two deletions [T12 and P134]) were strongly blocked in virion formation. The corresponding sites are located in the part of the protein forming the body of the capsid and not in the spike. These mutations may alter sites on the particle which contact surface proteins during envelopment, or they may block the appearance of a signal for the transport or the maturation of the capsid which is linked to viral DNA synthesis and required for envelopment. PMID:10590084

Holocene Accumulation and Ice Flow near the West Antarctic Ice Sheet Divide Ice Core Site

NASA Technical Reports Server (NTRS)

Koutnik, Michelle R.; Fudge, T.J.; Conway, Howard; Waddington, Edwin D.; Neumann, Thomas A.; Cuffey, Kurt M.; Buizert, Christo; Taylor, Kendrick C.

2016-01-01

The West Antarctic Ice Sheet Divide Core (WDC) provided a high-resolution climate record from near the Ross-Amundsen Divide in Central West Antarctica. In addition, radar-detected internal layers in the vicinity of the WDC site have been dated directly from the ice core to provide spatial variations in the age structure of the region. Using these two data sets together, we first infer a high-resolution Holocene accumulation-rate history from 9.2 thousand years of the ice-core timescale and then confirm that this climate history is consistent with internal layers upstream of the core site. Even though the WDC was drilled only 24 kilometers from the modern ice divide, advection of ice from upstream must be taken into account. We evaluate histories of accumulation rate by using a flowband model to generate internal layers that we compare to observed layers. Results show that the centennially averaged accumulation rate was over 20 percent lower than modern at 9.2 thousand years before present (B.P.), increased by 40 percent from 9.2 to 2.3 thousand years B.P., and decreased by at least 10 percent over the past 2 thousand years B.P. to the modern values; these Holocene accumulation-rate changes in Central West Antarctica are larger than changes inferred from East Antarctic ice-core records. Despite significant changes in accumulation rate, throughout the Holocene the regional accumulation pattern has likely remained similar to today, and the ice-divide position has likely remained on average within 5 kilometers of its modern position. Continent-scale ice-sheet models used for reconstructions of West Antarctic ice volume should incorporate this accumulation history.

The Hominin Sites and Paleolakes Drilling Project (HSPDP): Understanding the paleoenvironmental and paleoclimatic context of human origins through continental drilling

NASA Astrophysics Data System (ADS)

Cohen, Andrew S.; Campisano, Christopher; Asrat, Asfawossen; Arrowsmith, Ramon; Deino, Alan; Feibel, Craig; Hill, Andrew; Kingston, John; Lamb, Henry; Lowenstein, Tim; Olago, Daniel; Bernhart Owen, R.; Renaut, Robin; Schabitz, Frank; Trauth, Martin

2015-04-01

The influence of climate and environmental history on human evolution is an existential question that continues to be hotly debated, in part because of the paucity of high resolution records collected in close proximity to the key fossil and archaeological evidence. To address this issue and transform the scientific debate, the HSPDP was developed to collect lacustrine sediment drill cores from basins in Kenya and Ethiopia that collectively encompass critical time intervals and locations for Plio-Quaternary human evolution in East Africa. After a 17 month campaign, drilling was completed in November, 2014, with over 1750m of core collected from 11 boreholes from five areas (1930m total drilling length, avg. 91% recovery). The sites, from oldest to youngest, include 1) N. Awash, Ethiopia (~3.5-2.9Ma core interval); 2) Baringo-Tugen Hills, Kenya (~3.3-2.5Ma); 3) West Turkana, Kenya (~1.9-1.4Ma); L. Magadi, Kenya (0.8-0Ma) and the Chew Bahir Basin, Ethiopia (~0.5-0Ma). Initial core description (ICD) and sampling for geochronology, geochemistry and paleoecology studies had been completed by mid2014, with the two remaining sites (Magadi and Chew Bahir) scheduled for ICD work in early 2015. Whereas the primary scientific targets were the lacustrine deposits from the hominin-bearing basin depocenters, many intervals of paleosols (representative of low lake stands and probable arid periods) were also encountered in drill cores. Preliminary analyses of drill core sedimentology and geochemistry show both long-term lake level changes and cyclic variability in lake levels, both of which may be indicative of climatic forcing events of interest to paleoanthropologists. Authors of this abstract also include the entire HSPDP field team.

Historical record and fluxes of DDTs at the Palos Verdes Shelf Superfund site, California.

PubMed

Liao, Chunyang; Taylor, Allison R; Kenney, William F; Schlenk, Daniel; Gan, Jay

2017-03-01

Marine sediments at many locations in the world are contaminated with a wide range of persistent organic pollutants. The Palos Verdes Shelf (PVS) is located in the ocean off the coast of Los Angeles, California and has been listed as a Superfund site by the US EPA since 1997, because of heavy contamination of DDTs and PCBs. However, little is known about the historical trend in the deposition of DDTs as a result of decades-long discharge of wastewater effluents. In this study, sediment cores were taken from the PVS site and determined for DDT and its metabolites including DDE and DDD (denoted as DDTs). Individual DDTs were found in the majority (95%) of the samples analyzed. The highest ∑DDT concentrations were found in three cores along the 60-meter isobath with geometric means of 31300, 7490, and 5010ng/gdw and medians of 82400, 17300, and 5200ng/g dw, respectively. Among DDT congeners, p,p'-DDE, o,p'-DDE and p,p'-DDD were predominant, contributing to approximately 54%, 27%, and 14% of the ΣDDTs in sediment. The vertical profiles of concentrations of contaminants in the sediment cores were examined. For most of the cores, a steady increase in the concentrations of DDTs during 1940s to 1980s was observed, while the concentrations declined gradually toward the surficial layers. On the basis of the mass flux of DDTs calculated and the area of the PVS Superfund site, we estimated the total deposition amount of DDTs in sediment and the deposition amount of ΣDDTs in this region during 1947-1971 was 132 tons, which was fairly close to what was reported in earlier studies for industrial wastewater discharge in the PVS site (870-1450tons). Our findings suggest that the elevated levels of DDTs in sediment from the PVS site are linked to the discharge of these contaminants between the 1940s-1980s. Copyright © 2016 Elsevier B.V. All rights reserved.

An Early Pleistocene high-resolution paleoclimate reconstruction from the West Turkana (Kenya) HSPDP drill site

NASA Astrophysics Data System (ADS)

Stockhecke, Mona; Beck, Catherine; Brown, Erik T.; Cohen, Andrew; Deocampo, Daniel M.; Feibel, Craig S.; Pelletier, Jon D.; Rabideaux, Nathane M.; Sier, Mark

2016-04-01

The Hominin Sites and Paleolakes Drilling Project (HSPDP), and the related Olorgesailie Drilling Project (ODP), recovered ~2 km of drill core since 2012. At the HSPDP West Turkana Kaitio (WTK) site a 216 m-long core that covers the Early Pleistocene time window (1.3 to 1.87 Ma) during which hominids first expanded out of Africa and marine records document reorganization of tropical climate and the development of the strong Walker circulation. WTK carries particular interest for paleoclimate and paleoenvironmental reconstructions as it is located only 2.5 km from the location of one of the most complete hominin skeletons ever recovered (Nariokotome Boy). XRF core scanning data provide a means of evaluating records of past environmental conditions continuously and at high resolution. However, the record contains complex lithologies reflecting repeated episodes of inundation and desiccation along a dynamic lake margin. Here we present a methodological approach to address the highly variable lithostratigraphy of the East African records to establish comprehensive paleoclimate timeseries. The power spectrum of the presented hydroclimate record peaks at Milankovitch cycles, qualifying HSPDP drill cores from the Turkana Basin to be used as high-resolution Early Pleistocene paleoclimate archive. Comparing these data with marine climate reconstructions sheds light into athmospheric processes and continental climate dynamics.

Recognition of the Xenopus ribosomal core promoter by the transcription factor xUBF involves multiple HMG box domains and leads to an xUBF interdomain interaction.

PubMed

Leblanc, B; Read, C; Moss, T

1993-02-01

The interaction of the ribosomal transcription factor xUBF with the RNA polymerase I core promoter of Xenopus laevis has been studied both at the DNA and protein levels. It is shown that a single xUBF-DNA complex forms over the 40S initiation site (+1) and involves at least the DNA sequences between -20 and +60 bp. DNA sequences upstream of +10 and downstream of +18 are each sufficient to direct complex formation independently. HMG box 1 of xUBF independently recognizes the sequences -20 to -1 and +1 to +22 and the addition of the N-terminal dimerization domain to HMG box 1 stabilizes its interaction with these sequences approximately 10-fold. HMG boxes 2/3 interact with the DNA downstream of +22 and can independently position xUBF across the initiation site. The C-terminal segment of xUBF, HMG boxes 4, 5 or the acidic domain, directly or indirectly interact with HMG box 1, making the core promoter sequences between -11 and -15 hypersensitive to DNase. This interaction also requires the DNA sequences between +17 and +32, i.e. the HMG box 2/3 binding site. The data suggest extensive folding of the core promoter within the xUBF complex.

In-situ stress measurements using core-based methods in the vicinity of Nojima fault.

NASA Astrophysics Data System (ADS)

Yano, S.; Sugimoto, T.; Lin, W.; Lin, A.

2017-12-01

In the cycle of repeatable occurrence of earthquakes, stress accumulates at the source fault and its surroundings in an interseismic period until the next earthquake, and releases abruptly when the earthquake occurs. However, it is almost unknown that the quantitative relationship between stress change and earthquake occurrence. Hence, in order to improve our understanding on the mechanisms of the outbreak of earthquakes, it is important to grasp the stress states in the vicinity of the source fault and to evaluate its change over time. In this study, we carried out in-situ stress measurements by using core samples obtained from a scientific drilling penetrated through the Nojima fault which ruptured and caused the Hyogo-ken Nanbu earthquake, Japan in 1995. Our stress measurements were conducted from 2016 to 2017 when is 22 years after the earthquake. For this purpose, we applied the Anelastic Strain Recovery (ASR) method and Diametrical Core Deformation Analysis (DCDA). First, we measure the ASR change with time of the cores from stress releasing soon and calculate three-dimensional principal in-situ stress orientations and magnitudes from the ASR data. In this study, to ensure the enough amount of ASR, we conducted the measurements using the cores collected within a short time (e.g. 2.5 - 3.5 hours) after stress releasing by drilling at an on-site laboratory in the drilling site in Awaji island, Japan. The site locates at the south-west part of the Nojima fault. In DCDA, we measure the core diameters in all (360°) azimuths, and determine difference of the two horizontal principal stresses and their orientation by using the other cores as those used for ASR. DCDA experiments were conducted indoor and after a long time passed from core collecting. Lithology of all the core samples we used for ASR and DCDA are granite, and 19 and 7 cores were used for ASR and DCDA, respectively. As a result, it was found that the stress state in the depth range of 500 - 560 m and around 822 m were of normal faulting stress regime and that in 711 - 730 m was strike slip faulting type. As mentioned above, we have obtained a data set of the current state of stress around Nojima fault using the two core-based stress measurement methods. In future, we will determine the core orientations and restore the directions of the principal stress axes to the geologic coodinates.

Influenza A Virus Host Shutoff Disables Antiviral Stress-Induced Translation Arrest

PubMed Central

Khaperskyy, Denys A.; Emara, Mohamed M.; Johnston, Benjamin P.; Anderson, Paul; Hatchette, Todd F.; McCormick, Craig

2014-01-01

Influenza A virus (IAV) polymerase complexes function in the nucleus of infected cells, generating mRNAs that bear 5′ caps and poly(A) tails, and which are exported to the cytoplasm and translated by host machinery. Host antiviral defences include mechanisms that detect the stress of virus infection and arrest cap-dependent mRNA translation, which normally results in the formation of cytoplasmic aggregates of translationally stalled mRNA-protein complexes known as stress granules (SGs). It remains unclear how IAV ensures preferential translation of viral gene products while evading stress-induced translation arrest. Here, we demonstrate that at early stages of infection both viral and host mRNAs are sensitive to drug-induced translation arrest and SG formation. By contrast, at later stages of infection, IAV becomes partially resistant to stress-induced translation arrest, thereby maintaining ongoing translation of viral gene products. To this end, the virus deploys multiple proteins that block stress-induced SG formation: 1) non-structural protein 1 (NS1) inactivates the antiviral double-stranded RNA (dsRNA)-activated kinase PKR, thereby preventing eIF2α phosphorylation and SG formation; 2) nucleoprotein (NP) inhibits SG formation without affecting eIF2α phosphorylation; 3) host-shutoff protein polymerase-acidic protein-X (PA-X) strongly inhibits SG formation concomitant with dramatic depletion of cytoplasmic poly(A) RNA and nuclear accumulation of poly(A)-binding protein. Recombinant viruses with disrupted PA-X host shutoff function fail to effectively inhibit stress-induced SG formation. The existence of three distinct mechanisms of IAV-mediated SG blockade reveals the magnitude of the threat of stress-induced translation arrest during viral replication. PMID:25010204

Characterization of mRNA-Cytoskeleton Interactions In Situ Using FMTRIP and Proximity Ligation

PubMed Central

Jung, Jeenah; Lifland, Aaron W.; Alonas, Eric J.; Zurla, Chiara; Santangelo, Philip J.

2013-01-01

Many studies have demonstrated an association between the cytoskeleton and mRNA, as well as the asymmetric distribution of mRNA granules within the cell in response to various signaling events. It is likely that the extensive cytoskeletal network directs mRNA transport and localization, with different cytoskeletal elements having their own specific roles. In order to understand the spatiotemporal changes in the interactions between the mRNA and the cytoskeleton as a response to a stimulus, a technique that can visualize and quantify these changes across a population of cells while capturing cell-to-cell variations is required. Here, we demonstrate a method for imaging and quantifying mRNA-cytoskeleton interactions on a per cell basis with single-interaction sensitivity. Using a proximity ligation assay with flag-tagged multiply-labeled tetravalent RNA imaging probes (FMTRIP), we quantified interactions between mRNAs and β-tubulin, vimentin, or filamentous actin (F-actin) for two different mRNAs, poly(A) + and β-actin mRNA, in two different cell types, A549 cells and human dermal fibroblasts (HDF). We found that the mRNAs interacted predominantly with F-actin (>50% in HDF, >20% in A549 cells), compared to β-tubulin (<5%) and vimentin (11-13%). This likely reflects differences in mRNA management by the two cell types. We then quantified changes in these interactions in response to two perturbations, F-actin depolymerization and arsenite-induced oxidative stress, both of which alter either the cytoskeleton itself and mRNA localization. Both perturbations led to a decrease in poly(A) + mRNA interactions with F-actin and an increase in the interactions with microtubules, in a time dependent manner. PMID:24040294

Mass spectrometric identification of proteins that interact through specific domains of the poly(A) binding protein

PubMed Central

Zhang, Chongxu; Nielsen, Maria E. O.; Chiang, Yueh-Chin; Kierkegaard, Morten; Wang, Xin; Lee, Darren J.; Andersen, Jens S.; Yao, Gang

2013-01-01

Poly(A) binding protein (PAB1) is involved in a number of RNA metabolic functions in eukaryotic cells and correspondingly is suggested to associate with a number of proteins. We have used mass spectrometric analysis to identify 55 non-ribosomal proteins that specifically interact with PAB1 from Saccharomyces cerevisiae. Because many of these factors may associate only indirectly with PAB1 by being components of the PAB1-mRNP structure, we additionally conducted mass spectrometric analyses on seven metabolically defined PAB1 deletion derivatives to delimit the interactions between these proteins and PAB1. These latter analyses identified 13 proteins whose associations with PAB1 were reduced by deleting one or another of PAB1’s defined domains. Included in this list of 13 proteins were the translation initiation factors eIF4G1 and eIF4G2, translation termination factor eRF3, and PBP2, all of whose previously known direct interactions with specific PAB1 domains were either confirmed, delimited, or extended. The remaining nine proteins that interacted through a specific PAB1 domain were CBF5, SLF1, UPF1, CBC1, SSD1, NOP77, yGR250c, NAB6, and GBP2. In further study, UPF1, involved in nonsense-mediated decay, was confirmed to interact with PAB1 through the RRM1 domain. We additionally established that while the RRM1 domain of PAB1 was required for UPF1-induced acceleration of deadenylation during nonsense-mediated decay, it was not required for the more critical step of acceleration of mRNA decapping. These results begin to identify the proteins most likely to interact with PAB1 and the domains of PAB1 through which these contacts are made. PMID:22836166

Mass spectrometric identification of proteins that interact through specific domains of the poly(A) binding protein.

PubMed

Richardson, Roy; Denis, Clyde L; Zhang, Chongxu; Nielsen, Maria E O; Chiang, Yueh-Chin; Kierkegaard, Morten; Wang, Xin; Lee, Darren J; Andersen, Jens S; Yao, Gang

2012-09-01

Poly(A) binding protein (PAB1) is involved in a number of RNA metabolic functions in eukaryotic cells and correspondingly is suggested to associate with a number of proteins. We have used mass spectrometric analysis to identify 55 non-ribosomal proteins that specifically interact with PAB1 from Saccharomyces cerevisiae. Because many of these factors may associate only indirectly with PAB1 by being components of the PAB1-mRNP structure, we additionally conducted mass spectrometric analyses on seven metabolically defined PAB1 deletion derivatives to delimit the interactions between these proteins and PAB1. These latter analyses identified 13 proteins whose associations with PAB1 were reduced by deleting one or another of PAB1's defined domains. Included in this list of 13 proteins were the translation initiation factors eIF4G1 and eIF4G2, translation termination factor eRF3, and PBP2, all of whose previously known direct interactions with specific PAB1 domains were either confirmed, delimited, or extended. The remaining nine proteins that interacted through a specific PAB1 domain were CBF5, SLF1, UPF1, CBC1, SSD1, NOP77, yGR250c, NAB6, and GBP2. In further study, UPF1, involved in nonsense-mediated decay, was confirmed to interact with PAB1 through the RRM1 domain. We additionally established that while the RRM1 domain of PAB1 was required for UPF1-induced acceleration of deadenylation during nonsense-mediated decay, it was not required for the more critical step of acceleration of mRNA decapping. These results begin to identify the proteins most likely to interact with PAB1 and the domains of PAB1 through which these contacts are made.

The CCR4-NOT complex mediates deadenylation and degradation of stem cell mRNAs and promotes planarian stem cell differentiation.

PubMed

Solana, Jordi; Gamberi, Chiara; Mihaylova, Yuliana; Grosswendt, Stefanie; Chen, Chen; Lasko, Paul; Rajewsky, Nikolaus; Aboobaker, A Aziz

2013-01-01

Post-transcriptional regulatory mechanisms are of fundamental importance to form robust genetic networks, but their roles in stem cell pluripotency remain poorly understood. Here, we use freshwater planarians as a model system to investigate this and uncover a role for CCR4-NOT mediated deadenylation of mRNAs in stem cell differentiation. Planarian adult stem cells, the so-called neoblasts, drive the almost unlimited regenerative capabilities of planarians and allow their ongoing homeostatic tissue turnover. While many genes have been demonstrated to be required for these processes, currently almost no mechanistic insight is available into their regulation. We show that knockdown of planarian Not1, the CCR4-NOT deadenylating complex scaffolding subunit, abrogates regeneration and normal homeostasis. This abrogation is primarily due to severe impairment of their differentiation potential. We describe a stem cell specific increase in the mRNA levels of key neoblast genes after Smed-not1 knock down, consistent with a role of the CCR4-NOT complex in degradation of neoblast mRNAs upon the onset of differentiation. We also observe a stem cell specific increase in the frequency of longer poly(A) tails in these same mRNAs, showing that stem cells after Smed-not1 knock down fail to differentiate as they accumulate populations of transcripts with longer poly(A) tails. As other transcripts are unaffected our data hint at a targeted regulation of these key stem cell mRNAs by post-transcriptional regulators such as RNA-binding proteins or microRNAs. Together, our results show that the CCR4-NOT complex is crucial for stem cell differentiation and controls stem cell-specific degradation of mRNAs, thus providing clear mechanistic insight into this aspect of neoblast biology.

The CCR4-NOT Complex Mediates Deadenylation and Degradation of Stem Cell mRNAs and Promotes Planarian Stem Cell Differentiation

PubMed Central

Solana, Jordi; Gamberi, Chiara; Mihaylova, Yuliana; Grosswendt, Stefanie; Chen, Chen; Lasko, Paul; Rajewsky, Nikolaus; Aboobaker, A. Aziz

2013-01-01

Post-transcriptional regulatory mechanisms are of fundamental importance to form robust genetic networks, but their roles in stem cell pluripotency remain poorly understood. Here, we use freshwater planarians as a model system to investigate this and uncover a role for CCR4-NOT mediated deadenylation of mRNAs in stem cell differentiation. Planarian adult stem cells, the so-called neoblasts, drive the almost unlimited regenerative capabilities of planarians and allow their ongoing homeostatic tissue turnover. While many genes have been demonstrated to be required for these processes, currently almost no mechanistic insight is available into their regulation. We show that knockdown of planarian Not1, the CCR4-NOT deadenylating complex scaffolding subunit, abrogates regeneration and normal homeostasis. This abrogation is primarily due to severe impairment of their differentiation potential. We describe a stem cell specific increase in the mRNA levels of key neoblast genes after Smed-not1 knock down, consistent with a role of the CCR4-NOT complex in degradation of neoblast mRNAs upon the onset of differentiation. We also observe a stem cell specific increase in the frequency of longer poly(A) tails in these same mRNAs, showing that stem cells after Smed-not1 knock down fail to differentiate as they accumulate populations of transcripts with longer poly(A) tails. As other transcripts are unaffected our data hint at a targeted regulation of these key stem cell mRNAs by post-transcriptional regulators such as RNA-binding proteins or microRNAs. Together, our results show that the CCR4-NOT complex is crucial for stem cell differentiation and controls stem cell-specific degradation of mRNAs, thus providing clear mechanistic insight into this aspect of neoblast biology. PMID:24367277

Genome Modification Leads to Phenotype Reversal in Human Myotonic Dystrophy type 1 iPS-cell Derived Neural Stem Cells

PubMed Central

Xia, Guangbin; Gao, Yuanzheng; Jin, Shouguang; Subramony, SH.; Terada, Naohiro; Ranum, Laura P.W.; Swanson, Maurice S.; Ashizawa, Tetsuo

2015-01-01

Objective Myotonic dystrophy type 1 (DM1) is caused by expanded CTG repeats in the 3'-untranslated region (3’ UTR) of the DMPK gene. Correcting the mutation in DM1 stem cells would be an important step towards autologous stem cell therapy. The objective of this study is to demonstrate in vitro genome editing to prevent production of toxic mutant transcripts and reverse phenotypes in DM1 stem cells. Methods Genome editing was performed in DM1 neural stem cells (NSCs) derived from human DM1 iPS cells. An editing cassette containing SV40/bGH polyA signals was integrated upstream of the CTG repeats by TALEN-mediated homologous recombination (HR). The expression of mutant CUG repeats transcript was monitored by nuclear RNA foci, the molecular hallmarks of DM1, using RNA fluorescence in situ hybridization (RNA-FISH). Alternative splicing of microtubule-associated protein tau (MAPT) and muscleblind-like (MBNL) proteins were analyzed to further monitor the phenotype reversal after genome modification. Results The cassette was successfully inserted into DMPK intron 9 and this genomic modification led to complete disappearance of nuclear RNA foci. MAPT and MBNL 1, 2 aberrant splicing in DM1 NSCs was reversed to normal pattern in genome-modified NSCs. Interpretation Genome modification by integration of exogenous polyA signals upstream of the DMPK CTG repeat expansion prevents the production of toxic RNA and leads to phenotype reversal in human DM1 iPS-cells derived stem cells. Our data provide proof-of-principle evidence that genome modification may be used to generate genetically modified progenitor cells as a first step toward autologous cell transfer therapy for DM1. PMID:25702800

Complex alternative splicing of acetylcholinesterase transcripts in Torpedo electric organ; primary structure of the precursor of the glycolipid-anchored dimeric form.

PubMed Central

Sikorav, J L; Duval, N; Anselmet, A; Bon, S; Krejci, E; Legay, C; Osterlund, M; Reimund, B; Massoulié, J

1988-01-01

In this paper, we show the existence of alternative splicing in the 3' region of the coding sequence of Torpedo acetylcholinesterase (AChE). We describe two cDNA structures which both diverge from the previously described coding sequence of the catalytic subunit of asymmetric (A) forms (Schumacher et al., 1986; Sikorav et al., 1987). They both contain a coding sequence followed by a non-coding sequence and a poly(A) stretch. Both of these structures were shown to exist in poly(A)+ RNAs, by S1 mapping experiments. The divergent region encoded by the first sequence corresponds to the precursor of the globular dimeric form (G2a), since it contains the expected C-terminal amino acids, Ala-Cys. These amino acids are followed by a 29 amino acid extension which contains a hydrophobic segment and must be replaced by a glycolipid in the mature protein. Analyses of intact G2a AChE showed that the common domain of the protein contains intersubunit disulphide bonds. The divergent region of the second type of cDNA consists of an adjacent genomic sequence, which is removed as an intron in A and Ga mRNAs, but may encode a distinct, less abundant catalytic subunit. The structures of the cDNA clones indicate that they are derived from minor mRNAs, shorter than the three major transcripts which have been described previously (14.5, 10.5 and 5.5 kb). Oligonucleotide probes specific for the asymmetric and globular terminal regions hybridize with the three major transcripts, indicating that their size is determined by 3'-untranslated regions which are not related to the differential splicing leading to A and Ga forms. Images PMID:3181125

The dopamine precursor L-dihydroxyphenylalanine is transported by the amino acid transporters rBAT and LAT2 in renal cortex.

PubMed

Quiñones, Henry; Collazo, Roberto; Moe, Orson W

2004-07-01

The intrarenal autocrine-paracrine dopamine (DA) system is critical for Na(+) homeostasis. l-Dihydroxyphenylalanine (l-DOPA) uptake from the glomerular filtrate and plasma provides the substrate for DA generation by the renal proximal tubule. The transporter(s) responsible for proximal tubule l-DOPA uptake has not been characterized. Renal cortical poly-A(+) RNA injected into Xenopus laevis oocytes induced l-DOPA uptake in a time- and dose-dependent fashion with biphasic K(m)s in the millimolar and micromolar range and independent of inward Na(+), K(+), or H(+) gradients, suggesting the presence of low- and high-affinity l-DOPA carriers. Complementary RNA from two amino acid transporters yielded l-DOPA uptake significantly above water-injected controls the rBAT/b(0,+)AT dimer (rBAT) and the LAT2/4F2 dimer (LAT2). In contradistinction to renal cortical poly-A(+), l-DOPA kinetics of rBAT and LAT2 showed classic Michaelis-Menton kinetics with K(m)s in the micromolar and millimolar range, respectively. Sequence-specific antisense oligonucleotides to rBAT or LAT2 (AS) caused inhibition of rBAT and LAT2 cRNA-induced l-DOPA transport and cortical poly-A(+)-induced arginine and phenylalanine transport. However, the same ASs only partially blocked poly-A(+)-induced l-DOPA transport. In cultured kidney cells, silencing inhibitory RNA (siRNA) to rBAT significantly inhibited l-DOPA uptake. We conclude that rBAT and LAT2 can mediate apical and basolateral l-DOPA uptake into the proximal tubule, respectively. Additional l-DOPA transport mechanisms exist in the renal cortex that remain to be identified.

CCR4-NOT deadenylates mRNA associated with RNA-induced silencing complexes in human cells.

PubMed

Piao, Xianghua; Zhang, Xue; Wu, Ligang; Belasco, Joel G

2010-03-01

MicroRNAs (miRNAs) repress gene expression posttranscriptionally by inhibiting translation and by expediting deadenylation so as to trigger rapid mRNA decay. Their regulatory influence is mediated by the protein components of the RNA-induced silencing complex (RISC), which deliver miRNAs and siRNAs to their mRNA targets. Here, we present evidence that CCR4-NOT is the deadenylase that removes poly(A) from messages destabilized by miRNAs in human cells. Overproducing a mutationally inactivated form of either of the catalytic subunits of this deadenylase (CCR4 or CAF1/POP2) significantly impedes the deadenylation and decay of mRNA targeted by a partially complementary miRNA. The same deadenylase initiates the degradation of "off-target" mRNAs that are bound by an imperfectly complementary siRNA introduced by transfection. The greater inhibitory effect of inactive CAF1 or POP2 (versus inactive CCR4) suggests a predominant role for this catalytic subunit of CCR4-NOT in miRNA- or small interfering RNA (siRNA)-mediated deadenylation. These effects of mi/siRNAs and CCR4-NOT can be fully reproduced by directly tethering RISC to mRNA without the guidance of a small RNA, indicating that the ability of RISC to accelerate deadenylation is independent of RNA base pairing. Despite its importance for mi/siRNA-mediated deadenylation, CCR4-NOT appears not to associate significantly with RISC, as judged by the failure of CAF1 and POP2 to coimmunoprecipitate detectably with either the Ago or TNRC6 subunit of RISC, a finding at odds with deadenylase recruitment as the mechanism by which RISC accelerates poly(A) removal.

Determining miRNA Expression Levels in Degraded RNA Samples Using Real-Time RT-qPCR and Microarray Technologies

PubMed Central

Tighe, S.; Holbrook, J.; Nadella, V.; Carmical, R.; Sol-Church, K.; Yueng, A.T.; Chittur, S.

2011-01-01

The Nucleic Acid Research Group (NARG) has previously conducted studies evaluating the impact of RNA integrity and priming strategies on cDNA synthesis and real-time RT-qPCR. The results of last year's field study as it relates to degraded RNA will be presented. In continuation of the RNA integrity theme, this year's study was designed to evaluate the impact of RNA integrity on the analysis of miRNA expression using real-time RT-qPCR. Target section was based on data obtained by the Microarray Research Group (MARG) and other published data from next gen sequencing. These 9 miRNAs represent three groups of miRNA that are expressed at low, medium or high levels in the First Choice human brain reference RNA sample. Two popular RT priming strategies tested in this study include the Megaplex miRNA TaqMan assay (ABI) and the RT2 miRNA qPCR assay (Qiagen/SA Biosciences). The basis for the ABI assay design is a target-specific stem-loop structure and reverse-transcription primer, while the Qiagen design combines poly(A) tailing and a universal reverse transcription in one cDNA synthesis reaction. For this study, the human brain reference RNA was subject to controlled degradation using RNase A to RIN (RNA Integrity Number) values of 7 (good), 4 (moderately degraded), and 2 (severely degraded).These templates were then used to assess both RT methods. In addition to this real-time RT-qPCR data, the same RNA templates were further analyzed using universal poly(A) tailing and hybridization to Affymetrix miRNA GeneChips. This talk will provide insights into RT priming strategies for miRNA and contrast the qPCR results obtained using different technologies.

Remodeling of the pioneer translation initiation complex involves translation and the karyopherin importin β

PubMed Central

Sato, Hanae; Maquat, Lynne E.

2009-01-01

Mammalian mRNAs lose and acquire proteins throughout their life span while undergoing processing, transport, translation, and decay. How translation affects messenger RNA (mRNA)–protein interactions is largely unknown. The pioneer round of translation uses newly synthesized mRNA that is bound by cap-binding protein 80 (CBP80)–CBP20 (also known as the cap-binding complex [CBC]) at the cap, poly(A)-binding protein N1 (PABPN1) and PABPC1 at the poly(A) tail, and, provided biogenesis involves pre-mRNA splicing, exon junction complexes (EJCs) at exon–exon junctions. Subsequent rounds of translation engage mRNA that is bound by eukaryotic translation initiation factor 4E (eIF4E) at the cap and PABPC1 at the poly(A) tail, but that lacks detectable EJCs and PABPN1. Using the level of intracellular iron to regulate the translation of specific mRNAs, we show that translation promotes not only removal of EJC constituents, including the eIF4AIII anchor, but also replacement of PABPN1 by PABPC1. Remarkably, translation does not affect replacement of CBC by eIF4E. Instead, replacement of CBC by eIF4E is promoted by importin β (IMPβ): Inhibiting the binding of IMPβ to the complex of CBC–IMPα at an mRNA cap using the IMPα IBB (IMPβ-binding) domain or a RAN variant increases the amount of CBC-bound mRNA and decreases the amount of eIF4E-bound mRNA. Our studies uncover a previously unappreciated role for IMPβ and a novel paradigm for how newly synthesized messenger ribonucleoproteins (mRNPs) are matured. PMID:19884259

Characteristics and significance of intergenic polyadenylated RNA transcription in Arabidopsis.

PubMed

Moghe, Gaurav D; Lehti-Shiu, Melissa D; Seddon, Alex E; Yin, Shan; Chen, Yani; Juntawong, Piyada; Brandizzi, Federica; Bailey-Serres, Julia; Shiu, Shin-Han

2013-01-01

The Arabidopsis (Arabidopsis thaliana) genome is the most well-annotated plant genome. However, transcriptome sequencing in Arabidopsis continues to suggest the presence of polyadenylated (polyA) transcripts originating from presumed intergenic regions. It is not clear whether these transcripts represent novel noncoding or protein-coding genes. To understand the nature of intergenic polyA transcription, we first assessed its abundance using multiple messenger RNA sequencing data sets. We found 6,545 intergenic transcribed fragments (ITFs) occupying 3.6% of Arabidopsis intergenic space. In contrast to transcribed fragments that map to protein-coding and RNA genes, most ITFs are significantly shorter, are expressed at significantly lower levels, and tend to be more data set specific. A surprisingly large number of ITFs (32.1%) may be protein coding based on evidence of translation. However, our results indicate that these "translated" ITFs tend to be close to and are likely associated with known genes. To investigate if ITFs are under selection and are functional, we assessed ITF conservation through cross-species as well as within-species comparisons. Our analysis reveals that 237 ITFs, including 49 with translation evidence, are under strong selective constraint and relatively distant from annotated features. These ITFs are likely parts of novel genes. However, the selective pressure imposed on most ITFs is similar to that of randomly selected, untranscribed intergenic sequences. Our findings indicate that despite the prevalence of ITFs, apart from the possibility of genomic contamination, many may be background or noisy transcripts derived from "junk" DNA, whose production may be inherent to the process of transcription and which, on rare occasions, may act as catalysts for the creation of novel genes.

A single amino-acid substitution in the Ets domain alters core DNA binding specificity of Ets1 to that of the related transcription factors Elf1 and E74.

PubMed

Bosselut, R; Levin, J; Adjadj, E; Ghysdael, J

1993-11-11

Ets proteins form a family of sequence specific DNA binding proteins which bind DNA through a 85 aminoacids conserved domain, the Ets domain, whose sequence is unrelated to any other characterized DNA binding domain. Unlike all other known Ets proteins, which bind specific DNA sequences centered over either GGAA or GGAT core motifs, E74 and Elf1 selectively bind to GGAA corecontaining sites. Elf1 and E74 differ from other Ets proteins in three residues located in an otherwise highly conserved region of the Ets domain, referred to as conserved region III (CRIII). We show that a restricted selectivity for GGAA core-containing sites could be conferred to Ets1 upon changing a single lysine residue within CRIII to the threonine found in Elf1 and E74 at this position. Conversely, the reciprocal mutation in Elf1 confers to this protein the ability to bind to GGAT core containing EBS. This, together with the fact that mutation of two invariant arginine residues in CRIII abolishes DNA binding, indicates that CRIII plays a key role in Ets domain recognition of the GGAA/T core motif and lead us to discuss a model of Ets proteins--core motif interaction.

Application of individual foraminifera Mg/Ca and δ18O analyses for paleoceanographic reconstructions in the Bay of Bengal and other active depositional environments

NASA Astrophysics Data System (ADS)

Fritz-Endres, T.; Dekens, P.; Fehrenbacher, J. S.; Spero, H. J.; Stine, A.

2017-12-01

Paleoceanographic research traditionally focuses on regions where sediment deposition is minimally affected by transport. However, sediment fans near tectonically active regions provide an opportunity to link oceanographic climate to terrestrial processes. Sediment cores recovered during IODP Expedition 354 in the Bay of Bengal include hemipelagic sections that record the history of tectonic uplift and the development of the Indian Monsoon through the last 10 Ma. Although these cores provide a unique opportunity to link marine and terrestrial climate, the complex depositional environment requires that the source of foraminifera is carefully considered before using these proxies to reconstruct oceanographic conditions. Foraminifera in Bengal Fan sediments may have been transported via turbidity currents from the northern Bay of Bengal, where the seasonal variability of SST and SSS is larger compared to the southern Bay of Bengal. We measured single Globigerinoides sacculifer Mg/Ca and δ18O from mudline samples of IODP Site U1454 (8.4°N, 85.5°E, 3721 m water depth) near the modern active channel and Site U1449 (8.4°N, 88.7°E, 3653 m water depth) far from channel activity. We compare these sites to single G. sacculifer from the core-top sample of Site 342KL (20.6°N, 90.1°E, 1256 m water depth) located on the continental shelf. Each foraminifera lives 2-4 weeks and the distribution of 60 to 80 data points reflects the seasonal range of SST and SSS at the location where the foraminifera calcified. Measurements in foraminifera from Site U1449 (away from active channel) are statistically different from the site in the northern Bay of Bengal and more consistent with local conditions. Conversely, foraminifera from the site near the active channel reflect a combined signal of local conditions recorded from the site far from channel activity and those recorded from the continental shelf. This suggests a portion of foraminifera from the active channel site have been transported from the northern Bay of Bengal. Our data show that foraminifera can be used to reconstruct SST and δ18O in this complex depositional environment, but caution must be taken when the down-core lithology indicates turbidites and possible sediment transport

Ocean Drilling Program: Science Operator Site Index

Science.gov Websites

time estimator Long-Term Observatories and Legacy Holes (University of Miami site) Drilling Services systems Internet systems Help Desk Database services How to obtain ODP data Data types and examples Core

Trafficking of Hepatitis C Virus Core Protein during Virus Particle Assembly

PubMed Central

Counihan, Natalie A.; Rawlinson, Stephen M.; Lindenbach, Brett D.

2011-01-01

Hepatitis C virus (HCV) core protein is directed to the surface of lipid droplets (LD), a step that is essential for infectious virus production. However, the process by which core is recruited from LD into nascent virus particles is not well understood. To investigate the kinetics of core trafficking, we developed methods to image functional core protein in live, virus-producing cells. During the peak of virus assembly, core formed polarized caps on large, immotile LDs, adjacent to putative sites of assembly. In addition, LD-independent, motile puncta of core were found to traffic along microtubules. Importantly, core was recruited from LDs into these puncta, and interaction between the viral NS2 and NS3-4A proteins was essential for this recruitment process. These data reveal new aspects of core trafficking and identify a novel role for viral nonstructural proteins in virus particle assembly. PMID:22028650

Physical, Chemical, and Hydrologic Characteristics of Active Layer and Permafrost Soils of Arctic Polygonal Tundra, Barrow, Alaska, 2013-2016

DOE Data Explorer

Yuxin Wu; Craig Ulrich; Tim Kneafsey

2018-03-22

This dataset reports the results of physical, chemical, and hydrologic analyses of soil cores collected at the NGEE Arctic Intensive Sites 0 and 1 (Plots A, B, C, and D) during late winter seasons (April-May) from 2013 to 2016. Locations were selected across the polygonal ground to include low, flat, and high centered polygons (LCPs, FCPs and HCPs) and internal features (i.e. the trough, center, and rim). The data are depth-resolved and down to ~ 4m below ground surface for some cores. The specific depths for each core where samples were analyzed vary from core to core.

Interstitial water studies on small core samples, Deep Sea Drilling Project, Leg 8

USGS Publications Warehouse

Manheim, F.T.; Sayles, F.L.

1971-01-01

Leg 8 sites are dominated by siliceous-calcareous biogenic oozes having depositional rates of 0.1 to 1.5 cm/1000 years. Conservative constituents of pore fluids showed, as have cores from other pelagic areas of the Pacific, insignificant or marginally significant changes with depth and location. However, in Sites 70 and 71, calcium, magnesium and strontium showed major shifts in concentration with depth. These changes appear to be related to recrystallization phenomena in skeletal debris of nannoplankton and to the relative accumulation rate of the sediments. The chemical anomalies increase relatively smoothly with depth, demonstrating the effectiveness of vertical diffusional communication, and apparent lack of bulk fluid movement, as noted in Leg 7 and other sites.

Spatial variability of shelf sediments in the STRATAFORM natural laboratory, Northern California

USGS Publications Warehouse

Goff, J.A.; Wheatcroft, R.A.; Lee, H.; Drake, D.E.; Swift, D.J.P.; Fan, S.

2002-01-01

The "Correlation Length Experiment", an intensive box coring effort on the Eel River shelf (Northern California) in the summer of 1997, endeavored to characterize the lateral variability of near-surface shelf sediments over scales of meters to kilometers. Coring focused on two sites, K60 and S60, separated by ??? 15 km along the 60 m isobath. The sites are near the sand-to-mud transition, although K60 is sandier owing to its proximity to the Eel River mouth. Nearly 140 cores were collected on dip and strike lines with core intervals from < 10m to 1 km. Measurements on each core included bulk density computed from gamma-ray attenuation, porosity converted from resistivity measurements, and surficial grain size. Grain size was also measured over the full depth range within a select subset of cores. X-radiograph images were also examined. Semi-variograms were computed for strike, dip, and down-hole directions at each site. The sand-to-mud transition exerts a strong influence on all measurements: on average, bulk density increases and porosity decreases with regional increases in mean grain size. Analysis of bulk density measurements indicates very strong contrasts in the sediment variability at K60 and S60. No coherent bedding is seen at K60; in the strike direction, horizontal variability is "white" (fully uncorrelated) from the smallest scales examined (a few meters) to the largest (8 km), with a variance equal to that seen within the cores. In contrast, coherent bedding exists at S60 related to the preservation of the 1995 flood deposit. A correlatable structure is found in the strike direction with a decorrelation distance of ??? 800 m, and can be related to long-wavelength undulations in the topography and/or thickness of the flood layer or overburden. We hypothesize that the high degree of bulk density variability at K60 is a result of more intense physical reworking of the seabed in the sandier environment. Without significant averaging, the resistivity-based porosity measurements are only marginally correlated to gamma-ray-bulk density measurements, and are largely independent of mean grain size. Furthermore, porosity displays a high degree of incoherent variability at both sites. Porosity, with a much smaller sample volume than bulk density, may therefore resolve small-scale biogenic variability which is filtered out in the bulk density measurement. ?? 2002 Elsevier Science Ltd. All rights reserved.

Autopsy consent, brain collection, and standardized neuropathologic assessment of ADNI participants: The essential role of the Neuropathology Core

PubMed Central

Cairns, Nigel J.; Taylor-Reinwald, Lisa; Morris, John C.

2010-01-01

Background Our objectives are to facilitate autopsy consent, brain collection, and perform standardized neuropathologic assessments of all Alzheimer's Disease Neuroimaging Initiative (ADNI) participants who come to autopsy at the 58 ADNI sites in the USA and Canada. Methods Building on the expertise and resources of the existing Alzheimer's Disease Research Center (ADRC) at Washington University School of Medicine, St. Louis, MO, a Neuropathology Core (NPC) to serve ADNI was established with one new highly motivated research coordinator. The ADNI-NPC coordinator provides training materials and protocols to assist clinicians at ADNI sites in obtaining voluntary consent for brain autopsy in ADNI participants. Secondly, the ADNI-NPC maintains a central laboratory to provide uniform neuropathologic assessments using the operational criteria for the classification of AD and other pathologies defined by the National Alzheimer Coordinating Center (NACC). Thirdly, the ADNI-NPC maintains a state-of-the-art brain bank of ADNI-derived brain tissue to promote biomarker and multi-disciplinary clinicopathologic studies. Results During the initial year of funding of the ADNI Neuropathology Core, there was notable improvement in the autopsy rate to 44.4%. In the most recent year of funding (September 1st, 2008 to August 31st 2009), our autopsy rate improved to 71.5%. Although the overall numbers to date are small, these data demonstrate that the Neuropathology Core has established the administrative organization with the participating sites to harvest brains from ADNI participants who come to autopsy. Conclusions Within two years of operation, the Neuropathology Core has: (1) implemented a protocol to solicit permission for brain autopsy in ADNI participants at all 58 sites who die and (2) to send appropriate brain tissue from the decedents to the Neuropathology Core for a standardized, uniform, and state-of-the-art neuropathologic assessment. The benefit to ADNI of the implementation of the NPC is very clear. Prior to the establishment of the NPC in September 2007, there were 6 deaths but no autopsies in ADNI participants. Subsequent to the establishment of the Core there have been 17 deaths of ADNI participants and 10 autopsies. Hence, the autopsy rate has gone from 0% to 59%. The third major accomplishment is the detection of co-existent pathologies with AD in the autopsied cases. It is possible that these co-morbidities may contribute to any variance in ADNI data. PMID:20451876

Using Macrofossils to Reconstruct Paleoenvironmental History of Cedar Swamp, High Point NJ

NASA Astrophysics Data System (ADS)

Buhler, K. M.; Peteet, D. M.

2016-12-01

Macrofossils from deep sediment cores taken from bog environments can reveal much about the paleoclimate of a certain site. A research study conducted by Niering in 1953 focused on the pollen records from Cedar Bog that, although are able to indicate which species were present regionally, are not able to establish the specific local vegetation of the site. By identifying and counting macrofossils from the core, we can determine the species growing in Cedar Bog in High Point State Park, NJ and utilize that data to reconstruct a paleoclimatic history at the highest elevation in New Jersey. We took a 6.5m core from Cedar Bog and, from that, used 4cm³ samples taken throughout the entire depth to wash through a 250um sieve and sift through at 60x magnification, identifying the macrofossils using an extensive Lamont Doherty Earth Observatory reference collection. Additionally, we tested for the percentage of organic matter through the core using loss on ignition. We found 5 zones throughout the core depth that indicate altering habitats. Beginning with low organic matter, less than 5%, the deepest part of the core has little vegetation but high rock content. The next zone introduced Chamaecyparis thyoides in the region and a period of changing organic matter. Following this comes a domination of aquatic plants and byrozoan statoblast and, afterwards, a shift towards sedges. The top of the core shows present day vegetation types from Cedar Bog, such as sphagnum moss and Tsuga canadensis, and 96% organic matter content. The lack of vegetation and high rock content, around 6.5m deep, signal erosion that took place after deglaciation. The vegetation types in zone 2 indicate deep lake/open landscape conditions. High aquatic vegetation and bryozoan statoblasts suggest a shallow lake. The site filled in to become a fen wetland as evidenced by the abundance of Scirpus and Cladium. The final zone signifies a bog habitat represented by the domination of sphagnum and Chamaedaphne. These shifts signal an altering landscape of Cedar Bog, evolving after deglaciation from a deep lake to a shallow lake, followed by a fen wetland and eventually into a modern bog habitat. Certain vegetation and organism data indicate climatic shifts at Cedar Bog that can uncover the paleoclimate for a site located at the highest elevation in New Jersey.

Study of lime vs. no lime in cold in-place recycled asphalt concrete pavements : final report.

DOT National Transportation Integrated Search

1991-09-01

The resilient characteristics of cold in-place recycled asphalt concrete with and without lime were examined. Six core samples were obtained from a site two months after construction; six months later, six additional core samples were obtained from t...

A Comparison of Naturally-Occurring and Artificially Stimulated Uranium(VI) Bioreduction in Sediment from a Field-scale Experiment in Rifle, CO

NASA Astrophysics Data System (ADS)

Campbell, K. M.; Williams, K. H.; Lesher, E.; Davis, J. A.; Long, P. E.

2007-12-01

Long-term remediation of uranium (U)-contaminated groundwater poses one of the greatest challenges in the clean-up of impacted sites. One solution is to reduce dissolved U(VI) to insoluble U(IV) precipitates by stimulating indigenous metal reducing bacterial populations in situ. Contamination from a former U mine tailings repository (Rifle, CO) provides a research site to study the efficacy of biostimulated U(VI) reduction at the field scale. Several cores were drilled in June 2007 across a region of naturally-occurring U(VI) bioreduction. The cores represent a cross section of sediment that ranges from minimally reducing to highly reducing. Anaerobic sediment samples from the cores were analyzed for labile U(VI) content by carbonate extraction in anoxic conditions (pH 9.4, 14 mM NaHCO3, 2.8 mM Na2CO¬3). A subset of the same core sections were dried and oxidized by exposure to air for 2 weeks. The carbonate extraction was repeated, and the amount of U(IV) present in the anaerobic sample was calculated by difference between the anoxic and oxidized extractions. An acid extraction was also performed on the oxidized sediments to compare the carbonate extractable and the acid extractable U fractions. The highest U concentrations were found in the highly bioreduced sediment, with the majority of U present as U(IV) (66-92%). The regions of highest bioreduction also correspond to elevated concentrations of solid phase organic carbon, suggesting that natural bioreduction is stimulated by zones of increased organic carbon content. The same field site was then used for an artificially stimulated bioreduction experiment, where the indigenous bacterial community was stimulated by injecting acetate upgradient of the core collection location. Carbonate and acid extractions were performed on core samples taken after the completion of the acetate injection. This work evaluates the composition of the sediment before and after biostimulation as a way of directly comparing the extent of natural U(VI) bioreduction to acetate-stimulated bioreduction in order to facilitate the design of a more effective bioremediation strategy for this site.

Accelerated gravity testing of aquitard core permeability and implications at formation and regional scale

NASA Astrophysics Data System (ADS)

Timms, W. A.; Crane, R.; Anderson, D. J.; Bouzalakos, S.; Whelan, M.; McGeeney, D.; Rahman, P. F.; Acworth, R. I.

2016-01-01

Evaluating the possibility of leakage through low-permeability geological strata is critically important for sustainable water supplies, the extraction of fuels from coal and other strata, and the confinement of waste within the earth. The current work demonstrates that relatively rapid and realistic vertical hydraulic conductivity (Kv) measurements of aquitard cores using accelerated gravity can constrain and compliment larger-scale assessments of hydraulic connectivity. Steady-state fluid velocity through a low-K porous sample is linearly related to accelerated gravity (g level) in a centrifuge permeameter (CP) unless consolidation or geochemical reactions occur. A CP module was custom designed to fit a standard 2 m diameter geotechnical centrifuge (550 g maximum) with a capacity for sample dimensions up to 100 mm diameter and 200 mm length, and a total stress of ˜ 2 MPa at the base of the core. Formation fluids were used as influent to limit any shrink-swell phenomena, which may alter the permeability. Kv results from CP testing of minimally disturbed cores from three sites within a clayey-silt formation varied from 10-10 to 10-7 m s-1 (number of samples, n = 18). Additional tests were focussed on the Cattle Lane (CL) site, where Kv within the 99 % confidence interval (n = 9) was 1.1 × 10-9 to 2.0 × 10-9 m s-1. These Kv results were very similar to an independent in situ Kv method based on pore pressure propagation though the sequence. However, there was less certainty at two other core sites due to limited and variable Kv data. Blind standard 1 g column tests underestimated Kv compared to CP and in situ Kv data, possibly due to deionised water interactions with clay, and were more time-consuming than CP tests. Our Kv results were compared with the set-up of a flow model for the region, and considered in the context of heterogeneity and preferential flow paths at site and formation scale. Reasonable assessments of leakage and solute transport through aquitards over multi-decadal timescales can be achieved by accelerated core testing together with complimentary hydrogeological monitoring, analysis, and modelling.

Application of ground-penetrating radar imagery for three-dimensional visualisation of near-surface structures in ice-rich permafrost, Barrow, Alaska

USGS Publications Warehouse

Munroe, Jeffrey S.; Doolittle, James A.; Kanevskiy, Mikhail; Hinkel, Kenneth M.; Nelson, Frederick E.; Jones, Benjamin M.; Shur, Yuri; Kimble, John M.

2007-01-01

Three-dimensional ground-penetrating radar (3D GPR) was used to investigate the subsurface structure of ice-wedge polygons and other features of the frozen active layer and near-surface permafrost near Barrow, Alaska. Surveys were conducted at three sites located on landscapes of different geomorphic age. At each site, sediment cores were collected and characterised to aid interpretation of GPR data. At two sites, 3D GPR was able to delineate subsurface ice-wedge networks with high fidelity. Three-dimensional GPR data also revealed a fundamental difference in ice-wedge morphology between these two sites that is consistent with differences in landscape age. At a third site, the combination of two-dimensional and 3D GPR revealed the location of an active frost boil with ataxitic cryostructure. When supplemented by analysis of soil cores, 3D GPR offers considerable potential for imaging, interpreting and 3D mapping of near-surface soil and ice structures in permafrost environments.

Patterns and variability in geochemical signatures and microbial activity within and between diverse cold seep habitats along the lower continental slope, Northern Gulf of Mexico

NASA Astrophysics Data System (ADS)

Bowles, Marshall; Hunter, Kimberley S.; Samarkin, Vladimir; Joye, Samantha

2016-07-01

We collected 69 sediment cores from distinct ecological and geological settings along the deep slope in the Northern Gulf of Mexico to evaluate whether specific geochemical- or habitat-related factors correlated with rates of microbial processes and geochemical signatures. By collecting replicate cores from distinct habitats across multiple sites, we illustrate and quantify the heterogeneity of cold seep geochemistry and microbial activity. These data also document the factors driving unique aspects of the geochemistry of deep slope gas, oil and brine seeps. Surprisingly little variation was observed between replicate (n=2-5) cores within sites for most analytes (except methane), implying that the common practice of collecting one core for geochemical analysis can capture the signature of a habitat in most cases. Depth-integrated concentrations of methane, dissolved inorganic carbon (DIC), and calcium were the predominant geochemical factors that correlated with a site's ecological or geological settings. Pore fluid methane concentration was related to the phosphate and DIC concentration, as well as to rates of sulfate reduction. While distinctions between seep habitats were identified from geochemical signatures, habitat specific geochemistry varied little across sites. The relative concentration of dissolved inorganic nitrogen versus phosphorus suggests that phosphorus availability limits biomass production at cold seeps. Correlations between calcium, chloride, and phosphate concentrations were indicative of brine-associated phosphate transport, suggesting that in addition to the co-migration of methane, dissolved organic carbon, and ammonium with brine, phosphate delivery is also associated with brine advection.

Dating of 30m ice cores drilled by Japanese Antarctic Research Expedition and environmental change study

NASA Astrophysics Data System (ADS)

Motoyama, H.; Suzuki, T.; Fukui, K.; Ohno, H.; Hoshina, Y.; Hirabayashi, M.; Fujita, S.

2017-12-01

1. Introduction It is possible to reveal the past climate and environmental change from the ice core drilled in polar ice sheet and glaciers. The 54th Japanese Antarctic Research Expedition conducted several shallow core drillings up to 30 m depth in the inland and coastal areas of the East Antarctic ice sheet. Ice core sample was cut out at a thickness of about 5 cm in the cold room of the National Institute of Polar Research, and analyzed ion, water isotope, dust and so one. We also conducted dielectric profile measurement (DEP measurement). The age as a key layer of large-scale volcanic explosion was based on Sigl et al. (Nature Climate Change, 2014). 2. Inland ice core Ice cores were collected at the NDF site (77°47'14"S, 39°03'34"E, 3754 m.a.s.l.) and S80 site (80°00'00"S, 40°30'04"E, 3622 m.a.s.l.). Dating of ice core was done as follows. Calculate water equivalent from core density. Accumulate water equivalent from the surface. Approximate the relation of depth - cumulative water equivalent by a quartic equation. We determined the key layer with nssSO42 - peak corresponding to several large volcanic explosions. The accumulation rate was kept constant between the key layers. As a result, NDF was estimated to be around 1360 AD and S80 was estimated to be around 1400 AD in the deepest ice core. 3. Coastal ice core An ice core was collected at coastal H15 sites (69°04'10"S, 40°44'51"E, 1030 m.a.s.l.). Dating of ice core was done as follows. Calculate water equivalent from ice core density. Accumulate water equivalent from the surface. Approximate the relation of depth - cumulative water equivalent by a quartic equation. Basically we decided to summer (December) and winter (June) due to the seasonal change of the water isotope (δD or δ18O). In addition to the seasonal change of isotope, confirm the following. Maximum of SO42- / Na +, which is earlier in time than the maximum of water isotope. Maximum of MSA at about the same time as the maximum of the water isotope. Na+ is maximal immediately after the local maximum of the water isotope. The deepest age was estimated to be around 1940 AD. 4. Example of results In the inland area, the annual average surface mass balance decreased from 1450 to 1850 AD, but it has increased since 1850 AD. The annual mass balance of coastal H15 is consistent with the result of snow stake measurement.

Paleomagnetism of the Oman Ophiolite: New Results from Oman Drilling Project Cores

NASA Astrophysics Data System (ADS)

Horst, A. J.; Till, J. L.; Koornneef, L.; Usui, Y.; Kim, H.; Morris, A.

2017-12-01

The Oman Drilling Project drilled holes at four sites in a transect through the southern massifs of the Samail ophiolite, and recovered 1500 m of igneous and metamorphic rocks. We focus on three sites from the oceanic crustal section including lower layered gabbros (GT1A), the mid-crustal layered to foliated gabbro transition (GT2A), and the shallower transition from sheeted dikes to varitextured gabbros (GT3A). Detailed core descriptions, analyses, and paleomagnetic measurements, were made on D/V Chikyu from July to September 2017 to utilize the core laboratory facilities similar to IODP expeditions. Shipboard measurements included anisotropy of magnetic susceptibility (AMS) and alternating field and thermal demagnetization of 597 discrete samples. Sample demagnetization behavior is varied from each of the cores, with some revealing multiple components of magnetization, and others yielding nearly univectorial data. The interpretation of results from the lower crustal cores is complicated by the pervasive presence of secondary magnetite. In almost all samples, a stable component was resolved (interpreted as a characteristic remanent magnetization) after removal of a lower-coercivity or lower unblocking-temperature component. The inclinations of the stable components in the core reference frame are very consistent in Hole GT1A. However, a transition from negative to positive inclinations in GT2A suggests some structural complexity, possibly as a result of intense late faulting activity. Both abrupt and gradual transitions between multiple zones of negative and positive inclinations occur in Hole GT3A. Interpretation and direct comparison of remanence between drill sites is difficult as recovered core pieces currently remain azimuthally unoriented, and GT2A was drilled at a plunge of 60°, whereas GT1A and GT3A were both drilled vertically. Work is ongoing to use borehole imagery to reorient the core pieces and paleomagnetic data into a geographic in situ reference frame. We will present an overview of preliminary AMS and remanence data that will be used in the future to 1) document deformational histories, 2) characterize magmatic flow directions at different structural levels, and 3) identify the magnetic mineralogy of remanence carriers throughout the oceanic crustal section.

Fast elemental screening of soil and sediment profiles using small-spot energy-dispersive X-ray fluorescence: application to mining sediments geochemistry.

PubMed

Gonzalez-Fernandez, Oscar; Queralt, Ignacio

2010-09-01

Elemental analysis of different sediment cores originating from the Cartagena-La Union mining district in Spain was carried out by means of a programmable small-spot energy-dispersive X-ray fluorescence (EDXRF) spectrometer to study the distribution of heavy metals along soil profiles. Cores were obtained from upstream sediments of a mining creek, from the lowland sedimentation plain, and from a mining landfill dump (tailings pile). A programmable two-dimensional (2D) stage and a focal spot resolution of 600 μm allow us to obtain complete core mapping. Geochemical results were verified using a more powerful wavelength-dispersion X-ray fluorescence (WDXRF) technique. The data obtained was processed in order to study the statistical correlations within the elemental compositions. The results obtained allow us to observe the differential in-depth distribution of heavy metals among the sampled zones. Dump site cores exhibit a homogeneous distribution of heavy metals, whereas the alluvial plain core shows accumulation of heavy metals in the upper part. This approach can be useful for the fast screening of heavy metals in depositional environments around mining sites.

Systematic Approach to Identifying Deeply Buried Archeological Deposits

DOT National Transportation Integrated Search

2018-02-01

Traditional methods used to discover archeological sites include pedestrian surface surveys and relatively shallow hand-dug shovel or soil core testing. While these methods are appropriate for locating surface and near-surface sites on ridges, hillto...

Patched bimetallic surfaces are active catalysts for ammonia decomposition.

PubMed

Guo, Wei; Vlachos, Dionisios G

2015-10-07

Ammonia decomposition is often used as an archetypical reaction for predicting new catalytic materials and understanding the very reason of why some reactions are sensitive on material's structure. Core-shell or surface-segregated bimetallic nanoparticles expose outstanding activity for many heterogeneously catalysed reactions but the reasons remain elusive owing to the difficulties in experimentally characterizing active sites. Here by performing multiscale simulations in ammonia decomposition on various nickel loadings on platinum (111), we show that the very high activity of core-shell structures requires patches of the guest metal to create and sustain dual active sites: nickel terraces catalyse N-H bond breaking and nickel edge sites drive atomic nitrogen association. The structure sensitivity on these active catalysts depends profoundly on reaction conditions due to kinetically competing relevant elementary reaction steps. We expose a remarkable difference in active sites between transient and steady-state studies and provide insights into optimal material design.

Patched bimetallic surfaces are active catalysts for ammonia decomposition

NASA Astrophysics Data System (ADS)

Guo, Wei; Vlachos, Dionisios G.

2015-10-01

Ammonia decomposition is often used as an archetypical reaction for predicting new catalytic materials and understanding the very reason of why some reactions are sensitive on material's structure. Core-shell or surface-segregated bimetallic nanoparticles expose outstanding activity for many heterogeneously catalysed reactions but the reasons remain elusive owing to the difficulties in experimentally characterizing active sites. Here by performing multiscale simulations in ammonia decomposition on various nickel loadings on platinum (111), we show that the very high activity of core-shell structures requires patches of the guest metal to create and sustain dual active sites: nickel terraces catalyse N-H bond breaking and nickel edge sites drive atomic nitrogen association. The structure sensitivity on these active catalysts depends profoundly on reaction conditions due to kinetically competing relevant elementary reaction steps. We expose a remarkable difference in active sites between transient and steady-state studies and provide insights into optimal material design.

Deposition and chemistry of bottom sediments in Cochiti Lake, north-central New Mexico

USGS Publications Warehouse

Wilson, Jennifer T.; Van Metre, Peter C.

2000-01-01

Bottom sediments were sampled at seven sites in Cochiti Lake in September 1996. Sediment cores penetrating the entire lacustrine sediment sequence were collected at one site near the dam. Surficial sediments were sampled at the near-dam site and six other sites located along the length of the reservoir. Analyses included grain size, major and trace elements, organochlorine compounds, polycyclic aromatic hydrocarbons (PAH's), and radionuclides. Concentrations of trace elements, organic compounds, and radionuclides are similar to those in other Rio Grande reservoirs and are low compared to published sediment-quality guidelines. Most elements and compounds that were detected did not show trends in the age estimated sediment cores with the exception of a decreasing trend in total DDT concentrations from about 1980 to 1992. The mixture of PAH's suggests that the increase is caused by inputs of fuel-related PAH and not combustion- related PAH.

CT Scanning and Geophysical Measurements of the Marcellus Formation from the Tippens 6HS Well

DOE Office of Scientific and Technical Information (OSTI.GOV)

Crandall, Dustin; Paronish, Thomas; Brown, Sarah

The computed tomography (CT) facilities and the Multi-Sensor Core Logger (MSCL) at the National Energy Technology Laboratory (NETL) Morgantown, West Virginia site were used to characterize core of the Marcellus Shale from a vertical well drilled in Eastern Ohio. The core is from the Tippens 6HS Well in Monroe County, Ohio and is comprised primarily of the Marcellus Shale from depths of 5550 to 5663 ft.

Description, dissection, and subsampling of Apollo 14 core sample 14230

NASA Technical Reports Server (NTRS)

Fryxell, R.; Heiken, G.

1971-01-01

Core sample 14230, collected at Triplet Crater near the Fra Mauro landing site of the Apollo 14 mission, was dissected in greater detail than any previous core. Sediment from the actual lunar surface was missing, and 6.7 grams of sediment were removed from the base of the core for a portion of the biotest prime sample. Upper and lower portions of the original 70.7-gram core (12.5 centimeters long) were fractured excessively but not mixed stratigraphically. Three major morphologic units and 11 subdivisions were recognized. Dissection provided 55 subsamples in addition to three others made by removing longitudinal sections of the core impregnated with n-butyl methacrylate for use as a permanent documentary record and for studies requiring particles of known orientation.

Core-Crosslinked Polymeric Micelles: Principles, Preparation, Biomedical Applications and Clinical Translation

PubMed Central

Rijcken, Cristianne J.; Kiessling, Fabian; Hennink, Wim E.; Lammers, Twan

2015-01-01

Polymeric micelles (PM) are extensively used to improve the delivery of hydrophobic drugs. Many different PM have been designed and evaluated over the years, and some of them have steadily progressed through clinical trials. Increasing evidence suggests, however, that for prolonged circulation times and for efficient EPR-mediated drug targeting to tumors and to sites of inflammation, PM need to be stabilized, to prevent premature disintegration. Core-crosslinking is among the most popular methods to improve the in vivo stability of PM, and a number of core-crosslinked polymeric micelles (CCPM) have demonstrated promising efficacy in animal models. The latter is particularly true for CCPM in which (pro−) drugs are covalently entrapped. This ensures proper drug retention in the micelles during systemic circulation, efficient drug delivery to pathological sites via EPR, and tailorable drug release kinetics at the target site. We here summarize recent advances in the CCPM field, addressing the chemistry involved in preparing them, their in vitro and in vivo performance, potential biomedical applications, and guidelines for efficient clinical translation. PMID:25893004

A Nubian Complex Site from Central Arabia: Implications for Levallois Taxonomy and Human Dispersals during the Upper Pleistocene

PubMed Central

Crassard, Rémy; Hilbert, Yamandú Hieronymus

2013-01-01

Archaeological survey undertaken in central Saudi Arabia has revealed 29 surface sites attributed to the Arabian Middle Paleolithic based on the presence of Levallois blank production methods. Technological analyses on cores retrieved from Al-Kharj 22 have revealed specific reduction modalities used to produce flakes with predetermined shapes. The identified modalities, which are anchored within the greater Levallois concept of core convexity preparation and exploitation, correspond with those utilized during the Middle Stone Age Nubian Complex of northeast Africa and southern Arabia. The discovery of Nubian technology at the Al-Kharj 22 site represents the first appearance of this blank production method in central Arabia. Here we demonstrate how a rigorous use of technological and taxonomic analysis may enable intra-regional comparisons across the Arabian Peninsula. The discovery of Al-Kharj 22 increases the complexity of the Arabian Middle Paleolithic archaeological record and suggests new dynamics of population movements between the southern and central regions of the Peninsula. This study also addresses the dichotomy within Nubian core typology (Types 1 and 2), which was originally defined for African assemblages. PMID:23894434

Observations and model predictions of water skin temperatures at MTI core site lakes and reservoirs

NASA Astrophysics Data System (ADS)

Garrett, Alfred J.; Kurzeja, Robert J.; O'Steen, Byron L.; Parker, Matthew J.; Pendergast, Malcolm M.; Villa-Aleman, Eliel; Pagnutti, Mary A.

2001-08-01

The Savannah River Technology Center (SRTC) measured water skin temperatures at four of the Multi-spectral Thermal Imager (MTI) core sites. The depression of the skin temperature relative to the bulk water temperature ((Delta) T) a few centimeters below the surface is a complex function of the weather conditions, turbulent mixing in the water and the bulk water temperature. Observed skin temperature depressions range from near zero to more than 1.0 degree(s)C. Skin temperature depressions tend to be larger when the bulk water temperature is high, but large depressions were also observed in cool bodies of water in calm conditions at night. We compared (Delta) T predictions from three models (SRTC, Schlussel and Wick) against measured (Delta) T's from 15 data sets taken at the MTI core sites. The SRTC and Wick models performed somewhat better than the Schlussel model, with RMSE and average absolute errors of about 0.2 degree(s)C, relative to 0.4 degree(s)C for the Schlussel model. The average observed (Delta) T for all 15 databases was -0.7 degree(s)C.

Contemporary prostate biopsy reporting: insights from a survey of clinicians' use of pathology data.

PubMed

Varma, Murali; Narahari, Krishna; Mason, Malcolm; Oxley, Jon D; Berney, Daniel M

2018-05-02

To determine how clinicians use data in contemporary prostate biopsy reports. A survey was circulated to members of the British Association of Urological Surgeons and the British Uro-oncology Group. Responses were received from 114 respondents (88 urologists, 26 oncologists). Ninety-seven (94%) use the number of positive cores from each side and 43 (42%) use the % number of positive cores. When determining the number and percentage of positive cores, 72 (71%) would not differentiate between targeted and non-targeted samples. If multiple Gleason Scores (GS) were included in a report, 77 (78%) would use the worst GS even if present in a core with very little tumour, 12% would use the global GS and 10% the GS in the core most involved by tumour. Fifty-five (55%) either never or rarely used perineural invasion for patient management. The number of positive cores is an important parameter for patient management but may be difficult to determine in the laboratory due to core fragmentation so the biopsy taker must indicate the number of biopsies obtained. Multiple biopsies taken from a single site are often interpreted by clinicians as separate cores when determining the number of positive cores so pathologists should also report the number of sites positive. Clinicians have a non-uniform approach to the interpretation of multiple GS in prostate biopsy reports so we recommend that pathologists also include a single 'bottom-line' GS for each case to direct the clinician's treatment decision. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

Sign changes in sums of the Liouville function

NASA Astrophysics Data System (ADS)

Borwein, Peter; Ferguson, Ron; Mossinghoff, Michael J.

2008-09-01

The Liouville function λ(n) is the completely multiplicative function whose value is -1 at each prime. We develop some algorithms for computing the sum T(n)Dsum_{kD1}^n λ(k)/k , and use these methods to determine the smallest positive integer n where T(n)<0 . This answers a question originating in some work of Turan, who linked the behavior of T(n) to questions about the Riemann zeta function. We also study the problem of evaluating Polya's sum L(n)Dsum_{kD1}^nλ(k) , and we determine some new local extrema for this function, including some new positive values.

Plasmodium falciparum spliceosomal RNAs: 3' and 5' end processing.

PubMed

Eliana, Calvo; Javier, Escobar; Moisés, Wasserman

2011-02-01

The major spliceosomal small nuclear ribonucleoproteins (snRNPs) consist of snRNA (U1, U2, U4 or U5) and several proteins which can be unique or common to each snRNP particle. The common proteins are known as Sm proteins; they are crucial for RNP assembly and nuclear import of spliceosomal RNPs. This paper reports detecting the interaction between Plasmodium falciparum snRNAs and Sm proteins, and the usual 5' trimethylated caps on the snRNAs, by immunoprecipitation with specific antibodies. Furthermore, an unusual poly(A) tail was detected on these non-coding RNAs. 2010 Elsevier B.V. All rights reserved.

Microbial characterization of nitrification in a shallow, nitrogen-contaminated aquifer, Cape Cod, Massachusetts and detection of a novel cluster associated with nitrifying Betaproteobacteria

USGS Publications Warehouse

Miller, D.N.; Smith, R.L.

2009-01-01

Groundwater nitrification is a poorly characterized process affecting the speciation and transport of nitrogen. Cores from two sites in a plume of contamination were examined using culture-based and molecular techniques targeting nitrification processes. The first site, located beneath a sewage effluent infiltration bed, received treated effluent containing O2 (> 300????M) and NH4+ (51-800????M). The second site was 2.5??km down-gradient near the leading edge of the ammonium zone within the contaminant plume and featured vertical gradients of O2, NH4+, and NO3- (0-300, 0-500, and 100-200????M with depth, respectively). Ammonia- and nitrite-oxidizers enumerated by the culture-based MPN method were low in abundance at both sites (1.8 to 350??g- 1 and 33 to 35,000??g- 1, respectively). Potential nitrifying activity measured in core material in the laboratory was also very low, requiring several weeks for products to accumulate. Molecular analysis of aquifer DNA (nested PCR followed by cloning and 16S rDNA sequencing) detected primarily sequences associated with the Nitrosospira genus throughout the cores at the down-gradient site and a smaller proportion from the Nitrosomonas genus in the deeper anoxic, NH4+ zone at the down-gradient site. Only a single Nitrosospira sequence was detected beneath the infiltration bed. Furthermore, the majority of Nitrosospira-associated sequences represent an unrecognized cluster. We conclude that an uncharacterized group associated with Nitrosospira dominate at the geochemically stable, down-gradient site, but found little evidence for Betaproteobacteria nitrifiers beneath the infiltration beds where geochemical conditions were more variable.

Microbial characterization of nitrification in a shallow, nitrogen-contaminated aquifer, Cape Cod, Massachusetts and detection of a novel cluster associated with nitrifying Betaproteobacteria

NASA Astrophysics Data System (ADS)

Miller, Daniel N.; Smith, Richard L.

2009-01-01

Groundwater nitrification is a poorly characterized process affecting the speciation and transport of nitrogen. Cores from two sites in a plume of contamination were examined using culture-based and molecular techniques targeting nitrification processes. The first site, located beneath a sewage effluent infiltration bed, received treated effluent containing O 2 (> 300 µM) and NH 4+ (51-800 µM). The second site was 2.5 km down-gradient near the leading edge of the ammonium zone within the contaminant plume and featured vertical gradients of O 2, NH 4+, and NO 3- (0-300, 0-500, and 100-200 µM with depth, respectively). Ammonia- and nitrite-oxidizers enumerated by the culture-based MPN method were low in abundance at both sites (1.8 to 350 g - 1 and 33 to 35,000 g - 1 , respectively). Potential nitrifying activity measured in core material in the laboratory was also very low, requiring several weeks for products to accumulate. Molecular analysis of aquifer DNA (nested PCR followed by cloning and 16S rDNA sequencing) detected primarily sequences associated with the Nitrosospira genus throughout the cores at the down-gradient site and a smaller proportion from the Nitrosomonas genus in the deeper anoxic, NH 4+ zone at the down-gradient site. Only a single Nitrosospira sequence was detected beneath the infiltration bed. Furthermore, the majority of Nitrosospira-associated sequences represent an unrecognized cluster. We conclude that an uncharacterized group associated with Nitrosospira dominate at the geochemically stable, down-gradient site, but found little evidence for Betaproteobacteria nitrifiers beneath the infiltration beds where geochemical conditions were more variable.

Endogenous Hot Spots of De Novo Telomere Addition in the Yeast Genome Contain Proximal Enhancers That Bind Cdc13

PubMed Central

Obodo, Udochukwu C.; Epum, Esther A.; Platts, Margaret H.; Seloff, Jacob; Dahlson, Nicole A.; Velkovsky, Stoycho M.; Paul, Shira R.

2016-01-01

DNA double-strand breaks (DSBs) pose a threat to genome stability and are repaired through multiple mechanisms. Rarely, telomerase, the enzyme that maintains telomeres, acts upon a DSB in a mutagenic process termed telomere healing. The probability of telomere addition is increased at specific genomic sequences termed sites of repair-associated telomere addition (SiRTAs). By monitoring repair of an induced DSB, we show that SiRTAs on chromosomes V and IX share a bipartite structure in which a core sequence (Core) is directly targeted by telomerase, while a proximal sequence (Stim) enhances the probability of de novo telomere formation. The Stim and Core sequences are sufficient to confer a high frequency of telomere addition to an ectopic site. Cdc13, a single-stranded DNA binding protein that recruits telomerase to endogenous telomeres, is known to stimulate de novo telomere addition when artificially recruited to an induced DSB. Here we show that the ability of the Stim sequence to enhance de novo telomere addition correlates with its ability to bind Cdc13, indicating that natural sites at which telomere addition occurs at high frequency require binding by Cdc13 to a sequence 20 to 100 bp internal from the site at which telomerase acts to initiate de novo telomere addition. PMID:27044869

Transcriptomic and proteomic analysis reveals wall-associated and glucan-degrading proteins with potential roles in Phytophthora infestans sexual spore development.

PubMed

Niu, Xiaofan; Ah-Fong, Audrey M V; Lopez, Lilianna A; Judelson, Howard S

2018-01-01

Sexual reproduction remains an understudied feature of oomycete biology. To expand our knowledge of this process, we used RNA-seq and quantitative proteomics to examine matings in Phytophthora infestans. Exhibiting significant changes in mRNA abundance in three matings between different A1 and A2 strains compared to nonmating controls were 1170 genes, most being mating-induced. Rising by >10-fold in at least one cross were 455 genes, and 182 in all three crosses. Most genes had elevated expression in a self-fertile strain. Many mating-induced genes were associated with cell wall biosynthesis, which may relate to forming the thick-walled sexual spore (oospore). Several gene families were induced during mating including one encoding histidine, serine, and tyrosine-rich putative wall proteins, and another encoding prolyl hydroxylases which may strengthen the extracellular matrix. The sizes of these families vary >10-fold between Phytophthora species and one exhibits concerted evolution, highlighting two features of genome dynamics within the genus. Proteomic analyses of mature oospores and nonmating hyphae using isobaric tags for quantification identified 835 shared proteins, with 5% showing >2-fold changes in abundance between the tissues. Enriched in oospores were β-glucanases potentially involved in digesting the oospore wall during germination. Despite being dormant, oospores contained a mostly normal complement of proteins required for core cellular functions. The RNA-seq data generated here and in prior studies were used to identify new housekeeping controls for gene expression studies that are more stable than existing normalization standards. We also observed >2-fold variation in the fraction of polyA+ RNA between life stages, which should be considered when quantifying transcripts and may also be relevant to understanding translational control during development.

Late Pleistocene to Holocene environmental changes as recorded in the sulfur geochemistry of coastal plain sediments, southwestern Taiwan

USGS Publications Warehouse

Chen, Y.-G.; Liu, J.C.-L.; Shieh, Y.-N.; Liu, T.-K.

2004-01-01

A core, drilled at San-liao-wan in the southwestern coastal plain of Taiwan, has been analyzed for total sulfur contents, isotopic values, as well as ratios of pyritic sulfur to organic carbon. Our results demonstrate a close relationship between late Pleistocene sea-level change and the proxies generated in this study. The inorganic sulfur contents indicate that at our study site, the Holocene transgression started at ???11 ka and remained under seawater for thousands of years until the late Holocene, corresponding to a depth of 20 m in the study core. The uppermost 20 m of core shows relatively high total organic carbon (TOC) and ??34S of inorganic sulfur, suggesting a transitional environment such as muddy lagoon or marsh, before the site turned into a modern coastal plain. In the lower part of the core, at depths of 110-145 m (corresponding ages of ???12-30 ka), low sulfur contents are recorded, probably indicating fluvial sediments deposited during the oceanic isotope stage (OIS) 2, a sea-level lowstand. The lower part of the core, roughly within OIS 3, records at least two transgressions, although the transgressional signals may be somewhat obscured by subsequent weathering. The reworked origin of organic matter reported in previous studies is confirmed by our organic sulfur data; however, the marine organic source was periodically dominant. The modern high sulfate concentrations in pore water have no correlation to the other sulfur species in the sediments, probably indicating that the sulfate migrated into the site subsequent to early diagenesis. ?? 2003 Elsevier Ltd. All rights reserved.

Microbial colonization of Ca-sulfate crusts in the hyperarid core of the Atacama Desert: implications for the search for life on Mars.

PubMed

Wierzchos, J; Cámara, B; de Los Ríos, A; Davila, A F; Sánchez Almazo, I M; Artieda, O; Wierzchos, K; Gómez-Silva, B; McKay, C; Ascaso, C

2011-01-01

The scarcity of liquid water in the hyperarid core of the Atacama Desert makes this region one of the most challenging environments for life on Earth. The low numbers of microbial cells in the soils suggest that within the Atacama Desert lies the dry limit for life on our planet. Here, we show that the Ca-sulfate crusts of this hyperarid core are the habitats of lithobiontic micro-organisms. This microporous, translucent substrate is colonized by epilithic lichens, as well as endolithic free-living algae, fungal hyphae, cyanobacteria and non photosynthetic bacteria. We also report a novel type of endolithic community, "hypoendoliths", colonizing the undermost layer of the crusts. The colonization of gypsum crusts within the hyperarid core appears to be controlled by the moisture regime. Our data shows that the threshold for colonization is crossed within the dry core, with abundant colonization in gypsum crusts at one study site, while crusts at a drier site are virtually devoid of life. We show that the cumulative time in 1 year of relative humidity (RH) above 60% is the best parameter to explain the difference in colonization between both sites. This is supported by controlled humidity experiments, where we show that colonies of endolithic cyanobacteria in the Ca-sulfate crust undergo imbibition process at RH >60%. Assuming that life once arose on Mars, it is conceivable that Martian micro-organisms sought refuge in similar isolated evaporite microenvironments during their last struggle for life as their planet turned arid. © 2010 Blackwell Publishing Ltd.