[Construction of cTnC-linker-TnI (P) Genes, Expression of Fusion Protein and Preparation of Lyophilized Protein].

Science.gov (United States)

Song, Xiaoli; Liu, Xiaoyun; Cai, Lei; Wu, Jianwei; Wang, Jihua

2015-12-01

In order to construct and express human cardiac troponin C-linker-troponin I(P) [ cTnC-linker-TnI(P)] fusion protein, detect its activity and prepare lyophilized protein, we searched the CDs of human cTnC and cTnI from GenBank, synthesized cTnC and cTnI(30-110aa) into cloning vector by a short DNA sequence coding for 15 neutral amino acid residues. pCold I-cTnC-linker-TnI(P) was constructed and transformed into E. coli BL21(DE3). Then, cTnC-linker-TnI(P) fusion protein was induced by isopropyl-β-D-thiogalactopyranoside (IPTG). Soluable expression of cTnC-linker-TnI(P) in prokaryotic system was successfully obtained. The fusion protein was purified by Ni²⁺ Sepharose 6 Fast Flow affinity chromatography with over 95% purity and prepared into lyophilized protein. The activity of purified cTnC-linker-TnI(P) and its lyophilized protein were detected by Wondfo Finecare™ cTnI Test. Lyophilized protein of cTnC-linker-TnI(P) was stable for 10 or more days at 37 °C and 4 or more months at 25 °C and 4 °C. The expression system established in this research is feasible and efficient. Lyophilized protein is stable enough to be provided as biological raw materials for further research.

BWR spent fuel transport and storage system for KKL: TN trademark 52L, TN trademark 97L, TN trademark 24 BHL

International Nuclear Information System (INIS)

Sicard, D.; Verdier, A.; Monsigny, P.A.

2004-01-01

The LEIBSTADT (KKL) nuclear power plant in Switzerland has opted to ship spent fuel to a central facility called ZWILAG for interim storage. In the mid-nineties, COGEMA LOGISTICS was contracted by KKL for the supply of the TN trademark a52L and TN trademark 97L transport and storage casks for BWR fuel types. In 2003, KKL also ordered from COGEMA LOGISTICS the supply of six TNae24 BHL transport and storage casks. This paper shows how all the three cask designs have responded to the KKL needs to ship and store BWR spent fuel. In addition, it highlights the already significant operational feedback of the TN trademark 52L and TN trademark 97L casks by the KKL and ZWILAG operators

Fraction A of armadillo submandibular glycoprotein and its desialylated product as sialyl-Tn and Tn receptors for lectins.

Science.gov (United States)

Wu, A M; Shen, F; Herp, A; Song, S C; Wu, J H

1995-02-27

Fraction A of the armadillo submandibular glycoprotein (ASG-A) is one of the simplest glycoproteins among mammalian salivary mucins. The carbohydrate side chains of this mucous glycoprotein have one-third of the NeuAc alpha 2-->6GalNAc (sialyl-Tn) sequence and two thirds of Tn (GalNAc alpha-->Ser/Thr) residues. Those of the desialylated product (ASG-Tn) are almost exclusively unsubstituted GalNAc residues (Tn determinant). When the binding properties of these glycoproteins were tested by a precipitin assay with Gal, GalNAc and GlcNAc specific lectins, it was found that ASG-Tn reacted strongly with all of the Tn-active lectins and completely precipitated Vicia villosa (VVL both B4 and mixture of A and B), Maclura pomifera (MPA), and Artocarpus integrifolia (jacalin) lectins. However, it precipitated poorly or negligibly with Ricinus communis (RCA1); Dolichos biflorus (DBA); Viscum album, ML-I; Arachis hypogaea (PNA), and Triticum vulgaris (WGA). The reactivity of ASG-A (sialyl-Tn) was as active as that of ASG-Tn with MPA and less or slightly less active than that of ASG-Tn with VVL-A+B, VVL-B4, HPA, WFA, and jacalin, as one-third of its Tn was sialylated. These findings indicate that ASG-A and its desialylated product (ASG-Tn) are highly useful reagents for the differentiation of Tn, T (Gal beta 1-->3GalNAc), A (GalNAc alpha 1-->3Gal) or Gal specific lectins and monoclonal antibodies against such epitopes.

Interaction of hamster submaxillary sialyl-Tn and Tn glycoproteins with Gal, GalNAc and GlcNAc specific lectins.

Science.gov (United States)

Wu, A M; Shen, F; Herp, A; Wu, J H

1994-04-01

Hamster submaxillary glycoprotein (HSM), one of the simplest glycoproteins among mammalian salivary mucins, is composed of approximately equivalent amounts of protein, hexosamine and sialic acid. The Thr and Ser residues in the protein core account for more than half of all of the amino acid residues, while Lys, Glu, Pro and Ala are the major components of the remaining portion of amino acids. The carbohydrate side chains of this mucous glycoprotein have mainly the NeuAc-GalNAc-(sialyl-Tn) sequence (HSM), and those of the desialylated product (HSM-Tn) are almost exclusively unsubstituted GalNAc residues (Tn determinants). The binding properties of sialyl-Tn (HSM) and asialo-HSM (HSM-Tn) glycoproteins were tested by precipitin assay with Gal, GalNAc and GlcNAc specific lectins. The HSM-Tn completely precipitated Vicia villosa (VVL both B4 and mixture of A and B), Maclura pomifera (MPL), and Artocarpus integrifolia (Jacalin) lectins; less than 2 micrograms of HSM-Tn were required for precipitating 50% of 5.0-6.3 micrograms lectin nitrogen added. HSM-Tn also reacted well with Helix pomatia lectin (HPL), Wistaria floribunda lectin (WFL) and Abrus precatorius agglutinin (APA) and precipitated in each case over 81% of the lectin nitrogen added. The reactivity of HSM-Tn with other lectins (Ricinus communis, RCA1; Dolichol biflorus, DBL; Viscum album, ML-I; Arachis hypogaea, PNA, and Triticum vulgaris, WGA) was weak or negligible. The activity of sialyl-Tn (HSM) was more restricted; HSM reacted well with Jacalin, moderately with MPL and VVL-B4, but was inactive or only weakly with the other lectins used. These findings indicate that HSM and its desialylated product (HSM-Tn) are highly useful reagents for the differentiation of Tn and T/Gal specific lectins and for anti-T, Tn and Af monoclonal antibodies.

Preparation and Characterization of cis- and trans-[Ir(tn)2Cl2]CF3SO3 and of [Ir(tn)3]Cl3 (tn=propane-1,3-diamine)

DEFF Research Database (Denmark)

Brorson, Michael; Galsbøl, Frode; Simonsen, Kim

1998-01-01

for the preparation of [Rh(tn)3]Cl3 in quantitative yield from Rh(thtp)3Cl3 is also given. The complexes were characterized by 1H and 13C NMR and by UV/VIS spectroscopy. The conformation of the six-membered chelate rings of [Ir(tn)3]3+ in the solid state was determined by single-crystal X-ray diffraction of [Ir(tn)3......Procedures are given for the preparation and isolation of cis- and trans-[Ir(tn)2Cl2]CF3SO3 and of [Ir(tn)3]Cl3, (tn=propane-1,3-diamine). The compounds were prepared by the use of Ir(thtp)3Cl3 (thtp=tetrahydrothiophene) as starting material, using either DMSO or neat tn as solvent. A procedure......] [Co(CN)6] x 5H2O. The three chelate rings all adopt the energetically favoured chair conformation; however, the overall idealized symmetry is C1. A comparative ligand field analysis, based on Gaussian resolution of the solution UV/VIS spectra for a number of homoleptic [M(N6)]3+ (M=CoIII, RhIII, Ir...

BWR-spent fuel transport and storage with the TN trademark 9/4 and TN trademark 24BH casks

International Nuclear Information System (INIS)

Wattez, L.; Marguerat, Y.; Hoesli, C.

2004-01-01

The Swiss Nuclear Utilities have started in 2001 to store spent fuel in dry metallic dual-purpose casks in ZWILAG, the Swiss interim storage facility. BKW FMB Energy Ltd., as Muehleberg Nuclear Power Plant owner, is involved in this process and has selected to store its spent fuel, a new high capacity dual-purpose cask, the TN trademark 24BH. For the transport in a medium size cask, COGEMA LOGISTICS has developed a new cask, the TN trademark 9/4, to replace the NTL9 cask, which performed numerous transports of BWR spent fuel in the past decades. Licensed IAEA 1996, the TN trademark 9/4 is a 40 ton transport cask, for 7 BWR high burn-up spent fuel assemblies. The spent fuel assemblies can be transferred in the ZWILAG hot cell in the TN trademark 24BH cask. The first use of these casks took place in 2003. Ten TN trademark 9/4 transports were performed, and one TN trademark 24BH was loaded. After a brief presentation of the operational aspects, the paper will focus on the TN trademark 24BH high capacity dual purpose cask, the TN trademark 9/4 transport cask and describe in detail their characteristics and possibilities

78 FR 7993 - Amendment of Class D and E Airspace; Tri-Cities, TN; Revocation of Class E Airspace; Tri-City, TN

Science.gov (United States)

2013-02-05

...). The SNPRM would list Hawkins County Airport and Virginia Highlands Airport under their own designator... feet above the surface for Hawkins County Airport, Rogersville, TN, and Virginia Highlands Airport, VA... TN E5 Rogersville, TN [New] Hawkins County Airport, TN (Lat. 36[deg]27'27'' N., long. 82[deg]53'06...

Antibiotic and antiseptic resistance genes are linked on a novel mobile genetic element: Tn6087

Science.gov (United States)

Ciric, Lena; Mullany, Peter; Roberts, Adam P.

2011-01-01

Objectives Tn916-like elements are one of the most common types of integrative and conjugative element (ICE). In this study we aimed to determine whether novel accessory genes, i.e. genes whose products are not involved in mobility or regulation, were present on a Tn916-like element (Tn6087) isolated from Streptococcus oralis from the human oral cavity. Methods A minocycline-resistant isolate was analysed using restriction fragment length polymorphism (RFLP) analysis on amplicons derived from Tn916 and DNA sequencing to determine whether there were genetic differences in Tn6087 compared with Tn916. Mutational analysis was used to determine whether the novel accessory gene found was responsible for an observed extra phenotype. Results A novel Tn916-like element, Tn6087, is described that encodes both antibiotic and antiseptic resistance. The antiseptic resistance protein is encoded by a novel small multidrug resistance gene, designated qrg, that was shown to encode resistance to cetyltrimethylammonium bromide (CTAB), also known as cetrimide bromide. Conclusions This is the first Tn916-like element described that confers both antibiotic and antiseptic resistance, suggesting that selection of either antibiotic or antiseptic resistance will also select for the other and further highlights the need for prudent use of both types of compound. PMID:21816764

Simple mucins (T, sialosyl-T, Tn and sialosyl-Tn) are not diagnostic for malignant breast lesions

DEFF Research Database (Denmark)

Reed, W.; Bryne, M.; Clausen, Henrik

1994-01-01

positive for T antigen, 82% for s-T antigen, 66% for Tn antigen and 22% for s-Tn antigen. The staining pattern was nearly identical for carcinomas with and without lymph node metastases. In conclusion, immunostaining for simple mucins does not permit a clear distinction between benign and malignant breast...

Rapid and efficient introduction of a foreign gene into bacterial artificial chromosome-cloned varicella vaccine by Tn7-mediated site-specific transposition

International Nuclear Information System (INIS)

Somboonthum, Pranee; Koshizuka, Tetsuo; Okamoto, Shigefumi; Matsuura, Masaaki; Gomi, Yasuyuki; Takahashi, Michiaki; Yamanishi, Koichi; Mori, Yasuko

2010-01-01

Using a rapid and reliable system based on Tn7-mediated site-specific transposition, we have successfully constructed a recombinant Oka varicella vaccine (vOka) expressing the mumps virus (MuV) fusion protein (F). The backbone of the vector was our previously reported vOka-BAC (bacterial artificial chromosome) genome. We inserted the transposon Tn7 attachment sequence, LacZα-mini-attTn7, into the region between ORF12 and ORF13 to generate a vOka-BAC-Tn genome. The MuV-F expressing cassette was transposed into the vOka-BAC genome at the mini-attTn7 transposition site. MuV-F protein was expressed in recombinant virus, rvOka-F infected cells. In addition, the MuV-F protein was cleaved in the rvOka-F infected cells as in MuV-infected cells. The growth of rvOka-F was similar to that of the original recombinant vOka without the F gene. Thus, we show that Tn7-mediated transposition is an efficient method for introducing a foreign gene expression cassette into the vOka-BAC genome as a live virus vector.

Chromosomal insertion of the entire Escherichia coli lactose operon, into two strains of Pseudomonas, using a modified mini-Tn5 delivery system

DEFF Research Database (Denmark)

Hansen, L. H.; Sørensen, S. J.; Jensen, Lars Bogø

1997-01-01

A 12-kb PstI fragment including the entire E. coli lactose operon (lacIPOZYA) was inserted in one copy into the chromosome of Pseudomonas putida, Pseudomonas fluorescens and an E. coli strain with lac(-) phenotype. This was made possible by improvements of an already existing mini-Tn5 transposon...... flanked by NotI sites needed in the mini-Tn5 delivery system; (b) the generation of E. coli nonlysogenic strains expressing the pi protein thus being capable of maintaining and delivering R6K-based mini-Tn5 vectors to other E. coli strains; (c) the successful insertion of the E. coli lactose operon...... into the P. fluorescens chromosome giving P. fluorescens the ability to grow on lactose; (d) evidence from Southern blotting that contradicts the assumption that the mini-Tn5 delivery system always creates one-copy inserts. These improvements allow insertion of large DNA fragments encoding highly expressed...

Demonstration of criticality safety for the modified TN-REG and TN-BRP transport/storage casks

International Nuclear Information System (INIS)

Parks, C.V.; Fox, P.B.

1989-01-01

An inability to model the structural performance of borated steel baskets under accident conditions forced the specially designed TN-BRP and TN-REG casks to be modified for half-loaded shipments. This paper discusses the approach used to demonstrate that the half-loaded casks would remain safely subcritical even if no credit were taken for the borated basket. Normal and accident configurations were analyzed with the KENO V.a code. The strategy conceived and the analyses performed to demonstrate an acceptable margin of safety are discussed. 5 refs., 3 figs., 2 tabs

In-site interaction evaluation of Tn density by inhibition/competition assays

Energy Technology Data Exchange (ETDEWEB)

Robles, Ana [Radiopharmacy Department, Nuclear Research Center, Faculty of Sciences, University of the Republic, Montevideo (Uruguay)], E-mail: anamar@cin.edu.uy; Medeiros, Andrea [Biochemistry Department, Faculty of Medicine, University of the Republic, Montevideo (Uruguay); Berois, Nora [Laboratory of Glycobiology and Tumor Immunology, Pasteur Institute of Montevideo (Uruguay); Balter, Henia S. [Radiopharmacy Department, Nuclear Research Center, Faculty of Sciences, University of the Republic, Montevideo (Uruguay); Pauwels, Ernest K. [University Medical Center Leiden, Department of Radiology, Leiden (Netherlands); Osinaga, Eduardo [Laboratory of Glycobiology and Tumor Immunology, Pasteur Institute of Montevideo (Uruguay); Department of Immunobiology, Faculty of Medicine, University of the Republic, Montevideo (Uruguay)

2010-05-15

The tumor-associated structure N-acetyl-galactosamine-O-Ser/Thr (Tn antigen), which is overexpressed in various tumor cell types, notably of the breast, ovary and colon, is an interesting determinant that is useful for cancer diagnosis and follow-up. The aim of this research was to study different assay strategies in order to determine the most sensitive system for further application in epitope characterization and binding assessment. The tetrameric isolectin obtained from Vicia villosa seeds (VVLB{sub 4}) shows high affinity for the tumor-associated structure. A monoclonal antibody against VVLB{sub 4}, MabVV{sub 34}, was generated, and the interaction between MabVV{sub 34} and VVLB{sub 4} was studied by means of binding and inhibition assays. Several synthetic peptides (10 amino acid sequences) designed from the amino acid sequence of VVLB{sub 4} and obtained from trypsin digestion were tested to determine which amino acids were involved in the interaction between MabVV{sub 34} and VVLB{sub 4}. The further unraveling of this epitope was investigated by inhibition using designed synthetic peptides as well as mixtures mimicking variable density effect. Under the experimental circumstances, MabVV{sub 34} was able to inhibit the binding of VVLB{sub 4} to Tn. Two of the four peptide sequences assayed showed better inhibition properties. Finally, mixtures containing these selected sequences allowed the evaluation of binding and inhibition as a function of Tn density. We conclude that the present study facilitates the further development of a specific Tn marker and may contribute to the development of Tn-like radiolabelled peptides or Tn-specific radiolabelled fragments providing a highly selective tool for cancer diagnosis and treatment. This strategy may contribute to characterize the new generation of radiopharmaceuticals for diagnosis and therapy based on biomolecules like antibodies, fragments or peptides, whose application is directly guided by their specific

A highly efficient transposon mutagenesis system for the tomato pathogen Clavibacter michiganensis subsp. michiganensis.

Science.gov (United States)

Kirchner, O; Gartemann, K H; Zellermann, E M; Eichenlaub, R; Burger, A

2001-11-01

A transposon mutagenesis system for Clavibacter michiganensis subsp. michiganensis was developed based on antibiotic resistance transposons that were derived from the insertion element IS1409 from Arthrobacter sp. strain TM1 NCIB12013. As a prerequisite, the electroporation efficiency was optimized by using unmethylated DNA and treatment of the cells with glycine such that about 5 x 10(6) transformants per microg of DNA were generally obtained. Electroporation of C. michiganensis subsp. michiganensis with a suicide vector carrying transposon Tn1409C resulted in approximately 1 x 10(3) transposon mutants per pg of DNA and thus is suitable for saturation mutagenesis. Analysis of Tn1409C insertion sites suggests a random mode of transposition. Transposition of Tn1409C was also demonstrated for other subspecies of C. michiganensis.

Influence of local regulations on TN dual purpose BWR casks

International Nuclear Information System (INIS)

Samson, P.; Neider, T.

1999-01-01

Transnucleaire (Paris, France) and Transnuclear, Inc. (Hawthorne, New York, United Sates) have both developed Pressurized Water Reactor (PWR) spent fuel casks for storage and transport purposes. The products are supplied in Europe by Transnucleaire and in the United States by Transnuclear, Inc. Now the TN Group is working on a design for Boiling Water Reactor (BWR) spent fuel assemblies: the TN 52 L cask is designed for transport and storage is Switzerland, the TN 68 cask is designed for transport and storage in the United States. For storage purpose, national regulatory requirements have to be met: each country has specific demands and criteria. As a consequence, differences between the TN 52 L design and the TN 68 design for rather similar contents appear in several fields: the design work, the licensing process, the manufacturing and the operational life. (author)

TRANSIT--A Software Tool for Himar1 TnSeq Analysis.

Directory of Open Access Journals (Sweden)

Michael A DeJesus

2015-10-01

Full Text Available TnSeq has become a popular technique for determining the essentiality of genomic regions in bacterial organisms. Several methods have been developed to analyze the wealth of data that has been obtained through TnSeq experiments. We developed a tool for analyzing Himar1 TnSeq data called TRANSIT. TRANSIT provides a graphical interface to three different statistical methods for analyzing TnSeq data. These methods cover a variety of approaches capable of identifying essential genes in individual datasets as well as comparative analysis between conditions. We demonstrate the utility of this software by analyzing TnSeq datasets of M. tuberculosis grown on glycerol and cholesterol. We show that TRANSIT can be used to discover genes which have been previously implicated for growth on these carbon sources. TRANSIT is written in Python, and thus can be run on Windows, OSX and Linux platforms. The source code is distributed under the GNU GPL v3 license and can be obtained from the following GitHub repository: https://github.com/mad-lab/transit.

Prediction of Microcystis Blooms Based on TN:TP Ratio and Lake Origin

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Yoshimasa Amano

2008-01-01

Full Text Available We evaluated the relationship between TN:TP ratio and Microcystis growth via a database that includes worldwide lakes based on four types of lake origin (dammed, tectonic, coastal, and volcanic lakes. We used microcosm and mesocosm for the nutrient elution tests with lake water and four kinds of sediment (nontreated, MgO sprinkling treated, dissolved air flotation [DAF] treated, and combined treated sediment in order to control TN:TP ratio and to suppress Microcystis growth. Microcystis growth was related to TN:TP ratio, with the maximum value at an optimum TN:TP ratio and the minimum values when the TN:TP ratios reached to 0 or ∞. The kurtosis of the distribution curve varied with the type of lake origin; the lowest kurtosis was found in dammed lakes, while the highest was found in volcanic lakes. The lake trophic state could affect the change in the kurtosis, providing much lower kurtosis at eutrophic lakes (dammed lakes than that at oligotrophic lakes (volcanic lakes. The relationship between TN:TP ratio and Microcystis growth could be explained by the nutrient elution tests under controlled TN:TP ratios through the various sediment treatments. A significant suppression of Microcystis growth of 70% could be achieved when the TN:TP ratios exceeded 21. Lake origin could be regarded as an index including morphological and geographical factors, and controlling the trophic state in lakes. The origin rather than trophic state for lakes could be considered as an important factor of TN:TP influences on Microcystis growth.

An insight into the thermodynamic characteristics of human thrombopoietin complexation with TN1 antibody

Science.gov (United States)

Shibazaki, Chie; Adachi, Motoyasu; Honjo, Eijiro; Tamada, Taro; Maeda, Yoshitake; Tahara, Tomoyuki; Kato, Takashi; Miyazaki, Hiroshi; Blaber, Michael; Kuroki, Ryota

2016-01-01

Abstract Human thrombopoietin (hTPO) primarily stimulates megakaryocytopoiesis and platelet production and is neutralized by the mouse TN1 antibody. The thermodynamic characteristics of TN1 antibody–hTPO complexation were analyzed by isothermal titration calorimetry (ITC) using an antigen‐binding fragment (Fab) derived from the TN1 antibody (TN1‐Fab). To clarify the mechanism by which hTPO is recognized by TN1‐Fab the conformation of free TN1‐Fab was determined to a resolution of 2.0 Å using X‐ray crystallography and compared with the hTPO‐bound form of TN1‐Fab determined by a previous study. This structural comparison revealed that the conformation of TN1‐Fab does not substantially change after hTPO binding and a set of 15 water molecules is released from the antigen‐binding site (paratope) of TN1‐Fab upon hTPO complexation. Interestingly, the heat capacity change (ΔCp) measured by ITC (−1.52 ± 0.05 kJ mol−1 K−1) differed significantly from calculations based upon the X‐ray structure data of the hTPO‐bound and unbound forms of TN1‐Fab (−1.02 ∼ 0.25 kJ mol−1 K−1) suggesting that hTPO undergoes an induced‐fit conformational change combined with significant desolvation upon TN1‐Fab binding. The results shed light on the structural biology associated with neutralizing antibody recognition. PMID:27419667

Establishment of colloid gold immunity chromatography assay for cardiac troponin I (cTnI)

International Nuclear Information System (INIS)

Wang Dezhi; Chen Jiying; Qin Lili; Zhao Baojian; Zhang Chunming

2006-01-01

Objective: To establish the colloid gold Immunity chromatography assay for cardiac troponin I. Methods: To purify cTnI from human cardiac muscle and immunize rabbit with it. cTnI antibody of rabbit anti-human cardiac muscle has been prepared and colloid gold immunity chromatography assay was established by using immunity chromatography technology. Results: Anti-serum titles of cTnI were 1:100000, Ka=2.38 x 10 9 L/mol; Methodological index: Sensitivity: 5 ng/ml; Specificity: cTnI is no cross-reaction with cTnT, cTnC and CK-MB. conclusion: The assay is highly specific, quick and simple. It can be widely used for the early diagnosis of AMI and scientific research. (authors)

ST6GalNAc-I controls expression of sialyl-Tn antigen in gastrointestinal tissues

DEFF Research Database (Denmark)

Marcos, Nuno T; Bennett, Eric P; Gomes, Joana

2011-01-01

Sialyl-Tn is a simple mucin-type carbohydrate antigen aberrantly expressed in gastrointestinal adenocarcinomas and in the precursor lesion intestinal metaplasia. Sialyl-Tn tumour expression is an independent indicator of poor prognosis. We have previously shown in vitro that ST6GalNAc-I and ST6GalNAc......-II sialyltransferases can synthesize sialyl-Tn. The aim of the present study was to establish whether ST6GalNAc-I is the major enzyme responsible for the expression of sialyl-Tn. We used a model of CHO-ldlD cells producing only MUC1-Tn glycoform and showed that ST6GalNAc-I is the key-enzyme leading to sialyl......-Tn biosynthesis. We developed novel monoclonal antibodies specific for ST6GalNAc-I and evaluated its expression in gastrointestinal tissues. ST6GalNAc-I was detected in normal colon mucosa co-localized with O-acetylated sialyl-Tn. Expression was largely unaltered in colorectal adenocarcinomas. In contrast, we...

Development of stable reporter system cloning luxCDABE genes into chromosome of Salmonella enterica serotypes using Tn7 transposon

Directory of Open Access Journals (Sweden)

Lawrence Mark L

2010-07-01

Full Text Available Abstract Background Salmonellosis may be a food safety problem when raw food products are mishandled and not fully cooked. In previous work, we developed bioluminescent Salmonella enterica serotypes using a plasmid-based reporting system that can be used for real-time monitoring of the pathogen's growth on food products in short term studies. In this study, we report the use of a Tn7-based transposon system for subcloning of luxCDABE genes into the chromosome of eleven Salmonella enterica serotypes isolated from the broiler production continuum. Results We found that the lux operon is constitutively expressed from the chromosome post-transposition and the lux cassette is stable without external pressure, i.e. antibiotic selection, for all Salmonella enterica serotypes used. Bioluminescence expression is based on an active electron transport chain and is directly related with metabolic activity. This relationship was quantified by measuring bioluminescence against a temperature gradient in aqueous solution using a luminometer. In addition, bioluminescent monitoring of two serotypes confirmed that our chicken skin model has the potential to be used to evaluate pathogen mitigation strategies. Conclusions This study demonstrated that our new stable reporting system eliminates bioluminescence variation due to plasmid instability and provides a reliable real-time experimental system to study application of preventive measures for Salmonella on food products in real-time for both short and long term studies.

Myofilament Calcium Sensitivity: Mechanistic Insight into TnI Ser-23/24 and Ser-150 Phosphorylation Integration

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Hussam E Salhi

2016-12-01

Full Text Available Troponin I (TnI is a major regulator of cardiac muscle contraction and relaxation. During physiological and pathological stress, TnI is differentially phosphorylated at multiple residues through different signaling pathways to match cardiac function to demand. The combination of these TnI phosphorylations can exhibit an expected or unexpected functional integration, whereby the function of two phosphorylations are different than that predicted from the combined function of each individual phosphorylation alone. We have shown that TnI Ser-23/24 and Ser-150 phosphorylation exhibit functional integration and are simultaneously increased in response to cardiac stress. In the current study, we investigated the functional integration of TnI Ser-23/24 and Ser-150 to alter cardiac contraction. We hypothesized that Ser-23/24 and Ser-150 phosphorylation each utilize distinct molecular mechanisms to alter the TnI binding affinity within the thin filament. Mathematical modeling predicts that Ser-23/24 and Ser-150 phosphorylation affect different TnI affinities within the thin filament to distinctly alter the Ca2+-binding properties of troponin. Protein binding experiments validate this assertion by demonstrating pseudo-phosphorylated Ser-150 decreases the affinity of isolated TnI for actin, whereas Ser-23/24 pseudo-phosphorylation is not different from unphosphorylated. Thus, our data supports that TnI Ser-23/24 affects TnI-TnC binding, while Ser-150 phosphorylation alters TnI-actin binding. By measuring force development in troponin-exchanged skinned myocytes, we demonstrate that the Ca2+ sensitivity of force is directly related to the amount of phosphate present on TnI. Furthermore, we demonstrate that Ser-150 pseudo-phosphorylation blunts Ser-23/24-mediated decreased Ca2+-sensitive force development whether on the same or different TnI molecule. Therefore, TnI phosphorylations can integrate across troponins along the myofilament. These data demonstrate

Tennessee Comptroller of the Treasury (TN COMP)

Data.gov (United States)

Social Security Administration — The Tennessee Comptroller of the Treasury (TN COMP) uses SSA's assistance in administering the Tennessee Property Tax Rebate Program. SSA provides assistance through...

ETIOLOGY CLASSIFICATION AND TREATMENT NEEDS (TN FOR ORAL MALODOR

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Anton Raharjo

2015-08-01

Full Text Available Background: Oral malodor, a generic descriptor term for foul smells emanating from the mouth can be classified as either pathological or physiological halitosis. Some problems are often confounded by the clinician's mismanagement. Objective: This paper reviews the etiology of classification and determination of treatment needs (TN for oral malodor. Literature review and discussion: In the majority of cases the problem has been shown to originate in the oral cavity. Although oral malodor cases are often related to physiological aspects, sometimes they can be related to extra oral sources and psychological aspects. Classification methods of oral malodor with corresponding treatment needs (TN have already been established. Although PTC & tongue brushing and appropriate mouthrinses are both important and basic treatment measures for halitosis, other dental treatments are sometimes required. Conclusion: Accurate screening and diagnosis of halitosis followed by appropriate TN may give better results and consequently reduce the risk of mismanagement.

Experimental manipulation of TN:TP ratiossuppress cyanobacterial biovolume and microcystinconcentration in large-scale in situ mesocosms

Science.gov (United States)

Harris, Theodore D.; Wilhelm, Frank M.; Graham, Jennifer L.; Loftin, Keith A.

2014-01-01

A global dataset was compiled to examine relations between the total nitrogen to total phosphorus ratio (TN:TP) and microcystin concentration in lakes and reservoirs. Microcystin concentration decreased as TN:TP ratios increased, suggesting that manipulation of the TN:TP ratio may reduce microcystin concentrations. This relationship was experimentally tested by adding ammonium nitrate to increase the TN:TP ratio in large-scale (70 m3), in situ mesocosms located in a eutrophic reservoir that routinely experiences toxic blooms of cyanobacteria. At a TN:TP ratio >75:1, chlorophytes dominated the phytoplankton community in the mesocosms, while cyanobacterial biovolume was significantly reduced and microcystin was not detected. In contrast, the unmanipulated reservoir was dominated by cyanobacteria, and microcystin was detected. Secchi depths were 1.1 to 1.8 times greater in the mesocosms relative to the reservoir. Cladoceran zooplankton had a larger body size (0.14 mm on average) in the mesocosms compared to conspecifics in the reservoir, which was likely related to the higher quality food. Combined, these empirical and experimental data indicate that although nutrient addition is counterintuitive to current cyanobacteria management practices, increasing the TN:TP ratio by adding nitrogen may be a potential short-term management strategy to reduce cyanobacteria and cyanotoxins when other alternatives (e.g., phosphorus reduction) are not possible. Additional experimental studies with careful controls are needed to define best management practices and identify any potential unintended consequences before nitrogen addition is implemented as a lake and reservoir management practice.

Some reproductive performances of Thai native (TN and 50% TN x Anglo-Nubian crossbred does with different levels of concentrate supplementation

Directory of Open Access Journals (Sweden)

Choldumrongkul, S.

2007-05-01

Full Text Available A 2 x 2 x 2 factorial in completely randomized design was conducted to determine the effect of genotype of does (Thai native (TN or 50% Anglo-Nubian (AN crossbred, levels of concentrate supplementation(1 % body weight or ad libitum and body condition of does (poor or good on oestrus incidence, kidding rate and multiple birth rate of does. Does rotationally grazed on Paspalum plicatulum pasture for 4 weeks andwere supplemented with concentrate. The experiment was divided into 4 periods: 1 does run with a teaser for 105 days before mating to determine an oestrus incidence 2 does joined with bucks of the same genotypefor 45 days 3 does run with a teaser for 150 days during pregnancy and 4 does run with a teaser 90 days post-partum. Genotype, levels of concentrate supplementation and body condition of does did not significantlyaffect (P>0.05 kidding rate or multiple birth rate and average kidding rate and multiple birth rate of the does was 83.8 and 82.3%, respectively. Birth weight of kids from TN does was significantly lower (P<0.01 than that from 50% TN x AN crossbred does (1.9 and 2.5 kg, respectively. Number of days of the first oestrus incidence in post-partum period for TN does was significantly (P<0.01 less than that of 50% ANcrossbred does (59.1 days and 75.0 days, respectively. Number of days of the first oestrus incidence in postpartum for does supplemented with concentrate ad libitum was significantly (P<0.01 lower than that ofdoes supplemented with 1% BW of concentrate (60.5 and 73.6 days, respectively.

HTR-TN achievements and prospects for future developments

International Nuclear Information System (INIS)

Hittner, D.; Angulo, C.; Basini, V.; Bogusch, E.; Breuil, E.; Buckthorpe, D.; Chauvet, V.; Futterer, M.A.; Van Heek, A.; Von Lensa, W.; Yvon, P.

2011-01-01

It is already 10 years since the (European) High Temperature Reactor Technology Network (HTR-TN) launched a program for development of HTR technology, which expanded through three successive Euratom framework programs, with many projects in line with the network strategy. Widely relying in the beginning on the legacy of the former European HTR developments (DRAGON, AVR, THTR, etc.) that it contributed to safeguard, this program led to advances in HTR/VHTR technologies and produced significant results, which can contribute to the international cooperation through Euratom involvement in the Generation IV International Forum (GIF). the main achievements of the European program, performed in complement to efforts made in several European countries and other GIF partners, are presented: they concern the validation of computer codes (reactor physics, as well as system transient analysis from normal operation to air ingress accident and fuel performance in normal and accident conditions), materials (metallic materials for vessel, direct cycle turbines and intermediate heat exchanger, graphite, etc.), component development, fuel manufacturing and irradiation behavior, and specific HTR waste management (fuel and graphite). Key experiments have been performed or are still ongoing, like irradiation of graphite and of fuel material (PYCASSO experiment), high burn-up fuel PIE, safety test and isotopic analysis, IHX mock-up thermohydraulic test in helium atmosphere, air ingress experiment for a block type core, etc. Now HTR-TN partners consider that it is time for Europe to go a step forward toward industrial demonstration. In line with the orientations of the 'Strategic Energy Technology Plan (SET-Plan)' recently issued by the European Commission that promotes a strategy for development of low-carbon energy technologies and mentions Generation IV nuclear systems as part of key technologies, HTR-TN proposes to launch a program for extending the contribution of nuclear energy to

ST6GalNAc-I controls expression of sialyl-Tn antigen in gastrointestinal tissues

DEFF Research Database (Denmark)

Marcos, Nuno T; Bennett, Eric Paul; Gomes, Joana

2011-01-01

-Tn biosynthesis. We developed novel monoclonal antibodies specific for ST6GalNAc-I and evaluated its expression in gastrointestinal tissues. ST6GalNAc-I was detected in normal colon mucosa co-localized with O-acetylated sialyl-Tn. Expression was largely unaltered in colorectal adenocarcinomas. In contrast, we......NAc-I as the major enzyme controlling the expression of cancer-associated sialyl-Tn antigen in gastrointestinal tissues....

cTnT can be a useful marker for early detection of anthracycline cardiotoxicity.

Science.gov (United States)

Kilickap, S; Barista, I; Akgul, E; Aytemir, K; Aksoyek, S; Aksoy, S; Celik, I; Kes, S; Tekuzman, G

2005-05-01

The level of serum cardiac troponin-T (cTnT) increases with myocardial damage. We sought to assess whether cTnT level could be a useful marker for the early detection of anthracycline cardiotoxicity. Forty-one patients who had been scheduled to receive anthracycline-containing combination chemotherapy were included in the study. Serum cTnT levels were measured before (baseline) and after the first cycle of chemotherapy, and again, after the last cycle of chemotherapy. In all patients, the left ventricular ejection fraction (LVEF), fractional shortening (FS), early peak flow/atrial flow velocity (E/A) ratio, and the isovolemic relaxation time (IRT) were measured echocardiographically, both before and after the completion of chemotherapy. LVEF and FS did not change in any patients. In 21 patients (49%), the E/A ratio decreased after therapy as compared to the pre-treatment values. The decrease in E/A ratio was more prominent in patients who were older than the mean age of our study group, which was 44 years. The post-treatment IRT was prolonged compared with the pretreatment IRT (94.0 +/- 2.0 versus 85.6 +/- 10.5 ms, respectively). cTnT levels after completion of therapy were elevated in 14 (34%) patients, and exceeded the upper limit of the normal range (>0.1 ng/ml) in only one patient. cTnT levels measured after completion of therapy were significantly higher, compared with those measured at baseline and after the first cycle of therapy. In the younger age group (< or =44 years old), there was a two-fold decrease in the E/A ratio in those patients whose cTnT levels increased during the therapy, when compared with those whose cTnT levels did not change (21% versus 43%, respectively). Increased serum cTnT level can be detected in the early stages of anthracycline therapy and it is associated with diastolic dysfunction of the left ventricle. Therefore, serum cTnT level could be a useful measure for early detection of anthracycline-induced cardiotoxicity.

Identification and Characterization of Non-Cellulose-Producing Mutants of Gluconacetobacter hansenii Generated by Tn5 Transposon Mutagenesis

Science.gov (United States)

Deng, Ying; Nagachar, Nivedita; Xiao, Chaowen; Tien, Ming

2013-01-01

The acs operon of Gluconacetobacter is thought to encode AcsA, AcsB, AcsC, and AcsD proteins that constitute the cellulose synthase complex, required for the synthesis and secretion of crystalline cellulose microfibrils. A few other genes have been shown to be involved in this process, but their precise role is unclear. We report here the use of Tn5 transposon insertion mutagenesis to identify and characterize six non-cellulose-producing (Cel−) mutants of Gluconacetobacter hansenii ATCC 23769. The genes disrupted were acsA, acsC, ccpAx (encoding cellulose-complementing protein [the subscript “Ax” indicates genes from organisms formerly classified as Acetobacter xylinum]), dgc1 (encoding guanylate dicyclase), and crp-fnr (encoding a cyclic AMP receptor protein/fumarate nitrate reductase transcriptional regulator). Protein blot analysis revealed that (i) AcsB and AcsC were absent in the acsA mutant, (ii) the levels of AcsB and AcsC were significantly reduced in the ccpAx mutant, and (iii) the level of AcsD was not affected in any of the Cel− mutants. Promoter analysis showed that the acs operon does not include acsD, unlike the organization of the acs operon of several strains of closely related Gluconacetobacter xylinus. Complementation experiments confirmed that the gene disrupted in each Cel− mutant was responsible for the phenotype. Quantitative real-time PCR and protein blotting results suggest that the transcription of bglAx (encoding β-glucosidase and located immediately downstream from acsD) was strongly dependent on Crp/Fnr. A bglAx knockout mutant, generated via homologous recombination, produced only ∼16% of the wild-type cellulose level. Since the crp-fnr mutant did not produce any cellulose, Crp/Fnr may regulate the expression of other gene(s) involved in cellulose biosynthesis. PMID:24013627

Isolation and characterization of Tn917lac-generated competence mutants of Bacillus subtilis

International Nuclear Information System (INIS)

Hahn, J.; Albano, M.; Dubnau, D.

1987-01-01

The authors isolated 28 mutants of Bacillus subtilis deficient in the development of competence by using the transposon Tn917lacZ as a mutagen. The mutant strains were poorly transformable with plasmid and chromosomal DNAs but were normally transducible and exhibited wild-type resistance to DNA-damaging agents. The mutations were genetically mapped, and the mutants were characterized with respect to their abilities to bind and take up radiolabeled DNA. All were defective in uptake, and some failed to bind significantly amounts of DNA. The abilities of the mutant strains to resolve into two buoyant density classes on Renografin gradients were studied. Most resolved normally, but several banded in Renografin only at the buoyant density of noncompetent cells. The genetic mapping studies and the other analyses suggested that the mutations define a minimum of seven distinct com genes

Impacts of climate change on TN load and its control in a River Basin with complex pollution sources.

Science.gov (United States)

Yang, Xiaoying; Warren, Rachel; He, Yi; Ye, Jinyin; Li, Qiaoling; Wang, Guoqing

2018-02-15

It is increasingly recognized that climate change could affect the quality of water through complex natural and anthropogenic mechanisms. Previous studies on climate change and water quality have mostly focused on assessing its impact on pollutant loads from agricultural runoff. A sub-daily SWAT model was developed to simulate the discharge, transport, and transformation of nitrogen from all known anthropogenic sources including industries, municipal sewage treatment plants, concentrated and scattered feedlot operations, rural households, and crop production in the Upper Huai River Basin. This is a highly polluted basin with total nitrogen (TN) concentrations frequently exceeding Class V of the Chinese Surface Water Quality Standard (GB3838-2002). Climate change projections produced by 16 Global Circulation Models (GCMs) under the RCP 4.5 and RCP 8.5 scenarios in the mid (2040-2060) and late (2070-2090) century were used to drive the SWAT model to evaluate the impacts of climate change on both the TN loads and the effectiveness of three water pollution control measures (reducing fertilizer use, constructing vegetative filter strips, and improving septic tank performance) in the basin. SWAT simulation results have indicated that climate change is likely to cause an increase in both monthly average and extreme TN loads in February, May, and November. The projected impact of climate change on TN loads in August is more varied between GCMs. In addition, climate change is projected to have a negative impact on the effectiveness of septic tanks in reducing TN loads, while its impacts on the other two measures are more uncertain. Despite the uncertainty, reducing fertilizer use remains the most effective measure for reducing TN loads under different climate change scenarios. Meanwhile, improving septic tank performance is relatively more effective in reducing annual TN loads, while constructing vegetative filter strips is more effective in reducing annual maximum monthly

Comblike dendrimers containing Tn antigen modulate natural killing and induce the production of Tn specific antibodies

Czech Academy of Sciences Publication Activity Database

Vepřek, Pavel; Hajdúch, M.; Džubák, P.; Kulík, R.; Poláková, J.; Bezouška, Karel

2006-01-01

Roč. 49, č. 21 (2006), s. 6400-6407 ISSN 0022-2623 R&D Projects: GA AV ČR IAA4020213; GA AV ČR IAA5020403; GA ČR GA304/06/1691 Institutional research plan: CEZ:AV0Z40550506; CEZ:AV0Z50200510 Keywords : Tn antigen * glycodendrimer * NKP-P1 Subject RIV: CE - Biochemistry Impact factor: 5.115, year: 2006

Pharmaceutical significance of Leuconostoc mesenteroides KS-TN11 isolated from Nile Tilapia, Oreochromis niloticus

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Bilal Ahmad Paray

2018-05-01

Full Text Available Aquatic animals are known for their myriad of beneficial bacteria with diverse biologically active compounds. The current study was aimed to isolate and characterize potentially beneficial lactic acid bacteria from Nile Tilapia and evaluate their pharmaceutical applications. The fish samples were dissected and stomach, intestine, and gills were collected and serially diluted for the isolation of lactic acid bacteria (LAB on BCP agar media. Identification of isolate was carried by biochemical and molecular characterization using API kit and 16S rRNA gene sequencing analysis, respectively. Further, KS-TN11 was assessed for α-glucosidase inhibitory potential using the chromogenic method. A lactic acid bacterium KS-TN11 was isolated from the stomach of Nile Tilapia and identified as Leuconostoc mesenteroides. Effect of KS-TN11 on lipid accumulation in adipocytes was done by using Oil Red O staining. The isolate showed strong antibacterial activity against a number of pathogenic bacteria in vitro. In addition, L. mesenteroides KS-TN11 KS-TN11 (50 mg/ml and 100 mg/ml tends to inhibit adipogenesis in 3T3-L1 adipocytes and thus may have possible anti-obesity effects. Moreover, L. mesenteroides KS-TN11 exhibited substantial α–glucosidase inhibitory activities by 41.33% at 50 mg/ml and 64% at 100 mg/ml, respectively. The bacterium showed potent antibacterial activity against a number of pathogenic bacteria; in addition to alpha-glucosidase activity, and inhibition of lipid accumulation in 3T3-L1 cell line. These results reinforce KS-TN11 as a novel bacterium with an impending pharmaceutical application. Keywords: Lactic acid bacteria, Antimicrobial, Fish microflora, Anti-diabetic

Two-stage soil infiltration treatment system for treating ammonium wastewaters of low COD/TN ratios.

Science.gov (United States)

Lei, Zhongfang; Wu, Ting; Zhang, Yi; Liu, Xiang; Wan, Chunli; Lee, Duu-Jong; Tay, Joo-Hwa

2013-01-01

Soil infiltration treatment (SIT) is ineffective to treat ammonium wastewaters of total nitrogen (TN) > 100 mg l(-1). This study applied a novel two-stage SIT process for effective TN removal from wastewaters of TN>100 mg l(-1) and of chemical oxygen demand (COD)/TN ratio of 3.2-8.6. The wastewater was first fed into the soil column (stage 1) at hydraulic loading rate (HLR) of 0.06 m(3) m(-2) d(-1) for COD removal and total phosphorus (TP) immobilization. Then the effluent from stage 1 was fed individually into four soil columns (stage 2) at 0.02 m(3) m(-2) d(-1) of HLR with different proportions of raw wastewater as additional carbon source. Over the one-year field test, balanced nitrification and denitrification in the two-stage SIT revealed excellent TN removal (>90%) from the tested wastewaters. Copyright © 2012 Elsevier Ltd. All rights reserved.

76 FR 14855 - Television Broadcasting Services; Nashville, TN

Science.gov (United States)

2011-03-18

... FEDERAL COMMUNICATIONS COMMISSION 47 CFR Part 73 [MB Docket No. 11-29, RM-11622; DA 11-335] Television Broadcasting Services; Nashville, TN AGENCY: Federal Communications Commission. ACTION: Proposed... 73 Television, Television broadcasting. Federal Communications Commission. Kevin R. Harding...

76 FR 33656 - Television Broadcasting Services; Nashville, TN

Science.gov (United States)

2011-06-09

... FEDERAL COMMUNICATIONS COMMISSION 47 CFR Part 73 [MB Docket No. 11-29; RM-11622, DA 11-949] Television Broadcasting Services; Nashville, TN AGENCY: Federal Communications Commission. ACTION: Final rule... Part 73 Television. Federal Communications Commission. Barbara A. Kreisman, Chief, Video Division...

Role of Myofibril-Inducing RNA in cardiac TnT expression in developing Mexican axolotl

Science.gov (United States)

Sferrazza, Gian-Franco; Zhang, Chi; Jia, Pingping; Lemanski, Sharon L.; Athauda, Gagani; Stassi, Alyssa; Halager, Kristine; Maier, Jennifer A.; Rueda-de-Leon, Elena; Gupta, Amit; Dube, Syamalima; Huang, Xupei; Prentice, Howard M.; Dube, Dipak K.; Lemanski, Larry F.

2007-01-01

The Mexican axolotl, Ambystoma mexicanum, has been a useful animal model to study heart development and cardiac myofibrillogenesis. A naturally-occurring recessive mutant, gene “c”, for cardiac non-function in the Mexican axolotl causes a failure of myofibrillogenesis due to a lack of tropomyosin expression in homozygous mutant (c/c) embryonic hearts.. Myofibril-Inducing RNA (MIR) rescues mutant hearts in vitro by promoting tropomyosin expression and myofibril formation thereafter. We have studied the effect of MIR on the expression of various isoforms of cardiac Troponin-T (cTnT), a component of the thin filament that binds with tropomyosin. Four alternatively spliced cTnT isoforms have been characterized from developing axolotl heart. The expression of various cTnT isoforms in normal, mutant, and mutant hearts corrected with MIR, is evaluated by real-time RT-PCR using isoform specific primer pairs; MIR affects the total transcription as well as the splicing of the cTnT in axolotl heart PMID:17408593

Japanese version transport/storage packaging 'TN24'

International Nuclear Information System (INIS)

Kakunai, H.; Iida, T.; Tsuda, K.; Akamatsu, H.

1993-01-01

Since 1983, Kobe Steel has been engaged jointly with the French company Transnucleaire in the development of 'TN24', a dry-type transport and storage packaging for irradiated fuel elements. This report describes the packaging, which has been adapted for use in domestic power stations using BWRs on the basis of the results of this development. (J.P.N.)

Peri-operative troponin monitoring using a prototype high-sensitivity cardiac troponin I (hs-cTnI) assay: comparisons with hs-cTnT and contemporary cTnI assays.

LENUS (Irish Health Repository)

Lee, Graham R

2013-09-18

Non-cardiac surgery is associated with major vascular complications and higher incidences of elevated plasma troponin (cTn) concentration. Goal-directed therapy (GDT) is a stroke volume (SV)-guided approach to intravenous (IV) fluid therapy that improves tissue perfusion, oxygenation and reduces post-operative complications. In patients undergoing major gastro-intestinal surgery, we compared high sensitive and contemporary troponin assays and correlated results with patient outcome.

Efficient transformation of sucrose into high pullulan concentrations by Aureobasidium melanogenum TN1-2 isolated from a natural honey.

Science.gov (United States)

Jiang, Hong; Xue, Si-Jia; Li, Yan-Feng; Liu, Guang-Lei; Chi, Zhen-Ming; Hu, Zhong; Chi, Zhe

2018-08-15

A very high pullulan producing yeast-like fungus, Aureobasidium melanogenum TN1-2 isolated from a natural honey was found to be able to produce 97.0 g/L of pullulan from 140.0 g/L sucrose at a flask level while it could yield 114.0 g/L of pullulan within 134 h during a 10-liter fermentation, the yield was 0.81 g/g and the productivity was 0.86 g/L/h. The high ability to biosynthesize pullulan by this yeast-like fungal strain TN1-2 was related to high glucosyltransferase activity, high phosphofructo-2-kinase activity, high content of its cellular glycerol and low glucose repressor. The Mw of the produced pullulan was 1.42 × 10 5  g/mol. The low Mw may be due to the high α-amylase, glucoamylase and isopullulanase activities. The intracellular level of trehalose had no influence on high pullulan production by the yeast-like fungal strain TN1-2. Copyright © 2018. Published by Elsevier Ltd.

A NEW CELL CLONE DERIVED FROM TRICHOPLUSIA NI TN5B1-4 CELLS

Institute of Scientific and Technical Information of China (English)

Jian-xiaoTian; Chang-youLi; Gui-lingZheng; Guo-xunLi; PingWang; Granados

2004-01-01

The characteristics of a cultured cell line do not always remain stable and may change upon continuous passage. Most continuous cell lines, even after cloning, possess several genotypes that are constantly changing. There are numerous selective and adaptive culture processes, in addition to genetic instability, that may improve phenotypic change in cell growth, virus susceptibility, gene expression, and production of virus. Similar detrimental effects of long term passaging of insect cells have also been reported for continuous cell lines, for example, Tn5B 1-4 cells, which are the most widely used for the baculovirus expression vector system (BEVS), provide superior production of recombinant proteins,however, this high productivity may be more evident in low passage cells. In this paper, we describe the isolation of a cell clone, Tn5B-40, from low passage Tn5B 1-4 cells. The growth characteristics,productions of virus, and high level of recombinant protein productions were determined. The results showed the susceptibility of both clone and Tn5B 1-4 cells to wild-type AcNPV was approximately the same rate with over 95% of infection; when the cloned cells were infected with recombinant baculoviruses expressing β-galactosidase and secreted alkaline phosphatase (SEAP), expression of the recombinant proteins from the cloned cells exceeded that from the parental Tn5B 1-4 cells.

Novel Aminoglycoside Resistance Transposons and Transposon-Derived Circular Forms Detected in Carbapenem-Resistant Acinetobacter baumannii Clinical Isolates

Science.gov (United States)

Dwibedi, Chinmay Kumar; Sjöström, Karin; Edquist, Petra; Wai, Sun Nyunt; Uhlin, Bernt Eric

2016-01-01

Acinetobacter baumannii has emerged as an important opportunistic pathogen equipped with a growing number of antibiotic resistance genes. Our study investigated the molecular epidemiology and antibiotic resistance features of 28 consecutive carbapenem-resistant clinical isolates of A. baumannii collected throughout Sweden in 2012 and 2013. The isolates mainly belonged to clonal complexes (CCs) with an extensive international distribution, such as CC2 (n = 16) and CC25 (n = 7). Resistance to carbapenems was related to blaOXA-23 (20 isolates), blaOXA-24/40-like (6 isolates), blaOXA-467 (1 isolate), and ISAba1-blaOXA-69 (1 isolate). Ceftazidime resistance was associated with blaPER-7 in the CC25 isolates. Two classical point mutations were responsible for resistance to quinolones in all the isolates. Isolates with high levels of resistance to aminoglycosides carried the 16S rRNA methylase armA gene. The isolates also carried a variety of genes encoding aminoglycoside-modifying enzymes. Several novel structures involved in aminoglycoside resistance were identified, including Tn6279, ΔTn6279, Ab-ST3-aadB, and different assemblies of Tn6020 and TnaphA6. Importantly, a number of circular forms related to the IS26 or ISAba125 composite transposons were detected. The frequent occurrence of these circular forms in the populations of several isolates indicates a potential role of these circular forms in the dissemination of antibiotic resistance genes. PMID:26824943

Identification of novel genes responsible for ethanol and/or thermotolerance by transposon mutagenesis in Saccharomyces cerevisiae

Energy Technology Data Exchange (ETDEWEB)

Kim, Hyun-Soo [Ewha Womans Univ., Seoul (Korea, Republic of). Dept. of Life Sciences; Kim, Na-Rae [Ewha Womans Univ., Seoul (Korea, Republic of). Div. of Life and Pharmaceutical Sciences; Yang, Jungwoo [Ewha Womans Univ., Seoul (Korea, Republic of). Microbial Resources Research Center; Choi, Wonja [Ewha Womans Univ., Seoul (Korea, Republic of). Dept. of Life Sciences; Ewha Womans Univ., Seoul (Korea, Republic of). Div. of Life and Pharmaceutical Sciences; Ewha Womans Univ., Seoul (Korea, Republic of). Microbial Resources Research Center

2011-08-15

Saccharomyces cerevisiae strains tolerant to ethanol and heat stresses are important for industrial ethanol production. In this study, five strains (Tn 1-5) tolerant to up to 15% ethanol were isolated by screening a transposon-mediated mutant library. Two of them displayed tolerance to heat (42 C). The determination of transposon insertion sites and Northern blot analysis identified seven putative genes (CMP2, IMD4, SSK2, PPG1, DLD3, PAM1, and MSN2) and revealed simultaneous down-regulations of CMP2 and IMD4, and SSK2 and PPG1, down-regulation of DLD3, and disruptions of the open reading frame of PAM1 and MSN2, indicating that ethanol and/or heat tolerance can be conferred. Knockout mutants of these seven individual genes were ethanol tolerant and three of them (SSK2, PPG1, and PAM1) were tolerant to heat. Such tolerant phenotypes reverted to sensitive phenotypes by the autologous or overexpression of each gene. Five transposon mutants showed higher ethanol production and grew faster than the control strain when cultured in rich media containing 30% glucose and initial 6% ethanol at 30 C. Of those, two thermotolerant transposon mutants (Tn 2 and Tn 3) exhibited significantly enhanced growth and ethanol production compared to the control at 42 C. The genes identified in this study may provide a basis for the application in developing industrial yeast strains. (orig.)

Analysis of Tn antigenicity with a panel of new IgM and IgG1 monoclonal antibodies raised against leukemic cells

DEFF Research Database (Denmark)

Blixt, Klas Ola; Lavrova, Olga I; Mazurov, Dmitriy V

2012-01-01

antigenicity remain poorly understood. As a consequence, a broad variety of anti-Tn monoclonal antibodies (mAbs) have been generated. To further investigate the nature and complexity of the Tn antigen, we generated seven different anti-Tn mAbs of IgM and IgG classes raised against human Jurkat T cells, which...... are Tn-positive due to the low activity of T-synthase and mutation in specific chaperone Cosmc. The binding analysis of anti-Tn mAbs with the array of synthetic saccharides, glycopeptides and O-glycoproteins revealed unexpected differences in specificities of anti-Tn mAbs. IgM mAbs bound the terminal Gal...

Diversity of the Tetracycline Mobilome within a Chinese Pig Manure Sample.

Science.gov (United States)

Leclercq, Sébastien Olivier; Wang, Chao; Zhu, Yaxin; Wu, Hai; Du, Xiaochen; Liu, Zhipei; Feng, Jie

2016-11-01

Tetracycline antibiotics are widely used in livestock, and tetracycline resistance genes (TRG) are frequently reported in the manure of farmed animals. However, the diversity of TRG-carrying transposons in manure has still been rarely investigated. Using a culture-free functional metagenomic procedure, combined with large-insert library construction and sequencing, bioinformatic analyses, and functional experiments, we identified 17 distinct TRGs in a single pig manure sample, including two new tet genes: tet(59), encoding a tetracycline efflux pump, and tet(W/N/W), encoding mosaic ribosomal protection. Our study also revealed six new TRG-carrying putative nonconjugative transposons: Tn5706-like transposon Tn6298, IS200/605-related transposon Tn6303, Tn3 family transposon Tn6299, and three ISCR2-related transposons, Tn62300, Tn62301, and Tn62302 IMPORTANCE: Fertilization of agricultural fields with animal manure is believed to play a major role in antibiotic resistance dissemination in the environment. There is growing concern for the possible spread of antibiotic resistance from the environment to humans since genetic resistance determinants may be located in transposons and other mobile genetic elements potentially transferable to pathogens. Among the various antibiotic resistance genes found in manure, tetracycline resistance genes (TRGs) are some of the most common. The present study provides a detailed snapshot of the tetracycline mobilome in a single pig manure sample, revealing an unappreciated diversity of TRGs and potential TRG mobility vectors. Our precise identification of the TRG-carrying units will enable us to investigate in more details their mobility effectiveness. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

A inter-relação das sialomucinas (antígenos Tn e Stn com o adenocarcinoma no esôfago de Barrett Relationship of the sialomucins (Tn and Stn antigens with adenocarcinoma in Barrett's esophagus

Directory of Open Access Journals (Sweden)

Plínio Conte de Faria Júnior

2007-08-01

Full Text Available OBJETIVO: O esôfago de Barrett (EB é conseqüência do refluxo gastroesofágico crônico e considerado fator de risco para o desenvolvimento de adenocarcinoma. Estudos do muco, em especial das mucinas ácidas representadas pelas sialomucinas presentes nas células caliciformes, mostraram que na metaplasia do tipo intestinal, o epitélio do órgão pode expressar antígenos denominados Tn e Stn. Estes antígenos já foram analisados em tumores gástricos e colônicos, porém não foram encontradas referências à sua utilização no EB. Este trabalho objetivou analisar estes antígenos em doentes com EB e em adenocarcinoma associado ao EB. MÉTODOS: Foram estudados, utilizando testes imunohistoquímicos, os antígenos Tn e Stn, nas biópsias endoscópicas de 29 doentes com EB, sete com adenocarcinoma no EB, além de oito indivíduos com epitélio esofágico normal. RESULTADOS: Nas células caliciformes, foi observada positividade para Stn em 100% dos casos e para Tn em 48% dos casos. Nas células colunares, o Stn foi sempre negativo, enquanto o Tn foi positivo em 100% dos casos. Entretanto, nos doentes com adenocarcinoma no EB, a positividade para ambos os antígenos foi de 100%. Nos indivíduos normais, houve positividade para o antígeno Tn e negatividade para Stn em todos os casos (100%. CONCLUSÃO: É provável que nos doentes com EB a positividade para o Tn, à semelhança do ocorrido quanto à positividade do mesmo antígeno nos portadores de adenocarcinoma, possa significar maior suscetibilidade para ocorrência futura de câncer. Assim, a pesquisa das sialomucinas poderá ser rotineiramente utilizada, contribuindo como fator prognóstico para desenvolvimento de adenocarcinoma no EB.OBJECIVE: Barrett's esophagus (BE is a consequence of chronic gastroesophageal reflux and is considered a risk factor for adenocarcinoma. The study of the mucus, especially acid mucins, such as the sialomucins in the goblet cells which characterize BE, showed

Characterization of the Complete Nucleotide Sequences of IncA/C2 Plasmids Carrying In809-Like Integrons from Enterobacteriaceae Isolates of Wildlife Origin.

Science.gov (United States)

Papagiannitsis, Costas C; Kutilova, Iva; Medvecky, Matej; Hrabak, Jaroslav; Dolejska, Monika

2017-09-01

A total of 18 Enterobacteriaceae (17 from gulls and 1 from a clinical sample) collected from Australia, carrying IncA/C plasmids with the IMP-encoding In809-like integrons, were studied. Seven plasmids, being representatives of different origins, plasmid sizes, replicon combinations, and resistance genes, were completely sequenced. Plasmid pEc158, identified in a clinical Escherichia coli ST752 isolate, showed extensive similarity to type 2 IncA/C 2 plasmids. pEc158 carried none of the bla CMY-2 -like region or ARI-B and ARI-A regions, while it contained a hybrid transposon structure. The six remaining plasmids, which were of wildlife origin, were highly similar to each other and probably were fusion derivatives of type 1 and type 2 A/C 2 plasmids. The latter plasmids contained an ARI-B region and hybrid transposon structures. In all plasmids, hybrid transposon structures containing In809-like integrons were inserted 3,434 bp downstream of the rhs2 start codon. In all cases, the one outermost 38-bp inverted repeat (IR) of the transposon was associated with the Tn 1696 tnp module, while the other outermost 38-bp IR of the transposon was associated with either a Tn 6317 -like module or a Tn 21 mer module. However, the internal structure of the transposon and the resistance genes were different in each plasmid. These findings indicated that, for the specific periods of time and settings, different IncA/C 2 plasmid types carrying In809-like elements circulated among isolates of wildlife and clinical origins. Additionally, they provided the basis for speculations regarding the reshuffling of IncA/C 2 plasmids with In809-like integrons and confirmed the rapid evolution of IncA/C 2 plasmid lineages. Copyright © 2017 American Society for Microbiology.

The Breast Cancer-Associated Glycoforms of MUC1, MUC1-Tn and sialyl-Tn, Are Expressed in COSMC Wild-Type Cells and Bind the C-Type Lectin MGL.

Directory of Open Access Journals (Sweden)

Richard Beatson

Full Text Available Aberrant glycosylation occurs in the majority of human cancers and changes in mucin-type O-glycosylation are key events that play a role in the induction of invasion and metastases. These changes generate novel cancer-specific glyco-antigens that can interact with cells of the immune system through carbohydrate binding lectins. Two glyco-epitopes that are found expressed by many carcinomas are Tn (GalNAc-Ser/Thr and STn (NeuAcα2,6GalNAc-Ser/Thr. These glycans can be carried on many mucin-type glycoproteins including MUC1. We show that the majority of breast cancers carry Tn within the same cell and in close proximity to extended glycan T (Galβ1,3GalNAc the addition of Gal to the GalNAc being catalysed by the T synthase. The presence of active T synthase suggests that loss of the private chaperone for T synthase, COSMC, does not explain the expression of Tn and STn in breast cancer cells. We show that MUC1 carrying both Tn or STn can bind to the C-type lectin MGL and using atomic force microscopy show that they bind to MGL with a similar dead adhesion force. Tumour associated STn is associated with poor prognosis and resistance to chemotherapy in breast carcinomas, inhibition of DC maturation, DC apoptosis and inhibition of NK activity. As engagement of MGL in the absence of TLR triggering may lead to anergy, the binding of MUC1-STn to MGL may be in part responsible for some of the characteristics of STn expressing tumours.

76 FR 35909 - Temporary Concession Contract for Big South Fork National Recreation Area, TN/KY

Science.gov (United States)

2011-06-20

... Recreation Area, TN/KY. SUMMARY: Pursuant to 36 CFR 51.24, public notice is hereby given that the National... Concession Contract for Big South Fork National Recreation Area, TN/KY AGENCY: National Park Service... services within Big South Fork National Recreation Area, Tennessee and Kentucky, for a term not to exceed 3...

A critical evaluation of the Beckman Coulter Access hsTnI: Analytical performance, reference interval and concordance.

Science.gov (United States)

Pretorius, Carel J; Tate, Jillian R; Wilgen, Urs; Cullen, Louise; Ungerer, Jacobus P J

2018-05-01

We investigated the analytical performance, outlier rate, carryover and reference interval of the Beckman Coulter Access hsTnI in detail and compared it with historical and other commercial assays. We compared the imprecision, detection capability, analytical sensitivity, outlier rate and carryover against two previous Access AccuTnI assay versions. We established the reference interval with stored samples from a previous study and compared the concordances and variances with the Access AccuTnI+3 as well as with two commercial assays. The Access hsTnI had excellent analytical sensitivity with the calibration slope 5.6 times steeper than the Access AccuTnI+3. The detection capability was markedly improved with the SD of the blank 0.18-0.20 ng/L, LoB 0.29-0.33 ng/L and LoD 0.58-0.69 ng/L. All the reference interval samples had a result above the LoB value. At a mean concentration of 2.83 ng/L the SD was 0.28 ng/L (CV 9.8%). Carryover (0.005%) and outlier (0.046%) rates were similar to the Access AccuTnI+3. The combined male and female 99th percentile reference interval was 18.2 ng/L (90% CI 13.2-21.1 ng/L). Concordance amongst the assays was poor with only 16.7%, 19.6% and 15.2% of samples identified by all 4 assays as above the 99th, 97.5th and 95th percentiles. Analytical imprecision was a minor contributor to the observed variances between assays. The Beckman Coulter Access hsTnI assay has excellent analytical sensitivity and precision characteristics close to zero. This allows cTnI measurement in all healthy individuals and the capability to identify numerically small differences between serial samples as statistically significant. Concordance in healthy individuals remains poor amongst assays. Crown Copyright © 2018. Published by Elsevier Inc. All rights reserved.

The TN-GEMINI: experience on a versatile alpha waste transport container

International Nuclear Information System (INIS)

Roland, V.; Chanzy, Y.

2001-01-01

The present paper discusses experience gained in moving alpha wastes and its teachings regarding transport aspects of D and D. Alpha wastes are generated in fuel cycle facilities such as those involved in reprocessing, in manufacture of mixed oxide fuel, and by research laboratories. If a significant amount of wastes has to be transported, then a Type B packaging is required. Developed by Transnucleaire and COGEMA, the TN GEMINI container enables nuclear facilities operators to optimise their alpha waste transport management, and more generally contribute to their D and D projects. After describing succinctly the design of the TN GEMINI, the paper will explain how the packaging is being operated. Teachings from experience will be shared. (orig.)

Development Of Protocols For Simultaneous Measurements Of Rn, Tn Using Nuclear Track Detectors And Trial Application In Mining Areas Of Vietnam

International Nuclear Information System (INIS)

Bui Dac Dung; Trinh Van Giap; Le Dinh Cuong; Tran Khanh Minh; Nguyen Huu Quyet; Nguyen Van Khanh; Tran Ngoc Toan

2013-01-01

Radioactive gases Radon (Rn) and Thoron (Tn) contribute of more than 50% of natural radiation dose. However, separate measurements of Rn and Tn have not been paid enough attention. An Intergovernmental Cooperation Project was conducted at the Institute for Nuclear Science and Technology with the help from Hungarian experts. The main tasks of the project were to finalize 02 protocols for simultaneous measurements of Rn and Tn using nuclear track detectors and to test these protocols in investigating the concentrations of Rn and Tn, calculating the natural dose due to Rn and Tn, and evaluating the increased radiation dose and radiation safety due to mining activities. Main results of the project include 02 protocols for simultaneous measurements of Rn and Tn and the test results in the mining areas. Rn and Tn concentrations inside the coal mining tunnels are in the average level of Vietnam and the world. Tn concentration inside the factory for separating Zircon at the Ha Tinh Zircon Processing Plant was found to be very high, up to 2931 Bq/m 3 . Based on annual effective dose calculation, workers inside the factory for separating Zircon at the Ha Tinh Zircon Processing Plant could receive an annual effective dose due to Rn and Tn of 4.890 mSv/year, and the increasing dose of 4.710 mSv/year is for higher than 1 mSv/year recommended by the IAEA. (author)

The Breast Cancer-Associated Glycoforms of MUC1, MUC1-Tn and sialyl-Tn, Are Expressed in COSMC Wild-Type Cells and Bind the C-Type Lectin MGL

DEFF Research Database (Denmark)

Beatson, Richard; Maurstad, Gjertrud; Picco, Gianfranco

2015-01-01

Aberrant glycosylation occurs in the majority of human cancers and changes in mucin-type O-glycosylation are key events that play a role in the induction of invasion and metastases. These changes generate novel cancer-specific glyco-antigens that can interact with cells of the immune system through...... carbohydrate binding lectins. Two glyco-epitopes that are found expressed by many carcinomas are Tn (GalNAc-Ser/Thr) and STn (NeuAcα2,6GalNAc-Ser/Thr). These glycans can be carried on many mucin-type glycoproteins including MUC1. We show that the majority of breast cancers carry Tn within the same cell...

Induction of the Tn10 Precise Excision in E. coli Cells after Accelerated Heavy Ions Irradiation

CERN Document Server

Zhuravel, D V

2003-01-01

The influence of the irradiation of different kinds on the indication of the structural mutations in the bacteria Escherichia coli is considered. The regularities of the Tn10 precise excision after accelerated ^{4}He and ^{12}C ions irradiations with different linear energy transfer (LET) were investigated. Dose dependences of the survival and relative frequency of the Tn10 precise excision were obtained. It was shown, that the relative frequency of the Tn10 precise excision is the exponential function from the irradiation dose. Relative biological efficiency (RBE), and relative genetic efficiency (RGE) were calculated, and were treated as the function of the LET.

Tn7::In2-8 dispersion in multidrug resistant isolates of Acinetobacter baumannii from Chile Dispersión de Tn7::In2-8 en aislamientos multirresistentes de Acinetobacter baumannii de Chile

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M. S. Ramírez

2010-06-01

Full Text Available Acinetobacter baumannii is considered an important pathogen in our hospital environment having a well-known capacity to acquire different mechanisms of antibiotic resistance. Previous studies in our laboratory had exposed the high dispersion of class 2 integrons in this species. In the present study, we analyzed 7 multiresistant intI2 positive A. baumannii isolates, 6 of which were found to harbour the Tn7::In2-8 element. Our results demonstrate the unusually high distribution of Tn7::In2-8 among different A. baumannii clones from Chile, suggesting a particular behavior of these elements at geographical level.Acinetobacter baumannii, patógeno de importancia clínica en el ámbito hospitalario, es reconocido como un microorganismo que posee la capacidad de evolucionar rápidamente hacia la multirresistencia. Estudios previos efectuados en nuestro laboratorio han demostrado la alta dispersión de los integrones de clase 2 en aislamientos de esta especie. En el presente trabajo se analizaron 7 aislamientos de Acinetobacter baumannii multirresistentes portadores de la integrasa de clase 2, 6 de los cuales portaban el inusual arreglo Tn7::In2-8. Nuestros resultados muestran una elevada frecuencia de dispersión del elemento Tn7::In2-8 en diferentes clones circulantes en Chile, lo que sugiere un comportamiento geográfico particular.

Highly efficient full-length hepatitis C virus genotype 1 (strain TN) infectious culture system

DEFF Research Database (Denmark)

Li, Yi-Ping; Ramirez, Santseharay; Jensen, Sanne B

2012-01-01

Chronic infection with hepatitis C virus (HCV) is an important cause of end stage liver disease worldwide. In the United States, most HCV-related disease is associated with genotype 1 infection, which remains difficult to treat. Drug and vaccine development was hampered by inability to culture...... full-length TN infection dose-dependently. Given the unique importance of genotype 1 for pathogenesis, this infectious 1a culture system represents an important advance in HCV research. The approach used and the mutations identified might permit culture development for other HCV isolates, thus......) culture systems in Huh7.5 cells. Here, we developed a highly efficient genotype 1a (strain TN) full-length culture system. We initially found that the LSG substitutions conferred viability to an intergenotypic recombinant composed of TN 5' untranslated region (5'UTR)-NS5A and JFH1 NS5B-3'UTR; recovered...

Transitioning high sensitivity cardiac troponin I (hs-cTnI) into routine diagnostic use: More than just a sensitivity issue

LENUS (Irish Health Repository)

Lee, Graham R

2016-04-01

High sensitivity cardiac troponin T and I (hs-cTnT and hs-cTnI) assays show analytical, diagnostic and prognostic improvement over contemporary sensitive cTn assays. However, given the importance of troponin in the diagnosis of myocardial infarction, implementing this test requires rigorous analytical and clinical verification across the total testing pathway. This was the aim of this study.

Transposon mutations in the flagella biosynthetic pathway of the solvent-tolerant Pseudomonas putida S12 result in a decreased expression of solvent efflux genes

NARCIS (Netherlands)

Kieboom, J; Bruinenberg, R; Keizer-Gunnink, [No Value; de Bont, JAM

2001-01-01

Fourteen solvent-sensitive transposon mutants were generated from the solvent-tolerant Pseudomonas putida strain S12 by applying the TnMOD-KmO mutagenesis system. These mutants were unable to grow in the presence of octanol and toluene. By cloning the region flanking the transposon insertion point a

Establishing exchange bias below T-N with polycrystalline Ni0.52Co0.48O/Co bilayers

DEFF Research Database (Denmark)

Berkowitz, A.E.; Hansen, Mikkel Fougt; Tang, Y.J.

2005-01-01

Exchange-coupled bilayers of polycrystalline ferromagnetic (FM) Co on antiferromagnetic (AFM) Ni0.52Co0.48O were investigated with emphasis on the issue of establishing an exchange-bias field, H-E, below the AFM ordering temperature, T-N. It was found that field-cooling the bilayers through T-N p...

78 FR 54168 - Special Local Regulation, Cumberland River, Mile 157.0 to 159.0; Ashland City, TN

Science.gov (United States)

2013-09-03

... Local Regulation, Cumberland River, Mile 157.0 to 159.0; Ashland City, TN AGENCY: Coast Guard, DHS... Special local regulation; Cumberland River, Miles 157.0 to 159.0, Ashland City, TN. (a) Location. The... regulation for the waters of the Cumberland River beginning at mile marker 157.0 and ending at mile marker...

First detection of the staphylococcal trimethoprim resistance gene dfrK and the dfrK-carrying transposon Tn559 in enterococci.

Science.gov (United States)

López, María; Kadlec, Kristina; Schwarz, Stefan; Torres, Carmen

2012-02-01

The trimethoprim resistance gene dfrK has been recently described in Staphylococcus aureus, but so far has not been found in other bacteria. A total of 166 enterococci of different species (E. faecium, E. faecalis, E. hirae, E. durans, E. gallinarum, and E. casseliflavus) and origins (food, clinical diseases in humans, healthy humans or animals, and sewage) were studied for their susceptibility to trimethoprim as determined by agar dilution (European Committee on Antimicrobial Susceptibility Testing) and the presence of (a) the dfrK gene and its genetic environment and (b) other dfr genes. The dfrK gene was detected in 49% of the enterococci (64% and 42% of isolates with minimum inhibitory concentrations of ≥2 mg/L or ≤1 mg/L, respectively). The tet(L)-dfrK linkage was detected in 21% of dfrK-positive enterococci. The chromosomal location of the dfrK gene was identified in one E. faecium isolate in which the dfrK was not linked to tet(L) gene but was part of a Tn559 element, which was integrated in the chromosomal radC gene. This Tn559 element was also found in 14 additional isolates. All combinations of dfr genes were detected among the isolates tested (dfrK, dfrG, dfrF, dfrK+dfrG, dfrK+dfrF, dfrF+dfrG, and dfrF+dfrG+dfrK). The gene dfrK gene was found together with other dfr genes in 58% of the tested enterococci. This study suggested an exchange of the trimethoprim resistance gene dfrK between enterococci and staphylococci, as previously observed for the trimethoprim resistance gene dfrG.

Optimization of high-rate TN removal in a novel constructed wetland integrated with microelectrolysis system treating high-strength digestate supernatant.

Science.gov (United States)

Guo, Luchen; He, Keli; Wu, Shubiao; Sun, Hao; Wang, Yanfei; Huang, Xu; Dong, Renjie

2016-08-01

The potential of high-rate TN removal in three aerated horizontal subsurface-flow constructed wetlands to treat high-strength anaerobic digestate supernatant was evaluated. Different strategies of intermittent aeration and effluent recirculation were applied to compare their effect on nitrogen depuration performance. Additional glucose supply and iron-activated carbon based post-treatment systems were established and examined, respectively, to further remove nitrate that accumulated in the effluents from aerated wetlands. The results showed that intermittent aeration (1 h on:1 h off) significantly improved nitrification with ammonium removal efficiency of 90% (18.1 g/(m(2)·d)), but limited TN removal efficiency (53%). Even though effluent recirculation (a ratio of 1:1) increased TN removal from 53% to 71%, the effluent nitrate concentration was still high. Additional glucose was used as a post-treatment option and further increased the TN removal to 82%; however, this implementation caused additional organic pollution. Furthermore, the iron-activated carbon system stimulated with a microelectrolysis process achieved greater than 85% effluent nitrate removal and resulted in 86% TN removal. Considering the high TN removal rate, aerated constructed wetlands integrated with a microelectrolysis-driven system show great potential for treating high-strength digestate supernatant. Copyright © 2016 Elsevier Ltd. All rights reserved.

Genomic analysis of 495 vancomycin-resistant Enterococcus faecium reveals broad dissemination of a vanA plasmid in more than 19 clones from Copenhagen, Denmark

DEFF Research Database (Denmark)

Pinholt, Mette; Gumpert, Heidi; Bayliss, Sion

2017-01-01

. The majority of the isolates were located in three groups (n = 44, 100 and 218) and clonal spread of VREfm between wards and hospitals was identified. Five Tn1546-like transposon types were identified. A dominant truncated transposon (type 4, 92%) was spread across all but one VREfm group. The closed van......A plasmid was highly covered by reads from isolates containing the type 4 transposon. CONCLUSIONS: This study suggests that it was the dissemination of the type 4 Tn1546-like transposon and plasmid via horizontal transfer to multiple populations of E. faecium, followed by clonal spread of new VREfm clones...

Superior Predictive Value for NTproBNP Compared with High Sensitivity cTnT in Dialysis Patients: A Pilot Prospective Observational Study

Directory of Open Access Journals (Sweden)

Luminita Voroneanu

2014-12-01

Full Text Available Background/Aims: The clinical utility of the new biomarker, high sensitivity cardiac T troponin (hs-cTnT is still unclear in dialysis patients. Furthermore, the prognostic value of combining N-terminal pro-B-type natriuretic peptide (NT-pro-BNP and hs-cTnT has not been explored so far. The objective of this pilot study was to determine the utility of hs-cTnT alone versus hs-cTnT in combination with NT-proBNP for predicting death in a stable hemodialysis cohort. Methods: A prospective observational pilot study including 98 chronic asymptomatic hemodialysis patients with a follow up period of 24 months was designed. The cut-off values for NT-proBNP and hs-cTnT were calculated using receiver operating characteristic (ROC analysis, using mortality as an end-point. Based on the cut-off values, the cohort was divided into four groups. Group 1 - NT-proBNP 69.48 ng/l; group 3 - NT-proBNP > 14275 pg/ml and hs-cTnT 14275 pg/ml and hs-cTnT > 69.48 ng/l. Survival for each group was determined using the Kaplan-Meier method and Cox regression analysis. Results: During the follow-up period 16 patients died. According to the ROC curves analysis, the cut-off point for hs-cTnT and for NT-proBNP were 69.43 ng/l (AUC = 0.618; p = 0.04 and 14275 pg/ml (AUC = 0.722; p = 0.003, respectively. In univariate Cox analysis, both hs-cTnT (HR = 3.34; p = 0.016 and NT-proBNP (HR = 5.94; p = 0.01 were predictors of death. In the multivariable Cox proportional hazards model, only NT-pro-BNP levels above the cut-off value remained an independent predictor of all-cause mortality. The combined elevation of both biomarkers did not improve significantly the prognostic value compared with NT-proBNP alone (HR = 6.15 versus HR =4 .78; p = 0.338. Conclusion: NT-pro-BNP is a strong predictor of overall mortality in asymptomatic hemodialysis patients. The addition of hs-cTnT did not improve the prognostic accuracy compared with NT proBNP alone.

High-Level Heat Resistance of Spores of Bacillus amyloliquefaciens and Bacillus licheniformis Results from the Presence of a spoVA Operon in a Tn1546 Transposon

NARCIS (Netherlands)

Berendsen, Erwin M; Koning, Rosella A; Boekhorst, Jos; de Jong, Anne; Kuipers, Oscar P; Wells-Bennik, Marjon H J

2016-01-01

Bacterial endospore formers can produce spores that are resistant to many food processing conditions, including heat. Some spores may survive heating processes aimed at production of commercially sterile foods. Recently, it was shown that a spoVA operon, designated spoVA(2mob), present on a Tn1546

Clinical application of combined detection of serum hs-CRP, GMP-140 and cTnI in patients with coronary heart diseases

International Nuclear Information System (INIS)

Qin Jibao; Wu Zhaozeng

2010-01-01

Objective: To explore the clinical significance of changes of serum hs-CRP, GMP-140 and cTnI levels in patients with coronary heart diseases. Methods: Serum GMP-140 (with RIA), cTnI (with ELISA) and hs-CRP (with immuno turbidity method) levels were determined in 91 patients with coronary heart diseases (42 SAP, 34UAP, 15AMI) and 35 controls. Results: Serum hs-CRP, GMP-140, cTnI levels in patients with coronary heart diseases were significantly higher than those in controls (P <0.01). Among the patients with of coronary heart diseases, the magnitude of changes of the levels of serum hs-CRP, GMP-140 and cTnI levels in AMI and UAP groups were significantly larger than those in SAP group (P < 0.05). Serum hs-CRP levels were positively correlated with serum GMP-140 and cTnI levels (r = 0.6214, 0.6023, P < 0.01). Conclusion: Serum hs-CRP, GMP140 and cTnI levels were closely related to the diseases process of coronary heart diseases and were of great clinical importance for assessment of the disease and outcome prediction. (authors)

Interaction of a novel Tn (GalNAc alpha 1-->Ser/Thr) glycoprotein with Gal, GalNAc and GlcNAc specific lectins.

Science.gov (United States)

Wu, A M; Wu, J H; Shen, F

1994-01-14

A naturally occurring Tn glycoprotein (Native ASG-Tn) with GalNAc alpha 1-->Ser/Thr as the only carbohydrate side chains, has been prepared from armadillo submandibular glands. In a quantitative precipitin assay, this glycoprotein completely precipitated Maclura pomifera (MPA), Vicia villosa B4 (VVL-B4) and Artocarpus integrifolia (Jacalin, AIL). It also reacted well with Helix pomatia (HPL) and Wistaria floribunda (WFL) and precipitated over 75% of the lectin nitrogen added, but poorly with Ricinus communis agglutinin (RCA1), ricin, peanut (Arachis hypogaea, PNA), Abrus precatorius agglutinin (APA) and Triticum vulgaris (WGA). This finding suggests that this novel Tn-glycoprotein may serve as a useful reagent for differentiating Tn and T specific monoclonal antibodies and lectins.

Essential Genes for In Vitro Growth of the Endophyte Herbaspirillum seropedicae SmR1 as Revealed by Transposon Insertion Site Sequencing.

Science.gov (United States)

Rosconi, Federico; de Vries, Stefan P W; Baig, Abiyad; Fabiano, Elena; Grant, Andrew J

2016-11-15

The interior of plants contains microorganisms (referred to as endophytes) that are distinct from those present at the root surface or in the surrounding soil. Herbaspirillum seropedicae strain SmR1, belonging to the betaproteobacteria, is an endophyte that colonizes crops, including rice, maize, sugarcane, and sorghum. Different approaches have revealed genes and pathways regulated during the interactions of H. seropedicae with its plant hosts. However, functional genomic analysis of transposon (Tn) mutants has been hampered by the lack of genetic tools. Here we successfully employed a combination of in vivo high-density mariner Tn mutagenesis and targeted Tn insertion site sequencing (Tn-seq) in H. seropedicae SmR1. The analysis of multiple gene-saturating Tn libraries revealed that 395 genes are essential for the growth of H. seropedicae SmR1 in tryptone-yeast extract medium. A comparative analysis with the Database of Essential Genes (DEG) showed that 25 genes are uniquely essential in H. seropedicae SmR1. The Tn mutagenesis protocol developed and the gene-saturating Tn libraries generated will facilitate elucidation of the genetic mechanisms of the H. seropedicae endophytic lifestyle. A focal point in the study of endophytes is the development of effective biofertilizers that could help to reduce the input of agrochemicals in croplands. Besides the ability to promote plant growth, a good biofertilizer should be successful in colonizing its host and competing against the native microbiota. By using a systematic Tn-based gene-inactivation strategy and massively parallel sequencing of Tn insertion sites (Tn-seq), it is possible to study the fitness of thousands of Tn mutants in a single experiment. We have applied the combination of these techniques to the plant-growth-promoting endophyte Herbaspirillum seropedicae SmR1. The Tn mutant libraries generated will enable studies into the genetic mechanisms of H. seropedicae-plant interactions. The approach that we

Dynamic Equilibrium of Cardiac Troponin C's Hydrophobic Cleft and Its Modulation by Ca2+ Sensitizers and a Ca2+ Sensitivity Blunting Phosphomimic, cTnT(T204E).

Science.gov (United States)

Schlecht, William; Dong, Wen-Ji

2017-10-18

Several studies have suggested that conformational dynamics are important in the regulation of thin filament activation in cardiac troponin C (cTnC); however, little direct evidence has been offered to support these claims. In this study, a dye homodimerization approach is developed and implemented that allows the determination of the dynamic equilibrium between open and closed conformations in cTnC's hydrophobic cleft. Modulation of this equilibrium by Ca 2+ , cardiac troponin I (cTnI), cardiac troponin T (cTnT), Ca 2+ -sensitizers, and a Ca 2+ -desensitizing phosphomimic of cTnT (cTnT(T204E) is characterized. Isolated cTnC contained a small open conformation population in the absence of Ca 2+ that increased significantly upon the addition of saturating levels of Ca 2+ . This suggests that the Ca 2+ -induced activation of thin filament arises from an increase in the probability of hydrophobic cleft opening. The inclusion of cTnI increased the population of open cTnC, and the inclusion of cTnT had the opposite effect. Samples containing Ca 2+ -desensitizing cTnT(T204E) showed a slight but insignificant decrease in open conformation probability compared to samples with cardiac troponin T, wild type [cTnT(wt)], while Ca 2+ sensitizer treated samples generally increased open conformation probability. These findings show that an equilibrium between the open and closed conformations of cTnC's hydrophobic cleft play a significant role in tuning the Ca 2+ sensitivity of the heart.

EnviroAtlas - Memphis, TN - Meter-Scale Urban Land Cover (MULC) Data (2012)

Data.gov (United States)

U.S. Environmental Protection Agency — The Memphis, TN EnviroAtlas One Meter-scale Urban Land Cover (MULC) dataset comprises 2,733 km2 around the city of Memphis, surrounding towns, and rural areas. These...

Proteus genomic island 1 (PGI1), a new resistance genomic island from two Proteus mirabilis French clinical isolates.

Science.gov (United States)

Siebor, Eliane; Neuwirth, Catherine

2014-12-01

To analyse the genetic environment of the antibiotic resistance genes in two clinical Proteus mirabilis isolates resistant to multiple antibiotics. PCR, gene walking and whole-genome sequencing were used to determine the sequence of the resistance regions, the surrounding genetic structure and the flanking chromosomal regions. A genomic island of 81.1 kb named Proteus genomic island 1 (PGI1) located at the 3'-end of trmE (formerly known as thdF) was characterized. The large MDR region of PGI1 (55.4 kb) included a class 1 integron (aadB and aadA2) and regions deriving from several transposons: Tn2 (blaTEM-135), Tn21, Tn6020-like transposon (aphA1b), a hybrid Tn502/Tn5053 transposon, Tn501, a hybrid Tn1696/Tn1721 transposon [tetA(A)] carrying a class 1 integron (aadA1) and Tn5393 (strA and strB). Several ISs were also present (IS4321, IS1R and IS26). The PGI1 backbone (25.7 kb) was identical to that identified in Salmonella Heidelberg SL476 and shared some identity with the Salmonella genomic island 1 (SGI1) backbone. An IS26-mediated recombination event caused the division of the MDR region into two parts separated by a large chromosomal DNA fragment of 197 kb, the right end of PGI1 and this chromosomal sequence being in inverse orientation. PGI1 is a new resistance genomic island from P. mirabilis belonging to the same island family as SGI1. The role of PGI1 in the spread of antimicrobial resistance genes among Enterobacteriaceae of medical importance needs to be evaluated. © The Author 2014. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Clinical relevance of determination the changes of serum cTnI, NPY and PGMP contents after treatment in patients with coronary heart diseases

International Nuclear Information System (INIS)

Ge Hengsong; Ma Chao; Wu Kaixia

2011-01-01

Objective: To explore the clinical significance of changes of serum cTnI, NPY and PGMP levels in patients with coronary heart diseases after treatment. Methods: Serum NPY, PGMP (with RIA), serum cTnI (with Biochemistry) levels were measured in 41 patients with coronary heart diseases both before and after treatment as well as in 35 normal controls. Results: Before treatment, serum cTnI, NPY and PGMP levels were significantly higher than those in controls (P 0.05), but serum PGMP level remained prominently higher (P<0.05). Conclusion: Pathogenesis and progress of CHD, might be closely related to serum cTnI, NPY and PGMP levels. (authors)

Genome-wide transposon mutagenesis of Proteus mirabilis: Essential genes, fitness factors for catheter-associated urinary tract infection, and the impact of polymicrobial infection on fitness requirements

Science.gov (United States)

Smith, Sara N.; Zhao, Lili; Wu, Weisheng

2017-01-01

The Gram-negative bacterium Proteus mirabilis is a leading cause of catheter-associated urinary tract infections (CAUTIs), which are often polymicrobial. Numerous prior studies have uncovered virulence factors for P. mirabilis pathogenicity in a murine model of ascending UTI, but little is known concerning pathogenesis during CAUTI or polymicrobial infection. In this study, we utilized five pools of 10,000 transposon mutants each and transposon insertion-site sequencing (Tn-Seq) to identify the full arsenal of P. mirabilis HI4320 fitness factors for single-species versus polymicrobial CAUTI with Providencia stuartii BE2467. 436 genes in the input pools lacked transposon insertions and were therefore concluded to be essential for P. mirabilis growth in rich medium. 629 genes were identified as P. mirabilis fitness factors during single-species CAUTI. Tn-Seq from coinfection with P. stuartii revealed 217/629 (35%) of the same genes as identified by single-species Tn-Seq, and 1353 additional factors that specifically contribute to colonization during coinfection. Mutants were constructed in eight genes of interest to validate the initial screen: 7/8 (88%) mutants exhibited the expected phenotypes for single-species CAUTI, and 3/3 (100%) validated the expected phenotypes for polymicrobial CAUTI. This approach provided validation of numerous previously described P. mirabilis fitness determinants from an ascending model of UTI, the discovery of novel fitness determinants specifically for CAUTI, and a stringent assessment of how polymicrobial infection influences fitness requirements. For instance, we describe a requirement for branched-chain amino acid biosynthesis by P. mirabilis during coinfection due to high-affinity import of leucine by P. stuartii. Further investigation of genes and pathways that provide a competitive advantage during both single-species and polymicrobial CAUTI will likely provide robust targets for therapeutic intervention to reduce P. mirabilis

Genome-wide transposon mutagenesis of Proteus mirabilis: Essential genes, fitness factors for catheter-associated urinary tract infection, and the impact of polymicrobial infection on fitness requirements.

Science.gov (United States)

Armbruster, Chelsie E; Forsyth-DeOrnellas, Valerie; Johnson, Alexandra O; Smith, Sara N; Zhao, Lili; Wu, Weisheng; Mobley, Harry L T

2017-06-01

The Gram-negative bacterium Proteus mirabilis is a leading cause of catheter-associated urinary tract infections (CAUTIs), which are often polymicrobial. Numerous prior studies have uncovered virulence factors for P. mirabilis pathogenicity in a murine model of ascending UTI, but little is known concerning pathogenesis during CAUTI or polymicrobial infection. In this study, we utilized five pools of 10,000 transposon mutants each and transposon insertion-site sequencing (Tn-Seq) to identify the full arsenal of P. mirabilis HI4320 fitness factors for single-species versus polymicrobial CAUTI with Providencia stuartii BE2467. 436 genes in the input pools lacked transposon insertions and were therefore concluded to be essential for P. mirabilis growth in rich medium. 629 genes were identified as P. mirabilis fitness factors during single-species CAUTI. Tn-Seq from coinfection with P. stuartii revealed 217/629 (35%) of the same genes as identified by single-species Tn-Seq, and 1353 additional factors that specifically contribute to colonization during coinfection. Mutants were constructed in eight genes of interest to validate the initial screen: 7/8 (88%) mutants exhibited the expected phenotypes for single-species CAUTI, and 3/3 (100%) validated the expected phenotypes for polymicrobial CAUTI. This approach provided validation of numerous previously described P. mirabilis fitness determinants from an ascending model of UTI, the discovery of novel fitness determinants specifically for CAUTI, and a stringent assessment of how polymicrobial infection influences fitness requirements. For instance, we describe a requirement for branched-chain amino acid biosynthesis by P. mirabilis during coinfection due to high-affinity import of leucine by P. stuartii. Further investigation of genes and pathways that provide a competitive advantage during both single-species and polymicrobial CAUTI will likely provide robust targets for therapeutic intervention to reduce P. mirabilis

In-house preparation of lectin panel and detection of Tn polyagglutination

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Sudipta Sekhar Das

2015-01-01

Full Text Available Polyagglutination is a condition in which red cells are agglutinated by ABO-compatible adult human sera, but not by cord blood sera and may be acquired or inherited. Lectins are invaluable reagents in the investigation of red cells polyagglutination. We prepared in-house lectin panel and confirmed Tn polyagglutination in a pregnant lady. The lady was anemic and refused blood transfusion elsewhere due to serological discrepancy. We found ABO discrepancy and an incompatible minor cross-match in the initial investigation and suspected polyagglutination. Confirmation of polyagglutination was done using adult and cord sera. We then used the in-house lectin panels to detect the type of polyagglutination. The agglutination pattern with the various lectins was suggestive of Tn polyagglutination, which was further supported by the enzyme study. Most blood banks in India lack commercial lectin panels because of cost and procurement difficulty. Lectins play an important role in the diagnosis and differentiation of polyagglutination and immunohematological management of patient. The important and basic lectins can be prepared in-house using specific raw seeds following standardized protocol.

In-house preparation of lectin panel and detection of Tn polyagglutination.

Science.gov (United States)

Das, Sudipta Sekhar

2015-01-01

Polyagglutination is a condition in which red cells are agglutinated by ABO-compatible adult human sera, but not by cord blood sera and may be acquired or inherited. Lectins are invaluable reagents in the investigation of red cells polyagglutination. We prepared in-house lectin panel and confirmed Tn polyagglutination in a pregnant lady. The lady was anemic and refused blood transfusion elsewhere due to serological discrepancy. We found ABO discrepancy and an incompatible minor cross-match in the initial investigation and suspected polyagglutination. Confirmation of polyagglutination was done using adult and cord sera. We then used the in-house lectin panels to detect the type of polyagglutination. The agglutination pattern with the various lectins was suggestive of Tn polyagglutination, which was further supported by the enzyme study. Most blood banks in India lack commercial lectin panels because of cost and procurement difficulty. Lectins play an important role in the diagnosis and differentiation of polyagglutination and immunohematological management of patient. The important and basic lectins can be prepared in-house using specific raw seeds following standardized protocol.

Performances of TN {sup registered} 24 E. An AREVA used fuel transport and interim storage cask for the German market

Energy Technology Data Exchange (ETDEWEB)

Brion, Thomas [AREVA TN International, Montigny Le Bretonneux (France)

2013-07-01

Part of the AREVA Group, TN International offers a complete range of transport and interim storage solutions for radioactive materials throughout the entire nuclear fuel cycle. A world leader in its sector, TN International has supported for 50 years the expansion of the nuclear industry, in particular by providing expertise in secure packing systems for the storage of used fuel assemblies. As an answer to EON and EnBW, two German utilities, needs, TN International has designed and manufactured the TN {sup registered} 24E cask, offering the following high level performances: 1. transport and storage over a period of 40 years of up to 21 PWR spent nuclear fuel (SNF), allowing for example to load up to 17 MOX fuel assemblies and 4 UOX SNF. 2. high flexibility in the fuel assemblies loading plans, inducing no general predefined constraints with regards to the MOX or UOX fuel positions in the basket of the cask Safety margin related to radioprotection, thermal and mechanical behaviour of the fuel assemblies can be calculated loading plan per loading plan. (orig.)

Detección del antígeno Tn en tumores epiteliales con la lectina de Vicia villosa isolectina B4.

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Catalina Limpias

2010-10-01

Full Text Available Antecedentes. Los epítopes T, Tn y sTn, se expresan en un alto porcentaje de tumores epiteliales y pueden detectarse con anticuerpos monoclonales y lectinas. Objetivo. Evaluar diferencias de expresión del antígeno Tn en cortes histológicos de epitelios no neoplásicos y tumores epiteliales mediante isolectina B4 de Vicia villosa. Material y métodos. Se evaluaron semicuantitativamente localización, intensidad y porcentaje de expresión del antígeno en carcinomas in-situ e infiltrantes y epitelios no neoplásicos de cérvix, seno y urotelio, mediante isolectina B4. Resultados La expresión de Tn en cérvix predominó en membrana de células no neoplásicas y citoplasma de células tumorales; su intensidad fue mayor en carcinomas in-situ e infiltrantes comparado con epitelio no neoplásico aunque en este el porcentaje de expresión fue mayor. En seno, la expresión de Tn fue predominantemente citoplasmática con intensidad similar, el porcentaje de expresión fué mayor en carcinomas ductales in-situ e infiltrantes. En urotelio no neoplásico y tumoral la expresión de Tn predominó en citoplasma; la intensidad y el porcentaje de expresión fueron mayores en neoplasias no invasivas de bajo y alto grado, mientras que en urotelio no neoplásico fue baja y no hubo tendencia definida en tumores infiltrantes. Conclusiones. La detección del antígeno Tn mediante la lectina VVB4 mostró una mayor extensión de marcación en carcinomas ductales de seno en relación con el epitelio no neoplásico, pero no mostró una tendencia definida entre el tejido normal, ni diferentes etapas del desarrollo de los tumores de cérvix y urotelio. Estos hallazgos pueden atribuirse a la heterogeneidad de los procesos carcinogénicos o a que la especificidad de la lectina VVB4 no está restringida a este antígeno.

75 FR 69398 - Foreign-Trade Zone 78-Nashville, TN; Application for Expansion

Science.gov (United States)

2010-11-12

... Company, 1800 Zinc Plant Road, Clarksville; and Proposed Site 12 (451.0 acres)--Gallatin Industrial Center..., TN; Application for Expansion An application has been submitted to the Foreign-Trade Zones Board (the... application was submitted pursuant to the provisions of the Foreign- Trade Zones Act, as amended (19 U.S.C...

Integrative Gene Cloning and Expression System for Streptomyces sp. US 24 and Streptomyces sp. TN 58 Bioactive Molecule Producing Strains

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Samiha Sioud

2009-01-01

Full Text Available Streptomyces sp. US 24 and Streptomyces sp. TN 58, two strains producing interesting bioactive molecules, were successfully transformed using E. coli ET12567 (pUZ8002, as a conjugal donor, carrying the integrative plasmid pSET152. For the Streptomyces sp. US 24 strain, two copies of this plasmid were tandemly integrated in the chromosome, whereas for Streptomyces sp. TN 58, the integration was in single copy at the attB site. Plasmid pSET152 was inherited every time for all analysed Streptomyces sp. US 24 and Streptomyces sp. TN 58 exconjugants under nonselective conditions. The growth, morphological differentiation, and active molecules production of all studied pSET152 integrated exconjugants were identical to those of wild type strains. Consequently, conjugal transfer using pSET152 integration system is a suitable means of genes transfer and expression for both studied strains. To validate the above gene transfer system, the glucose isomerase gene (xylA from Streptomyces sp. SK was expressed in strain Streptomyces sp. TN 58. Obtained results indicated that heterologous glucose isomerase could be expressed and folded effectively. Glucose isomerase activity of the constructed TN 58 recombinant strain is of about eighteenfold higher than that of the Streptomyces sp. SK strain. Such results are certainly of importance due to the potential use of improved strains in biotechnological process for the production of high-fructose syrup from starch.

A comprehensive analysis of in vitro and in vivo genetic fitness of Pseudomonas aeruginosa using high-throughput sequencing of transposon libraries.

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David Skurnik

Full Text Available High-throughput sequencing of transposon (Tn libraries created within entire genomes identifies and quantifies the contribution of individual genes and operons to the fitness of organisms in different environments. We used insertion-sequencing (INSeq to analyze the contribution to fitness of all non-essential genes in the chromosome of Pseudomonas aeruginosa strain PA14 based on a library of ∼300,000 individual Tn insertions. In vitro growth in LB provided a baseline for comparison with the survival of the Tn insertion strains following 6 days of colonization of the murine gastrointestinal tract as well as a comparison with Tn-inserts subsequently able to systemically disseminate to the spleen following induction of neutropenia. Sequencing was performed following DNA extraction from the recovered bacteria, digestion with the MmeI restriction enzyme that hydrolyzes DNA 16 bp away from the end of the Tn insert, and fractionation into oligonucleotides of 1,200-1,500 bp that were prepared for high-throughput sequencing. Changes in frequency of Tn inserts into the P. aeruginosa genome were used to quantify in vivo fitness resulting from loss of a gene. 636 genes had <10 sequencing reads in LB, thus defined as unable to grow in this medium. During in vivo infection there were major losses of strains with Tn inserts in almost all known virulence factors, as well as respiration, energy utilization, ion pumps, nutritional genes and prophages. Many new candidates for virulence factors were also identified. There were consistent changes in the recovery of Tn inserts in genes within most operons and Tn insertions into some genes enhanced in vivo fitness. Strikingly, 90% of the non-essential genes were required for in vivo survival following systemic dissemination during neutropenia. These experiments resulted in the identification of the P. aeruginosa strain PA14 genes necessary for optimal survival in the mucosal and systemic environments of a mammalian

Glycan elongation beyond the mucin associated Tn antigen protects tumor cells from immune-mediated killing

DEFF Research Database (Denmark)

Madsen, Caroline B; Lavrsen, Kirstine; Steentoft, Catharina

2013-01-01

are recognized as cancer associated truncated glycans, and are expressed in many adenocarcinomas, e.g. breast- and pancreatic cancer cells. To investigate the role of the cancer associated glycan truncations in immune-mediated killing we created glyco-engineered breast- and pancreatic cancer cells expressing...... only the shortest possible mucin-like glycans (Tn and STn). Glyco-engineering was performed by zinc finger nuclease (ZFN) knockout (KO) of the Core 1 enzyme chaperone COSMC, thereby preventing glycan elongation beyond the initial GalNAc residue in O-linked glycans. We find that COSMC KO in the breast...

Design analysis report for the TN-WHC cask and transportation system

Energy Technology Data Exchange (ETDEWEB)

Brisbin, S.A., Fluor Daniel Hanford

1997-02-13

This document presents the evaluation of the Spent Nuclear Fuel Cask and Transportation System. The system design was developed by Transnuclear, Inc. and its team members NAC International, Nelson Manufacturing, Precision Components Corporation, and Numatec, Inc. The cask is designated the TN-WHC cask. This report describes the design features and presents preliminary analyses performed to size critical dimensions of the system while meeting the requirements of the performance specification.

Glycan elongation beyond the mucin associated Tn antigen protects tumor cells from immune-mediated killing.

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Caroline B Madsen

Full Text Available Membrane bound mucins are up-regulated and aberrantly glycosylated during malignant transformation in many cancer cells. This results in a negatively charged glycoprotein coat which may protect cancer cells from immune surveillance. However, only limited data have so far demonstrated the critical steps in glycan elongation that make aberrantly glycosylated mucins affect the interaction between cancer cells and cytotoxic effector cells of the immune system. Tn (GalNAc-Ser/Thr, STn (NeuAcα2-6GalNAc-Ser/Thr, T (Galβ1-3GalNAc-Ser/Thr, and ST (NeuAcα2-6Galβ1-3GalNAc-Ser/Thr antigens are recognized as cancer associated truncated glycans, and are expressed in many adenocarcinomas, e.g. breast- and pancreatic cancer cells. To investigate the role of the cancer associated glycan truncations in immune-mediated killing we created glyco-engineered breast- and pancreatic cancer cells expressing only the shortest possible mucin-like glycans (Tn and STn. Glyco-engineering was performed by zinc finger nuclease (ZFN knockout (KO of the Core 1 enzyme chaperone COSMC, thereby preventing glycan elongation beyond the initial GalNAc residue in O-linked glycans. We find that COSMC KO in the breast and pancreatic cancer cell lines T47D and Capan-1 increases sensitivity to both NK cell mediated antibody-dependent cellular-cytotoxicity (ADCC and cytotoxic T lymphocyte (CTL-mediated killing. In addition, we investigated the association between total cell surface expression of MUC1/MUC16 and NK or CTL mediated killing, and observed an inverse correlation between MUC16/MUC1 expression and the sensitivity to ADCC and CTL-mediated killing. Together, these data suggest that up-regulation of membrane bound mucins protects cells from immune mediated killing, and that particular glycosylation steps, as demonstrated for glycan elongation beyond Tn and STn, can be important for fine tuning of the immune escape mechanisms in cancer cells.

Enhanced nitrogen removal from piggery wastewater with high NH4+ and low COD/TN ratio in a novel upflow microaerobic biofilm reactor.

Science.gov (United States)

Meng, Jia; Li, Jiuling; Li, Jianzheng; Antwi, Philip; Deng, Kaiwen; Nan, Jun; Xu, Pianpian

2018-02-01

To enhance nutrient removal more cost-efficiently in microaerobic process treating piggery wastewater characterized by high ammonium (NH 4 + -N) and low chemical oxygen demand (COD) to total nitrogen (TN) ratio, a novel upflow microaerobic biofilm reactor (UMBR) was constructed and the efficiency in nutrient removal was evaluated with various influent COD/TN ratios and reflux ratios. The results showed that the biofilm on the carriers had increased the biomass in the UMBR and enhanced the enrichment of slow-growth-rate bacteria such as nitrifiers, denitrifiers and anammox bacteria. The packed bed allowed the microaerobic biofilm process perform well at a low reflux ratio of 35 with a NH 4 + -N and TN removal as high as 93.1% and 89.9%, respectively. Compared with the previously developed upflow microaerobic sludge reactor, the UMBR had not changed the dominant anammox approach to nitrogen removal, but was more cost-efficiently in treating organic wastewater with high NH 4 + -N and low COD/TN ratio. Copyright © 2017 Elsevier Ltd. All rights reserved.

ORF Alignment: NC_003384 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available [Zymomonas mobilis] gb|AAQ67239.1| ... Bla [Transposon delivery vector pUT-miniTn5-gfp-tet] ... ... ... 3.5.2.6) precursor - Escherichia coli plasmids ... gb|AAC79082.1| beta lactamase [Transposon delivery...AG47772.1| beta-lactamase ... TEM-1 [Klebsiella pneumoniae] gb|AAG37887.1| ampicillin ... resistance [Tn10 delivery... vector pHV1249] gb|AAG37882.1| ... ampicillin resistance [Tn10 delivery

Dynamic changes of serum TNF-α and cTnI levels in BALB/c Mouse models of autoimmune myocarditis

International Nuclear Information System (INIS)

Han Li'na

2007-01-01

Objective: To prepare mouse models of experimental autoimmune myocarditis (EAM) with subcutaneous injection of synthesized peptide (containing heavy chain segment of cardiac myosin) and examine the dynamic changes of cardiac pathology as well as TNF -α, cTnI and autoimmune antibody contents. Methods: BALB/c mice were subcutaneously injected with 0.8 mg of the synthesized peptide to produce the experimental autoimmune myocarditis (EAM) models. The animals were sacrificed 14, 21, 30 and 60d after establishment of the models, heart muscle was examined pathologically to assess the severity of myocarditis and blood were examined for the plasma contents of TNF-α (with RIA) cTnI (with ELISA) and anti-cardiac myosin heavy chain (CMHC) antibody titier (with ELISA). Results: Evidence of myocardial damage appeared on d14 specimens and progressed there after with interstitial infiltration of macrophages and myocardial necrosis. In the 60d specimens, fibrosis was obvious. The plasma TNF-α and cTnI levels were highest in the d30 specimens and declined in the d60 specimens but still remained much higher than normal. Most of the models demonstrated anti-CMHC antibodies in the 21d and 30d specimens. Conclusion: It is feasible to prepare EMC mouse models with subcutaneous injection of synthesized peptide containing heavy chain segment of cardiac myosin and the plasma levels of TNF-α, cTnI in the models are well correlated to the pathologic change of myocarditis. (authors)

Association of Elevated High Sensitivity Cardiac Troponin T(hs-cTnT Levels with Hemorrhagic Transformation and 3-Month Mortality in Acute Ischemic Stroke Patients with Rheumatic Heart Disease in China.

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Junfeng Liu

Full Text Available Elevated levels of high sensitivity cardiac troponin T (hs-cTnT occur in a substantial proportion of patients with acute ischemic stroke (AIS and can predict poor outcome and mortality after stroke. Whether elevated hs-cTnT levels can also predict hemorrhagic transformation (HT or prognosis in AIS patients with rheumatic heart disease (RHD remains unclear.Data from the Chengdu Stroke Registry on consecutive AIS patients with RHD admitted to West China Hospital within 1 month of stroke onset from October 2011 to February 2014 were examined. Clinico-demographic characteristics, HT, functional outcomes and stroke recurrence were compared between patients with elevated hs-cTnT levels (≥14 ng/L and patients with normal hs-cTnT levels (<14 ng/L.The final analysis involved 84 patients (31 males; mean age, 61.6±12.2 years, of whom serum hs-cTnT levels were elevated in 58.3%. Renal impairment was independently associated with elevated hs-cTnT levels (OR 4.184, 95%CI 1.17 to 15.01, P = 0.028, and patients with elevated hs-cTnT levels were at significantly higher risk of HT, 3-month mortality and 3-month disability/mortality (all P≤0.029. After controlling for age, sex, hypertension, renal impairment and National Institutes of Health Stroke Scale score on admission, the risk of HT and 3-month mortality was, respectively, 4.0- and 5.5-fold higher in patients with elevated hs-cTnT levels than in patients with normal hs-cTnT levels.Elevated hs-cTnT levels are independently associated with HT and 3-month mortality in AIS patients with RHD. These results with a small cohort should be verified and extended in large studies.

Literary Depictions i n Ghazavâtnâmahs That Adress the Crimean War

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Kürşat Şamil ŞAHİN

2015-07-01

Full Text Available The Crimean War that started in 1853 between the Ottoman Empire and Russia lasted until 1856. It ended with the defeat of the Russians as England and France sided with the Ottoman Empire. A great number of work has been written then and since about this war which deeply affect our social and politica l life. Among these, there are ghazavâtnâmahs that describe what happened down - to - line, usually by the pen of the poets and writers who closely witnessed the war. The causes of war, the preparations, what happened at the time of expedition and measures tha t were taken, the outcome of the events during the war and afterwards are all brought into sharp relief in most of these works. Whether in verse, prose or mixed typed, these works of art have gradually increased after 15 th century in Turkish literature. Th is genre has decreased by the Ottoman Empire began to decline and the raids were scarce; and it totally disappeared after the tradition of ghaza were ceased. In this study the literary depictions in - the last examples of the genre - Salih Hayri’s Kırım Zafe rnamesi (Hayrâbât, Ahmed Rızâ Trabzonî’s Manzume - i Sivastopol and Süleyman Şâdî’s Muzaffernâme are presented. There are not many studies that focus on literary depictions in ghazavâtnâmahs, particularly on literary war depictions. The characteristics of t hese literary depictions are tried to be explained with reference to ghazavâtnâmahs belonging to the Empire's last era.

The L‐type Ca2+ channel facilitates abnormal metabolic activity in the cTnI‐G203S mouse model of hypertrophic cardiomyopathy

Science.gov (United States)

Viola, Helena; Johnstone, Victoria; Cserne Szappanos, Henrietta; Richman, Tara; Tsoutsman, Tatiana; Filipovska, Aleksandra; Semsarian, Christopher

2016-01-01

Key points Genetic mutations in cardiac troponin I (cTnI) are associated with development of hypertrophic cardiomyopathy characterized by myocyte remodelling, disorganization of cytoskeletal proteins and altered energy metabolism.The L‐type Ca2+ channel is the main route for calcium influx and is crucial to cardiac excitation and contraction. The channel also regulates mitochondrial function in the heart by a functional communication between the channel and mitochondria via the cytoskeletal network.We find that L‐type Ca2+ channel kinetics are altered in cTnI‐G203S cardiac myocytes and that activation of the channel causes a significantly greater increase in mitochondrial membrane potential and metabolic activity in cTnI‐G203S cardiac myocytes.These responses occur as a result of impaired communication between the L‐type Ca2+ channel and cytoskeletal protein F‐actin, involving decreased movement of actin–myosin and block of the mitochondrial voltage‐dependent anion channel, resulting in a ‘hypermetabolic’ mitochondrial state.We propose that L‐type Ca2+ channel antagonists, such as diltiazem, might be effective in reducing the cardiomyopathy by normalizing mitochondrial metabolic activity. Abstract Genetic mutations in cardiac troponin I (cTnI) account for 5% of families with hypertrophic cardiomyopathy. Hypertrophic cardiomyopathy is associated with disorganization of cytoskeletal proteins and altered energy metabolism. The L‐type Ca2+ channel (ICa‐L) plays an important role in regulating mitochondrial function. This involves a functional communication between the channel and mitochondria via the cytoskeletal network. We investigate the role of ICa‐L in regulating mitochondrial function in 25‐ to 30‐week‐old cardiomyopathic mice expressing the human disease‐causing mutation Gly203Ser in cTnI (cTnI‐G203S). The inactivation rate of ICa‐L is significantly faster in cTnI‐G203S myocytes [cTnI‐G203S: τ1 = 40.68 ± 3.22, n

Random insertion and gene disruption via transposon mutagenesis of Ureaplasma parvum using a mini-transposon plasmid.

Science.gov (United States)

Aboklaish, Ali F; Dordet-Frisoni, Emilie; Citti, Christine; Toleman, Mark A; Glass, John I; Spiller, O Brad

2014-11-01

While transposon mutagenesis has been successfully used for Mycoplasma spp. to disrupt and determine non-essential genes, previous attempts with Ureaplasma spp. have been unsuccessful. Using a polyethylene glycol-transformation enhancing protocol, we were able to transform three separate serovars of Ureaplasma parvum with a Tn4001-based mini-transposon plasmid containing a gentamicin resistance selection marker. Despite the large degree of homology between Ureaplasma parvum and Ureaplasma urealyticum, all attempts to transform the latter in parallel failed, with the exception of a single clinical U. urealyticum isolate. PCR probing and sequencing were used to confirm transposon insertion into the bacterial genome and identify disrupted genes. Transformation of prototype serovar 3 consistently resulted in transfer only of sequence between the mini-transposon inverted repeats, but some strains showed additional sequence transfer. Transposon insertion occurred randomly in the genome resulting in unique disruption of genes UU047, UU390, UU440, UU450, UU520, UU526, UU582 for single clones from a panel of screened clones. An intergenic insertion between genes UU187 and UU188 was also characterised. Two phenotypic alterations were observed in the mutated strains: Disruption of a DEAD-box RNA helicase (UU582) altered growth kinetics, while the U. urealyticum strain lost resistance to serum attack coincident with disruption of gene UUR10_137 and loss of expression of a 41 kDa protein. Transposon mutagenesis was used successfully to insert single copies of a mini-transposon into the genome and disrupt genes leading to phenotypic changes in Ureaplasma parvum strains. This method can now be used to deliver exogenous genes for expression and determine essential genes for Ureaplasma parvum replication in culture and experimental models. Copyright © 2014 Elsevier GmbH. All rights reserved.

Application value of combined measurement of serum sTn, CA242, CA19-9 and CEA in the diagnosis of gastroenterological neoplasm

International Nuclear Information System (INIS)

Zhang Wanzhong; Chen Zhizhou; Fan Zhenfu

2007-01-01

To determine the application value of four serum tumor markers sTn, CA242, CA 19-9 and CEA in the diagnosis of gastroenterological neoplasm, the serum sTn, CA242, CA19-9 and CEA in 30 normal adult controls and 60 patients with gastroenterological neoplasm were measured by IRMA. The results showed that the serum sTn, CA242, CA19-9 and CEA levels in patients with gastric carcinoma or colorectal carcinoma were much higher than those in control group (P<0.01). The serum CEA, CA19-9 and CA242 levels in patients with colorectal carcinoma were significantly higher than those in patients with gastric carcinoma (P<0.01), but the serum sTn level in the former was markedly lower (P<0.01) than that in the latter. The sensitivity of tumor marker increased with the progress of clinical stages, with a considerably higher sensitivity for stage IV compared with stage I-II (P<0.01). The combined test of four tumor markers could be more sensitive than single test in detecting gastric carcinoma and colorectal carcinoma (P<0.05). Four tumor markers are useful for diagnosing gastroenterological neoplasm, and the combined measurement of 4 tumor markers could increase the sensitivity of detecting gastric carcinoma. (authors)

Insertion of transposon in the vicinity of SSK2 confers enhanced tolerance to furfural in Saccharomyces cerevisiae

Energy Technology Data Exchange (ETDEWEB)

Kim, Hyun-Soo [Ewha Womans Univ., Seoul (Korea, Republic of). Dept. of Life Science; Kim, Na-Rae [Ewha Womans Univ., Seoul (Korea, Republic of). Div. of Life and Pharmaceutical Sciences; Kim, Wankee [Ewha Womans Univ., Seoul (Korea, Republic of). Dept. of Life Science; Ajou Univ., Suwon (Korea, Republic of). Inst. for Medical Sciences; Choi, Wonja [Ewha Womans Univ., Seoul (Korea, Republic of). Dept. of Life Science; Ewha Womans Univ., Seoul (Korea, Republic of). Microbial Resources Research Center

2012-07-15

Furfural is one of the major inhibitors generated during sugar production from cellulosic materials and, as an aldehyde, inhibits various cellular activities of microorganisms used, leading to prolonged lag time during ethanologenic fermentation. Since Saccharomyces cerevisiae strains tolerant to furfural are of great economic benefit in producing bioethanol, much effort to obtain more efficient strains continues to be made. In this study, we examined the furfural tolerance of transposon mutant strains (Tn 1-5) with enhanced ethanol tolerance and found that one of them (Tn 2), in which SSK2 is downregulated at the transcriptional level, displayed improved furfural tolerance. Such phenotype was abolished by complementation of the entire open reading frame of SSK2, which encodes a mitogen-activated protein (MAP) kinase kinase kinase of the high osmolarity glycerol (HOG) signaling pathway, suggesting an inhibitory effect of SSK2 in coping with furfural stress. Tn 2 showed a significant decrease in the intracellular level of reactive oxygen species (ROS) and early and high activation of Hog1p, a MAP kinase integral to the HOG pathway in response to furfural. The transcriptional levels of CTT1 and GLR1, two of known Hog1p downstream target genes whose protein products are involved in reducing ROS, were increased by 43 % and 56 % respectively compared with a control strain, probably resulting in the ROS decrease. Tn 2 also showed a shortened lag time during fermentation in the presence of furfural, resulting from efficient conversion of furfural to non-toxic (or less toxic) furfuryl alcohol. Taken together, the enhanced furfural tolerance of Tn 2 is suggested to be conferred by the combined effect of an early event of less ROS accumulation and a late event of efficient detoxification of furfural. (orig.)

Insertion of transposon in the vicinity of SSK2 confers enhanced tolerance to furfural in Saccharomyces cerevisiae.

Science.gov (United States)

Kim, Hyun-Soo; Kim, Na-Rae; Kim, Wankee; Choi, Wonja

2012-07-01

Furfural is one of the major inhibitors generated during sugar production from cellulosic materials and, as an aldehyde, inhibits various cellular activities of microorganisms used, leading to prolonged lag time during ethanologenic fermentation. Since Saccharomyces cerevisiae strains tolerant to furfural are of great economic benefit in producing bioethanol, much effort to obtain more efficient strains continues to be made. In this study, we examined the furfural tolerance of transposon mutant strains (Tn 1-5) with enhanced ethanol tolerance and found that one of them (Tn 2), in which SSK2 is downregulated at the transcriptional level, displayed improved furfural tolerance. Such phenotype was abolished by complementation of the entire open reading frame of SSK2, which encodes a mitogen-activated protein (MAP) kinase kinase kinase of the high osmolarity glycerol (HOG) signaling pathway, suggesting an inhibitory effect of SSK2 in coping with furfural stress. Tn 2 showed a significant decrease in the intracellular level of reactive oxygen species (ROS) and early and high activation of Hog1p, a MAP kinase integral to the HOG pathway in response to furfural. The transcriptional levels of CTT1 and GLR1, two of known Hog1p downstream target genes whose protein products are involved in reducing ROS, were increased by 43 % and 56 % respectively compared with a control strain, probably resulting in the ROS decrease. Tn 2 also showed a shortened lag time during fermentation in the presence of furfural, resulting from efficient conversion of furfural to non-toxic (or less toxic) furfuryl alcohol. Taken together, the enhanced furfural tolerance of Tn 2 is suggested to be conferred by the combined effect of an early event of less ROS accumulation and a late event of efficient detoxification of furfural.

Complete Sequence of p07-406, a 24,179-base-pair plasmid harboring the blaVIM-7 metallo-beta-lactamase gene in a Pseudomonas aeruginosa isolate from the United States.

Science.gov (United States)

Li, Hongyang; Toleman, Mark A; Bennett, Peter M; Jones, Ronald N; Walsh, Timothy R

2008-09-01

An outbreak involving a Pseudomonas aeruginosa strain that was resistant to all tested antimicrobials except polymyxin B occurred in a hospital in Houston, TX. Previous studies on this strain showed that it possesses a novel mobile metallo-beta-lactamase (MBL) gene, designated bla(VIM-7), located on a plasmid (p07-406). Here, we report the complete sequence, annotation, and functional characterization of this plasmid. p07-406 is 24,179 bp in length, and 29 open reading frames were identified related to known or putatively recognized proteins. Analysis of this plasmid showed it to be comprised of four distinct regions: (i) a region of 5,200 bp having a Tn501-like mercuric resistance (mer) transposon upstream of the replication region; (ii) a Tn3-like transposon carrying a truncated integron with a bla(VIM-7) gene and an insertion sequence inserted at the other end of this transposon; (iii) a region of four genes, upstream of the Tn3-like transposon, possessing very high similarity to plasmid pXcB from Xanthomonas campestris pv. citri commonly associated with plants; (iv) a backbone sequence similar to the backbone structure of the IncP group plasmid Rms149, pB10, and R751. This is the first plasmid to be sequenced carrying an MBL gene and highlights the amelioration of DNA segments from disparate origins, most noticeably from plant pathogens.

Association of Elevated High Sensitivity Cardiac Troponin T(hs-cTnT) Levels with Hemorrhagic Transformation and 3-Month Mortality in Acute Ischemic Stroke Patients with Rheumatic Heart Disease in China.

Science.gov (United States)

Liu, Junfeng; Wang, Deren; Xiong, Yao; Liu, Bian; Hao, Zilong; Tao, Wendan; Liu, Ming

2016-01-01

Elevated levels of high sensitivity cardiac troponin T (hs-cTnT) occur in a substantial proportion of patients with acute ischemic stroke (AIS) and can predict poor outcome and mortality after stroke. Whether elevated hs-cTnT levels can also predict hemorrhagic transformation (HT) or prognosis in AIS patients with rheumatic heart disease (RHD) remains unclear. Data from the Chengdu Stroke Registry on consecutive AIS patients with RHD admitted to West China Hospital within 1 month of stroke onset from October 2011 to February 2014 were examined. Clinico-demographic characteristics, HT, functional outcomes and stroke recurrence were compared between patients with elevated hs-cTnT levels (≥14 ng/L) and patients with normal hs-cTnT levels (mortality and 3-month disability/mortality (all P≤0.029). After controlling for age, sex, hypertension, renal impairment and National Institutes of Health Stroke Scale score on admission, the risk of HT and 3-month mortality was, respectively, 4.0- and 5.5-fold higher in patients with elevated hs-cTnT levels than in patients with normal hs-cTnT levels. Elevated hs-cTnT levels are independently associated with HT and 3-month mortality in AIS patients with RHD. These results with a small cohort should be verified and extended in large studies.

77 FR 17408 - Foreign-Trade Zone 204-Tri-Cities Area, TN/VA; Application for Reorganization Under Alternative...

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2012-03-26

... Technology Park, 4509 West Stone Drive, Kingsport, Hawkins County, TN; Site 9 (134 acres, expires 6/30/15.... The grantee's proposed service area under the ASF would be the Counties of Sullivan, Hawkins, Greene...

Dengue virus inactivation by minipool TnBP/Triton X-45 treatment of plasma and cryoprecipitate.

Science.gov (United States)

Burnouf, T; Chou, M-L; Cheng, L-H; Li, Z-R; Wu, Y-W; El-Ekiaby, M; Tsai, K-H

2013-01-01

A minipool solvent/detergent (S/D; 1% TnBP/1% Triton X-45; 31°C) process was developed for viral inactivation of plasma and cryoprecipitate used for transfusion. The goal of this study was to determine the rate and extent of inactivation of dengue virus (DENV) during this process. DENV-1 was propagated using C6/36 mosquito cells to an infectivity titre close to 9 log and spiked (10% v/v) into individual plasma and cryoprecipitate samples from two distinct donors. Samples were taken right after spiking and during viral inactivation treatment by 1% TnBP-1% Triton X-45 at 31°C. DENV-1 infectivity was assessed on Vero E6 cells by a focus-forming assay (FFA). Culture medium and complement-inactivated plasma were used as experimental controls. Experiments were done in duplicate. DENV-1 infectivity was 7·5 log in spiked plasma and 7·1 and 7·3 log in spiked cryoprecipitate. There was no loss of DENV-1 infectivity in the spiked materials, nor in the controls not subjected to S/D treatment. No infectivity was found in plasma and cryoprecipitate subjected to S/D treatment at the first time-point evaluated (10 min). DENV-1 was strongly inactivated in plasma and cryoprecipitate, respectively, within 10 min of 1% TnBP/1% Triton X-45 treatment at 31°C. These data provide a reassurance of the safety of such S/D-treated plasma and cryoprecipitate with regard to the risk of transmission of all DENV serotypes and other flaviviruses. © 2012 The Author(s). Vox Sanguinis © 2012 International Society of Blood Transfusion.

On the almost everywhere convergence of the eigenfunction expansions from Liouville classes L_1^\\alpha ({T^N})

Science.gov (United States)

Ahmedov, Anvarjon; Materneh, Ehab; Zainuddin, Hishamuddin

2017-09-01

The relevance of waves in quantum mechanics naturally implies that the decomposition of arbitrary wave packets in terms of monochromatic waves plays an important role in applications of the theory. When eigenfunction expansions does not converge, then the expansions of the functions with certain smoothness should be considered. Such functions gained prominence primarily through their application in quantum mechanics. In this work we study the almost everywhere convergence of the eigenfunction expansions from Liouville classes L_p^α ({T^N}), related to the self-adjoint extension of the Laplace operator in torus TN . The sufficient conditions for summability is obtained using the modified Poisson formula. Isomorphism properties of the elliptic differential operators is applied in order to obtain estimation for the Fourier series of the functions from the classes of Liouville L_p^α .

76 FR 53115 - Foreign-Trade Zone 77-Memphis, TN; Application for Temporary/Interim Manufacturing Authority...

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2011-08-25

... Phone/Mobile Handset Kitting); Memphis, TN An application has been submitted to the Executive Secretary... authority to produce cell phones/mobile handsets kits (HTSUS 8517.12, HTSUS 8517.62, HTSUS 8517.69, duty... cell phone/mobile handset kits (duty free) for the foreign inputs noted above. In accordance with the...

The TN-RAM - a new cask for shipping high activity irradiated hardware

International Nuclear Information System (INIS)

Neider, T.; Hanson, A.S.

1993-01-01

Transnuclear, Inc. has developed a Type B(U) radioactive material transport packaging designed specifically for the transport of dry, irradiated, non-fuel bearing components (NFBC). The TN-RAM is a transport cask in the configuration of a right circular cylinder, fabricated from lead and stainless steel, with wood-filled impact limiters attached at both ends. The lead and stainless steel construction of the lid, walls, and bottom provides a shielding effectiveness of approximately 7.1 inches (18 cm) lead equivalent. (J.P.N.)

HTR fuel research in the HTR-TN network on the high flux reactor

Energy Technology Data Exchange (ETDEWEB)

Guidez, J.; Conrad, R.; Sevini, P.; Burghartz, M. [HFR Unit, Institute for Advanced Materials, European Commission, Joint Research Centre, Petten (Netherlands); Languille, A. [CEA Cadarache, 13 - Saint Paul lez Durance (France); Guillermier, P. [FRAMATOME ANP, 69 - Lyon (France); Bakker, K. [Nuclear Research and Consultancy Group, Petten (Netherlands); Nabielek, H. [Forschungszentrum Juelich (Germany)

2001-07-01

Foremost, this paper explains the economic and strategic reasons for the comeback of the HTR reactor as one of the most promising reactors in the future. To study all the points related to HTR technology, a European network called HTR-TN was created in April 2000, with actually twenty European companies involved. This paper explains the organisation of the network and the related task-groups. In the field of fuel, one of these task-groups works on the fuel cycle and another works on the fuel itself in order to validate by testing HTR fuel possibilities. To this aim, an experimental loop is under construction in the HFR reactor to test full-size pebble type fuel elements and another under study to test compact fuel possibilities. These loops are based on all the experience accumulated by the High Flux Reactor in the years 70-90, when a lot of test were performed for fuel and material for the HTR technology and the facility design uses all the existing HFR knowledge. In conclusion, a host of research work, co-ordinated in the frame of a European network HTR-TN has begun. and should allow in the near future a substantial progress in the knowledge of this very promising fuel. (author)

HTR fuel research in the HTR-TN network on the high flux reactor

International Nuclear Information System (INIS)

Guidez, J.; Conrad, R.; Sevini, P.; Burghartz, M.; Languille, A.; Guillermier, P.; Bakker, K.; Nabielek, H.

2001-01-01

Foremost, this paper explains the economic and strategic reasons for the comeback of the HTR reactor as one of the most promising reactors in the future. To study all the points related to HTR technology, a European network called HTR-TN was created in April 2000, with actually twenty European companies involved. This paper explains the organisation of the network and the related task-groups. In the field of fuel, one of these task-groups works on the fuel cycle and another works on the fuel itself in order to validate by testing HTR fuel possibilities. To this aim, an experimental loop is under construction in the HFR reactor to test full-size pebble type fuel elements and another under study to test compact fuel possibilities. These loops are based on all the experience accumulated by the High Flux Reactor in the years 70-90, when a lot of test were performed for fuel and material for the HTR technology and the facility design uses all the existing HFR knowledge. In conclusion, a host of research work, co-ordinated in the frame of a European network HTR-TN has begun. and should allow in the near future a substantial progress in the knowledge of this very promising fuel. (author)

Effect of PCI on inflammatory factors, cTnI, MMP-9 and NT-pro BNP in patients with unstable angina pectoris

Directory of Open Access Journals (Sweden)

Ke-Tong Liu

2016-05-01

Full Text Available Objective: To investigate the effect of PCI on inflammatory factors, cTnI, MMP-9and NTpro BNP in patients with unstable angina pectoris. Methods: A total of 80 unstable angina pectoris patients were divided into observation group (40 cases and control group (40 cases. The observation group was given the therapy of PCI, and the control group was given coronary angiography. To observe the of inflammatory factors, cTnI, MMP-9 and NT-pro BNP were tested and compared before and after operation. Results: At 24 h after operation, CRP and IL-18 levels were increased significantly after treatment inoperation groups, there was no difference on inflammatory factors in control group, and had significant difference on inflammatory factors in two groups; At 24 h after operation, cTnI, MMP-9 and NT-pro BNP levels were increased significantly after treatment inoperation groups, there was no difference on inflammatory factors in control group, and had significant difference on inflammatory factors in two groups. Conclusion: PCI therapy can induce inflammation and myocardial injury in patients with unstable angina pectoris.

The Nucleoid Binding Protein H-NS Biases Genome-Wide Transposon Insertion Landscapes

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Satoshi Kimura

2016-08-01

Full Text Available Transposon insertion sequencing (TIS; also known as TnSeq is a potent approach commonly used to comprehensively define the genetic loci that contribute to bacterial fitness in diverse environments. A key presumption underlying analyses of TIS datasets is that loci with a low frequency of transposon insertions contribute to fitness. However, it is not known whether factors such as nucleoid binding proteins can alter the frequency of transposon insertion and thus whether TIS output may systematically reflect factors that are independent of the role of the loci in fitness. Here, we investigated whether the histone-like nucleoid structuring (H-NS protein, which preferentially associates with AT-rich sequences, modulates the frequency of Mariner transposon insertion in the Vibrio cholerae genome, using comparative analysis of TIS results from wild-type (wt and Δhns V. cholerae strains. These analyses were overlaid on gene classification based on GC content as well as on extant genome-wide identification of H-NS binding loci. Our analyses revealed a significant dearth of insertions within AT-rich loci in wt V. cholerae that was not apparent in the Δhns insertion library. Additionally, we observed a striking correlation between genetic loci that are overrepresented in the Δhns insertion library relative to their insertion frequency in wt V. cholerae and loci previously found to physically interact with H-NS. Collectively, our findings reveal that factors other than genetic fitness can systematically modulate the frequency of transposon insertions in TIS studies and add a cautionary note to interpretation of TIS data, particularly for AT-rich sequences.

76 FR 60802 - Foreign-Trade Zone 77-Memphis, TN; Withdrawal of Request for Temporary/Interim Manufacturing...

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2011-09-30

..., Inc., (Cell Phone/Mobile Handset Kitting). Memphis, TN Notice is hereby given of the withdrawal of the application of the City of Memphis, grantee of FTZ 77, requesting temporary/interim manufacturing authority on behalf of Flextronics Logistics USA, Inc. in Memphis Tennessee. The application was filed on August 19...

Heidegeer a Kierkegaard: pojetí boha a posvátného

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Jiří Olšovský

2010-05-01

Full Text Available Text je zaměřen zejména na posvátný charakter světa a přírody, jak se zračí v pojetí bytí a božského u Heideggera a Kierkegaarda. Bytí zde zůstává tématem, spolu se základní péčí a starostí o svět a věci v něm. Ukazuje se, že spodní proud Heideggerova myšlení je znovu-probuzení „zbožného myšlení“ a rezervovanost. Zejména článek chce upozornit na Heideggerovy odkazy k mysticismu a Kierkegaardův „pramen víry“. V úvahách nad Heideggerovým dílem je třeba se zamyslet, zda otázka Boha může být rozhodnuta jen v blízkosti bytí a svatosti. O noumenální charakter naší „zahrady“ je nezbytné znovu náležitě pečovat, a zaposlouchat se proto do řeči těch, kteří byli s to se přiblížit k bytí a božskému, k posvátnému pobývání na zemi.

Anal Carcinoma: Impact of TN Category of Disease on Survival, Disease Relapse, and Colostomy Failure in US Gastrointestinal Intergroup RTOG 98-11 Phase 3 Trial

Science.gov (United States)

Gunderson, Leonard L.; Moughan, Jennifer; Ajani, Jaffer A.; Pedersen, John E.; Winter, Kathryn A.; Benson, Al B.; Thomas, Charles R.; Mayer, Robert J.; Haddock, Michael G.; Rich, Tyvin A.; Willett, Christopher G.

2013-01-01

Purpose The long-term update of US GI Intergroup RTOG 98-11 anal cancer trial found that concurrent chemoradiation (CCRT) with fluorouracil (5-FU) plus mitomycin had a significant impact on disease-free survival (DFS) and overall survival (OS) compared with induction plus concurrent 5-FU plus cisplatin. The intent of the current analysis was to determine the impact of tumor node (TN) category of disease on survival (DFS and OS), colostomy failure (CF), and relapse (local-regional failure [LRF] and distant metastases [DM]) in this patient group. Methods and Materials DFS and OS were estimated univariately by using the Kaplan-Meier method, and 6 TN categories were compared by the log–rank test (T2N0, T3N0, T4N0, T2N1-3, T3N1-3, and T4N1-3). Time to relapse and colostomy were estimated by the cumulative incidence method, and TN categories were compared using Gray’s test. Results Of 682 patients, 620 were analyzable for outcomes by TN category. All endpoints showed statistically significant differences among the TN categories of disease (OS, P<.0001; DFS, P<.0001; LRF, P<.0001; DM, P=.0011; CF, P=.01). Patients with the poorest OS, DFS, and LRF outcomes were those with T3-4N-positive (+) disease. CF was lowest for T2N0 and T2N+ (11%, 11%, respectively) and worst for the T4N0, T3N+, and T4N+ categories (26%, 27%, 24%, respectively). Conclusions TN category of disease has a statistically significant impact on OS, DFS, LRF, DM, and CF in patients treated with CCRT and provides excellent prognostic information for outcomes in patients with anal carcinoma. Significant challenges remain for patients with T4N0 and T3-4N+ categories of disease with regard to survival, relapse, and CF and lesser challenges for T2-3N0/T2N+ categories. PMID:24035327

Anal Carcinoma: Impact of TN Category of Disease on Survival, Disease Relapse, and Colostomy Failure in US Gastrointestinal Intergroup RTOG 98-11 Phase 3 Trial

Energy Technology Data Exchange (ETDEWEB)

Gunderson, Leonard L., E-mail: gunderson.leonard@mayo.edu [Mayo Clinic Cancer Center, Scottsdale, Arizona (United States); Moughan, Jennifer [Radiation Therapy Oncology Group, Philadelphia, Pennsylvania (United States); Ajani, Jaffer A. [The University of Texas MD Anderson Cancer Center, Houston, Texas (United States); Pedersen, John E. [Cross Cancer Institute, Edmonton, Alberta (Canada); Winter, Kathryn A. [Radiation Therapy Oncology Group, Philadelphia, Pennsylvania (United States); Benson, Al B. [Northwestern University, Chicago, Illinois (United States); Thomas, Charles R. [Knight Cancer Institute/Oregon Health and Science University, Portland, Oregon (United States); Mayer, Robert J. [Dana-Farber Cancer Institute, Boston, Massachusetts (United States); Haddock, Michael G. [Mayo Clinic Cancer Center, Rochester, Minnesota (United States); Rich, Tyvin A. [University of Virginia, Charlottesville, Virginia (United States); Willett, Christopher G. [Duke University, Durham, North Carolina (United States)

2013-11-15

Purpose: The long-term update of US GI Intergroup RTOG 98-11 anal cancer trial found that concurrent chemoradiation (CCRT) with fluorouracil (5-FU) plus mitomycin had a significant impact on disease-free survival (DFS) and overall survival (OS) compared with induction plus concurrent 5-FU plus cisplatin. The intent of the current analysis was to determine the impact of tumor node (TN) category of disease on survival (DFS and OS), colostomy failure (CF), and relapse (local-regional failure [LRF] and distant metastases [DM]) in this patient group. Methods and Materials: DFS and OS were estimated univariately by using the Kaplan-Meier method, and 6 TN categories were compared by the log–rank test (T2N0, T3N0, T4N0, T2N1-3, T3N1-3, and T4N1-3). Time to relapse and colostomy were estimated by the cumulative incidence method, and TN categories were compared using Gray's test. Results: Of 682 patients, 620 were analyzable for outcomes by TN category. All endpoints showed statistically significant differences among the TN categories of disease (OS, P<.0001; DFS, P<.0001; LRF, P<.0001; DM, P=.0011; CF, P=.01). Patients with the poorest OS, DFS, and LRF outcomes were those with T3-4N-positive (+) disease. CF was lowest for T2N0 and T2N+ (11%, 11%, respectively) and worst for the T4N0, T3N+, and T4N+ categories (26%, 27%, 24%, respectively). Conclusions: TN category of disease has a statistically significant impact on OS, DFS, LRF, DM, and CF in patients treated with CCRT and provides excellent prognostic information for outcomes in patients with anal carcinoma. Significant challenges remain for patients with T4N0 and T3-4N+ categories of disease with regard to survival, relapse, and CF and lesser challenges for T2-3N0/T2N+ categories.

Anal Carcinoma: Impact of TN Category of Disease on Survival, Disease Relapse, and Colostomy Failure in US Gastrointestinal Intergroup RTOG 98-11 Phase 3 Trial

International Nuclear Information System (INIS)

Gunderson, Leonard L.; Moughan, Jennifer; Ajani, Jaffer A.; Pedersen, John E.; Winter, Kathryn A.; Benson, Al B.; Thomas, Charles R.; Mayer, Robert J.; Haddock, Michael G.; Rich, Tyvin A.; Willett, Christopher G.

2013-01-01

Purpose: The long-term update of US GI Intergroup RTOG 98-11 anal cancer trial found that concurrent chemoradiation (CCRT) with fluorouracil (5-FU) plus mitomycin had a significant impact on disease-free survival (DFS) and overall survival (OS) compared with induction plus concurrent 5-FU plus cisplatin. The intent of the current analysis was to determine the impact of tumor node (TN) category of disease on survival (DFS and OS), colostomy failure (CF), and relapse (local-regional failure [LRF] and distant metastases [DM]) in this patient group. Methods and Materials: DFS and OS were estimated univariately by using the Kaplan-Meier method, and 6 TN categories were compared by the log–rank test (T2N0, T3N0, T4N0, T2N1-3, T3N1-3, and T4N1-3). Time to relapse and colostomy were estimated by the cumulative incidence method, and TN categories were compared using Gray's test. Results: Of 682 patients, 620 were analyzable for outcomes by TN category. All endpoints showed statistically significant differences among the TN categories of disease (OS, P<.0001; DFS, P<.0001; LRF, P<.0001; DM, P=.0011; CF, P=.01). Patients with the poorest OS, DFS, and LRF outcomes were those with T3-4N-positive (+) disease. CF was lowest for T2N0 and T2N+ (11%, 11%, respectively) and worst for the T4N0, T3N+, and T4N+ categories (26%, 27%, 24%, respectively). Conclusions: TN category of disease has a statistically significant impact on OS, DFS, LRF, DM, and CF in patients treated with CCRT and provides excellent prognostic information for outcomes in patients with anal carcinoma. Significant challenges remain for patients with T4N0 and T3-4N+ categories of disease with regard to survival, relapse, and CF and lesser challenges for T2-3N0/T2N+ categories

The androgen receptor controls expression of the cancer-associated sTn antigen and cell adhesion through induction of ST6GalNAc1 in prostate cancer

Science.gov (United States)

Munkley, Jennifer; Oltean, Sebastian; Vodák, Daniel; Wilson, Brian T.; Livermore, Karen E.; Zhou, Yan; Star, Eleanor; Floros, Vasileios I.; Johannessen, Bjarne; Knight, Bridget; McCullagh, Paul; McGrath, John; Crundwell, Malcolm; Skotheim, Rolf I.; Robson, Craig N.; Leung, Hing Y.; Harries, Lorna W.; Rajan, Prabhakar; Mills, Ian G.; Elliott, David J.

2015-01-01

Patterns of glycosylation are important in cancer, but the molecular mechanisms that drive changes are often poorly understood. The androgen receptor drives prostate cancer (PCa) development and progression to lethal metastatic castration-resistant disease. Here we used RNA-Seq coupled with bioinformatic analyses of androgen-receptor (AR) binding sites and clinical PCa expression array data to identify ST6GalNAc1 as a direct and rapidly activated target gene of the AR in PCa cells. ST6GalNAc1 encodes a sialytransferase that catalyses formation of the cancer-associated sialyl-Tn antigen (sTn), which we find is also induced by androgen exposure. Androgens induce expression of a novel splice variant of the ST6GalNAc1 protein in PCa cells. This splice variant encodes a shorter protein isoform that is still fully functional as a sialyltransferase and able to induce expression of the sTn-antigen. Surprisingly, given its high expression in tumours, stable expression of ST6GalNAc1 in PCa cells reduced formation of stable tumours in mice, reduced cell adhesion and induced a switch towards a more mesenchymal-like cell phenotype in vitro. ST6GalNAc1 has a dynamic expression pattern in clinical datasets, being significantly up-regulated in primary prostate carcinoma but relatively down-regulated in established metastatic tissue. ST6GalNAc1 is frequently upregulated concurrently with another important glycosylation enzyme GCNT1 previously associated with prostate cancer progression and implicated in Sialyl Lewis X antigen synthesis. Together our data establishes an androgen-dependent mechanism for sTn antigen expression in PCa, and are consistent with a general role for the androgen receptor in driving important coordinate changes to the glycoproteome during PCa progression. PMID:26452038

Troponin C Mutations Partially Stabilize the Active State of Regulated Actin and Fully Stabilize the Active State When Paired with Δ14 TnT.

Science.gov (United States)

Baxley, Tamatha; Johnson, Dylan; Pinto, Jose R; Chalovich, Joseph M

2017-06-13

Striated muscle contraction is regulated by the actin-associated proteins tropomyosin and troponin. The extent of activation of myosin ATPase activity is lowest in the absence of both Ca 2+ and activating cross-bridges (i.e., S1-ADP or rigor S1). Binding of activating species of myosin to actin at a saturating Ca 2+ concentration stabilizes the most active state (M state) of the actin-tropomyosin-troponin complex (regulated actin). Ca 2+ binding alone produces partial stabilization of the active state. The extent of stabilization at a saturating Ca 2+ concentration depends on the isoform of the troponin subunits, the phosphorylation state of troponin, and, in the case of cardiac muscle, the presence of hypertrophic cardiomyopathy-producing mutants of troponin T and troponin I. Cardiac dysfunction is also associated with mutations of troponin C (TnC). Troponin C mutants A8V, C84Y, and D145E increase the Ca 2+ sensitivity of ATPase activity. We show that these mutants change the distribution of regulated actin states. The A8V and C84Y TnC mutants decreased the inactive B state distribution slightly at low Ca 2+ concentrations, but the D145E mutants had no effect on that state. All TnC mutants increased the level of the active M state compared to that of the wild type, at a saturating Ca 2+ concentration. Troponin complexes that contained two mutations that stabilize the active M state, A8V TnC and Δ14 TnT, appeared to be completely in the active state in the presence of only Ca 2+ . Because Ca 2+ gives full activation, in this situation, troponin must be capable of positioning tropomyosin in the active M state without the need for rigor myosin binding.

CREATINE PHOSPHOKINASE ISOENZYME-MB MASS (CK-MB MASS AND TROPONIN I (cTnI IN DOGS (Canis familiaris

Directory of Open Access Journals (Sweden)

Millena Vidal Freitas

2015-07-01

Full Text Available Cardiac markers, especially CK-MB mass and cTnI, have an essential role in both human and veterinary clinical and surgical cardiology, allowing a more precise and accurate diagnosis in myocardial lesions. The goal of this work was to measure these cardiac markers in veterinary medicine, improve their use and provide information about these laboratory methods that allow non-invasive health monitoring of the myocardial cell. Parameters quantification was obtained from sera of healthy dogs examined during routine procedures at the Small Animal Veterinary Hospital of Universidade Estadual do Norte Fluminense Darcy Ribeiro. The human chemiluminescence essay turbo kit (IMMULITE Turbo, Siemens ® test proved to be effective in canine species for both CK-MB mass and cTnI. In addition, the values found for CK-MB will significantly contribute to clinical routine or experimental work with dogs.

The VanE operon in Enterococcus faecalis N00-410 is found on a putative integrative and conjugative element, Tn6202.

Science.gov (United States)

Boyd, D A; Mulvey, M R

2013-02-01

To date no complete genetic structure of acquired DNA harbouring a d-Ala-d-Ser operon in an Enterococcus is known. We wished to characterize the acquired DNA harbouring the vanE operon located in the Enterococcus faecalis N00-410 chromosome. Whole genome sequencing of E. faecalis N00-410 was conducted by massively parallel sequencing. Two sequence contigs harbouring the vanE region were linked by PCR and the acquired DNA harbouring the vanE operon was completely characterized. Excision/integration of the region was determined by PCR and transfer attempted by conjugation. The regions flanking the vanE operon were analysed and a total of 42 open reading frames were identified in a region flanked by inverted terminal and direct repeats (Tn6202). Tn6202 could be excised from the chromosome, circularized and the target site rejoined, but transfer could not be demonstrated. The vanE operon was found on the putative integrative and conjugative element Tn6202 in the E. faecalis N00-410 chromosome. This represents the first characterization of acquired DNA harbouring a D-Ala-D-Ser operon.

Mutations in the Histone-Like Nucleoid Structuring Regulatory Gene (hns) Decrease the Adherence of Shiga Toxin-Producing Escherichia coli 091:H21 Strain B2F1 to Human Colonic Epithelial Cells and Increase the Production of Hemolysin

National Research Council Canada - National Science Library

Scott, Maria

1999-01-01

...:H21 to human colonic epithelial cells. Transposon mutagenesis of B2F1 was accomplished with the mini-Tn5phoACm mobile element and a mutant bank of B2F1 colonies that carried putative in-frame PhoA-positive transposon insertions was isolated...

Distribution of the multidrug resistance gene cfr in Staphylococcus species isolates from swine farms in China.

Science.gov (United States)

Wang, Yang; Zhang, Wanjiang; Wang, Juan; Wu, Congming; Shen, Zhangqi; Fu, Xiao; Yan, Yang; Zhang, Qijing; Schwarz, Stefan; Shen, Jianzhong

2012-03-01

A total of 149 porcine Staphylococcus isolates with florfenicol MICs of ≥ 16 μg/ml were screened for the presence of the multiresistance gene cfr, its location on plasmids, and its genetic environment. In total, 125 isolates carried either cfr (16 isolates), fexA (92 isolates), or both genes (17 isolates). The 33 cfr-carrying staphylococci, which included isolates of the species Staphylococcus cohnii, S. arlettae, and S. saprophyticus in which the cfr gene has not been described before, exhibited a wide variety of SmaI pulsed-field gel electrophoresis patterns. In 18 cases, the cfr gene was located on plasmids. Four different types of cfr-carrying plasmids--pSS-01 (n = 2; 40 kb), pSS-02 (n = 3; 35.4 kb), pSS-03 (n = 10; 7.1 kb), and pBS-01 (n = 3; 16.4 kb)--were differentiated on the basis of their sizes, restriction patterns, and additional resistance genes. Sequence analysis revealed that in plasmid pSS-01, the cfr gene was flanked in the upstream part by a complete aacA-aphD-carrying Tn4001-like transposon and in the downstream part by a complete fexA-carrying transposon Tn558. In plasmid pSS-02, an insertion sequence IS21-558 and the cfr gene were integrated into transposon Tn558 and thereby truncated the tnpA and tnpB genes. The smallest cfr-carrying plasmid pSS-03 carried the macrolide-lincosamide-streptogramin B resistance gene erm(C). Plasmid pBS-01, previously described in Bacillus spp., harbored a Tn917-like transposon, including the macrolide-lincosamide-streptogramin B resistance gene erm(B) in the cfr downstream region. Plasmids, which in part carry additional resistance genes, seem to play an important role in the dissemination of the gene cfr among porcine staphylococci.

Production of diketopiperazine derivative cyclo (l-leu-l-arg) by streptomyces sp. tn262 after exposure to heat-killed fungus fusarium sp

International Nuclear Information System (INIS)

Elleuch, L.; Smaoui, S.; Najah, S.; Sellem, I

2013-01-01

In a screening program for new active secondary metabolites producers, a strain of Streptomyces called TN262 was isolated from Tunisian soil and selected for its ability to produce eleven active compounds in pure culture conditions. In this work, the effect of different concentrations of heat-killed fungus Fusarium sp. on the production of active compounds by TN262 strain was studied. The ethyl acetate extract from the culture of Streptomyces sp. TN262 combined with heat-killed Fusarium sp. at 50 micro g/ml inhibited the growth of the three used indicator microorganisms. In fact, an increase of 36%, 21% and 20% in inhibitory activity was obtained against Micrococcus luteus LB 14110, Escherichia coli ATCC 8739 and Fusarium sp. respectively. The HPLC chromatographic profiles of the ethyl acetate extracts from both culture conditions were different and an additional active compound was produced only under induced conditions. This active component was isolated and identified as Cyclo (L-Leu-L-Arg) (1), a diketopiperazine derivative, possessing antibacterial and antifungal activity. Consequently, this study showed that the addition of heat-killed fungus is a useful method for inducing the production of bioactive compounds. (author)

Construction of a large-scale Burkholderia cenocepacia J2315 transposon mutant library

Science.gov (United States)

Wong, Yee-Chin; Pain, Arnab; Nathan, Sheila

2014-09-01

Burkholderia cenocepacia, a pathogenic member of the Burkholderia cepacia complex (Bcc), has emerged as a significant threat towards cystic fibrosis patients, where infection often leads to the fatal clinical manifestation known as cepacia syndrome. Many studies have investigated the pathogenicity of B. cenocepacia as well as its ability to become highly resistant towards many of the antibiotics currently in use. In addition, studies have also been undertaken to understand the pathogen's capacity to adapt and survive in a broad range of environments. Transposon based mutagenesis has been widely used in creating insertional knock-out mutants and coupled with recent advances in sequencing technology, robust tools to study gene function in a genome-wide manner have been developed based on the assembly of saturated transposon mutant libraries. In this study, we describe the construction of a large-scale library of B. cenocepacia transposon mutants. To create transposon mutants of B. cenocepacia strain J2315, electrocompetent bacteria were electrotransformed with the EZ-Tn5 transposome. Tetracyline resistant colonies were harvested off selective agar and pooled. Mutants were generated in multiple batches with each batch consisting of ˜20,000 to 40,000 mutants. Transposon insertion was validated by PCR amplification of the transposon region. In conclusion, a saturated B. cenocepacia J2315 transposon mutant library with an estimated total number of 500,000 mutants was successfully constructed. This mutant library can now be further exploited as a genetic tool to assess the function of every gene in the genome, facilitating the discovery of genes important for bacterial survival and adaptation, as well as virulence.

Complete sequence of the IncA/C1 plasmid pCf587 carrying blaPER-2 from Citrobacter freundii.

Science.gov (United States)

Ruggiero, Melina; Girlich, Delphine; Dabos, Laura; Power, Pablo; Naas, Thierry; Gutkind, Gabriel

2018-02-20

The bla PER-2 harboring plasmid pCf587 (191,541 bp) belongs to lineage IncA/C 1 and is closely related to pRA1. It contains a large resistance island including the bla PER-2 gene between two copies of IS Kox2 -like elements, the toxin-antitoxin module pemK-pemI, several other resistance genes inserted within a Tn 2 transposon, a Tn 21 -like structure, and a class 1 integron. pCf587 belongs into ST 13, a new pMLST. Copyright © 2018 American Society for Microbiology.

Physicochemical and toxicological profiling of ash from the 2010 and 2011 eruptions of Eyjafjallajökull and Grímsvötn volcanoes, Iceland using a rapid respiratory hazard assessment protocol.

Science.gov (United States)

Horwell, C J; Baxter, P J; Hillman, S E; Calkins, J A; Damby, D E; Delmelle, P; Donaldson, K; Dunster, C; Fubini, B; Kelly, F J; Le Blond, J S; Livi, K J T; Murphy, F; Nattrass, C; Sweeney, S; Tetley, T D; Thordarson, T; Tomatis, M

2013-11-01

The six week eruption of Eyjafjallajökull volcano in 2010 produced heavy ash fall in a sparsely populated area of southern and south eastern Iceland and disrupted European commercial flights for at least 6 days. We adopted a protocol for the rapid analysis of volcanic ash particles, for the purpose of informing respiratory health risk assessments. Ash collected from deposits underwent a multi-laboratory physicochemical and toxicological investigation of their mineralogical parameters associated with bio-reactivity, and selected in vitro toxicology assays related to pulmonary inflammatory responses. Ash from the eruption of Grímsvötn, Iceland, in 2011 was also studied. The results were benchmarked against ash from Soufrière Hills volcano, Montserrat, which has been extensively studied since the onset of eruptive activity in 1995. For Eyjafjallajökull, the grain size distributions were variable: 2-13 vol% of the bulk samples were <4 µm, with the most explosive phases of the eruption generating abundant respirable particulate matter. In contrast, the Grímsvötn ash was almost uniformly coarse (<3.5 vol%<4 µm material). Surface area ranged from 0.3 to 7.7 m2 g(-1) for Eyjafjallajökull but was very low for Grímsvötn (<0.6 m2 g(-1)). There were few fibre-like particles (which were unrelated to asbestos) and the crystalline silica content was negligible in both eruptions, whereas Soufrière Hills ash was cristobalite-rich with a known potential to cause silicosis. All samples displayed a low ability to deplete lung antioxidant defences, showed little haemolysis and low acute cytotoxicity in human alveolar type-1 like epithelial cells (TT1). However, cell-free tests showed substantial hydroxyl radical generation in the presence of hydrogen peroxide for Grímsvötn samples, as expected for basaltic, Fe-rich ash. Cellular mediators MCP-1, IL-6, and IL-8 showed chronic pro-inflammatory responses in Eyjafjallajökull, Grímsvötn and Soufrière Hills samples

IHF-independent assembly of the Tn10 strand transfer transpososome: implications for inhibition of disintegration

OpenAIRE

Stewart, Barry J.; Wardle, Simon J.; Haniford, David B.

2002-01-01

The frequency of DNA transposition in transposition systems that employ a strand transfer step may be significantly affected by the occurrence of a disintegration reaction, a reaction that reverses the strand transfer event. We have asked whether disintegration occurs in the Tn10 transposition system. We show that disintegration substrates (substrates constituting one half of the strand transfer product) are assembled into a transpososome that mimics the strand transfer intermediate. This str...

Studies on the process of attachment of diazotroph alcaligenes faecalis and its Tn5 mutants to rice roots using 15N-labelling technique

International Nuclear Information System (INIS)

Fang Xuanjun; Lin Min; You Chongbiao

1993-09-01

By using 15 N-labelling technique and Tn5-induced mutants the attachment of associative diazotroph Alcaligenes faecalis to intact rice plants was examined in vitro. Three distinguished modes of attachment of Alcaligenes faecalis: adsorption, anchoring and colonization were proposed by using 15 N-labelling bacterial cells and Tn5-induced mutants. Che - mutants affected on adsorption, but not on anchoring. Exo - Che - mutant is defective in both adsorption and anchoring. Exo - or exo ++ mutants are only defective in anchoring. Effective colonization is benefit for establishment on the associative system. The data also indicated that EPS (exopolysaccharide) play rather important roles in the association between the host plant and bacteria

Exact comparison of dose rate measurements and calculation of TN12/2 packages

International Nuclear Information System (INIS)

Taniuchi, H.; Matsuda, F.

1998-01-01

Both of dose rate measurements of TN 12/2 package and calculations by Monte Carlo code MORSE in SCALE code system and MCNP were performed to evaluate the difference between the measurement and the calculation and finding out the cause of the difference. The calculated gamma-ray dose rates agreed well with measured ones, but calculated neutron dose rates overestimated more than a factor of 1.7. When considering the cause of the difference and applying the modification into the neutron calculation, the calculated neutron dose rates become to agree well, and the factor decreased to around 1.3. (authors)

Panovnické postavy v závěru schodištních maleb

Czech Academy of Sciences Publication Activity Database

Kubínová, Kateřina

2006-01-01

Roč. 13, příl. (2006), s. 23-36 ISSN 1212-1487. [Schodištní cykly Velké věže hradu Karlštejna. Stav po restaurování. Praha, 05.05.2004-07.05.2004] R&D Projects: GA AV ČR(CZ) IAA8033301 Institutional research plan: CEZ:AV0Z80330511 Keywords : Bohemian medieval wall painting * Karlštejn * Charles IV Subject RIV: AL - Art, Architecture, Cultural Heritage

TN-68 Spent Fuel Transport Cask Analytical Evaluation for Drop Events

International Nuclear Information System (INIS)

Shah, M.J.; Klymyshyn, Nicholas A.; Adkins, Harold E.; Koeppel, Brian J.

2007-01-01

The U.S. Nuclear Regulatory Commission (NRC) is responsible for licensing commercial spent nuclear fuel transported in casks certified by NRC under the Code of Federal Regulations (10 CFR), Title 10, Part 71 (1). Both the International Atomic Energy Agency regulations for transporting radioactive materials (2, paragraph 727), and 10 CFR 71.73 require casks to be evaluated for hypothetical accident conditions, which includes a 9-meter (m) (30-ft) drop-impact event onto a flat, essentially unyielding, horizontal surface, in the most damaging orientation. This paper examines the behavior of one of the NRC certified transportation casks, the TN-68 (3), for drop-impact events. The specific area examined is the behavior of the bolted connections in the cask body and the closure lid, which are significantly loaded during the hypothetical drop-impact event. Analytical work to evaluate the NRC-certified TN-68 spent fuel transport cask (3) for a 9-m (30-ft) drop-impact event on a flat, unyielding, horizontal surface, was performed using the ANSYS (4) and LS DYNA (5) finite-element analysis codes. The models were sufficiently detailed, in the areas of bolt closure interfaces and containment boundaries, to evaluate the structural integrity of the bolted connections under 9-m (30-ft) free-drop hypothetical accident conditions, as specified in 10 CFR 71.73. Evaluation of the cask for puncture, caused by a free drop through a distance of 1-m (40-in.) onto a mild steel bar mounted on a flat, essentially unyielding, horizontal surface, required by 10 CFR 71.73, was not included in the current work, and will have to be addressed in the future. Based on the analyses performed to date, it is concluded that, even though brief separation of the flange and the lid surfaces may occur under some conditions, the seals would close at the end of the drop events, because the materials remain elastic during the duration of the event

Green digital signage using nanoparticle embedded narrow-gap field sequential TN-LCDs

Science.gov (United States)

Kobayashi, Shunsuke; Shiraishi, Yukihide; Sawai, Hiroya; Toshima, Naoki; Okita, Masaya; Takeuchi, Kiyofumi; Takatsu, Haruyoshi

2012-03-01

We have fabricated field sequential color (FSC)-LCDs using cells and modules of narrow-gap TN-LCDs with and without doping the nanoparticles of PCyD-ZrO2 and AF-SiO2. It is shown that the FSC-LCD exhibits a high optical efficiency of OE=4.5 that is defined as OE=[Luminance]/[W/m2]=(cd/W). This figure may provide us a good reference or to clear the Energy Star Program Version 5-3 that issues a guideline: LCD with 50 inch on the diagonal consumes the energy of 108W. Through this research it is claimed that our FSC=LCD may be a novel green digital signage.

Assessment of regeneration of bone in the extracted third molar sockets augmented using xenograft (CollaPlugTN Zimmer in comparison with the normal healing on the contralateral side

Directory of Open Access Journals (Sweden)

Murugan Ranganathan

2017-01-01

Full Text Available Introduction: Alveolar bone resorption is a significant clinical problem. Bone loss in third molar region following extraction or surgical removal not only leads to periodontal problems in second molar region but also it may lead to some serious problems like increased incidence of angle fractures. In order to reduce the risks following third molar surgery, the socket should be augmented with bone grafts. In recent days guided tissue regeneration is the most accepted and successful technique followed many authors and its efficacy has been proved. Materials and Methods: Based upon our clinical experience, the use of bio absorbable collagen wound dressing such as CollaPlugTN has achieved quick healing and more primary wound coverage. Amongst the graft materials collagen is preferable due to its high biocompatibility and hemostatic ability. This study was done to assess the regeneration of bone in the extracted third molar sockets using xenograft (CollaPlugTN-Zimmer which was compared with the normal healing on the contra lateral side. The assessment was done to analyze post-operative healing complications and to compare the bone density formed between control site and implant site radiologically. Conclusion: On this basis of this study, the use of collaplugTN appears to be beneficial to the patient in postoperative wound healing and also for better bone formation. The use of this material was advantageous because of its simplicity of application cost effectiveness and availability. There is enhanced wound healing and early bone formation.

Assessment of Regeneration of Bone in the Extracted Third Molar Sockets Augmented Using Xenograft (CollaPlugTN Zimmer) in Comparison with the Normal Healing on the Contralateral Side.

Science.gov (United States)

Ranganathan, Murugan; Balaji, M; Krishnaraj, R; Narayanan, Vivek; Thangavelu, Annamalai

2017-11-01

Alveolar bone resorption is a significant clinical problem. Bone loss in third molar region following extraction or surgical removal not only leads to periodontal problems in second molar region but also it may lead to some serious problems like increased incidence of angle fractures. In order to reduce the risks following third molar surgery, the socket should be augmented with bone grafts. In recent days guided tissue regeneration is the most accepted and successful technique followed many authors and its efficacy has been proved. Based upon our clinical experience, the use of bio absorbable collagen wound dressing such as CollaPlug TN has achieved quick healing and more primary wound coverage. Amongst the graft materials collagen is preferable due to its high biocompatibility and hemostatic ability. This study was done to assess the regeneration of bone in the extracted third molar sockets using xenograft (CollaPlug TN -Zimmer) which was compared with the normal healing on the contra lateral side. The assessment was done to analyze post-operative healing complications and to compare the bone density formed between control site and implant site radiologically. On this basis of this study, the use of collaplugTN appears to be beneficial to the patient in postoperative wound healing and also for better bone formation. The use of this material was advantageous because of its simplicity of application cost effectiveness and availability. There is enhanced wound healing and early bone formation.

'Test n Treat (TnT)': a cluster-randomised feasibility trial of frequent, rapid-testing and same-day, on-site treatment to reduce rates of chlamydia in high-risk further education college students: statistical analysis plan.

Science.gov (United States)

Phillips, Rachel; Oakeshott, Pippa; Kerry-Barnard, Sarah; Reid, Fiona

2018-06-05

There are high rates of sexually transmitted infections (STIs) in ethnically diverse, sexually active students aged 16-24 years attending London further education (FE) colleges. However, uptake of chlamydia screening remains low. The TnT study aims to assess the feasibility of conducting a future trial in FE colleges to investigate if frequent, rapid, on-site testing and treatment (TnT) reduces chlamydia rates. This article presents the statistical analysis plan for the main study publication as approved and signed off by the Trial Management Group prior to the first data extraction for the final report. TnT is a cluster-randomised feasibility trial conducted over 7 months with parallel qualitative and economic assessments. Colleges will be randomly allocated into the intervention (TnT) or the control group (no TnT). Six FE colleges in London will be included. At each college for 2 days, 80 consecutive sexually active students aged 16-24 years (total 480 students across all six colleges) will be recruited from public areas and asked to provide baseline samples. One and 4 months after recruitment intervention colleges will be visited on two consecutive days by the TnT team where participating students will be texted and invited to come for same-day, on-site, rapid chlamydia testing and, if positive, treatment. Participants in the control colleges will receive 'thank you' texts 1 and 4 months after recruitment. Seven months after recruitment, participants from both groups will be invited to complete questionnaires and provide samples for TnT. All samples will be tested, and same-day treatment offered to participants with positive results. Key feasibility outcomes include: recruitment rates, testing and treatment uptake rates (at 1 and 4 months) and follow-up rates (at 7 months). ISRCTN 58038795 . Registered on 31 August 2016.

77 FR 59573 - Proposed Amendment of Class D and E Airspace; Tri-Cities, TN; Revocation of Class E Airspace; Tri...

Science.gov (United States)

2012-09-28

... at Hawkins County Airport, Rogersville, TN, and Virginia Highlands Airport, Abington, VA. The Tri... separation of existing Class E airspace surrounding Virginia Highlands Airport, Abingdon, VA, and Hawkins... publication, the FAA reassessed the proposal to show the separation of Hawkins County Airport, and Virginia...

Genotypic and phenotypic characterization of the Sdccag8Tn(sb-Tyr2161B.CA1C2Ove mouse model.

Directory of Open Access Journals (Sweden)

Katie Weihbrecht

Full Text Available Nephronophthisis-related ciliopathies (NPHP-RC are a group of disorders that present with end-stage renal failure in childhood/adolescence, kidney cysts, retinal degeneration, and cerebellar hypoplasia. One disorder that shares clinical features with NPHP-RC is Bardet-Biedl Syndrome (BBS. Serologically defined colon cancer antigen 8 (SDCCAG8; also known as NPHP10 and BBS16 is an NPHP gene that is also associated with BBS. To better understand the patho-mechanisms of NPHP and BBS caused by loss of SDCCAG8 function, we characterized an SDCCAG8 mouse model (Sdccag8Tn(sb-Tyr2161B.CA1C2Ove generated by Sleeping Beauty Transposon (SBT-mediated insertion mutagenesis. Consistent with the previously reported, independent SDCCAG8 mouse models, our mutant mice display pre-axial polydactyly in their hind limbs. In addition, we report patterning defects in the secondary palate, brain abnormalities, as well as neonatal lethality associated with developmental defects in the lung in our mouse model. The neonatal lethality phenotype is genetic background dependent and rescued by introducing 129S6/SvEvTac background. Genetic modifier(s responsible for this effect were mapped to a region between SNPs rs3714172 and rs3141832 on chromosome 11. While determining the precise genetic lesion in our mouse model, we found that SBT insertion resulted in a deletion of multiple exons from both Sdccag8 and its neighboring gene Akt3. We ascribe the patterning defects in the limb and the secondary palate as well as lung abnormalities to loss of SDCCAG8, while the developmental defects in the brain are likely due to the loss of AKT3. This mouse model may be useful to study features not observed in other SDCCAG8 models but cautions are needed in interpreting data.

77 FR 21505 - Proposed Amendment of Class D and E Airspace; Blountville, TN, and Revocation of Class E Airspace...

Science.gov (United States)

2012-04-10

... Airport, VA (Lat. 36[deg]41'14'' N., long. 82[deg]02'00'' W.) Rogersville, Hawkins County Airport, TN (Lat... Highlands Airport and within a 7-mile radius of Hawkins County Airport, and within 7 miles each side of Runway 07/25 centerline, extending from the 7-mile radius to 12 miles east of Hawkins County Airport...

Distinguishing Truncated and Normal MUC1 Glycoform Targeting from Tn-MUC1-Specific CAR T Cells

DEFF Research Database (Denmark)

Posey, Avery D; Clausen, Henrik; June, Carl H

2016-01-01

Genetically modified T cells expressing chimeric antigen receptors (CARs) demonstrate potent clinical antitumor effects in a variety of blood cancers. However, clinical activity in solid tumors has been disappointing and toxicity has been a serious concern (Lamers et al., 2013; Morgan et al., 2010......). We recently found that a CAR composed of a scFv antibody fragment specific for the Tn-glycoform of MUC1 had potent activity in preclinical models of blood cancer and adenocarcinoma (Posey et al., 2016)....

CARACTERIZACIÓN DE IGYS ANTI-LECTINA DE Salvia bogotensis Y SU APLICACIÓN EN ESTUDIOS CITOQUÍMICOS PARA LA DETECCIÓN DEL ANTÍGENO TN

Directory of Open Access Journals (Sweden)

Nohora Vega

2010-02-01

Full Text Available Las inmunoglobulinas aisladas de la yema de huevo (IgY son muy utilizadas actualmente en diversos campos de las ciencias biológicas, dadas sus ventajas frente a las IgG séricas de mamíferos. En un trabajo previo establecimos las condiciones de obtención de IgY dirigidas contra la lectinas de Salvia bogotensis; su utilización en estudios inmunocitoquímicos requiere conocer sus principales características moleculares y las condiciones para la interacción IgY-lectina. La lectina de Salvia bogotensis (SBoL reconoce específicamente el antígeno Tn, marcador tumoral en muchos tipos de cáncer, pero se requieren herramientas adicionales para evidenciar esta interacción a nivel celular. Dada la disponibilidad de IgY anti-lectina de S. bogotensis, se realizó este trabajo con el objeto de caracterizar molecularmente estas IgY y evaluar su utilización en estudios inmunocitoquímicos para la detección del antígeno Tn en células tumorales. A las IgY purificadas se les determinó su punto isoeléctrico, peso molecular y contenido de carbohidratos. Para establecer la especificidad de interacción IgY-SBoL se obtuvieron lectinas homólogas y heterólogas y se ensayaron por ELLSA. La detección del antígeno Tn en las líneas celulares MCF-7 y HeLa con la lectina y las IgYs marcadas con biotina o peroxidasa se realizó por CELISA e inmunocitoquímica. Los resultados mostraron que los anticuerpos IgY anti-SBoL son una herramienta de una alta sensibilidad para los ensayos de reconocimiento específico del antígeno Tn.

Tumor-associated Tn-MUC1 glycoform is internalized through the macrophage galactose-type C-type lectin and delivered to the HLA class I and II compartments in dendritic cells

DEFF Research Database (Denmark)

Napoletano, Chiara; Rughetti, Aurelia; Agervig Tarp, Mads P

2007-01-01

. This results in the expression of tumor-associated glycoforms and in MUC1 carrying the tumor-specific glycan Tn (GalNAcalpha1-O-Ser/Thr). Glycopeptides corresponding to three tandem repeats of MUC1, enzymatically glycosylated with 9 or 15 mol of GalNAc, were shown to specifically bind and to be internalized...... and ELISA done on subcellular fractions of iDCs showed that the Tn-MUC1 glycopeptides colocalized with HLA class I and II compartments after internalization. Importantly, although Tn-MUC1 recombinant protein was bound and internalized by MGL, the glycoprotein entered the HLA class II compartment......, but not the HLA class I pathway. These data indicate that MGL expressed on iDCs is an optimal receptor for the internalization of short GalNAcs carrying immunogens to be delivered into HLA class I and II compartments. Such glycopeptides therefore represent a new way of targeting the HLA class I and II pathways...

Fingerprints of field-induced Berezinskii–Kosterlitz–Thouless transition in quasi-two-dimensional S=1/2 Heisenberg magnets Cu(en)(H2O)2SO4 and Cu(tn)Cl2

International Nuclear Information System (INIS)

Baranová, Lucia; Orendáčová, Alžbeta; Čižmár, Erik; Tarasenko, Róbert; Tkáč, Vladimír; Orendáč, Martin; Feher, Alexander

2016-01-01

Organo-metallic compounds Cu(en)(H 2 O) 2 SO 4 (en=C 2 H 8 N 2 ) and Cu(tn)Cl 2 (tn=C 3 H 10 N 2 ) representing S=1/2 quasi-two-dimensional Heisenberg antiferromagnets with an effective intra-layer exchange coupling J/k B ≈3 K, have been examined by specific heat measurements at temperatures down to nominally 50 mK and magnetic fields up to 14 T. A comparative analysis of magnetic specific heat in zero magnetic field revealed nearly identical contribution of short-range magnetic correlations and significant differences were observed at lowest temperatures. A phase transition to long-range order was observed in Cu(en)(H 2 O) 2 SO 4 at T C =0.9 K while hidden in Cu(tn)Cl 2 . A response of both compounds to the application of magnetic field has rather universal features characteristic for a field-induced Berezinskii–Kosterlitz–Thouless transition theoretically predicted for ideal two-dimensional magnets. - Highlights: • Magnetic specific heat of Cu(en)(H 2 O) 2 SO 4 (1) and Cu(tn)Cl 2 (2) was analysed. • In zero magnetic field, (1) and (2) behave as quasi-two-dimensional magnets. • We observed universal thermodynamic response of (1) and (2) to applied field. • Features of field-induced Berezinskii–Kosterlitz–Thouless transition were detected.

[Establishment of Reference Value of Hs-cTnT in Sichuan Region and Its Diagnostic Value in Patients with Chest Pain].

Science.gov (United States)

Guo, Ying; Du, Xiang-Yang; Huang, Hua-Lan; Wang, Wei-Qing; Nie, Xin; Li, Gui-Xing

2017-11-01

To establish the reference value of high sensitive cardiac troponin T (hs-cTnT) and the efficiency of reference value in the diagnosis of chest pain. Volunteers from eight independent communities in Chengdu,Sichuan were selected with detailed records of physical examination,electrocardiogram,ultrasound examination. The level of hs-cTnT for healthy volunteers was tested to determine ninety-ninth percentile references according to sex and ages. 2 249 patients with chest pain in the emergency department of Western China Hospital from July 2009 to July 2014 were enrolled to measure the efficiency of reference value for diagnosing acute myocardial infarction (AMI). There were 1 305 volunteers included finally. Among them,the mean hs-cTnT level of male was 4.3 (3.2-5.9) ng/L,which was significantly higher than that of female 3.0 (3.0-3.1) ng/L ( P value was less than 0.01. The 99th percentile values of male were 10.8 ng/L,15.4 ng/L and 19.7 ng/L for values of female were 4.6 ng/L,8.9 ng/L,18.8 ng/L,respectively. There was no difference in sensitivity and specificity between the value we figured out and manufactures provided (14.0 ng/L) for thosevalue did not show diversity ( P >0.05) but the specificity and positive predictive value showed significant difference (male: 0.67 vs. 0.56 and 0.83 vs. 0.79, P value of patients<60 yr. should be 14.0 ng/L,while 20.0 ng/L for those patients≥60 yr.

Elaboration and quality control of the inoculum of the experimental vaccine Brucella S19-tn7-GFP for use in white animals and associated serological test for the detection of anti-GFP antibodies

International Nuclear Information System (INIS)

Salas Alfaro, Dariana

2014-01-01

The preparation of the inoculum of the experimental vaccine Brucella S19-Tn7-GFP is optimized for application in white animals. An associated serological test has allowed differentiating infected animals from those vaccinated with the experimental strain. The same bacteriological and biological properties of the B. abortus S19-Tn7-GFP strain have maintained in the parental vaccine strain S19 and is stable over time. A protocol for the inoculums of strain S19-Tn7-GFP is established for its preparation and use in white animals and quality control. The inoculum stability is evaluated through the simulation of conditions that can be presented in the transportation and application process in the field. An enzyme immunoassay ELISA is optimized for the detection of anti-GFP antibodies in cattle [es

42 CFR 60.14 - The insurance premium.

Science.gov (United States)

2010-10-01

... 42 Public Health 1 2010-10-01 2010-10-01 false The insurance premium. 60.14 Section 60.14 Public... LOAN PROGRAM The Loan § 60.14 The insurance premium. (a) General. (1) The Secretary insures each lender... lender an insurance premium. The insurance premium is due to the Secretary on the date of disbursement of...

Immunization of breast cancer patients using a synthetic sialyl-Tn glycoconjugate plus Detox adjuvant.

Science.gov (United States)

MacLean, G D; Reddish, M; Koganty, R R; Wong, T; Gandhi, S; Smolenski, M; Samuel, J; Nabholtz, J M; Longenecker, B M

1993-01-01

We have synthesized various formulations that have potential for active specific immunotherapy (ASI) of human cancers. Sialyl-Tn (STn) is a potentially important target structure for ASI because its expression on mucins is a strong, independent predictor of poor prognosis, suggesting that it may have functional significance in the metastatic process. In this first pilot study of synthetic sialyl-Tn hapten conjugated to keyhole limpet hemocyanin (STn-KLH), with Detox adjuvant, toxicity and humoral immunogenicity were assessed in 12 patients with metastatic breast cancer. Toxicity was minimal, restricted to local cutaneous reactions (apart from transient nausea and vomiting following single low-dose cyclophosphamide treatment). Using STn-conjugated human serum albumin in a solid-phase enzyme-linked immunosorbent assay, it was shown that all patients developed IgM and IgG specific for the synthetic STn hapten. Following immunization, most patients were shown to develop increased titres of complement-mediated cytotoxic antibodies, partially inhibited by synthetic STn hapten, but not by the related TF hapten. We also detected IgM and IgG antibodies reactive with natural STn determinants expressed on ovine submaxillary mucin, the STn specificity of this reactivity being confirmed by hapten inhibition. Evaluation of clinical efficacy in a small pilot study is difficult. Five patients are alive 12 or more months after entry, and another 4 patients are alive 6 or more months after entry into the study. All 3 patients with known widespread bulky disease progressed despite ASI, 2 having died from widespread cancer. Two patients had partial responses, each lasting 6 months. While several patients had disease stability for 3-10 months, 1 patient with pulmonary metastases remains stable 15 months after entry into the program.

Volcanic ash leaching as a means of tracing the environmental impact of the 2011 Grímsvötn eruption, Iceland.

Science.gov (United States)

Cabré, J; Aulinas, M; Rejas, M; Fernandez-Turiel, J L

2016-07-01

The Grímsvötn volcanic eruption, from 21 to 28 May, 2011, was the largest eruption of the Grímsvötn Volcanic System since 1873, with a Volcanic Explosivity Index (VEI) of magnitude 4. The main geochemical features of the potential environmental impact of the volcanic ash-water interaction were determined using two different leaching methods as proxies (batch and vertical flow-through column experiments). Ash consists of glass with minor amounts of plagioclase, clinopyroxene, diopside, olivine and iron sulphide; this latter mineral phase is very rare in juvenile ash. Ash grain morphology and size reflect the intense interaction of magma and water during eruption. Batch and column leaching tests in deionised water indicate that Na, K, Ca, Mg, Si, Cl, S and F had the highest potential geochemical fluxes to the environment. Release of various elements from volcanic ash took place immediately through dissolution of soluble salts from the ash surface. Element solubilities of Grímsvötn ash regarding bulk ash composition were <1 %. Combining the element solubilities and the total estimated mass of tephra (7.29 × 10(14) g), the total inputs of environmentally important elements were estimated to be 8.91 × 10(9) g Ca, 7.02 × 10(9) g S, 1.10 × 10(9) g Cl, 9.91 × 10(8) g Mg, 9.91 × 10(8) g Fe and 1.45 × 10(8) g P The potential environmental problems were mainly associated with the release of F (5.19 × 10(9) g).

Effect of the polymer concentration on the ON/OFF states of a TN-LCD: polyvinyl alcohol vs. soy lecithin

Science.gov (United States)

de Coss Martinez, Romeo; Gonzalez Murguia, Jose Luis

2011-03-01

In this work we study the response of a Twisted Nematic Liquid Crystal Display (TN-LCD) by varying both the concentration and the polymer used for the microgroove. We compare the performance of two polymers: polyvinyl alcohol and soy lecithin. In particular, the light transmission for the ON/OFF states is evaluated. The polyvinyl alcohol is a polymer widely used in LCDs while lecithin soy is a natural polymer.

20 CFR 601.4 - Certification for tax credit.

Science.gov (United States)

2010-04-01

... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false Certification for tax credit. 601.4 Section 601.4 Employees' Benefits EMPLOYMENT AND TRAINING ADMINISTRATION, DEPARTMENT OF LABOR ADMINISTRATIVE... and Additional Tax Credit and Grant Purposes § 601.4 Certification for tax credit. (a) Within 30 days...

Characterization and protective property of Brucella abortus cydC and looP mutants.

Science.gov (United States)

Truong, Quang Lam; Cho, Youngjae; Barate, Abhijit Kashinath; Kim, Suk; Hahn, Tae-Wook

2014-11-01

Brucella abortus readily multiplies in professional or nonprofessional phagocytes in vitro and is highly virulent in mice. Isogenic mutants of B. abortus biovar 1 strain IVKB9007 lacking the ATP/GDP-binding protein motif A (P-loop) (named looP; designated here the IVKB9007 looP::Tn5 mutant) and the ATP-binding/permease protein (cydC; designated here the IVKB9007 cydC::Tn5 mutant) were identified and characterized by transposon mutagenesis using the mini-Tn5Km2 transposon. Both mutants were found to be virtually incapable of intracellular replication in both murine macrophages (RAW264.7) and the HeLa cell line, and their virulence was significantly impaired in BALB/c mice. Respective complementation of the IVKB9007 looP::Tn5 and IVKB9007 cydC::Tn5 mutants restored their ability to survive in vitro and in vivo to a level comparable with that of the wild type. These findings indicate that the cydC and looP genes play important roles in the virulence of B. abortus. In addition, intraperitoneal immunization of mice with a dose of the live IVKB9007 looP::Tn5 and IVKB9007 cydC::Tn5 mutants provided a high degree of protection against challenge with pathogenic B. abortus strain 544. Both mutants should be evaluated further as a live attenuated vaccine against bovine brucellosis for their ability to stimulate a protective immune response. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

In vivo study of the HC-TN strain of hepatitis C virus recovered from a patient with fulminant hepatitis: RNA transcripts of a molecular clone (pHC-TN) are infectious in chimpanzees but not in Huh7.5 cells

DEFF Research Database (Denmark)

Sakai, Akito; Takikawa, Shingo; Thimme, Robert

2007-01-01

Both viral and host factors are thought to influence the pathogenesis of hepatitis C virus (HCV) infection. We studied strain HC-TN (genotype 1a), which caused fulminant hepatic failure in a patient and, subsequently, severe hepatitis in a chimpanzee (CH1422), to analyze the relationship between...... persistently infected. CH1579 and CH1581, despite their differing outcomes, both developed significant intrahepatic cellular immune responses, but not antibodies to the envelope glycoproteins or neutralizing antibodies, during the acute infection. We analyzed the polyprotein sequences of virus recovered...

Two types of phytases (histidine acid phytase and β-propeller phytase) in Serratia sp. TN49 from the gut of Batocera horsfieldi (coleoptera) larvae.

Science.gov (United States)

Zhang, Rui; Yang, Peilong; Huang, Huoqing; Shi, Pengjun; Yuan, Tiezheng; Yao, Bin

2011-11-01

Microbial phytases play a major role in the mineralization of organic phosphorous, especially in symbiotic plants and animals. In this study, we identified two types of phytases in Serratia sp. TN49 that was harbored in the gut of Batocera horsfieldi (Coleoptera) larvae. The two phytases, an acidic histidine acid phosphatase (PhyH49) and an alkaline β-propeller phytase (PhyB49), shared low identities with known phytases (61% at most). PhyH49 and PhyB49 produced in Escherichia coli exhibited maximal activities at pH 5.0 (60°C) and pH 7.5-8.0 (45°C), respectively, and are complementary in phytate degradation over the pH range 2.0-9.0. Serratia sp. TN49 harboring both PhyH49 and PhyB49 might make it more adaptive to environment change, corresponding to the evolution trend of microorganism.

Discrimination between Native and Tn6010-Associated oqxAB in Klebsiella spp., Raoultella spp., and other Enterobacteriaceae by Using a Two-Step Strategy

DEFF Research Database (Denmark)

Guillard, Thomas; Lebreil, Anne-Laure; Hansen, Lars Hestbjerg

2015-01-01

ABSTRACT We developed a two-step PCR-based strategy to detect genes encoding OqxAB, allowing a specific assignment of Tn6010-associated oqxAB in Enterobacteriaceae. Chromosomal location in this setup was confirmed by hybridization with I-CeuI-restricted genomes. This approach led us to find...

Loading and transport of high-active waste (HAW) with the TN85 flask in 2008

International Nuclear Information System (INIS)

Rys, Michael; Horn, Thomas; Graf, Wilhelm; Bonface, Jean-Michael

2009-01-01

As a part of the operation of nuclear power plants, it is essential to safely manage the radioactive waste. With new developments in science and technology, it is a dynamic process to adapt procedures, equipment and flasks to be used in the future. This is a task for specialists - a task for GNS Gesellschaft fuer Nuklear-Service mbH and for TN International. Until 1994 reprocessing of spent fuel from German nuclear power plants was mandatory for the Utilities (EVU) in Germany. Basis for the reprocessing was the German Atomic Act. The German Utilities concluded contracts on reprocessing with Compagnie Generale des Matieres Nucleaires (COGEMA, now AREVA NC) in France and British Nuclear Fuels plc (BNFL, now INS) in England. The total amount to be reprocessed comes to 5309 t HM contracted to AREVA NC and 768 t HM contracted to INS. The waste generated from reprocessing - or an equivalent amount of radioactive material - has to be returned to the country of origin. In 1979 already an exchange of notes took place between the German and the French government with the obligation of both sides to enable and support the return of reprocessing residues or equivalents. The return of high-active waste (HAW) from France has started in 1996 with the first attribution of 28 glass canisters (one flask) to German Utilities by AREVA NC. Until 2007, 75 flasks loaded with vitrified residue (VR) canisters have been transported to Gorleben. For these transports CASTOR registered HAW 20/28 CG flasks have been used. This presentation will give some background information about the last HAW transport in 2008 with the new flask generation of the type TN85. It will also describe the assembly of the new flask, the preparation of the flask for the loading campaign as well as the loading procedure. (orig.)

Loading and transport of high-active waste (HAW) with the TN85 flask in 2008

Energy Technology Data Exchange (ETDEWEB)

Rys, Michael; Horn, Thomas; Graf, Wilhelm [GNS Gesellschaft fuer Nuklear-Service mbH (Germany); Bonface, Jean-Michael [TN International, Montigny-le-Bretonneux (France)

2009-07-01

As a part of the operation of nuclear power plants, it is essential to safely manage the radioactive waste. With new developments in science and technology, it is a dynamic process to adapt procedures, equipment and flasks to be used in the future. This is a task for specialists - a task for GNS Gesellschaft fuer Nuklear-Service mbH and for TN International. Until 1994 reprocessing of spent fuel from German nuclear power plants was mandatory for the Utilities (EVU) in Germany. Basis for the reprocessing was the German Atomic Act. The German Utilities concluded contracts on reprocessing with Compagnie Generale des Matieres Nucleaires (COGEMA, now AREVA NC) in France and British Nuclear Fuels plc (BNFL, now INS) in England. The total amount to be reprocessed comes to 5309 t HM contracted to AREVA NC and 768 t HM contracted to INS. The waste generated from reprocessing - or an equivalent amount of radioactive material - has to be returned to the country of origin. In 1979 already an exchange of notes took place between the German and the French government with the obligation of both sides to enable and support the return of reprocessing residues or equivalents. The return of high-active waste (HAW) from France has started in 1996 with the first attribution of 28 glass canisters (one flask) to German Utilities by AREVA NC. Until 2007, 75 flasks loaded with vitrified residue (VR) canisters have been transported to Gorleben. For these transports CASTOR {sup registered} HAW 20/28 CG flasks have been used. This presentation will give some background information about the last HAW transport in 2008 with the new flask generation of the type TN85. It will also describe the assembly of the new flask, the preparation of the flask for the loading campaign as well as the loading procedure. (orig.)

Characterization of the Pathogenicity of Streptococcus intermedius TYG1620 Isolated from a Human Brain Abscess Based on the Complete Genome Sequence with Transcriptome Analysis and Transposon Mutagenesis in a Murine Subcutaneous Abscess Model.

Science.gov (United States)

Hasegawa, Noriko; Sekizuka, Tsuyoshi; Sugi, Yutaka; Kawakami, Nobuhiro; Ogasawara, Yumiko; Kato, Kengo; Yamashita, Akifumi; Takeuchi, Fumihiko; Kuroda, Makoto

2017-02-01

Streptococcus intermedius is known to cause periodontitis and pyogenic infections in the brain and liver. Here we report the complete genome sequence of strain TYG1620 (genome size, 2,006,877 bp; GC content, 37.6%; 2,020 predicted open reading frames [ORFs]) isolated from a brain abscess in an infant. Comparative analysis of S. intermedius genome sequences suggested that TYG1620 carries a notable type VII secretion system (T7SS), two long repeat regions, and 19 ORFs for cell wall-anchored proteins (CWAPs). To elucidate the genes responsible for the pathogenicity of TYG1620, transcriptome analysis was performed in a murine subcutaneous abscess model. The results suggest that the levels of expression of small hypothetical proteins similar to phenol-soluble modulin β1 (PSMβ1), a staphylococcal virulence factor, significantly increased in the abscess model. In addition, an experiment in a murine subcutaneous abscess model with random transposon (Tn) mutant attenuation suggested that Tn mutants with mutations in 212 ORFs in the Tn mutant library were attenuated in the murine abscess model (629 ORFs were disrupted in total); the 212 ORFs are putatively essential for abscess formation. Transcriptome analysis identified 37 ORFs, including paralogs of the T7SS and a putative glucan-binding CWAP in long repeat regions, to be upregulated and attenuated in vivo This study provides a comprehensive characterization of S. intermedius pathogenicity based on the complete genome sequence and a murine subcutaneous abscess model with transcriptome and Tn mutagenesis, leading to the identification of pivotal targets for vaccines or antimicrobial agents for the control of S. intermedius infections. Copyright © 2017 American Society for Microbiology.

Sbírka rukopisných zlomků ve Státním okresním archivu v Kutné Hoře

Czech Academy of Sciences Publication Activity Database

Brodský, Pavel; Hradilová, Marta; Hrdinová, Martina; Petr, Stanislav; Tošnerová, Marie

2013-01-01

Roč. 43, č. 0 (2013), s. 207-248 ISSN 0585-5691 R&D Projects: GA ČR GAP405/10/0027 Institutional support: RVO:67985921 Keywords : codicology * manuscripts fragments * Státní okresní archiv Kutná Hora Subject RIV: AB - History

Evolution of Regions Containing Antibiotic Resistance Genes in FII-2-FIB-1 ColV-Colla Virulence Plasmids.

Science.gov (United States)

Moran, Robert A; Hall, Ruth M

2018-05-01

Three ColV virulence plasmids carrying antibiotic resistance genes were assembled from draft genome sequences of commensal ST95, ST131, and ST2705 Escherichia coli isolates from healthy Australians. Plasmids pCERC4, pCERC5, and pCERC9 include almost identical backbones containing FII-2 and FIB-1 replicons and the conserved ColV virulence region with an additional ColIa determinant. Only pCERC5 includes a complete, uninterrupted F-like transfer region and was able to conjugate. pCERC5 and pCERC9 contain Tn1721, carrying the tet(A) tetracycline resistance determinant in the same location, with Tn2 (bla TEM ; ampicillin resistance) interrupting the Tn1721 in pCERC5. pCERC4 has a Tn1721/Tn21 hybrid transposon carrying dfrA5 (trimethoprim resistance) and sul1 (sulfamethoxazole resistance) in a class 1 integron. Four FII-2:FIB-1 ColV-ColIa plasmids in the GenBank nucleotide database have a related transposon in the same position, but an IS26 has reshaped the resistance gene region, deleting 2,069 bp of the integron 3'-CS, including sul1, and serving as a target for IS26 translocatable units containing bla TEM , sul2 and strAB (streptomycin resistance), or aphA1 (kanamycin/neomycin resistance). Another ColV-ColIa plasmid containing a related resistance gene region has lost the FII replicon and acquired a unique transfer region via recombination within the resistance region and at oriT. Eighteen further complete ColV plasmid sequences in GenBank contained FIB-1, but the FII replicons were of three types, FII-24, FII-18, and a variant of FII-36.

Effects of short-term tocopherol (T) feeding on structure-localized protein tyrosine nitration (pTN) patterns of mitochondrial ATPase following endotoxin (LPS) challenge in beef calves.

Science.gov (United States)

Mitochondrial ATPase/Complex-V (MCV) is an electron transport chain (ETC) component needed for ATP synthesis. The ETC, exquisitely sensitive to proinflammatory mediators (PIM), generates oxynitrogen reactants leading to pTN formation as mitochondrial membrane leakage occurs. Immunohistochemical loca...

HTR-TN a European network for the development of HTR technology

International Nuclear Information System (INIS)

Von Lensa, W.

2001-01-01

A network called High-temperature reactor technology network (HTR-TN) has been created at a European level to coordinate works and knowledge on the subject with a long-term perspective and to serve as a channel for international collaboration. An analysis confirmed that the obvious economic penalty of HTR due to its low density power could be compensated by the combination of recent advances that may completely change the positioning of HTR on the energy market: -) the modular concept allowed to get a reactor free from core melt risk without intervention of any active safety system, implying a drastic simplification of the design of the reactor and the safety systems as well as a standardisation and potential for shop fabrication in series; -) the development of gas turbines, the efficiency of which increased, in 10 years, from 35% till 50% and more, enabling to consider suppression of the secondary system; -) the ultra high burn-up potential of HTR fuel and the possibility for direct disposal of spent HTR fuel elements that may reduce cost of the fuel cycle and contribute to the reduction of civil and military plutonium stockpiles. (A.C.)

Fingerprints of field-induced Berezinskii–Kosterlitz–Thouless transition in quasi-two-dimensional S=1/2 Heisenberg magnets Cu(en)(H{sub 2}O){sub 2}SO{sub 4} and Cu(tn)Cl{sub 2}

Energy Technology Data Exchange (ETDEWEB)

Baranová, Lucia [Civil Engineering Faculty, Department of Applied Mathematics, Technical University of Košice, Vysokoškolská 4 SK-042 00, Košice (Slovakia); Orendáčová, Alžbeta, E-mail: alzbeta.orendacova@upjs.sk [Center of Low Temperature Physics, Faculty of Science, P. J. Šafárik University, Park Angelinum 9 SK-041 54, Košice (Slovakia); Čižmár, Erik [Center of Low Temperature Physics, Faculty of Science, P. J. Šafárik University, Park Angelinum 9 SK-041 54, Košice (Slovakia); Tarasenko, Róbert; Tkáč, Vladimír [Center of Low Temperature Physics, Faculty of Science, P. J. Šafárik University, Park Angelinum 9 SK-041 54, Košice (Slovakia); Charles University, Faculty of Mathematics and Physics, Ke Karlovu 5 12116, Prague (Czech Republic); Orendáč, Martin; Feher, Alexander [Center of Low Temperature Physics, Faculty of Science, P. J. Šafárik University, Park Angelinum 9 SK-041 54, Košice (Slovakia)

2016-04-15

Organo-metallic compounds Cu(en)(H{sub 2}O){sub 2}SO{sub 4} (en=C{sub 2}H{sub 8}N{sub 2}) and Cu(tn)Cl{sub 2} (tn=C{sub 3}H{sub 10}N{sub 2}) representing S=1/2 quasi-two-dimensional Heisenberg antiferromagnets with an effective intra-layer exchange coupling J/k{sub B}≈3 K, have been examined by specific heat measurements at temperatures down to nominally 50 mK and magnetic fields up to 14 T. A comparative analysis of magnetic specific heat in zero magnetic field revealed nearly identical contribution of short-range magnetic correlations and significant differences were observed at lowest temperatures. A phase transition to long-range order was observed in Cu(en)(H{sub 2}O){sub 2}SO{sub 4} at T{sub C}=0.9 K while hidden in Cu(tn)Cl{sub 2}. A response of both compounds to the application of magnetic field has rather universal features characteristic for a field-induced Berezinskii–Kosterlitz–Thouless transition theoretically predicted for ideal two-dimensional magnets. - Highlights: • Magnetic specific heat of Cu(en)(H{sub 2}O){sub 2}SO{sub 4} (1) and Cu(tn)Cl{sub 2} (2) was analysed. • In zero magnetic field, (1) and (2) behave as quasi-two-dimensional magnets. • We observed universal thermodynamic response of (1) and (2) to applied field. • Features of field-induced Berezinskii–Kosterlitz–Thouless transition were detected.

Rethinking Modal Gender in the Context of the Universe of Tn-Types: Definitions and Mathematical Models for Tonicity and Phonicity

Directory of Open Access Journals (Sweden)

Marcus Alessi Bittencourt

2016-07-01

Full Text Available An indisputable cornerstone of the Western music tradition, the dialectic opposition between the major and minor grammatical modal genders has always been present in the imagination of musicians and music theorists for centuries. Such dialectics of opposition is especially important in the context of nineteenth-century harmonic dualism, with its ideas of tonicity and phonicity. These concepts serve as the main foundation for the way harmonic dualism conceives the major and minor worlds: two worlds with equivalent rights and properties, but with opposed polarities. This paper presents a redefinition of the terms tonicity and phonicity, translating those concepts to the context of post-tonal music theory. The terminologies of generatrix, tonicity, root, phonicity, vertex, and azimuth are explained in this paper, followed by propositions of mathematical models for those concepts, which spring from Richard Parncutt’s root-salience model for pitch-class sets. In order to demonstrate the possibilities of using modal gender as a criterion for the study and classification of the universe of Tn-types, we will present a taxonomy of the 351 transpositional set types, which comprises the categories of tonic (major, phonic (minor and neutral (genderless. In addition, there will be a small discussion on the effect of set symmetries and set asymmetries on the tonic/phonic properties of a Tn-type.

Shielding calculations with SCALE/MAVRIC and comparison with measurements for the TN85 cask with vitrified high level radioactive waste

Science.gov (United States)

Thiele, Holger; Börst, Frank-Michael

2017-09-01

A series of dose rate/spectra measurements in the German interim storage facility Gorleben was carried out at a TN85 cask in April 2009. This type of cask is used for the transport and interim storage of vitrified high level radioactive waste (HAW) from reprocessing. The aim of this work is to assess the shielding component MAVRIC of the SCALE code system with these measurements for the use in the German Bundesamt für Kerntechnische Entsorgungssicherheit (BfE).

Statement of Roy F. Pruett, Mayor, City of Oak Ridge, TN

International Nuclear Information System (INIS)

Anon.

1987-01-01

Thank you very much, Mr. Chairman, and Members of the Committee. My name is Roy Pruett, mayor of the city of Oak Ridge, TN, and an executive committee member on the Clinch River MRS task force. It is my pleasure to be here today to testify before this distinguished Committee and present the findings of the Clinch River task force as they relate to subtitle C of the Nuclear Waste Policy Act of 1982, and to the Department of Energy proposal on the Monitored Retrievable Storage facility, the MRS. I represent the city of Oak Ridge, which has been selected for the primary and the first alternative for the siting of an integrated MRS facility for the preparation and packaging of high-level nuclear waste and spent reactor fuel. Of civic and governmental leaders, we concluded that an MRS could safely be built and operated in Oak Ridge. We further concluded, however, that the facility would not be generally perceived as being safe unless the recommendations of the task force were adopted to address concerns and help mitigate impact. Indeed the MRS would not be viewed as a net economic benefit to the site's community, the region, or the State of Tennessee without such appropriate conditions

Formation of ethanol from lactose by Zymomonas mobilis

Energy Technology Data Exchange (ETDEWEB)

Goodman, A E; Strzelecki, A T; Rogers, P L

1984-01-01

The lactose transposon Tn951 on the IncP plasmid RP1, was introduced into a strain of Zymomonas mobilis. The lac Z gene was expressed and US -galactosidase was produced. The synthesis of US -galactosidase was induced by either isopropyl-US -D-thiogalactopyranoside or lactose, and was not sensitive to catabolite repression. Cells of Zymomonas mobilis containing Tn951 were unable to form colonies on lactose plates, but in liquid medium produced ethanol from lactose as the sole carbon source. The original strain of Zymomonas mobilis without Tn951 produced little or no US -galactosidase and was unable to produce ethanol from lactose in liquid medium.

Ionothermal synthesis of discrete supertetrahedral Tn (n = 4, 5) clusters with tunable components, band gaps, and fluorescence properties.

Science.gov (United States)

Yang, Dan-Dan; Li, Wei; Xiong, Wei-Wei; Li, Jian-Rong; Huang, Xiao-Ying

2018-05-01

The preparation of crystalline molecularly supertetrahedral Tn clusters with variable sizes and components is of vital importance for the fundamental study of their physicochemical properties. However, setting up an efficient method to stabilize large discrete Tn clusters is a challenge due to their high negative charges and polymerization nature. In this work, we report on the ionothermal synthesis of three discrete T4 cluster compounds, namely [Bmmim]5[(CH3)2NH2]4[NH4][M4In16S31(SH)4]·6H2O (M = Mn (1), Zn (2), Cd (3), Bmmim = 1-buty-2,3-dimethyl-imidazolium), and four discrete T5 cluster compounds, namely [Bmmim]10[NH4]3[Cu5Ga30-xInxS52(SH)4] (x = 6.6 (5), 14.5 (6), 23.8 (7), and 30 (8)). The compound [Bmmim]10[NH4]3[Cu5Ga30S52(SH)4] (4) previously reported by us features a discrete T5 cluster. The steep UV-Vis absorption edges indicate band gaps of 2.20 eV for 1, 2.64 eV for 2, 2.69 eV for 3, 3.04 eV for 4, 2.65 eV for 5, 2.48 eV for 6, 2.32 eV for 7, and 2.30 eV for 8. The compositions of T5 clusters could be varied with the ratios of Ga : In in the starting reagents, providing an opportunity to systematically control the band gaps and fluorescence performances of T5 cluster-based compounds. This research might advance the understanding of the ionothermal preparation and functionality tuning of crystalline chalcogenides.

[Differential expression of DTSsa4 Tc1-like transposons in closely related populations of Baikal ciscoes].

Science.gov (United States)

Bychenko, O S; Sukhanova, L V; Azhikina, T L; Sverdlov, E D

2009-01-01

Two representatives of Baikal ciscoes - lake cisco and omul - diverged from a common ancestor as recently as 10-20 thousand years ago. We have found an increasing expression level of DTSsa4 Tc1-like DNA transposons in cisco and omul brains. The mapping of the sequences of these transposons from Salmo salar and Danio rerio genomes has shown that in some cases, these transposons are located in the 5' and 3' regions, as well as in the promoter regions of various genes. Probably, Tc1-like transposons affect the activity of neighboring genes, providing the adaptive divergence of the cisco population.

Statement of Kenneth E. Yager, County Executive, Roane County, TN

International Nuclear Information System (INIS)

Anon.

1987-01-01

I am Ken Yager, I am the county executive for Roane County, TN which is the preferred site for the proposed Monitored Retrieval Storage facility, or the MRS. and I am submitting into the record additional written testimony which was written largely by the Honorable Robert Peele, who is sitting here, who is a member of the Roane County Board of Commissioners, our local legislative body. The premise of my remarks is that the Department of Energy proposal for the MRS in our county in unacceptable unless the Congress and the Department of Energy are to be willing partners with the local communities. Our considerations have been limited to: one, can an MRS be operated safely, and if so under what conditions; and what would be the impacts of the facility and how might they be mitigated. Our recommendations on those two points lead me to my earlier comment about being partners with the Congress, because our recommendations can be summed up simply, Mr. Chairman, as local control. We are exceedingly concerned about the prospect of being trapped in a national squeeze play, caught between a recognized need for a way to deal with nuclear waste and intense political efforts to put that problem in somebody else's back yard. And if we are to be that back yard, we would ask that the Congress and the Department of Energy recognize our stake in this national effort. We must have an equal role in the process

Postoperative change of anti-Thomsen-Friedenreich and Tn IgG level: the follow-up study of gastrointestinal cancer patients.

Science.gov (United States)

Smorodin, Eugeniy-P; Kurtenkov, Oleg-A; Sergeyev, Boris-L; Kodar, Kristel-E; Chuzmarov, Valentin-I; Afanasyev, Vladimir-P

2008-07-21

To study the influence of tumor removal on the serum level of IgG antibodies to tumor-associated Thomsen-Friedenreich (TF), Tn carbohydrate epitopes and xenogeneic alphaGal, and to elucidate on the change of the level during the follow-up as well as its association with the stage and morphology of the tumor and the values of blood parameters in gastrointestinal cancer. Sixty patients with gastric cancer and 34 patients with colorectal cancer in stages I-IV without distant metastases were subjected to follow-up. The level of antibodies in serum was determined by the enzyme-linked immunosorbent assay (ELISA) using synthetic polyacrylamide (PAA) glycoconjugates. Biochemical and haematological analyses were performed using automated equipment. In gastrointestinal cancer, the TF antibody level was found to have elevated significantly after the removal of G3 tumors as compared with the preoperative level (u = 278.5, P gastric cancer patients (sign test, 20 vs 8, P cancer, the elevated postoperative level of TF, Tn and alphaGal antibodies was noted in most patients with G3 tumors (sign test, 22 vs 5, P stage I, whose mean anti-TF IgG level remained above the cut-off value (1.26), was significantly higher than that of patients with the cancer in stages III-IV (c2 = 4.71, gastric cancer; c2 = 4.11, gastrointestinal cancer; P stage but remains low in its terminal stages. The stage- and morphology-dependent immunosuppression affects the TF-antibody response and may be one of the reasons for unresponsiveness to the immunization with TF-antigens.

Tn5-induced pBS286 plasmid mutations blocking early stages of napthalene oxidation

International Nuclear Information System (INIS)

Kosheleva, I.A.; Tsoi, T.V.; Ivashina, T.V.; Selifonov, S.A.; Starovoitov, I.I.; Boronin, A.M.

1988-01-01

The authors present data on the further analysis of the structural and functional organization of the nah region of plasmid pBS286 controlling the constitutive oxidation of naphthalene by Pseudomonas putida cells. They have studied Tn5-induced mutations blocking early stages of naphthalene oxidation. They present and discuss data providing evidence that, in contrast to plasmid NAH7, the mechanism of regulation of the nahl operon of plasmid NPL-1, the parent plasmid of plasmid pBS286, with inducible synthesis of naphthalene dioxygenase can include elements of a negative control with participation of the regulatory locus R, located proximal to the structural nah genes and closely linked to or overlapped by the inverted control DNA segment (4.2 kb). They also present data on the possibility of regulation of the activity of the catechol-splitting meta-pathway genes with the participation of products of early stages of naphthalene oxidation

Thermal-hydraulic analyses of the TN-24P cask loaded with consolidated and unconsolidated spent nuclear fuel

International Nuclear Information System (INIS)

Michener, T.E.; McKinnon, M.A.; Rector, D.R.; Creer, J.M.

1989-06-01

This paper presents the results of comparisons of COBRA-SFS (spent fuel storage) temperature predictions with experimental data from the TN-24P (Transnuclear) spent fuel storage cask loaded with unconsolidated and consolidated spent PWR fuel. Peak cladding temperature predictions using the COBRA-SFS code are compared with test data and predicted axial and radial temperature distributions are compared with measured temperature profiles. The pre-test accuracy of the COBRA-SFS code in predicting temperature distributions is discussed, along with the effect of post-test model improvements on temperature predictions. This paper also briefly describes the COBRA-SFS code, which is designed to accurately predict flow and temperature distributions in spent nuclear fuel storage and transportation systems. 6 refs., 14 figs

Evaluation of terrestrial microcosms for assessing the fate and effects of genetically engineered microorganisms on ecological processes

Energy Technology Data Exchange (ETDEWEB)

Fredrickson, J.K.; Bentjen, S.A.; Bolton, H. Jr.; Li, S.W.; Ligotke, M.W.; McFadden, K.M.; Van Voris, P.

1989-04-01

This project evaluates and modifies the existing US Environmental Protection Agency's Office of Pesticides and Toxic Substances (EPA/OPTS) terrestrial microcosm test system and test protocols such that they can be used to determine the environmental fate and ecological hazards of genetically engineered microorganisms (GEMs). The intact soil-core microcosm represents terrestrial ecosystems, and when coupled with appropriate test protocols, such microcosms may be appropriate to define and limit risks associated with the intentional release of GEMs. The terrestrial microcosm test system was used to investigate the survival and transport of two model GEMs (Azospirillum lipoferum and Pseudomonas sp. Tn5 mutants) to various trophic levels and niches and through intact soil cores. Subsequent effects on nutrient cycling and displacement of indigenous microorganisms were evaluated. The model organisms were a diazotrophic root-colonizing bacterium (A. lipoferum) and a wheat root growth-inhibiting rhizobacterium (Pseudomonas sp.). The transposable element Tn5 was used as a genetic marker for both microorganisms in two separate experiments. The organisms were subjected to transposon mutagenesis using a broad host-range-mobilizable suicide plasmid. The transposon Tn5 conferred levels of kanamycin resistance up to 500 ..mu..g/ml (Pseudomonas sp.), which allowed for selection of the bacteria from environmental samples. The presence of Tn5 DNA in the genome of the model GEMs also allowed the use of Tn5 gene probes to confirm and enumerate the microorganisms in different samples from the microcosms. Two types of root growth-inhibiting Pseudomonas sp. Tn5 mutants were obtained and used in microcosm studies: those that lacked the ability to inhibit either wheat root growth or the growth of other microorganisms in vitro (tox/sup /minus//) and those which retained these properties (tox/sup +/). 53 refs., 7 figs., 6 tabs.

The Identification of Genes Important in Pseudomonas syringae pv. phaseolicola Plant Colonisation Using In Vitro Screening of Transposon Libraries.

Directory of Open Access Journals (Sweden)

Bharani Manoharan

Full Text Available The bacterial plant pathogen Pseudomonas syringae pv. phaseolicola (Pph colonises the surface of common bean plants before moving into the interior of plant tissue, via wounds and stomata. In the intercellular spaces the pathogen proliferates in the apoplastic fluid and forms microcolonies (biofilms around plant cells. If the pathogen can suppress the plant's natural resistance response, it will cause halo blight disease. The process of resistance suppression is fairly well understood, but the mechanisms used by the pathogen in colonisation are less clear. We hypothesised that we could apply in vitro genetic screens to look for changes in motility, colony formation, and adhesion, which are proxies for infection, microcolony formation and cell adhesion. We made transposon (Tn mutant libraries of Pph strains 1448A and 1302A and found 106/1920 mutants exhibited alterations in colony morphology, motility and biofilm formation. Identification of the insertion point of the Tn identified within the genome highlighted, as expected, a number of altered motility mutants bearing mutations in genes encoding various parts of the flagellum. Genes involved in nutrient biosynthesis, membrane associated proteins, and a number of conserved hypothetical protein (CHP genes were also identified. A mutation of one CHP gene caused a positive increase in in planta bacterial growth. This rapid and inexpensive screening method allows the discovery of genes important for in vitro traits that can be correlated to roles in the plant interaction.

TN trademark FLEX: a new generation of fluorocarbon o-rings developed by COGEMA logistics with enhanced characteristics at low temperature (-40 C)

International Nuclear Information System (INIS)

Issard, H.; Andre, R.

2004-01-01

Three main types of elastomers are used for the sealing of radioactive material transport casks with elastomeric gaskets: EPDM, fluorocarbons type Viton registered (standard designation: FKM) and silicon rubbers. Each rubber has specific characteristics in terms of temperature range, permeability, coefficient of expansion.. For the casks where high temperatures can be reached (200 C in continuous using), FKM gaskets are generally used. The problem is that this type of gasket does not guarantee the leaktightness at -40 C, which is a regulatory requirement. Two solutions are generally used: to specify a minimum heat load or a minimum ambient temperature. The direct consequence is that it is impossible to get B(U) approvals on the new concepts when FKM gaskets are used but only B(M) approvals, which generate significant additional justification costs (multiple submittals of Safety Analysis Reports, calculation of the minimum heat load or of the minimum ambient temperature..). Thus, it is important to develop gaskets with the same performance as FKM gaskets at high temperature but with enhanced performance at low temperature (and mainly, which guarantee the leaktightness at -40 C). COGEMA LOGISTICS has qualified a new generation of fluorocarbon O-rings (TN trademark FLEX gaskets) which can be used in continuous service on a -47 C/+200 C temperature range. TN trademark FLEX gaskets will be implemented on new casks designs

Molecular characterisation of Vibrio cholerae O1 strains carrying an SXT/R391-like element from cholera outbreaks in Kenya: 1994-2007

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Goddeeris Bruno M

2009-12-01

Full Text Available Abstract Background Over the last decade, cholera outbreaks in parts of Kenya have become common. Although a number of recent studies describe the epidemiology of cholera in Kenya, there is paucity of information concerning the diversity and occurrence of mobile genetic elements in Vibrio cholerae strains implicated in these outbreaks. A total of 65 Vibrio cholerae O1 El Tor serotype Inaba isolated between 1994 and 2007 from various outbreaks in Kenya were investigated for mobile genetic elements including integrons, transposons, the integrating conjugative elements (ICEs, conjugative plasmids and for their genotypic relatedness. Results All the strains were haemolytic on 5% sheep blood and positive for the Vibrio cholerae El Tor-specific haemolysin toxin gene (hylA by PCR. They all contained strB, sulII, floR and the dfrA1 genes encoding resistance to streptomycin, sulfamethoxazole, chloramphenicol and trimethoprim respectively. These genes, together with an ICE belonging to the SXT/R391 family were transferable to the rifampicin-resistant E. coli C600 en bloc. All the strains were negative for integron class 1, 2 and 3 and for transposase gene of transposon Tn7 but were positive for integron class 4 and the trpM gene of transposon Tn21. No plasmids were isolated from any of the 65 strains. All the strains were also positive for all V. cholera El Tor pathogenic genes except the NAG- specific heat-stable toxin (st gene. None of the strains were positive for virulence genes associated with the V. cholerae classical biotype. All the strains were positive for El Tor-specific CTXphi bacteriophage rstrR repressor gene (CTXETΦ but negative for the Classical, Calcutta, and the Environmental repressor types. Pulse Field Gel Electrophoresis (PFGE showed that regardless of the year of isolation, all the strains bearing the SXT element were clonally related. Conclusions This study demonstrates that the V. cholerae O1 strains carrying an SXT/R391-like

Insights into the transposable mobilome of Paracoccus spp. (Alphaproteobacteria).

Science.gov (United States)

Dziewit, Lukasz; Baj, Jadwiga; Szuplewska, Magdalena; Maj, Anna; Tabin, Mateusz; Czyzkowska, Anna; Skrzypczyk, Grazyna; Adamczuk, Marcin; Sitarek, Tomasz; Stawinski, Piotr; Tudek, Agnieszka; Wanasz, Katarzyna; Wardal, Ewa; Piechucka, Ewa; Bartosik, Dariusz

2012-01-01

Several trap plasmids (enabling positive selection of transposition events) were used to identify a pool of functional transposable elements (TEs) residing in bacteria of the genus Paracoccus (Alphaproteobacteria). Complex analysis of 25 strains representing 20 species of this genus led to the capture and characterization of (i) 37 insertion sequences (ISs) representing 9 IS families (IS3, IS5, IS6, IS21, IS66, IS256, IS1182, IS1380 and IS1634), (ii) a composite transposon Tn6097 generated by two copies of the ISPfe2 (IS1634 family) containing two predicted genetic modules, involved in the arginine deiminase pathway and daunorubicin/doxorubicin resistance, (iii) 3 non-composite transposons of the Tn3 family, including Tn5393 carrying streptomycin resistance and (iv) a transposable genomic island TnPpa1 (45 kb). Some of the elements (e.g. Tn5393, Tn6097 and ISs of the IS903 group of the IS5 family) were shown to contain strong promoters able to drive transcription of genes placed downstream of the target site of transposition. Through the application of trap plasmid pCM132TC, containing a promoterless tetracycline resistance reporter gene, we identified five ways in which transposition can supply promoters to transcriptionally silent genes. Besides highlighting the diversity and specific features of several TEs, the analyses performed in this study have provided novel and interesting information on (i) the dynamics of the process of transposition (e.g. the unusually high frequency of transposition of TnPpa1) and (ii) structural changes in DNA mediated by transposition (e.g. the generation of large deletions in the recipient molecule upon transposition of ISPve1 of the IS21 family). We also demonstrated the great potential of TEs and transposition in the generation of diverse phenotypes as well as in the natural amplification and dissemination of genetic information (of adaptative value) by horizontal gene transfer, which is considered the driving force of bacterial

Insights into the transposable mobilome of Paracoccus spp. (Alphaproteobacteria.

Directory of Open Access Journals (Sweden)

Lukasz Dziewit

Full Text Available Several trap plasmids (enabling positive selection of transposition events were used to identify a pool of functional transposable elements (TEs residing in bacteria of the genus Paracoccus (Alphaproteobacteria. Complex analysis of 25 strains representing 20 species of this genus led to the capture and characterization of (i 37 insertion sequences (ISs representing 9 IS families (IS3, IS5, IS6, IS21, IS66, IS256, IS1182, IS1380 and IS1634, (ii a composite transposon Tn6097 generated by two copies of the ISPfe2 (IS1634 family containing two predicted genetic modules, involved in the arginine deiminase pathway and daunorubicin/doxorubicin resistance, (iii 3 non-composite transposons of the Tn3 family, including Tn5393 carrying streptomycin resistance and (iv a transposable genomic island TnPpa1 (45 kb. Some of the elements (e.g. Tn5393, Tn6097 and ISs of the IS903 group of the IS5 family were shown to contain strong promoters able to drive transcription of genes placed downstream of the target site of transposition. Through the application of trap plasmid pCM132TC, containing a promoterless tetracycline resistance reporter gene, we identified five ways in which transposition can supply promoters to transcriptionally silent genes. Besides highlighting the diversity and specific features of several TEs, the analyses performed in this study have provided novel and interesting information on (i the dynamics of the process of transposition (e.g. the unusually high frequency of transposition of TnPpa1 and (ii structural changes in DNA mediated by transposition (e.g. the generation of large deletions in the recipient molecule upon transposition of ISPve1 of the IS21 family. We also demonstrated the great potential of TEs and transposition in the generation of diverse phenotypes as well as in the natural amplification and dissemination of genetic information (of adaptative value by horizontal gene transfer, which is considered the driving force of

A novel, simple, high-throughput method for isolation of genome-wide transposon insertion mutants of Escherichia coli K-12.

Science.gov (United States)

Miki, Takeyoshi; Yamamoto, Yoshihiro; Matsuda, Hideo

2008-01-01

We developed a novel, simple, high-throughput method for isolation of genome-wide transposon insertion mutants of Escherichia coli K-12. The basic idea of the method is to randomly disrupt the genes on the DNA fragments cloned on the Kohara library by inserting a mini-transposon first, and then transfer the disrupted genes from the lambda vector to the E. coli chromosome by homologous recombination. Using this method, we constructed a set of 8402 Km(r) cis-diploid mutants harboring a mini-Tn10 insertion mutation and the corresponding wild-type gene on a chromosome, as well as a set of 6954 haploid mutants derived from the cis-diploid mutants. The major advantage of the strategy used is that the indispensable genes or sites for growth can be identified. Preliminary results suggest that 415 open reading frames are indispensable for growth in E. coli cells. A total of 6404 haploid mutants were deposited to Genetic Strains Research Center, National Institute of Genetics, Japan (Chapter 26) and are available for public distribution upon request (http://shigen.lab.nig.ac.jp/ecoli/strain/nbrp/resource.jsp).

A putative ABC transporter is involved in negative regulation of biofilm formation by Listeria monocytogenes

DEFF Research Database (Denmark)

Zhu, Xinna; Long, Fei; Chen, Yonghui

2008-01-01

Listeria monocytogenes may persist for long periods in food processing environments. In some instances, this may be due to aggregation or biofilm formation. To investigate the mechanism controlling biofilm formation in the food-borne pathogen L. monocytogenes, we characterized LM-49, a mutant...... with enhanced ability of biofilm-formation generated via transposon Tn917 mutagenesis of L. monocytogenes 4b G. In this mutant, a Tn917 insertion has disrupted the coding region of the gene encoding a putative ATP binding cassette (ABC) transporter permease identical to Lmof2365_1771 (a putative ABC...... the same amount of biofilm biomass as the wild-type strain. Furthermore, transcription of the downstream lm.G_1770 was not influenced by the upstream Tn917 insertion, and the presence of Tn917 has no effect on biofilm formation. These results suggest that lm.G_1771 was solely responsible for the negative...

Chromosomally Encoded mcr-5 in Colistin non-susceptible Pseudomonas aeruginosa.

Science.gov (United States)

Snesrud, Erik; Maybank, Rosslyn; Kwak, Yoon I; Jones, Anthony R; Hinkle, Mary K; Mc Gann, Patrick

2018-05-29

Whole genome sequencing (WGS) of historical Pseudomonas aeruginosa clinical isolates identified a chromosomal copy of mcr-5 within a Tn 3 -like transposon in P. aeruginosa MRSN 12280. The isolate was non-susceptible to colistin by broth microdilution and genome analysis revealed no mutations known to confer colistin resistance. To the best of our knowledge, this is the first report of mcr in colistin non-susceptible P. aeruginosa .

Effects of dexmedetomidine on H-FABP, CK-MB, cTnI levels, neurological function and near-term prognosis in patients undergoing heart valve replacement.

Science.gov (United States)

Wang, Zhi; Chen, Qiang; Guo, Hao; Li, Zhishan; Zhang, Jinfeng; Lv, Lei; Guo, Yongqing

2017-12-01

This study investigated the effects of dexmedetomidine on heart-type fatty acid binding protein (H-FABP), creatine kinase isoenzymes (CK-MB), and troponin I (cTnI) levels, neurological function and near-term prognosis in patients undergoing heart valve replacement. Patients undergoing heart valve replacement were randomly allocated to remifentanil anesthesia (control group, n=48) or dexmedetomidine anesthesia (observation group, n=48). Hemodynamic parameters were measured before anesthesia induction (T1), 1 min after intubation (T2), 10 min after start of surgery (T3), and on completion of surgery (T4). Levels of plasma H-FABP, CK-MB and cTnI were measured 10 min before anesthesia induction (C1), 10 min after start of surgery (C2), on completion of surgery (C3), 6 h after surgery (C4), and 24 h after surgery (C5). S100β protein and serum neuron-specific enolase (NSE) were detected 10 min before anesthesia induction (C1), and 24 h after surgery (C5). Neurological and cardiac function was evaluated 24 h after surgery. Incidence of cardiovascular adverse events was recorded for 1 year of follow-up. There were no significant differences in the average heart rate between the two groups during the perioperative period. The mean arterial pressure in the observation group was significantly lower than control group (PH-FABP, CK-MB and cTnI at C2, C3, C4 and C5, were significantly higher than C1, but significantly lower in the observation versus control group (P<0.05). Twenty-four hours after surgery, levels of S100β and NSE in both groups were higher than those before induction (P<0.05), but significantly lower in the observation versus control group (P<0.05). Twenty-four hours after surgery, neurological function scores were better, and myocardial contractility and arrhythmia scores significantly lower in the observation versus control group (P<0.05 for all). After follow-up for 1 year, incidence of cardiovascular adverse events was significantly lower in the observation

Mn porphyrin-based SOD mimic, MnTnHex-2-PyP(5+), and non-SOD mimic, MnTBAP(3-), suppressed rat spinal cord ischemia/reperfusion injury via NF-κB pathways.

Science.gov (United States)

Celic, T; Španjol, J; Bobinac, M; Tovmasyan, A; Vukelic, I; Reboucas, J S; Batinic-Haberle, I; Bobinac, D

2014-12-01

Herein we have demonstrated that both superoxide dismutase (SOD) mimic, cationic Mn(III) meso-tetrakis(N-n-hexylpyridinium-2-yl)porphyrin (MnTnHex-2-PyP(5+)), and non-SOD mimic, anionic Mn(III) meso-tetrakis(4-carboxylatophenyl)porphyrin (MnTBAP(3-)), protect against oxidative stress caused by spinal cord ischemia/reperfusion via suppression of nuclear factor kappa B (NF-κB) pro-inflammatory pathways. Earlier reports showed that Mn(III) N-alkylpyridylporphyrins were able to prevent the DNA binding of NF-κB in an aqueous system, whereas MnTBAP(3-) was not. Here, for the first time, in a complex in vivo system-animal model of spinal cord injury-a similar impact of MnTBAP(3-), at a dose identical to that of MnTnHex-2-PyP(5+), was demonstrated in NF-κB downregulation. Rats were treated subcutaneously at 1.5 mg/kg starting at 30 min before ischemia/reperfusion, and then every 12 h afterward for either 48 h or 7 days. The anti-inflammatory effects of both Mn porphyrins (MnPs) were demonstrated in the spinal cord tissue at both 48 h and 7 days. The downregulation of NF-κB, a major pro-inflammatory signaling protein regulating astrocyte activation, was detected and found to correlate well with the suppression of astrogliosis (as glial fibrillary acidic protein) by both MnPs. The markers of oxidative stress, lipid peroxidation and protein carbonyl formation, were significantly reduced by MnPs. The favorable impact of both MnPs on motor neurons (Tarlov score and inclined plane test) was assessed. No major changes in glutathione peroxidase- and SOD-like activities were demonstrated, which implies that none of the MnPs acted as SOD mimic. Increasing amount of data on the reactivity of MnTBAP(3-) with reactive nitrogen species (RNS) (.NO/HNO/ONOO(-)) suggests that RNS/MnTBAP(3-)-driven modification of NF-κB protein cysteines may be involved in its therapeutic effects. This differs from the therapeutic efficacy of MnTnHex-2-PyP(5+) which presumably occurs via reactive

Mycothiol-Deficient Mycobacterium smegmatis Mutants Are Hypersensitive to Alkylating Agents, Free Radicals, and Antibiotics

Science.gov (United States)

Rawat, Mamta; Newton, Gerald L.; Ko, Mary; Martinez, Gladys J.; Fahey, Robert C.; Av-Gay, Yossef

2002-01-01

Mycothiol (MSH; 1d-myo-inosityl 2-[N-acetyl-l-cysteinyl]amido-2-deoxy-α-d-glucopyranoside) is the major low-molecular-weight thiol produced by mycobacteria. Mutants of Mycobacterium smegmatis mc2155 deficient in MSH production were produced by chemical mutagenesis as well as by transposon mutagenesis. One chemical mutant (mutant I64) and two transposon mutants (mutants Tn1 and Tn2) stably deficient in MSH production were isolated by screening for reduced levels of MSH content. The MSH contents of transposon mutants Tn1 and Tn2 were found to be less than 0.1% that of the parent strain, and the MSH content of I64 was found to be 1 to 5% that of the parent strain. All three strains accumulated 1d-myo-inosityl 2-deoxy-α-d-glucopyranoside to levels 20- to 25-fold the level found in the parent strain. The cysteine:1d-myo-inosityl 2-amino-2-deoxy-α-d-glucopyranoside ligase (MshC) activities of the three mutant strains were ≤2% that of the parent strain. Phenotypic analysis revealed that these MSH-deficient mutants possess increased susceptibilities to free radicals and alkylating agents and to a wide range of antibiotics including erythromycin, azithromycin, vancomycin, penicillin G, rifamycin, and rifampin. Conversely, the mutants possess at least 200-fold higher levels of resistance to isoniazid than the wild type. We mapped the mutation in the chemical mutant by sequencing the mshC gene and showed that a single amino acid substitution (L205P) is responsible for reduced MSH production and its associated phenotype. Our results demonstrate that there is a direct correlation between MSH depletion and enhanced sensitivity to toxins and antibiotics. PMID:12384335

ISAba1 and Tn6168 acquisition by natural transformation leads to third-generation cephalosporins resistance in Acinetobacter baumannii.

Science.gov (United States)

Domingues, Sara; Rosário, Natasha; Ben Cheikh, Hadhemi; Da Silva, Gabriela Jorge

2018-05-15

Acinetobacter baumannii has intrinsic beta-lactamase genes, namely ampC and bla OXA-51 -like, which are only strongly expressed when the ISAba1 insertion sequence is upstream the 5' end of the genes. A second ampC gene has also been identified in some clinical A. baumannii strains. The increased expression of these genes leads to resistance to beta-lactams, including third-generation cephalosporins and/or carbapenems. The aim of this work was to assess the involvement of natural transformation in the transfer of chromosomal ampC-associated mobile elements, and related changes in the resistance profile of recipient cells. Natural transformation assays with the naturally competent A. baumannii A118 clinical isolate as recipient cell and the multidrug resistant A. baumannii Ab51 clinical isolate as the source of donor DNA produced transformants. All tested transformants showed integration of the ISAba1 close to the ampC gene. In two transformants, the ISAba1 was acquired by transposition and inserted between the usual folE and the ampC genes. The remaining transformants acquired the ISAba1 adjacent to a second ampC gene, as part of Tn6168, likely by homologous recombination. Our study demonstrates that natural transformation can contribute to the widespread of beta-lactams resistance, and acquisition of non-resistant determinants can lead to changes in the susceptibility profile of A. baumannii strains. Copyright © 2018 Elsevier B.V. All rights reserved.

Physical, chemical and biological CTD and bottle data from R/V Thomas G. Thompson cruise TN278 in eastern tropical North Pacific Ocean from March 19 to April 20, 2012 (NODC Accession 0109846)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — This report contains data from R/V Thomas G. Thompson cruise TN278 to the eastern tropical north pacific oxygen deficient zone. The objective of the cruise was to...

45 CFR 60.14 - Fees applicable to requests for information.

Science.gov (United States)

2010-10-01

... 45 Public Welfare 1 2010-10-01 2010-10-01 false Fees applicable to requests for information. 60.14 Section 60.14 Public Welfare DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION NATIONAL PRACTITIONER DATA BANK FOR ADVERSE INFORMATION ON PHYSICIANS AND OTHER HEALTH CARE PRACTITIONERS Disclosure of...

A retrospective investigation of HLA-B*5801 in hyperuricemia patients in a Han population of China.

Science.gov (United States)

Cheng, Heng; Yan, Dewen; Zuo, Xin; Liu, Junying; Liu, Wenlan; Zhang, Youming

2018-05-01

Hyperuricemia and gout have become increasingly prevalent in China. Allopurinol is an effective urate-lowering therapy, but it has severe side effects. HLA-B*5801 is highly associated with the allopurinol-induced toxic epidermal necrolysis and Stevens-Johnson syndrome. In this retrospective report, we had genotyped HLA-B*5801 in 253 cases of hyperuricemia and gout patients in a Han population in Shenzhen and analyzed the clinical management of medications. We found 30 carriers of the HLA-B*5801 allele in 253 cases of hyperuricemia or gout patients in the population (11.9%). Allopurinol was prescribed in both HLA-B*5801-positive and HLA-B*5801-negative groups. The evaluation of four models with or without genetic screening and management of allopurinol or febuxostat indicated that the HLA-B*5801 screening had significant cost benefit for clinical management. For appropriate management and cost-effectiveness, the HLA-B*5801 allele should be screened in all patients with hyperuricemia and gout in the Chinese population.

Zbiornik w wyrobisku końcowym Odkrywki "Pątnów" Kopalni Węgla Brunatnego "Konin" i jego bilans wodny za okres 2003-2004

OpenAIRE

Wachowiak, Grzegorz; Wachowiak, Agata

2005-01-01

The idea of water recultivation and management of the closing open pits in the Lignite Opencast Mine "Konin" is of a growing importance. Hence, the necessity to explore water relations conditioned by SLich undertakings. Recultivation scheme of the closing open pit "Pątnów" of Lignite Opencast Mine "Konin" includes the construction of a 346 ha reservoir with the capacity of over 38 mln m. So far, it is the biggest reservoir built as a result of an open pit reculti...

Mobile genetic elements, a key to microbial adaptation in extreme environments

Science.gov (United States)

van Houdt, Rob; Mijnendonckx, Kristel; Provoost, Ann; Monsieurs, Pieter; Mergeay, Max; Leys, Natalie

hydrogenotrophy. Five transposons were identified in CH34. Two mercury transposons Tn4378 and Tn4380, respectively located on pMOL28 and pMOL30, were previously described. In addition, 3 novel transposons were identified. The large Tn6048 transposon contained 8 genes highly induced by zinc and lead. Transposon Tn6049 was found in twelve copies in the genome of strain CH34 and is seem-ingly often associated with genomic islands. Finally, Tn6050 was observed twice in the second chromosome with accessory genes not classically associated with transposons, namely a sulfate permease, a universal stress protein (UspA) and a DksA-like DnaK suppressor protein. The role of these genes is unclear but a functional association could be assumed. Finally, a set of 21 IS elements was found, counting in total for 57 copies dispersed over the genome. The number of copies ranged from 1 to 9 (IS1088 ) and 10 (ISRme3 ). The 21 IS elements could be divided into 10 different families. The elements ISRme5 and IS1071 were putatively involved in the recruitment of the genes for hydrogenotrophy and CO2 fixation, with mutants unable to grow on H2 and CO2 appeared to have lost these genes by IS1071 -mediated excision. These data clearly showed that the C. metallidurans CH34 model bacterium and some of it's Cupriavidus and Ralstonia relatives carry a multitude of tools that allowed them to genetically adapt to polluted and extreme soil environments, and that are a key in their success to adapt to the new anthropogenic spacecraft environments. Furthermore, these tools could be exploited to assess and measure genetic effects of spaceflight conditions. Acknowledgements This work was supported by the European Space Agency (ESA-PRODEX) and the Belgian Science Policy (Belspo) through the MISSEX and COMICS projects.

Bleomycin resistance : a new dominant selectable marker for plant cell transformation

NARCIS (Netherlands)

Hille, Jacques; Verheggen, Frank; Roelvink, Peter; Franssen, Henk; Kammen, Ab van; Zabel, Pim

1986-01-01

Plant cells are sensitive to the antibiotic bleomycin, a DNA damaging glycopeptide. A bleomycin resistance determinant, located on transposon Tn5 and functional in bacteria, has been cloned in a plant expression vector and introduced into Nicotiana plumbaginifolia using Agrobacterium tumefaciens.

Novel anti-Sialyl-Tn monoclonal antibodies and antibody-drug conjugates demonstrate tumor specificity and anti-tumor activity.

Science.gov (United States)

Prendergast, Jillian M; Galvao da Silva, Ana Paula; Eavarone, David A; Ghaderi, Darius; Zhang, Mai; Brady, Dane; Wicks, Joan; DeSander, Julie; Behrens, Jeff; Rueda, Bo R

Targeted therapeutics that can differentiate between normal and malignant tumor cells represent the ideal standard for the development of a successful anti-cancer strategy. The Sialyl-Thomsen-nouveau antigen (STn or Sialyl-Tn, also known as CD175s) is rarely seen in normal adult tissues, but it is abundantly expressed in many types of human epithelial cancers. We have identified novel antibodies that specifically target with high affinity the STn glycan independent of its carrier protein, affording the potential to recognize a wider array of cancer-specific sialylated proteins. A panel of murine monoclonal anti-STn therapeutic antibodies were generated and their binding specificity and efficacy were characterized in vitro and in in vivo murine cancer models. A subset of these antibodies were conjugated to monomethyl auristatin E (MMAE) to generate antibody-drug conjugates (ADCs). These ADCs demonstrated in vitro efficacy in STn-expressing cell lines and significant tumor growth inhibition in STn-expressing tumor xenograft cancer models with no evidence of overt toxicity.

Identification of the genes required for the culture of Liberibacter crescens, the closest cultured relative of the uncultured Liberibacter plant pathogens

Directory of Open Access Journals (Sweden)

Kin-Kwan eLai

2016-04-01

Full Text Available Here Tn5 random transposon mutagenesis was used to identify the essential elements for culturing Liberibacter crescens BT-1 that can serve as antimicrobial targets for the closely related pathogens of citrus, Candidatus Liberibacter asiaticus (Las and tomato and potato, Candidatus Liberibacter solanacearum (Lso. In order to gain insight on the virulence, metabolism, and culturability of the pathogens within the genus Liberibacter, a mini-Tn5 transposon derivative system consisting of a gene specifying resistance to kanamycin, flanked by a 19-base-pair terminal repeat sequence of Tn5, was used for the genome-wide mutagenesis of L. crescens BT-1 and created an insertion mutant library. By analyzing the location of insertions using Sanger and Illumina Mi-Seq sequencing, 314 genes are proposed as essential for the culture of L. crescens BT-1 on BM-7 medium. Of those genes, 76 are not present in the uncultured Liberibacter pathogens and, as a result, suggest molecules necessary for the culturing these pathogens. Those molecules include the aromatic amino acids, several vitamins, histidine, cysteine, lipopolysaccharides, and fatty acids. In addition, the 238 essential genes of L. crescens in common with L. asiaticus are potential targets for the development of therapeutics against the disease.

IS21-558 insertion sequences are involved in the mobility of the multiresistance gene cfr

DEFF Research Database (Denmark)

Kehrenberg, Corinna; Aarestrup, Frank Møller; Schwarz, Stefan

2007-01-01

During a study of florfenicol-resistant porcine staphylococci from Denmark, the genes cfr and fexA were detected in the chromosomal DNA or on plasmids of Staphylococcus hyicus, Staphylococcus warneri, and Staphylococcus simulans. A novel variant of the phenicol resistance transposon Tn558...

Some transition metal ions complexes of tricine (Tn and amino acids: pH-titration, synthesis and antimicrobial activity

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M.E. Zayed

2014-12-01

Full Text Available Equilibrium studies have been carried out on complex formation of M(II (M = Co(II, Cu(II and Zn(II with tricine (Tn and L = amino acids in aqueous solution, at 25 °C and ionic strength of I = 0.1 M (NaNO3. The ternary complexes of amino acids are formed by simultaneous reactions. The concentration distribution of the complexes is evaluated. The solid complexes of [M(II–Tn–Histidine (Hist] have been synthesized and characterized by elemental analysis, infrared, magnetic and conductance measurements. The synthesized complexes have been screened for their antibacterial activities and the complexes show a significant antibacterial activity against four bacterial species: Staphylococcus aureus (Gram +ve, Streptococcus pyogenesr (Gram +ve, Serratia marcescens (Gram −ve and Escherichia coli (Gram −ve. The activity increases by increasing the concentration of the complexes.

An additional regulator, TsaQ, is involved with TsaR in regulation of transport during the degradation of p-toluenesulfonate in Comamonas testosteroni T-2

OpenAIRE

Tralau, Tewes; Cook, Alasdair M.; Ruff, Jürgen

2003-01-01

The degradation of p-toluenesulfonate (TSA) by Comamonas testosteroni T-2 is initiated by a transport system (TsaST) and enzymes (TsaMBCD) encoded on the tsa transposon, Tn tsa, on the TSA plasmid (pTSA). Tn tsa comprises an insert of 15 kb between two IS 1071 elements. The left-hand 6 kb and the right-hand 6 kb are nearly mirror images. The regulator of the tsaMBCD1 genes (right-hand side) is the centrally located LysR-type TsaR, which is encoded upstream of tsaMBCD1 on the reverse strand. T...

Complete nucleotide sequence of the multidrug resistance IncA/C plasmid pR55 from Klebsiella pneumoniae isolated in 1969.

Science.gov (United States)

Doublet, Benoît; Boyd, David; Douard, Gregory; Praud, Karine; Cloeckaert, Axel; Mulvey, Michael R

2012-10-01

To determine the complete nucleotide sequence of the multidrug resistance IncA/C plasmid pR55 from a clinical Klebsiella pneumoniae strain that was isolated from a urinary tract infection in 1969 in a French hospital and compare it with those of contemporary emerging IncA/C plasmids. The plasmid was purified and sequenced using a 454 sequencing approach. After draft assembly, additional PCRs and walking reads were performed for gap closure. Sequence comparisons and multiple alignments with other IncA/C plasmids were done using the BLAST algorithm and CLUSTAL W, respectively. Plasmid pR55 (170 810 bp) revealed a shared plasmid backbone (>99% nucleotide identity) with current members of the IncA/C(2) multidrug resistance plasmid family that are widely disseminating antibiotic resistance genes. Nevertheless, two specific multidrug resistance gene arrays probably acquired from other genetic elements were identified inserted at conserved hotspot insertion sites in the IncA/C backbone. A novel transposon named Tn6187 showed an atypical mixed transposon configuration composed of two mercury resistance operons and two transposition modules that are related to Tn21 and Tn1696, respectively, and an In0-type integron. IncA/C(2) multidrug resistance plasmids have a broad host range and have been implicated in the dissemination of antibiotic resistance among Enterobacteriaceae from humans and animals. This typical IncA/C(2) genetic scaffold appears to carry various multidrug resistance gene arrays and is now also a successful vehicle for spreading AmpC-like cephalosporinase and metallo-β-lactamase genes, such as bla(CMY) and bla(NDM), respectively.

Combinatorial events of insertion sequences and ICE in Gram-negative bacteria.

Science.gov (United States)

Toleman, Mark A; Walsh, Timothy R

2011-09-01

The emergence of antibiotic and antimicrobial resistance in Gram-negative bacteria is incremental and linked to genetic elements that function in a so-called 'one-ended transposition' manner, including ISEcp1, ISCR elements and Tn3-like transposons. The power of these elements lies in their inability to consistently recognize one of their own terminal sequences, while recognizing more genetically distant surrogate sequences. This has the effect of mobilizing the DNA sequence found adjacent to their initial location. In general, resistance in Gram-negatives is closely linked to a few one-off events. These include the capture of the class 1 integron by a Tn5090-like transposon; the formation of the 3' conserved segment (3'-CS); and the fusion of the ISCR1 element to the 3'-CS. The structures formed by these rare events have been massively amplified and disseminated in Gram-negative bacteria, but hitherto, are rarely found in Gram-positives. Such events dominate current resistance gene acquisition and are instrumental in the construction of large resistance gene islands on chromosomes and plasmids. Similar combinatorial events appear to have occurred between conjugative plasmids and phages constructing hybrid elements called integrative and conjugative elements or conjugative transposons. These elements are beginning to be closely linked to some of the more powerful resistance mechanisms such as the extended spectrum β-lactamases, metallo- and AmpC type β-lactamases. Antibiotic resistance in Gram-negative bacteria is dominated by unusual combinatorial mistakes of Insertion sequences and gene fusions which have been selected and amplified by antibiotic pressure enabling the formation of extended resistance islands. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

Nucleotide sequence of pOLA52: a conjugative IncX1 plasmid from Escherichia coli which enables biofilm formation and multidrug efflux

DEFF Research Database (Denmark)

Norman, Anders; Hansen, Lars H.; She, Qunxin

2008-01-01

. The plasmid was also classified as IncX1 with incompatibility testing. The conjugal transfer and plasmid maintenance regions of pOLA52 therefore seem to represent IncX1 orthologues of the well-characterized IncX2 plasmid R6K. Sequence homology searches in GenBank also suggested a considerably higher...... of type 3 fimbriae (mrkABCDF). The plasmid was found to be 51,602 bp long with 68 putative genes. About half of the plasmid constituted a conserved IncX1-type backbone with predicted regions for conjugation, replication and partitioning, as well as a toxin/antitoxin (TA) plasmid addiction system...... prevalence of IncX1 group plasmids than IncX2. The 21 kb 'genetic load' region of pOLA52 was shown to consist of a mosaic, among other things a fragmented Tn3 transposon encoding ampicillin resistance. Most notably the oqxAB and mrkABCDF cassettes were contained within two composite transposons (Tn6010...

Insight into the mobilome of Aeromonas strains.

Science.gov (United States)

Piotrowska, Marta; Popowska, Magdalena

2015-01-01

The mobilome is a pool of genes located within mobile genetic elements (MGE), such as plasmids, IS elements, transposons, genomic/pathogenicity islands, and integron-associated gene cassettes. These genes are often referred to as "flexible" and may encode virulence factors, toxic compounds as well as resistance to antibiotics. The phenomenon of MGE transfer between bacteria, known as horizontal gene transfer (HGT), is well documented. The genes present on MGE are subject to continuous processes of evolution and environmental changes, largely induced or significantly accelerated by man. For bacteria, the only chance of survival in an environment contaminated with toxic chemicals, heavy metals and antibiotics is the acquisition of genes providing the ability to survive in such conditions. The process of acquiring and spreading antibiotic resistance genes (ARG) is of particular significance, as it is important for the health of humans and animals. Therefore, it is important to thoroughly study the mobilome of Aeromonas spp. that is widely distributed in various environments, causing many diseases in fishes and humans. This review discusses the recently published information on MGE prevalent in Aeromonas spp. with special emphasis on plasmids belonging to different incompatibility groups, i.e., IncA/C, IncU, IncQ, IncF, IncI, and ColE-type. The vast majority of plasmids carry a number of different transposons (Tn3, Tn21, Tn1213, Tn1721, Tn4401), the 1st, 2nd, or 3rd class of integrons, IS elements (e.g., IS26, ISPa12, ISPa13, ISKpn8, ISKpn6) and encode determinants such as antibiotic and mercury resistance genes, as well as virulence factors. Although the actual role of Aeromonas spp. as a human pathogen remains controversial, species of this genus may pose a serious risk to human health. This is due to the considerable potential of their mobilome, particularly in terms of antibiotic resistance and the possibility of the horizontal transfer of resistance genes.

Insight into the mobilome of Aeromonas strains

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Marta ePiotrowska

2015-05-01

Full Text Available The mobilome is a pool of genes located within mobile genetic elements (MGE, such as plasmids, IS elements, transposons, genomic/pathogenicity islands, and integron-associated gene cassettes. These genes are often referred to as flexible and may encode virulence factors, toxic compounds as well as resistance to antibiotics. The phenomenon of MGE transfer between bacteria, known as horizontal gene transfer (HGT, is well documented. The genes present on MGE are subject to continuous processes of evolution and environmental changes, largely induced or significantly accelerated by man. For bacteria, the only chance of survival in an environment contaminated with toxic chemicals, heavy metals and antibiotics is the acquisition of genes providing the ability to survive in such conditions. The process of acquiring and spreading antibiotic resistance genes (ARG is of particular significance, as it is important for the health of humans and animals. Therefore, it is important to thoroughly study the mobilome of Aeromonas spp. that is widely distributed in various environments, causing many diseases in fishes and humans. This review discusses the recently published information on MGE prevalent in Aeromonas spp. with special emphasis on plasmids belonging to different incompatibility groups, i.e., IncA/C, IncU, IncQ, IncF, IncI and ColE-type. The vast majority of plasmids carry a number of different transposons (Tn3, Tn21, Tn1213, Tn1721, Tn4401, the 1st, 2nd or 3rd class of integrons, IS elements (e.g. IS26, ISPa12, ISPa13, ISKpn8, ISKpn6 and encode determinants such as antibiotic and mercury resistance genes, as well as virulence factors. Although the actual role of Aeromonas spp. as a human pathogen remains controversial, species of this genus may pose a serious risk to human health. This is due to the considerable potential of their mobilome, particularly in terms of antibiotic resistance and the possibility of the horizontal transfer of resistance

Construction of an efficient biologically contained Pseudomonas putida strain and its survival in outdoor assays

DEFF Research Database (Denmark)

Molina, Lazaro; Rodriguez, Cayo Juan Ramos; Ronchel, Maria C.

1998-01-01

Active biological containment systems consist of two components, a killing element designed to induce cell death and a control element which modulates the expression of the killing function. We constructed a mini-Tn5 transposon bearing a fusion of the P(lac) promoter to the gef killing gene...

Double Copies of bla(KPC-3)::Tn4401a on an IncX3 Plasmid in Klebsiella pneumoniae Successful Clone ST512 from Italy.

Science.gov (United States)

Fortini, Daniela; Villa, Laura; Feudi, Claudia; Pires, João; Bonura, Celestino; Mammina, Caterina; Endimiani, Andrea; Carattoli, Alessandra

2016-01-01

A carbapenem-resistant sequence type 512 (ST512) Klebsiella pneumoniae carbapenemase 3 (KPC-3)-producing K. pneumoniae strain showing a novel variant plasmid content was isolated in Palermo, Italy, in 2014. ST512 is a worldwide successful clone associated with the spread of bla(KPC) genes located on the IncFIIk pKpQIL plasmid. In our ST512 strain, the bla(KPC-3) gene was unusually located on an IncX3 plasmid, whose complete sequence was determined. Two copies of bla(KPC-3)::Tn4401a caused by intramolecular transposition events were detected in the plasmid. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

CNS bioavailability and radiation protection of normal hippocampal neurogenesis by a lipophilic Mn porphyrin-based superoxide dismutase mimic, MnTnBuOE-2-PyP5+

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David Leu

2017-08-01

Full Text Available Although radiation therapy can be effective against cancer, potential damage to normal tissues limits the amount that can be safely administered. In central nervous system (CNS, radiation damage to normal tissues is presented, in part, as suppressed hippocampal neurogenesis and impaired cognitive functions. Mn porphyrin (MnP-based redox active drugs have demonstrated differential effects on cancer and normal tissues in experimental animals that lead to protection of normal tissues and radio- and chemo-sensitization of cancers. To test the efficacy of MnPs in CNS radioprotection, we first examined the tissue levels of three different MnPs – MnTE-2-PyP5+(MnE, MnTnHex-2-PyP5+(MnHex, and MnTnBuOE-2-PyP5+(MnBuOE. Nanomolar concentrations of MnHex and MnBuOE were detected in various brain regions after daily subcutaneous administration, and MnBuOE was well tolerated at a daily dose of 3 mg/kg. Administration of MnBuOE for one week before cranial irradiation and continued for one week afterwards supported production and long-term survival of newborn neurons in the hippocampal dentate gyrus. MnP-driven S-glutathionylation in cortex and hippocampus showed differential responses to MnP administration and radiation in these two brain regions. A better understanding of how preserved hippocampal neurogenesis correlates with cognitive functions following cranial irradiation will be helpful in designing better MnP-based radioprotection strategies. Keywords: Mn porphyrin, Bioavailability, BMX-001, Hippocampus, Neurogenesis, Radioprotection

Significant reduction of peripheral blood interleukin-35 and CD4+EBI3+ T cells, which are negatively correlated with an increase in the plasma IL-17 and cTnI level, in viral myocarditis patients

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Han Ouyang

2017-02-01

Full Text Available Introduction: Viral myocarditis (VMC has become an increasingly common heart disease that endangers human health. In the present study, the plasma interleukin-35 (IL-35 level and the percentage of CD4 + EBI3 + T cells in VMC patients were detected to investigate the significance of changes in these parameters in the plasma of VMC patients and their association with the disease. Material and methods: ELISA was performed to detect the plasma IL-35 level and the percentage of peripheral blood CD4 + EBI3 + T cells in 40 VMC patients and in 20 healthy individuals. Moreover, the plasma IL-17 levels in the VMC patients and in the healthy individuals were detected using an ELISA, and the cardiac Troponin-I (cTnI levels were detected using a chemiluminescent microparticle immunoassay to compare the differences in the groups. Results : Plasma IL-35 level and the percentage of CD4 + EBI3 + T cells in acute phase VMC patients was lower than that in the healthy control group and the convalescent phase VMC patients. Additionally, the plasma IL-35 level in the VMC patients exhibited a negative correlation with the levels of cTnI and IL-17. The percentage of CD4 + EBI3 + T cells also showed a negative correlation with the levels of cTnI and IL-17. Conclusions : The plasma IL-35 level and the percentage of CD4 + EBI3 + T cells in VMC patients was reduced, and the amount of the decrease was associated with the severity of the disease. These results suggest that IL-35 and CD4 + EBI3 + T might play important roles in the progression of VMC and could be used as indictors of the disease.

Selective methanol or formate production during continuous CO₂ fermentation by the acetogen biocatalysts engineered via integration of synthetic pathways using Tn7-tool.

Science.gov (United States)

Tyurin, Michael; Kiriukhin, Michael

2013-09-01

Methanol-resistant mutant acetogen Clostridium sp. MT1424 originally producing only 365 mM acetate from CO₂/CO was engineered to eliminate acetate production and spore formation using Cre-lox66/lox71-system to power subsequent methanol production via expressing synthetic methanol dehydrogenase, formaldehyde dehydrogenase and formate dehydrogenase, three copies of each, assembled in cluster and integrated to chromosome using Tn7-based approach. Production of 2.2 M methanol was steady (p integrated cluster comprised only three copies of formate dehydrogenase the respective recombinants produced 95 mM formate (p < 0.005) under the same conditions. For commercialization, the suggested source of inorganic carbon would be CO₂ waste of IGCC power plant. Hydrogen may be produced in situ via powered by solar panels electrolysis.

24 CFR 5.801 - Uniform financial reporting standards.

Science.gov (United States)

2010-04-01

... 24 Housing and Urban Development 1 2010-04-01 2010-04-01 false Uniform financial reporting... and Urban Development GENERAL HUD PROGRAM REQUIREMENTS; WAIVERS Uniform Financial Reporting Standards § 5.801 Uniform financial reporting standards. (a) Applicability. This subpart H implements uniform...

pGLO Mutagenesis: A Laboratory Procedure in Molecular Biology for Biology Students

Science.gov (United States)

Bassiri, Eby A.

2011-01-01

A five-session laboratory project was designed to familiarize or increase the laboratory proficiency of biology students and others with techniques and instruments commonly used in molecular biology research laboratories and industries. In this project, the EZ-Tn5 transposon is used to generate and screen a large number of cells transformed with…

Detection of Sleeping Beauty transposition in the genome of host cells by non-radioactive Southern blot analysis

Energy Technology Data Exchange (ETDEWEB)

Aravalli, Rajagopal N., E-mail: aravalli@umn.edu [Department of Radiology, University of Minnesota Medical School, MMC 292, 420 Delaware Street SE, Minneapolis, MN 55455 (United States); Park, Chang W. [Department of Medicine, University of Minnesota Medical School, MMC 36, 420 Delaware Street SE, Minneapolis, MN 55455 (United States); Steer, Clifford J., E-mail: steer001@umn.edu [Department of Medicine, University of Minnesota Medical School, MMC 36, 420 Delaware Street SE, Minneapolis, MN 55455 (United States); Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455 (United States)

2016-08-26

The Sleeping Beauty transposon (SB-Tn) system is being used widely as a DNA vector for the delivery of therapeutic transgenes, as well as a tool for the insertional mutagenesis in animal models. In order to accurately assess the insertional potential and properties related to the integration of SB it is essential to determine the copy number of SB-Tn in the host genome. Recently developed SB100X transposase has demonstrated an integration rate that was much higher than the original SB10 and that of other versions of hyperactive SB transposases, such as HSB3 or HSB17. In this study, we have constructed a series of SB vectors carrying either a DsRed or a human β-globin transgene that was encompassed by cHS4 insulator elements, and containing the SB100X transposase gene outside the SB-Tn unit within the same vector in cis configuration. These SB-Tn constructs were introduced into the K-562 erythroid cell line, and their presence in the genomes of host cells was analyzed by Southern blot analysis using non-radioactive probes. Many copies of SB-Tn insertions were detected in host cells regardless of transgene sequences or the presence of cHS4 insulator elements. Interestingly, the size difference of 2.4 kb between insulated SB and non-insulated controls did not reflect the proportional difference in copy numbers of inserted SB-Tns. We then attempted methylation-sensitive Southern blots to assess the potential influence of cHS4 insulator elements on the epigenetic modification of SB-Tn. Our results indicated that SB100X was able to integrate at multiple sites with the number of SB-Tn copies larger than 6 kb in size. In addition, the non-radioactive Southern blot protocols developed here will be useful to detect integrated SB-Tn copies in any mammalian cell type.

Detection of Sleeping Beauty transposition in the genome of host cells by non-radioactive Southern blot analysis

International Nuclear Information System (INIS)

Aravalli, Rajagopal N.; Park, Chang W.; Steer, Clifford J.

2016-01-01

The Sleeping Beauty transposon (SB-Tn) system is being used widely as a DNA vector for the delivery of therapeutic transgenes, as well as a tool for the insertional mutagenesis in animal models. In order to accurately assess the insertional potential and properties related to the integration of SB it is essential to determine the copy number of SB-Tn in the host genome. Recently developed SB100X transposase has demonstrated an integration rate that was much higher than the original SB10 and that of other versions of hyperactive SB transposases, such as HSB3 or HSB17. In this study, we have constructed a series of SB vectors carrying either a DsRed or a human β-globin transgene that was encompassed by cHS4 insulator elements, and containing the SB100X transposase gene outside the SB-Tn unit within the same vector in cis configuration. These SB-Tn constructs were introduced into the K-562 erythroid cell line, and their presence in the genomes of host cells was analyzed by Southern blot analysis using non-radioactive probes. Many copies of SB-Tn insertions were detected in host cells regardless of transgene sequences or the presence of cHS4 insulator elements. Interestingly, the size difference of 2.4 kb between insulated SB and non-insulated controls did not reflect the proportional difference in copy numbers of inserted SB-Tns. We then attempted methylation-sensitive Southern blots to assess the potential influence of cHS4 insulator elements on the epigenetic modification of SB-Tn. Our results indicated that SB100X was able to integrate at multiple sites with the number of SB-Tn copies larger than 6 kb in size. In addition, the non-radioactive Southern blot protocols developed here will be useful to detect integrated SB-Tn copies in any mammalian cell type.

Development of a high-frequency in vivo transposon mutagenesis system for Synechocystis sp. PCC 6803 and Synechococcus elongatus PCC 7942.

Science.gov (United States)

Watabe, Kazuyuki; Mimuro, Mamoru; Tsuchiya, Tohru

2014-11-01

Synechocystis sp. PCC 6803 (Synechocystis) is the first sequenced photosynthetic organism and has two advantages: natural transformation and light-activated heterotrophic growth. Such characteristics have mainly promoted reverse genetic analysis in this organism, however, to date approximately 50% of genes are still annotated as 'unknown protein' or 'hypothetical protein'. Therefore, forward genetic analysis is required for the identification of significant genes responsible for photosynthesis and other physiological phenomena among the genes of unknown function. The in vivo transposon mutagenesis system is one of the major methods for random mutagenesis. However, present in vivo transposon mutagenesis systems for cyanobacteria face problems such as relatively low frequency of transposition and repeated transposition in the host cells. In this study, we constructed vectors based on a mini-Tn5-derived vector that was designed to prevent repeated transposition. Our vectors carry a hyperactive transposase and optimized recognition sequence of transposase, which were reported to enhance frequency of transposition. Using the vector, we succeeded in highly frequent transposition (9×10(-3) per recipient cell) in Synechocystis. Transposon insertion sites of 10 randomly selected mutants indicated that the insertion sites spread throughout the genome with low sequence dependency. Furthermore, one of the 10 mutants exhibited the slow-growing phenotype, and the mutant was functionally complemented by using our expression vector. Our system also worked with another model cyanobacterium, Synechococcus elongatus PCC 7942, with high frequency. These results indicate that the developed system can be applied to the forward genetic analysis of a broad range of cyanobacteria. © The Author 2014. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Myofibrillar troponin exists in three states and there is signal transduction along skeletal myofibrillar thin filaments.

Science.gov (United States)

Swartz, Darl R; Yang, Zhenyun; Sen, Asok; Tikunova, Svetlana B; Davis, Jonathan P

2006-08-18

Activation of striated muscle contraction is a highly cooperative signal transduction process converting calcium binding by troponin C (TnC) into interactions between thin and thick filaments. Once calcium is bound, transduction involves changes in protein interactions along the thin filament. The process is thought to involve three different states of actin-tropomyosin (Tm) resulting from changes in troponin's (Tn) interaction with actin-Tm: a blocked (B) state preventing myosin interaction, a closed (C) state allowing weak myosin interactions and favored by calcium binding to Tn, and an open or M state allowing strong myosin interactions. This was tested by measuring the apparent rate of Tn dissociation from rigor skeletal myofibrils using labeled Tn exchange. The location and rate of exchange of Tn or its subunits were measured by high-resolution fluorescence microscopy and image analysis. Three different rates of Tn exchange were observed that were dependent on calcium concentration and strong cross-bridge binding that strongly support the three-state model. The rate of Tn dissociation in the non-overlap region was 200-fold faster at pCa 4 (C-state region) than at pCa 9 (B-state region). When Tn contained engineered TnC mutants with weakened regulatory TnI interactions, the apparent exchange rate at pCa 4 in the non-overlap region increased proportionately with TnI-TnC regulatory affinity. This suggests that the mechanism of calcium enhancement of the rate of Tn dissociation is by favoring a TnI-TnC interaction over a TnI-actin-Tm interaction. At pCa 9, the rate of Tn dissociation in the overlap region (M-state region) was 100-fold faster than the non-overlap region (B-state region) suggesting that strong cross-bridges increase the rate of Tn dissociation. At pCa 4, the rate of Tn dissociation was twofold faster in the non-overlap region (C-state region) than the overlap region (M-state region) that likely involved a strong cross-bridge influence on Tn

Emamectin benzoate induces ROS-mediated DNA damage and apoptosis in Trichoplusia Tn5B1-4 cells.

Science.gov (United States)

Luan, Shaorong; Yun, Xinming; Rao, Wenbing; Xiao, Ciying; Xu, Zhikang; Lang, Jialin; Huang, Qingchun

2017-08-01

Emamectin benzoate (EMB), a novel macrocyclic lactone insecticide, possesses high efficacy and beneficial selective toxicity in agriculture, but so far the EMB-induced cytotoxic action in arthropod insect remains unclear. The present studies were carried out to characterize the property of EMB on the induction of reactive oxygen species (ROS)-mediated DNA damage and apoptosis in Trichoplusia Tn5B1-4 cell model. Following the exposure to EMB at 2.5, 5, 10 or 15 μM, the cells changed to be round, suspended and aggregated, and the decline of cell proliferating ability and cell viability was positively related with the exposure time. Median inhibitory concentration (IC 50 ) of EMB on cell viability was 3.72 μM during 72 h exposure. Apoptosis was induced in 29.8% (24 h) and 39.5% (48 h) of the cells by EMB at 15 μM, showing chromatin condensation in nuclei. The content of ROS in the cells increased rapidly as the concentration of EMB increased, and the pre-incubation of the cells with vitamin E significantly reduced the ROS accumulation. In the treatment of 15 μM EMB, the migrated cell nucleus with DNA strand breaks appeared a teardrop, pear-shaped, or large fan-like tail, and 63.1% of γH2AX-positive cells contained more than four foci, accompanying with high expression level of caspase-3 in time-dependent manner, which consequently led to cell apoptotic death. These evidences in ROS-mediated DNA damage and cell apoptosis induced by EMB may be helpful for deep understanding the cytotoxic action of EMB based on cell model. Copyright © 2017 Elsevier B.V. All rights reserved.

The cell envelope subtilisin-like proteinase is a virulence determinant for Streptococcus suis

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Gottschalk Marcelo

2010-02-01

Full Text Available Abstract Background Streptococcus suis is a major swine pathogen and zoonotic agent that mainly causes septicemia, meningitis, and endocarditis. It has recently been suggested that proteinases produced by S. suis (serotype 2 are potential virulence determinants. In the present study, we screened a S. suis mutant library created by the insertion of Tn917 transposon in order to isolate a mutant deficient in a cell surface proteinase. We characterized the gene and assessed the proteinase for its potential as a virulence factor. Results Two mutants (G6G and M3G possessing a single Tn917 insertion were isolated. The affected gene coded for a protein (SSU0757 that shared a high degree of identity with Streptococccus thermophilus PrtS (95.9% and, to a lesser extent, with Streptococcus agalactiae CspA (49.5%, which are cell surface serine proteinases. The SSU0757 protein had a calculated molecular mass of 169.6 kDa and contained the catalytic triad characteristic of subtilisin family proteinases: motif I (Asp200, motif II (His239, and motif III (Ser568. SSU0757 also had the Gram-positive cell wall anchoring motif (Leu-Pro-X-Thr-Gly at the carboxy-terminus, which was followed by a hydrophobic domain. All the S. suis isolates tested, which belonged to different serotypes, possessed the gene encoding the SSU0757 protein. The two mutants devoid of subtilisin-like proteinase activity had longer generation times and were more susceptible to killing by whole blood than the wild-type parent strain P1/7. The virulence of the G6G and M3G mutants was compared to the wild-type strain in the CD1 mouse model. Significant differences in mortality rates were noted between the P1/7 group and the M3G and G6G groups (p Conclusion In summary, we identified a gene coding for a cell surface subtilisin-like serine proteinase that is widely distributed in S. suis. Evidences were brought for the involvement of this proteinase in S. suis virulence.

A randomised phase II study of sialyl-Tn and DETOX-B adjuvant with or without cyclophosphamide pretreatment for the active specific immunotherapy of breast cancer.

Science.gov (United States)

Miles, D W; Towlson, K E; Graham, R; Reddish, M; Longenecker, B M; Taylor-Papadimitriou, J; Rubens, R D

1996-10-01

Studies in animal models of mouse mammary carcinoma have shown that ovine submaxillary mucin, which carries multiple sialyl-Tn (STn) epitopes, is effective in stimulating an immune response and inhibiting tumour growth. In similar studies using carbohydrate antigens, pretreatment with low-dose cyclophosphamide has been shown to be important in modulating the immune response to antigen possibly by inhibiting suppresser T-cell activity. In a clinical trial assessing the efficacy and toxicity of synthetic STn, patients with metastatic breast cancer were randomised to receive 100 micrograms STn linked to keyhole limpet haemocyanin (KLH) with DETOX-B adjuvant given by subcutaneous injection at weeks 0, 2, 5 and 9 with or without low-dose cyclophosphamide (CTX, 300 mg m-2) pretreatment, 3 days before the start of immunotherapy. Patients with responding or stable disease after the first four injections were eligible to receive STn-KLH at 4 week intervals. The main toxicity noted was the development of subcutaneous granulomata at injection sites. Of 23 patients randomised, 18 received four injections, 5 patients having developed progressive disease during the initial 12 week period. Two minor responses were noted in the 18 patients who received four active specific immunotherapy (ASI) injections and a further five patients had stable disease. Six patients continued ASI at 4 week intervals and a partial response was noted in a patient who had previously had stable disease. All patients developed IgG and IgM responses to sialyl-Tn and levels of IgM antibodies were significantly higher in those patients who were pretreated with CTX. Measurable tumour responses have been recorded following ASI with STn-KLH plus DETOX and the immunomodulatory properties of low-dose CTX have been confirmed.

Molecular characterization of Vulmar1, a complete mariner transposon of sugar beet and diversity of mariner- and En/Spm-like sequences in the genus Beta.

Science.gov (United States)

Jacobs, Gunnar; Dechyeva, Daryna; Menzel, Gerhard; Dombrowski, Cora; Schmidt, Thomas

2004-12-01

Transposons of the Tc1-mariner superfamily are widespread in eukaryotic genomes. We have isolated the mariner element Vulmar1 from Beta vulgaris L., which is 3909 bp long and bordered by perfect terminal inverted repeats of 32 bp with homology to terminal inverted repeats of transposons from soybean and rice. According to a characteristic amino acid signature, Vulmar1 can be assigned to the DD39D group of mariner transposons. Vulmar1 is flanked by a 5'-TA-3' target site duplication that is typical for mariner transposons. Southern hybridization revealed that mariner-like copies are highly abundant in Beta species, and sequence analysis of 10 transposase fragments from representative species of the four Beta sections revealed an identity between 34% and 100% after conceptual translation. By fluorescent in situ hybridization, Vulmar1 was detected in distal euchromatin as well as in some intercalary and pericentromeric regions of all B. vulgaris chromosomes. In addition, using PCR, we were able to amplify fragments of the transposase gene of En/Spm-like transposons in the genus Beta. En/Spm-like transposase sequences are highly amplified in four Beta sections and showed a considerable degree of conservation (88.5-100%) at the protein level, while the homology to corresponding regions of En/Spm transposons of other plant species ranges from 49.5% to 62.5%. By fluorescent in situ hybridization, En/Spm-like transposon signals of strong intensity were detected on all chromosomes of B. vulgaris.

Development of in vitro transposon assisted signal sequence trapping and its use in screening Bacillus halodurans C125 and Sulfolobus solfataricus P2 gene libraries

DEFF Research Database (Denmark)

Becker, F.; Schnorr, K.; Wilting, R.

2004-01-01

To identify genes encoding extracytosolic proteins, a minitransposon, TnSig, containing a signal-less beta-lactamase ('bla) as reporter gene, was constructed and used for in vitro transposition of genomic libraries made in Escherichia coli. The 'bla gene was cloned into a bacteriophage MU...... minitransposon enabling translational fusions between 'bla and target genes. Fusion of TnSig in the correct reading frame to a protein carrying transmembrane domains or signal peptides resulted in ampicillin resistance of the corresponding clone. Prokaryotic gene libraries from the alkaliphilic bacterium...... Bacillus halodurans C125 and the hyperthermophilic archaeon Sulfolobus solfataricus P2 were tagged with TnSig. The genomic sequences, which are publicly available (EMBL BA000004 and EMBL AE006641), were used for rapid open reading frame (ORF) identification and prediction of protein localisation...

Structure of the plasminogen kringle 4 binding calcium-free form of the C-type lectin-like domain of tetranectin

DEFF Research Database (Denmark)

Nielbo, Steen Günther; Thomsen, J.K.; Graversen, J. H.

2004-01-01

. A conserved proline, which was found to be in the cis conformation in holoTN3, is in apoTN3 predominantly in the trans conformation. Backbone dynamics indicate that, in apoTN3 especially, two of the three calcium-binding loops and two of the three K4-binding residues exhibit increased flexibility, whereas...

Xanthomonas oryzae pv oryzae the Causal Agent of Bacterial Leaf Blight of rice: Isolation, Characterization, and Study of Transposon Mutagenesis

Directory of Open Access Journals (Sweden)

Abdjad Asih Nawangsih

2011-04-01

Full Text Available Xanthomonas oryzae pv oryzae the Causal Agent of Bacterial Leaf Blight of rice: Isolation, Characterization, and Study of Transposon Mutagenesis. X. oryzae pv. oryzae (Xoo causes bacterial leaf blight (BLB of rice (Oryza sativa L., a major disease that constrains production of the staple crop in many countries of the world. Identification of X. oryzae pv. oryzae (Xoo was conducted based on the disease symptoms, pathogenicity, morphological, physiological, and genetic characteristics of bacterial cultures isolated from the infected plants. Fifty bacterial isolates predicted as Xoo have been successfully isolated. They are aerobic, rod shaped, and Gram negative bacteria. The isolates were evaluated for their hypersensitivity in tobacco and pathogenicity in rice plant. Fifty isolates induced hypersensitive reaction in tobacco and showed pathogenicity symptom in rice in different length. Based on physiological test, hypersensitivity and pathogenicity reactions, three bacterial isolates strongly predicted as Xoo, i.e. STG21, STG42, and STG46, were non indole formation, non pigment fluorescent, hydrolyzed casein, catalase activity positive, but negative oxidase. Partial sequencing of 16S rRNA genes of STG21 and STG42 showed 80% and 82% homology with X. oryzae, respectively, while STG46 showed 84% homology with X. campestris. Mini-Tn5 transposon mutagenesis of STG21 generated one of the mutants (M5 lossed it’s ability to induce hypersensitive reaction in tobacco plant and deficient in pathogenicity on rice. The lesion length of rice leaf caused by the mutant M5 decreased up to 80%.

Exploring the Molecular Basis for Selective Binding of Homoserine Dehydrogenase from Mycobacterium leprae TN toward Inhibitors: A Virtual Screening Study

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Dongling Zhan

2014-01-01

Full Text Available Homoserine dehydrogenase (HSD from Mycobacterium leprae TN is an antifungal target for antifungal properties including efficacy against the human pathogen. The 3D structure of HSD has been firmly established by homology modeling methods. Using the template, homoserine dehydrogenase from Thiobacillus denitrificans (PDB Id 3MTJ, a sequence identity of 40% was found and molecular dynamics simulation was used to optimize a reliable structure. The substrate and co-factor-binding regions in HSD were identified. In order to determine the important residues of the substrate (l-aspartate semialdehyde (l-ASA binding, the ASA was docked to the protein; Thr163, Asp198, and Glu192 may be important because they form a hydrogen bond with HSD through AutoDock 4.2 software. neuraminidaseAfter use of a virtual screening technique of HSD, the four top-scoring docking hits all seemed to cation–π ion pair with the key recognition residue Lys107, and Lys207. These ligands therefore seemed to be new chemotypes for HSD. Our results may be helpful for further experimental investigations.

Transportation of a 451 ton generator stator and a 234 ton generator rotor from Hartsville, TN, to Los Alamos, NM

International Nuclear Information System (INIS)

Boenig, H.J.; Rogers, J.D.; McLelland, G.R.; Pelts, C.T.

1989-01-01

A 1430 MVA steam turbine generator was acquired from a cancelled nuclear power plant in Tennessee to be used as the pulsed power and energy storage unit for the Confinement Physics Research Facility being built at Los Alamos, NM. The transportation from Hartsville, near Nashville, TN, to Los Alamos, NM, of the two largest single pieces of the generator, a 451 t stator and a 234 t rotor presented a special challenge. Details of the move, by barge from Hartsville to Catoosa, near Tulsa, OK, by rail from Catoosa to Lamy, near Santa Fe, NM, and by road from Lamy to Los Alamos are described. The greatest difficulty of the successful move was the crossing of the Rio Grande river on an existing reinforced concrete bridge. The two-lane wide road transporters for the stator and rotor were fitted with outriggers to provide a four-lane wide vehicle, thus spreading the load over the entire bridge width and meeting acceptable load distribution and bridge safety factors. 2 refs., 6 figs

HrcQ is necessary for Xanthomonas oryzae pv. oryzae HR-induction in non-host tobacco and pathogenicity in host rice

Directory of Open Access Journals (Sweden)

Xiaoping Zhang

2013-12-01

Full Text Available Bacterial blight, caused by Xanthomonas oryzae pv. oryzae (Xoo, is one of the most destructive diseases of rice (Oryza sativa L. worldwide. The type III secretion system (T3SS of Xoo, encoded by the hrp (hypersensitive response and pathogenicity genes, plays critical roles in conferring pathogenicity in host rice and triggering a hypersensitive response (HR in non-host plants. To investigate the major genes conferring the pathogenicity and avirulence of Xoo, we previously constructed a random Tn5-insertion mutant library of Xoo strain PXO99A. We report here the isolation and characterization of a Tn5-insertion mutant PXM69. Tn5-insertion mutants were screened on indica rice JG30, which is highly susceptible to PXO99A, by leaf-cutting inoculation. Four mutants with reduced virulence were obtained after two rounds of screening. Among them, the mutant PXM69 had completely lost virulence to the rice host and ability to elicit HR in non-host tobacco. Southern blotting analysis showed a single copy of a Tn5-insertion in the genome of PXM69. PCR walking and sequencing analysis revealed that the Tn5 transposon was inserted at nucleotide position 70,192–70,201 in the genome of PXO99A, disrupting the type III hrc (hrp-conserved gene hrcQ, the first gene in the D operon of the hrp cluster in Xoo. To confirm the relationship between the Tn5-insertion and the avirulence phenotype of PXM69, we used the marker exchange mutagenesis to create a PXO99A mutant, ΔhrcQ::KAN, in which the hrcQ was disrupted by a kanamycin-encoding gene cassette at the same site as that of the Tn5-insertion. ΔhrcQ::KAN showed the same phenotype as mutant PXM69. Reintroduction of the wild-type hrcQ gene partially complemented the pathogenic function of PXM69. RT-PCR and cellulase secretion assays showed that the Tn5-disruption of hrcQ did not affect transcription of downstream genes in the D operon and function of the type II secretion system. Our results provide new insights into

HrcQ is necessary for Xanthomonas oryzae pv. oryzae HR-induction in non-host tobacco and pathogenicity in host rice

Institute of Scientific and Technical Information of China (English)

Xiaoping; Zhang; Chunlian; Wang; Chongke; Zheng; Jinying; Che; Yanqiang; Li; Kaijun; Zhao

2013-01-01

Bacterial blight, caused by Xanthomonas oryzae pv. oryzae(Xoo), is one of the most destructive diseases of rice(Oryza sativa L.) worldwide. The type III secretion system(T3SS) of Xoo, encoded by the hrp(hypersensitive response and pathogenicity) genes, plays critical roles in conferring pathogenicity in host rice and triggering a hypersensitive response(HR) in non-host plants. To investigate the major genes conferring the pathogenicity and avirulence of Xoo, we previously constructed a random Tn5-insertion mutant library of Xoo strain PXO99A. We report here the isolation and characterization of a Tn5-insertion mutant PXM69. Tn5-insertion mutants were screened on indica rice JG30, which is highly susceptible to PXO99A, by leaf-cutting inoculation.Four mutants with reduced virulence were obtained after two rounds of screening. Among them, the mutant PXM69 had completely lost virulence to the rice host and ability to elicit HR in non-host tobacco. Southern blotting analysis showed a single copy of a Tn5-insertion in the genome of PXM69. PCR walking and sequencing analysis revealed that the Tn5 transposon was inserted at nucleotide position 70,192–70,201 in the genome of PXO99A, disrupting the type III hrc(hrp-conserved) gene hrcQ, the first gene in the D operon of the hrp cluster in Xoo. To confirm the relationship between the Tn5-insertion and the avirulence phenotype of PXM69, we used the marker exchange mutagenesis to create a PXO99Amutant, ΔhrcQ::KAN, in which the hrcQ was disrupted by a kanamycin-encoding gene cassette at the same site as that of the Tn5-insertion. ΔhrcQ::KAN showed the same phenotype as mutant PXM69. Reintroduction of the wild-type hrcQ gene partially complemented the pathogenic function of PXM69. RT-PCR and cellulase secretion assays showed that the Tn5-disruption of hrcQ did not affect transcription of downstream genes in the D operon and function of the type II secretion system. Our results provide new insights into the pathogenic

Coagulase-negative staphylococci (CoNS) isolated from ready-to-eat food of animal origin--phenotypic and genotypic antibiotic resistance.

Science.gov (United States)

Chajęcka-Wierzchowska, Wioleta; Zadernowska, Anna; Nalepa, Beata; Sierpińska, Magda; Łaniewska-Trokenheim, Łucja

2015-04-01

The aim of this work was to study the pheno- and genotypical antimicrobial resistance profile of coagulase negative staphylococci (CoNS) isolated from 146 ready-to-eat food of animal origin (cheeses, cured meats, sausages, smoked fishes). 58 strains were isolated, they were classified as Staphylococcus xylosus (n = 29), Staphylococcus epidermidis (n = 16); Staphylococcus lentus (n = 7); Staphylococcus saprophyticus (n = 4); Staphylococcus hyicus (n = 1) and Staphylococcus simulans (n = 1) by phenotypic and genotypic methods. Isolates were tested for resistance to erythromycin, clindamycin, gentamicin, cefoxitin, norfloxacin, ciprofloxacin, tetracycline, tigecycline, rifampicin, nitrofurantoin, linezolid, trimetoprim, sulphamethoxazole/trimethoprim, chloramphenicol, quinupristin/dalfopristin by the disk diffusion method. PCR was used for the detection of antibiotic resistance genes encoding: methicillin resistance--mecA; macrolide resistance--erm(A), erm(B), erm(C), mrs(A/B); efflux proteins tet(K) and tet(L) and ribosomal protection proteins tet(M). For all the tet(M)-positive isolates the presence of conjugative transposons of the Tn916-Tn1545 family was determined. Most of the isolates were resistant to cefoxitin (41.3%) followed by clindamycin (36.2%), tigecycline (24.1%), rifampicin (17.2%) and erythromycin (13.8%). 32.2% staphylococcal isolates were multidrug resistant (MDR). All methicillin resistant staphylococci harboured mecA gene. Isolates, phenotypic resistant to tetracycline, harboured at least one tetracycline resistance determinant on which tet(M) was most frequent. All of the isolates positive for tet(M) genes were positive for the Tn916-Tn1545 -like integrase family gene. In the erythromycin-resistant isolates, the macrolide resistance genes erm(C) or msr(A/B) were present. Although coagulase-negative staphylococci are not classical food poisoning bacteria, its presence in food could be of public health significance due to the possible spread of

Relations between bilinear multipliers on Rn,Tn Rn,Tn Rn,Tn and Zn

Indian Academy of Sciences (India)

Since then the study of bilinear multiplier operators which commute with simultaneous translations have attracted a great deal of ... Unlike in the linear case, the boundedness of the symbol ψC is not known. In this article we will be dealing with .... For the converse, let ψ ∈ M p3 p1,p2 (Z). For f, g ∈ C∞ c (R), we have.

Tenascin-C in peripheral nerve morphogenesis.

Science.gov (United States)

Chiquet, M; Wehrle-Haller, B

1994-01-01

The extracellular matrix (ECM) molecule tenascin/cytotactin (TN-C) is expressed at a high level by satellite (glial precursor) cells in developing peripheral nerves of the chick embryo; synthesis of its mRNA peaks at the time period when axonal growth is maximal. When offered as a substrate in vitro, TN-C mediates neurite outgrowth by both motor and sensory neurons. The ability to grow neurites on TN-C is developmentally regulated: sensory neurons from 4-day chick embryos (the stage at which peripheral nerves start to develop) grow immediately and rapidly, whereas neurons from older embryos respond with a long delay. A TN-C domain responsible for this activity is located within the C-terminal (distal) portion of TN-C subunits. Integrin receptors seem to be involved on peripheral neurites because their growth on TN-C is completely blocked by antibodies to beta 1 integrins. In striking contrast to neuronal processes, nerve satellite cells can attach to a TN-C substrate but are completely inhibited in their migratory activity. Artificial substrate borders between tenascin and fibronectin or laminin act as selective barriers that allow neurites to pass while holding up satellite cells. The repulsive action of TN-C on satellite cells is similar to that observed for other cell types and is likely to be mediated by additional TN-C domains. In view of these data, it is surprising that mice seem to develop normally without a functional TN-C gene. TN-C is likely to be redundant, that is, its dual action on cell adhesion is shared by other molecules.(ABSTRACT TRUNCATED AT 250 WORDS)

The dog as a naturally-occurring model for insulin-like growth factor type 1 receptor-overexpressing breast cancer: an observational cohort study

International Nuclear Information System (INIS)

Jaillardon, Laetitia; Abadie, Jérome; Godard, Tiffanie; Campone, Mario; Loussouarn, Delphine; Siliart, Brigitte; Nguyen, Frédérique

2015-01-01

Dogs spontaneously develop invasive mammary carcinoma with a high prevalence of the triple-negative (TN) subtype (lack of ER-Estrogen Receptor and PR-Progesterone Receptor expression, lack of HER2-Human Epidermal Growth Factor Receptor 2 overexpression), making this animal model relevant for investigating new therapeutic pathways. Insulin-like growth factor Type-1 receptor (IGF1R) is frequently overexpressed in primary human breast cancers, with a growing role in the TN phenotype. The purpose of this study was to investigate the Dog as a candidate model for IGF1R-overexpressing mammary carcinoma. 150 bitches with canine mammary carcinoma (CMC) and a known 2-year follow-up were retrospectively included. IGF1R expression was assessed by immunohistochemistry (IHC) using a similar scoring system as for HER2 in breast cancer. The prognostic value of the IGF1R expression was assessed in terms of overall and specific survival as well as disease-free interval (DFI). 47 CMC (31 %) were classified as luminal and 103 (69 %) as triple-negative (TN-CMC). 41 % of CMC overexpressed IGF1R (IHC score 3+) of which 76 % were TN-CMC and 62 % grade III. IGF1R overexpression was associated with aggressive features including lymphovascular invasion, histological grade III, low ER expression and the TN phenotype. Univariate and multivariate analyses revealed that IGF1R overexpression was associated with shorter overall and specific survivals and shorter DFI in TN-CMC. IGF1R overexpression is common and related to a poor outcome in canine invasive mammary carcinoma, particularly in the triple negative subtype, as in human breast cancer. Preclinical studies using the Dog as a spontaneous animal model could be considered to investigate new therapies targeting IGF1R in triple-negative breast cancer. The online version of this article (doi:10.1186/s12885-015-1670-6) contains supplementary material, which is available to authorized users

Development of a new neutron shielding material, TN trademark Resin Vyal for transport/storage casks for radioactive materials

Energy Technology Data Exchange (ETDEWEB)

Abadie, P. [COGEMA Logistics (AREVA Group), Saint-Quentin-en-Yvelines (France)

2004-07-01

TN trademark Resin Vyal, a patent pending composite, is a new neutron shielding material developed by COGEMA LOGISTICS for transport/storage casks of radioactive materials (spent fuel, reprocessed fuel,..). This material is composed of a thermosetting resin (vinylester resin in solution of styrene) and two mineral fillers (alumine hydrate and zinc borate). Its shielding ability for neutron radiation is related to a high hydrogen content (for slowing down neutrons) and a high boron content (for absorbing neutrons). Source of hydrogen is organic matrix (resin) and alumine hydrate; source of boron is zinc borate. Atomic concentrations are equal to 5.10{sup 22} at/cm{sup 3} for hydrogen and 9.10{sup 20} at/cm{sup 3} for boron. Due to the flame retardant character of components, the final material has a good fire resistance (it is auto-extinguishable). Its density is equal to 1,8. The manufacturing process of these material is easy: it consists in mixing the fillers and pouring in-situ (in cask); so, the curing of this composite, which leads to a three-dimensional structure, takes place at ambient temperature. Temperature resistance of this material was evaluated by performing exposition tests of samples at different temperatures (150 C to 170 C). According to tests results, its maximal temperature of use was confirmed equal to 160 C.

Development of a new neutron shielding material, TN trademark Resin Vyal for transport/storage casks for radioactive materials

International Nuclear Information System (INIS)

Abadie, P.

2004-01-01

TN trademark Resin Vyal, a patent pending composite, is a new neutron shielding material developed by COGEMA LOGISTICS for transport/storage casks of radioactive materials (spent fuel, reprocessed fuel,..). This material is composed of a thermosetting resin (vinylester resin in solution of styrene) and two mineral fillers (alumine hydrate and zinc borate). Its shielding ability for neutron radiation is related to a high hydrogen content (for slowing down neutrons) and a high boron content (for absorbing neutrons). Source of hydrogen is organic matrix (resin) and alumine hydrate; source of boron is zinc borate. Atomic concentrations are equal to 5.10 22 at/cm 3 for hydrogen and 9.10 20 at/cm 3 for boron. Due to the flame retardant character of components, the final material has a good fire resistance (it is auto-extinguishable). Its density is equal to 1,8. The manufacturing process of these material is easy: it consists in mixing the fillers and pouring in-situ (in cask); so, the curing of this composite, which leads to a three-dimensional structure, takes place at ambient temperature. Temperature resistance of this material was evaluated by performing exposition tests of samples at different temperatures (150 C to 170 C). According to tests results, its maximal temperature of use was confirmed equal to 160 C

Novel plasmids and resistance phenotypes in Yersinia pestis: unique plasmid inventory of strain Java 9 mediates high levels of arsenic resistance.

Science.gov (United States)

Eppinger, Mark; Radnedge, Lyndsay; Andersen, Gary; Vietri, Nicholas; Severson, Grant; Mou, Sherry; Ravel, Jacques; Worsham, Patricia L

2012-01-01

Growing evidence suggests that the plasmid repertoire of Yersinia pestis is not restricted to the three classical virulence plasmids. The Java 9 strain of Y. pestis is a biovar Orientalis isolate obtained from a rat in Indonesia. Although it lacks the Y. pestis-specific plasmid pMT, which encodes the F1 capsule, it retains virulence in mouse and non-human primate animal models. While comparing diverse Y. pestis strains using subtractive hybridization, we identified sequences in Java 9 that were homologous to a Y. enterocolitica strain carrying the transposon Tn2502, which is known to encode arsenic resistance. Here we demonstrate that Java 9 exhibits high levels of arsenic and arsenite resistance mediated by a novel promiscuous class II transposon, named Tn2503. Arsenic resistance was self-transmissible from Java 9 to other Y. pestis strains via conjugation. Genomic analysis of the atypical plasmid inventory of Java 9 identified pCD and pPCP plasmids of atypical size and two previously uncharacterized cryptic plasmids. Unlike the Tn2502-mediated arsenic resistance encoded on the Y. enterocolitica virulence plasmid; the resistance loci in Java 9 are found on all four indigenous plasmids, including the two novel cryptic plasmids. This unique mobilome introduces more than 105 genes into the species gene pool. The majority of these are encoded by the two entirely novel self-transmissible plasmids, which show partial homology and synteny to other enterics. In contrast to the reductive evolution in Y. pestis, this study underlines the major impact of a dynamic mobilome and lateral acquisition in the genome evolution of the plague bacterium.

Novel plasmids and resistance phenotypes in Yersinia pestis: unique plasmid inventory of strain Java 9 mediates high levels of arsenic resistance.

Directory of Open Access Journals (Sweden)

Mark Eppinger

Full Text Available Growing evidence suggests that the plasmid repertoire of Yersinia pestis is not restricted to the three classical virulence plasmids. The Java 9 strain of Y. pestis is a biovar Orientalis isolate obtained from a rat in Indonesia. Although it lacks the Y. pestis-specific plasmid pMT, which encodes the F1 capsule, it retains virulence in mouse and non-human primate animal models. While comparing diverse Y. pestis strains using subtractive hybridization, we identified sequences in Java 9 that were homologous to a Y. enterocolitica strain carrying the transposon Tn2502, which is known to encode arsenic resistance. Here we demonstrate that Java 9 exhibits high levels of arsenic and arsenite resistance mediated by a novel promiscuous class II transposon, named Tn2503. Arsenic resistance was self-transmissible from Java 9 to other Y. pestis strains via conjugation. Genomic analysis of the atypical plasmid inventory of Java 9 identified pCD and pPCP plasmids of atypical size and two previously uncharacterized cryptic plasmids. Unlike the Tn2502-mediated arsenic resistance encoded on the Y. enterocolitica virulence plasmid; the resistance loci in Java 9 are found on all four indigenous plasmids, including the two novel cryptic plasmids. This unique mobilome introduces more than 105 genes into the species gene pool. The majority of these are encoded by the two entirely novel self-transmissible plasmids, which show partial homology and synteny to other enterics. In contrast to the reductive evolution in Y. pestis, this study underlines the major impact of a dynamic mobilome and lateral acquisition in the genome evolution of the plague bacterium.

Robustness encoded across essential and accessory replicons of the ecologically versatile bacterium Sinorhizobium meliloti

Science.gov (United States)

Walker, Graham C.; Finan, Turlough M.; Mengoni, Alessio; Griffitts, Joel S.

2018-01-01

Bacterial genome evolution is characterized by gains, losses, and rearrangements of functional genetic segments. The extent to which large-scale genomic alterations influence genotype-phenotype relationships has not been investigated in a high-throughput manner. In the symbiotic soil bacterium Sinorhizobium meliloti, the genome is composed of a chromosome and two large extrachromosomal replicons (pSymA and pSymB, which together constitute 45% of the genome). Massively parallel transposon insertion sequencing (Tn-seq) was employed to evaluate the contributions of chromosomal genes to growth fitness in both the presence and absence of these extrachromosomal replicons. Ten percent of chromosomal genes from diverse functional categories are shown to genetically interact with pSymA and pSymB. These results demonstrate the pervasive robustness provided by the extrachromosomal replicons, which is further supported by constraint-based metabolic modeling. A comprehensive picture of core S. meliloti metabolism was generated through a Tn-seq-guided in silico metabolic network reconstruction, producing a core network encompassing 726 genes. This integrated approach facilitated functional assignments for previously uncharacterized genes, while also revealing that Tn-seq alone missed over a quarter of wild-type metabolism. This work highlights the many functional dependencies and epistatic relationships that may arise between bacterial replicons and across a genome, while also demonstrating how Tn-seq and metabolic modeling can be used together to yield insights not obtainable by either method alone. PMID:29672509

Assessing total nitrogen in surface-water samples--precision and bias of analytical and computational methods

Science.gov (United States)

Rus, David L.; Patton, Charles J.; Mueller, David K.; Crawford, Charles G.

2013-01-01

percent. However, because particulate nitrogen constituted only 14 percent, on average, of TN-C, the precision of the TN-C method approached that of the method for dissolved nitrogen (2.3 percent). On the other hand, total Kjeldahl nitrogen (having a variability of 7.6 percent) constituted an average of 40 percent of TN-K, suggesting that the reduced precision of the Kjeldahl digestion may affect precision of the TN-K estimates. For most samples, the precision of TN computed as TN-C would be better (lower variability) than the precision of TN-K. In general, TN-A precision (having a variability of 2.1 percent) was superior to TN-C and TN-K methods. The laboratory experiment indicated that negative bias in TN-A was present across the entire range of sediment concentration and increased as sediment concentration increased. This suggested that reagent limitation was not the predominant cause of observed bias in TN-A. Furthermore, analyses of particulate nitrogen present in digest residues provided an almost complete accounting for the nitrogen that was underestimated by alkaline-persulfate digestion. This experiment established that, for the reference materials at least, negative bias in TN-A was caused primarily by the sequestration of some particulate nitrogen that was refractory to the digestion process. TN-K biases varied between positive and negative values in the laboratory experiment. Positive bias in TN-K is likely the result of the unintended reduction of a small and variable amount of nitrate to ammonia during the Kjeldahl digestion process. Negative TN-K bias may be the result of the sequestration of a portion of particulate nitrogen during the digestion process. Negative bias in TN-A was present across the entire range of suspended-sediment concentration (1 to 14,700 milligrams per liter [mg/L]) in the synoptic-field study, with relative bias being nearly as great at sediment concentrations below 10 mg/L (median of -3.5 percent) as that observed at sediment

An Enterobacter Plasmid as a New Genetic Background for the Transposon Tn1331

Science.gov (United States)

2011-11-25

determined to be 99% similar to E. cloacae by both 16S rDNA and Phoenix analysis and was designated Enterobacter sp W001. Enterobacter sp W001 was...adolescents. JAMA. 2002;287(23):3096–3102. 9. Foster TJ. Plasmid- determined resistance to antimicrobial drugs and toxic metal ions in bacteria. Microbiol...mediated type II dihydrofolate reductase gene among trimethoprim -resistant urinary pathogens in Greek hospitals. J Antimicrob Chemother. 1992;29

Naringenin Regulates Expression of Genes Involved in Cell Wall Synthesis in Herbaspirillum seropedicae▿

Science.gov (United States)

Tadra-Sfeir, M. Z.; Souza, E. M.; Faoro, H.; Müller-Santos, M.; Baura, V. A.; Tuleski, T. R.; Rigo, L. U.; Yates, M. G.; Wassem, R.; Pedrosa, F. O.; Monteiro, R. A.

2011-01-01

Five thousand mutants of Herbaspirillum seropedicae SmR1 carrying random insertions of transposon pTnMod-OGmKmlacZ were screened for differential expression of LacZ in the presence of naringenin. Among the 16 mutants whose expression was regulated by naringenin were genes predicted to be involved in the synthesis of exopolysaccharides, lipopolysaccharides, and auxin. These loci are probably involved in establishing interactions with host plants. PMID:21257805

Naringenin regulates expression of genes involved in cell wall synthesis in Herbaspirillum seropedicae.

Science.gov (United States)

Tadra-Sfeir, M Z; Souza, E M; Faoro, H; Müller-Santos, M; Baura, V A; Tuleski, T R; Rigo, L U; Yates, M G; Wassem, R; Pedrosa, F O; Monteiro, R A

2011-03-01

Five thousand mutants of Herbaspirillum seropedicae SmR1 carrying random insertions of transposon pTnMod-OGmKmlacZ were screened for differential expression of LacZ in the presence of naringenin. Among the 16 mutants whose expression was regulated by naringenin were genes predicted to be involved in the synthesis of exopolysaccharides, lipopolysaccharides, and auxin. These loci are probably involved in establishing interactions with host plants.

Transposon domestication versus mutualism in ciliate genome rearrangements.

Directory of Open Access Journals (Sweden)

Alexander Vogt

Full Text Available Ciliated protists rearrange their genomes dramatically during nuclear development via chromosome fragmentation and DNA deletion to produce a trimmer and highly reorganized somatic genome. The deleted portion of the genome includes potentially active transposons or transposon-like sequences that reside in the germline. Three independent studies recently showed that transposase proteins of the DDE/DDD superfamily are indispensible for DNA processing in three distantly related ciliates. In the spirotrich Oxytricha trifallax, high copy-number germline-limited transposons mediate their own excision from the somatic genome but also contribute to programmed genome rearrangement through a remarkable transposon mutualism with the host. By contrast, the genomes of two oligohymenophorean ciliates, Tetrahymena thermophila and Paramecium tetraurelia, encode homologous PiggyBac-like transposases as single-copy genes in both their germline and somatic genomes. These domesticated transposases are essential for deletion of thousands of different internal sequences in these species. This review contrasts the events underlying somatic genome reduction in three different ciliates and considers their evolutionary origins and the relationships among their distinct mechanisms for genome remodeling.

Staphylococcus aureus virulence factors identified by using a high-throughput Caenorhabditis elegans-killing model.

Science.gov (United States)

Begun, Jakob; Sifri, Costi D; Goldman, Samuel; Calderwood, Stephen B; Ausubel, Frederick M

2005-02-01

Staphylococcus aureus is an important human pathogen that is also able to kill the model nematode Caenorhabditis elegans. We constructed a 2,950-member Tn917 transposon insertion library in S. aureus strain NCTC 8325. Twenty-one of these insertions exhibited attenuated C. elegans killing, and of these, 12 contained insertions in different genes or chromosomal locations. Ten of these 12 insertions showed attenuated killing phenotypes when transduced into two different S. aureus strains, and 5 of the 10 mutants correspond to genes that have not been previously identified in signature-tagged mutagenesis studies. These latter five mutants were tested in a murine renal abscess model, and one mutant harboring an insertion in nagD exhibited attenuated virulence. Interestingly, Tn917 was shown to have a very strong bias for insertions near the terminus of DNA replication.

Transposon characterization of vancomycin-resistant Enterococcus faecium (VREF) and dissemination of resistance associated with transferable plasmids

DEFF Research Database (Denmark)

Migura, Lourdes Garcia; Liebana, Ernesto; Jensen, Lars Bogø

2007-01-01

Objectives: VanA glycopeptide resistance has persisted on broiler farms in the UK despite the absence of the antimicrobial selective pressure, avoparcin. This study aimed to investigate the contribution of horizontal gene transfer of Tn 1546 versus clonal spread in the dissemination of the resist......Objectives: VanA glycopeptide resistance has persisted on broiler farms in the UK despite the absence of the antimicrobial selective pressure, avoparcin. This study aimed to investigate the contribution of horizontal gene transfer of Tn 1546 versus clonal spread in the dissemination...... plasmid replicons, associated with antimicrobial resistance on several unrelated farms. Conclusions: Horizontal transfer of vancomycin resistance may play a more important role in the persistence of antimicrobial resistance than clonal spread. The presence of different plasmid replicons, associated...... with antimicrobial resistance on several unrelated farms, illustrates the ability of these enterococci to acquire and disseminate mobile genetic elements within integrated livestock systems....

Trigeminal neuralgia

DEFF Research Database (Denmark)

Maarbjerg, Stine; Di Stefano, Giulia; Bendtsen, Lars

2017-01-01

-occupying lesion affecting the trigeminal nerve. Differential diagnosis and treatment Important differential diagnoses include trigeminal autonomic cephalalgias, posttraumatic or postherpetic pain and other facial pains. First line treatment is prophylactic medication with sodium channel blockers, and second line......Introduction Trigeminal neuralgia (TN) is characterized by touch-evoked unilateral brief shock-like paroxysmal pain in one or more divisions of the trigeminal nerve. In addition to the paroxysmal pain, some patients also have continuous pain. TN is divided into classical TN (CTN) and secondary TN...

[Active miniature inverted-repeat transposable elements transposon in plants: a review].

Science.gov (United States)

Hu, Bingjie; Zhou, Mingbing

2018-02-25

Miniature inverted-repeat transposable elements transposon is a special transposon that could transpose by "cut-paste" mechanism, which is one of characteristics of DNA transposons. Otherwise, the copy number of MITEs is very high, which is one of characteristics of RNA transposons. Many MITE families have been reported, but little about active MITEs. We summarize recent advances in studying active MITEs. Most the MITEs belong to the Tourist-like family, such as mPing, mGing, PhTourist1, Tmi1 and PhTst-3. Additionally, DTstu1 and MITE-39 belong to Stowaway-like family, and AhMITEs1 belongs to Mutator-like family. Moreover, we summarize the structure (terminal inverse repeats and target site duplications), copy number, evolution pattern and transposition characteristics of these active MITEs, to provide the foundation for the identification of other active MITEs and subsequent research on MITE transposition and amplification mechanism.

A newly emerging HIV-1 recombinant lineage (CRF58_01B) disseminating among people who inject drugs in Malaysia.

Science.gov (United States)

Chow, Wei Zhen; Takebe, Yutaka; Syafina, Nur Ezreen; Prakasa, Malarvelli Soorya; Chan, Kok Gan; Al-Darraji, Haider Abdulrazzaq Abed; Koh, Clayton; Kamarulzaman, Adeeba; Tee, Kok Keng

2014-01-01

The HIV epidemic is primarily characterised by the circulation of HIV-1 group M (main) comprising of 11 subtypes and sub-subtypes (A1, A2, B-D, F1, F2, G, H, J, and K) and to date 55 circulating recombinant forms (CRFs). In Southeast Asia, active inter-subtype recombination involving three main circulating genotypes--subtype B (including subtype B', the Thai variant of subtype B), CRF01_AE, and CRF33_01B--have contributed to the emergence of novel unique recombinant forms. In the present study, we conducted the molecular epidemiological surveillance of HIV-1 gag-RT genes among 258 people who inject drugs (PWIDs) in Kuala Lumpur, Malaysia, between 2009 and 2011 whereby a novel CRF candidate was recently identified. The near full-length genome sequences obtained from six epidemiologically unlinked individuals showed identical mosaic structures consisting of subtype B' and CRF01_AE, with six unique recombination breakpoints in the gag-RT, pol, and env regions. Among the high-risk population of PWIDs in Malaysia, which was predominantly infected by CRF33_01B (>70%), CRF58_01B circulated at a low but significant prevalence (2.3%, 6/258). Interestingly, the CRF58_01B shared two unique recombination breakpoints with other established CRFs in the region: CRF33_01B, CRF48_01B, and CRF53_01B in the gag gene, and CRF15_01B (from Thailand) in the env gene. Extended Bayesian Markov chain Monte Carlo sampling analysis showed that CRF58_01B and other recently discovered CRFs were most likely to have originated in Malaysia, and that the recent spread of recombinant lineages in the country had little influence from neighbouring countries. The isolation, genetic characterization, and evolutionary features of CRF58_01B among PWIDs in Malaysia signify the increasingly complex HIV-1 diversity in Southeast Asia that may hold an implication on disease treatment, control, and prevention.

A newly emerging HIV-1 recombinant lineage (CRF58_01B disseminating among people who inject drugs in Malaysia.

Directory of Open Access Journals (Sweden)

Wei Zhen Chow

Full Text Available The HIV epidemic is primarily characterised by the circulation of HIV-1 group M (main comprising of 11 subtypes and sub-subtypes (A1, A2, B-D, F1, F2, G, H, J, and K and to date 55 circulating recombinant forms (CRFs. In Southeast Asia, active inter-subtype recombination involving three main circulating genotypes--subtype B (including subtype B', the Thai variant of subtype B, CRF01_AE, and CRF33_01B--have contributed to the emergence of novel unique recombinant forms. In the present study, we conducted the molecular epidemiological surveillance of HIV-1 gag-RT genes among 258 people who inject drugs (PWIDs in Kuala Lumpur, Malaysia, between 2009 and 2011 whereby a novel CRF candidate was recently identified. The near full-length genome sequences obtained from six epidemiologically unlinked individuals showed identical mosaic structures consisting of subtype B' and CRF01_AE, with six unique recombination breakpoints in the gag-RT, pol, and env regions. Among the high-risk population of PWIDs in Malaysia, which was predominantly infected by CRF33_01B (>70%, CRF58_01B circulated at a low but significant prevalence (2.3%, 6/258. Interestingly, the CRF58_01B shared two unique recombination breakpoints with other established CRFs in the region: CRF33_01B, CRF48_01B, and CRF53_01B in the gag gene, and CRF15_01B (from Thailand in the env gene. Extended Bayesian Markov chain Monte Carlo sampling analysis showed that CRF58_01B and other recently discovered CRFs were most likely to have originated in Malaysia, and that the recent spread of recombinant lineages in the country had little influence from neighbouring countries. The isolation, genetic characterization, and evolutionary features of CRF58_01B among PWIDs in Malaysia signify the increasingly complex HIV-1 diversity in Southeast Asia that may hold an implication on disease treatment, control, and prevention.

Troponin T isoform expression is modulated during Atlantic Halibut metamorphosis

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Llewellyn Lynda

2007-06-01

Full Text Available Abstract Background Flatfish metamorphosis is a thyroid hormone (TH driven process which leads to a dramatic change from a symmetrical larva to an asymmetrical juvenile. The effect of THs on muscle and in particular muscle sarcomer protein genes is largely unexplored in fish. The change in Troponin T (TnT, a pivotal protein in the assembly of skeletal muscles sarcomeres and a modulator of calcium driven muscle contraction, during flatfish metamophosis is studied. Results In the present study five cDNAs for halibut TnT genes were cloned; three were splice variants arising from a single fast TnT (fTnT gene; a fourth encoded a novel teleost specific fTnT-like cDNA (AfTnT expressed exclusively in slow muscle and the fifth encoded the teleost specific sTnT2. THs modified the expression of halibut fTnT isoforms which changed from predominantly basic to acidic isoforms during natural and T4 induced metamorphosis. In contrast, expression of red muscle specific genes, AfTnT and sTnT2, did not change during natural metamorphosis or after T4 treatment. Prior to and after metamorphosis no change in the dorso-ventral symmetry or temporal-spatial expression pattern of TnT genes and muscle fibre organization occurred in halibut musculature. Conclusion Muscle organisation in halibut remains symmetrical even after metamorphosis suggesting TH driven changes are associated with molecular adaptations. We hypothesize that species specific differences in TnT gene expression in teleosts underlies different larval muscle developmental programs which better adapts them to the specific ecological constraints.

The identification of a tetracycline resistance gene tet(M), on a Tn916-like transposon, in the Bacillus cereus group

DEFF Research Database (Denmark)

Agersø, Yvonne; Jensen, Lars Bogø; Givskov, Michael Christian

2002-01-01

In order to investigate whether resistance genes present in bacteria in manure could transfer to indigenous soil bacteria, resistant isolates belonging to the Bacillus cereus group (Bacillus cereus, Bacillus anthracis and Bacillus thuringiensis) were isolated from farm soil (72 isolates) and manure...

Simultaneous Identification of Potential Pathogenicity Factors of Mycoplasma agalactiae in the Natural Ovine Host by Negative Selection

OpenAIRE

Hegde, Shivanand; Hegde, Shrilakshmi; Zimmermann, Martina; Flöck, Martina; Spergser, Joachim; Rosengarten, Renate; Chopra-Dewasthaly, Rohini

2015-01-01

Mycoplasmas possess complex pathogenicity determinants that are largely unknown at the molecular level. Mycoplasma agalactiae serves as a useful model to study the molecular basis of mycoplasma pathogenicity. The generation and in vivo screening of a transposon mutant library of M. agalactiae were employed to unravel its host colonization factors. Tn4001mod mutants were sequenced using a novel sequencing method, and functionally heterogeneous pools containing 15 to 19 selected mutants were sc...

Mapping the resistance-associated mobilome of a carbapenem-resistant Klebsiella pneumoniae strain reveals insights into factors shaping these regions and facilitates generation of a 'resistance-disarmed' model organism.

Science.gov (United States)

Bi, Dexi; Jiang, Xiaofei; Sheng, Zi-Ke; Ngmenterebo, David; Tai, Cui; Wang, Minggui; Deng, Zixin; Rajakumar, Kumar; Ou, Hong-Yu

2015-10-01

This study aims to investigate the landscape of the mobile genome, with a focus on antibiotic resistance-associated factors in carbapenem-resistant Klebsiella pneumoniae. The mobile genome of the completely sequenced K. pneumoniae HS11286 strain (an ST11, carbapenem-resistant, near-pan-resistant, clinical isolate) was annotated in fine detail. The identified mobile genetic elements were mapped to the genetic contexts of resistance genes. The blaKPC-2 gene and a 26 kb region containing 12 clustered antibiotic resistance genes and one biocide resistance gene were deleted, and the MICs were determined again to ensure that antibiotic resistance had been lost. HS11286 contains six plasmids, 49 ISs, nine transposons, two separate In2-related integron remnants, two integrative and conjugative elements (ICEs) and seven prophages. Sixteen plasmid-borne resistance genes were identified, 14 of which were found to be directly associated with Tn1721-, Tn3-, Tn5393-, In2-, ISCR2- and ISCR3-derived elements. IS26 appears to have actively moulded several of these genetic regions. The deletion of blaKPC-2, followed by the deletion of a 26 kb region containing 12 clustered antibiotic resistance genes, progressively decreased the spectrum and level of resistance exhibited by the resultant mutant strains. This study has reiterated the role of plasmids as bearers of the vast majority of resistance genes in this species and has provided valuable insights into the vital role played by ISs, transposons and integrons in shaping the resistance-coding regions in this important strain. The 'resistance-disarmed' K. pneumoniae ST11 strain generated in this study will offer a more benign and readily genetically modifiable model organism for future extensive functional studies. © The Author 2015. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Molecular characterization of antibiotic resistance in enterococci recovered from seagulls (Larus cachinnans) representing an environmental health problem.

Science.gov (United States)

Radhouani, Hajer; Igrejas, Gilberto; Pinto, Luís; Gonçalves, Alexandre; Coelho, Céline; Rodrigues, Jorge; Poeta, Patrícia

2011-08-01

Antimicrobial resistance and the mechanisms implicated were studied in 54 enterococci recovered from 57 seagull fecal samples. Almost 78% of the recovered enterococci showed resistance against one or more antibiotics and these isolates were identified to the species level. E. faecium was the most prevalent species (52.4%). High percentages of erythromycin and tetracycline resistances were found among our isolates (95.2%), and lower percentages were identified to other antibiotics. Most of the tetracycline-resistant strains carried the tet(M) and/or tet(L) genes. Genes associated with Tn916/Tn1545 and/or Tn5397 transposons were detected in 45% of tetracycline-resistant isolates. The erm(B) gene was detected in 65% of erythromycin-resistant isolates. The vat(D) and vat(E) genes were present in 5.9% and 11.8% of quinupristin/dalfopristin-resistant isolates, respectively. The ant(6)-Ia gene was identified in 57.1% of streptomycin-resistant isolates. All nine kanamycin-resistant isolates carried the aph(3)'-IIIa gene. The cat(A) gene was found in two chloramphenicol-resistant isolates. Seagulls should be considered a risk species for spreading in the environment antimicrobial resistant enterococci and can serve as a sentinel for antibiotic pressure from the surrounding farm and urban setting.

Complete genome sequence of hypervirulent and outbreak-associated Acinetobacter baumannii strain LAC-4: epidemiology, resistance genetic determinants and potential virulence factors

Science.gov (United States)

Ou, Hong-Yu; Kuang, Shan N.; He, Xinyi; Molgora, Brenda M.; Ewing, Peter J.; Deng, Zixin; Osby, Melanie; Chen, Wangxue; Xu, H. Howard

2015-01-01

Acinetobacter baumannii is an important human pathogen due to its multi-drug resistance. In this study, the genome of an ST10 outbreak A. baumannii isolate LAC-4 was completely sequenced to better understand its epidemiology, antibiotic resistance genetic determinants and potential virulence factors. Compared with 20 other complete genomes of A. baumannii, LAC-4 genome harbors at least 12 copies of five distinct insertion sequences. It contains 12 and 14 copies of two novel IS elements, ISAba25 and ISAba26, respectively. Additionally, three novel composite transposons were identified: Tn6250, Tn6251 and Tn6252, two of which contain resistance genes. The antibiotic resistance genetic determinants on the LAC-4 genome correlate well with observed antimicrobial susceptibility patterns. Moreover, twelve genomic islands (GI) were identified in LAC-4 genome. Among them, the 33.4-kb GI12 contains a large number of genes which constitute the K (capsule) locus. LAC-4 harbors several unique putative virulence factor loci. Furthermore, LAC-4 and all 19 other outbreak isolates were found to harbor a heme oxygenase gene (hemO)-containing gene cluster. The sequencing of the first complete genome of an ST10 A. baumannii clinical strain should accelerate our understanding of the epidemiology, mechanisms of resistance and virulence of A. baumannii. PMID:25728466

Tunneling nanotubes: A versatile target for cancer therapy.

Science.gov (United States)

Sahoo, Pragyaparamita; Jena, Soumya Ranjan; Samanta, Luna

2017-11-29

Currently Cancer is the leading cause of death worldwide. Malignancy or cancer is a class of diseases characterized by uncontrolled cell growth that eventually invade other tissues and dvelop secondary malignant growth at other sites by metastasis. Intercellular communication plays a major in cancer, particularly in the process of cell proliferation and coordination which in turn leads to tumor invasion, metastasis and development of resistance to therapy. Cells communicate among themselves in a variety of ways, namely, i) via gap junctions with adjacent cells, ii) via exosomes with nearby cells and iii) via chemical messengers with distant cells. Besides, cell - cell connection by tunneling nanotubes (TnTs) is recently gaining importance where intercellular components are transferred between cells. In general cell organelles like Golgi vesicle and mitochondria; and biomolecules like nucleic acids and proteins are transferred through these TnTs. These TnTs are long cytoplasmic extensions made up of actin that function as intercellular bridge and connect a wide verity of cell types. Malignant cells form TnTs with either another malignant cells or cells of the surrounding tumor matrix. These TnTs help in the process of initiation of tumor formation, its organization and propagation. The current review focuses on the role of TnTs mediated cell – cell signaling in cancer micro-environment. Drugs that inhibit TnT-formation such as metformin and everolimus can be targeted towards TnTs in the management of cancer growth, proliferation, tumor invasion and metastasis. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Cost-effectiveness analysis of HLA-B*5801 testing in preventing allopurinol-induced SJS/TEN in Thai population.

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Surasak Saokaew

Full Text Available BACKGROUND: Stevens-Johnson syndrome (SJS and Toxic Epidermal Necrolysis (TEN, caused by allopurinol therapy, are strongly associated with the human leukocyte antigen (HLA, HLA-B*5801. Identification of HLA-B*5801 genotype before prescribing allopurinol offers the possibility of avoiding allopurinol-induced SJS/TEN. As there is a paucity of evidence about economic value of such testing, this study aims to determine the cost-effectiveness of HLA-B*5801 testing compared with usual care (no genetic testing before allopurinol administration in Thailand. METHODS AND FINDING: A decision analytical and Markov model was used to estimate life time costs and outcomes represented as quality adjusted life years (QALYs gained. The model was populated with relevant information of the association between gene and allopurinol-induced SJS/TEN, test characteristics, costs, and epidemiologic data for Thailand from a societal perspective. Input data were obtained from the literature and a retrospective database analysis. The results were expressed as incremental cost per QALY gained. A base-case analysis was performed for patients at age 30. A series of sensitivity analyses including scenario, one-way, and probabilistic sensitivity analyses were constructed to explore the robustness of the findings. Based on a hypothetical cohort of 1,000 patients, the incremental total cost was 923,919 THB (USD 29,804 and incremental QALY was 5.89 with an ICER of 156,937.04 THB (USD 5,062 per QALY gained. The cost of gout management, incidence of SJS/TEN, case fatality rate of SJS/TEN, and cost of genetic testing are considered very influential parameters on the cost-effectiveness value of HLA-B*5801 testing. CONCLUSIONS: The genetic testing for HLA-B*5801 before allopurinol administration is considered a highly potential cost-effective intervention in Thailand. The findings are sensitive to a number of factors. In addition to cost-effectiveness findings, consideration of other

Tailed nuclei and dicentric chromosomes in irradiated subjects

International Nuclear Information System (INIS)

Kravtsov, V.Yu.; Fedortseva, R.F.; Starkova, Ye.V.; Yartseva, N.M.; Nikiforov, A.M.

2000-01-01

A study of peripheral blood smears from irradiated Chernobyl liquidators and other subjects has shown nuclei of some lymphocytes to have a protrusion into the cytoplasm. Such abnormal nuclei are called 'tailed' nuclei (TN). Sixteen main morphologic types, observed in human peripheral blood lymphocytes, are described. The frequency of appearance of lymphocytes with TN in a group of Chernobyl liquidators was significantly higher than in control groups (p<0.001, in all cases). A positive correlation was found between the TN frequency in lymphocytes and dicentric chromosomes in lymphocytes cultured to metaphase (p<0.001). Elevated frequencies of dicentrics, higher than 0.1%, were found in 17 out of 22 subjects in whom the frequency of lymphocytes with TN was 0.8% and more. Abnormalities of the TN type in lymphocytes are likely to result from breakdown of chromosome bridges formed by dicentrics. The TN can be considered as a possible marker of irradiation and, therefore, their detailed study is important

Two weather radar time series of the altitude of the volcanic plume during the May 2011 eruption of Grímsvötn, Iceland

Directory of Open Access Journals (Sweden)

G. N. Petersen

2012-10-01

Full Text Available The eruption of Grímsvötn volcano in Iceland in 2011 lasted for a week, 21–28 May. The eruption was explosive and peaked during the first hours, with the eruption plume reaching 20–25 km altitude. The height of the plume was monitored every 5 min with a C-band weather radar located at Keflavík International Airport and a mobile X-band radar, 257 km and 75 km distance from the volcano respectively. In addition, photographs taken during the first half-hour of the eruption give information regarding the initial rise. Time series of the plume-top altitude were constructed from the radar observations. This paper presents the two independent radar time series. The series have been cross validated and there is a good agreement between them. The echo top radar series of the altitude of the volcanic plume are publicly available from the Pangaea Data Publisher (doi:10.1594/PANGAEA.778390.

Elevated Cardiac Troponin T in Patients With Skeletal Myopathies.

Science.gov (United States)

Schmid, Johannes; Liesinger, Laura; Birner-Gruenberger, Ruth; Stojakovic, Tatjana; Scharnagl, Hubert; Dieplinger, Benjamin; Asslaber, Martin; Radl, Roman; Beer, Meinrad; Polacin, Malgorzata; Mair, Johannes; Szolar, Dieter; Berghold, Andrea; Quasthoff, Stefan; Binder, Josepha S; Rainer, Peter P

2018-04-10

Cardiac troponins are often elevated in patients with skeletal muscle disease who have no evidence of cardiac disease. The goal of this study was to characterize cardiac troponin concentrations in patients with myopathies and derive insights regarding the source of elevated troponin T measurements. Cardiac troponin T (cTnT) and cardiac troponin I (cTnI) concentrations were determined by using high sensitivity assays in 74 patients with hereditary and acquired skeletal myopathies. Patients underwent comprehensive cardiac evaluation, including 12-lead electrocardiogram, 24-h electrocardiogram, cardiac magnetic resonance imaging, and coronary artery computed tomography. cTnT and cTnI protein expression was determined in skeletal muscle samples of 9 patients and in control tissues derived from autopsy using antibodies that are used in commercial assays. Relevant Western blot bands were subjected to liquid chromatography tandem mass spectrometry for protein identification. Levels of cTnT (median: 24 ng/l; interquartile range: 11 to 54 ng/l) were elevated (>14 ng/l) in 68.9% of patients; cTnI was elevated (>26 ng/l) in 4.1% of patients. Serum cTnT levels significantly correlated with creatine kinase and myoglobin (r = 0.679 and 0.786, respectively; both p < 0.001). Based on cTnT serial testing, 30.1% would have fulfilled current rule-in criteria for myocardial infarction. Noncoronary cardiac disease was present in 23%. Using cTnT antibodies, positive bands were found in both diseased and healthy skeletal muscle at molecular weights approximately 5 kDa below cTnT. Liquid chromatography tandem mass spectrometry identified the presence of skeletal troponin T isoforms in these bands. Measured cTnT concentrations were chronically elevated in the majority of patients with skeletal myopathies, whereas cTnI elevation was rare. Our data indicate that cross-reaction of the cTnT immunoassay with skeletal muscle troponin isoforms was the likely cause. Copyright © 2018 The

Guatemala: Ozbrojený konflikt 1960-1996 a současná situace: se zvláštním zaměřením na Máje

OpenAIRE

Vilímková, Olga

2009-01-01

Tématem disertační práce je Guatemala: ozbrojený konflikt v letech 1960 - 1996 a současná situace se zvláštním zaměřením na Máje. Příčiny ozbrojeného konfliktu druhé ploviny XX. století (a mnoha současných problémů) v Guatemale se nacházejí v hluboké minulosti země. Už v době koloniální se začaly formovat ekonomicko - sociální vztahy mezi dvěma antagonistickými vrstvami společnosti: kolonizátory a původním obyvatelstvem, které bylo porobeno, vytlačeno na okraj společnosti a využíváno jako pra...

Global mapping of transposon location.

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Abram Gabriel

2006-12-01

Full Text Available Transposable genetic elements are ubiquitous, yet their presence or absence at any given position within a genome can vary between individual cells, tissues, or strains. Transposable elements have profound impacts on host genomes by altering gene expression, assisting in genomic rearrangements, causing insertional mutations, and serving as sources of phenotypic variation. Characterizing a genome's full complement of transposons requires whole genome sequencing, precluding simple studies of the impact of transposition on interindividual variation. Here, we describe a global mapping approach for identifying transposon locations in any genome, using a combination of transposon-specific DNA extraction and microarray-based comparative hybridization analysis. We use this approach to map the repertoire of endogenous transposons in different laboratory strains of Saccharomyces cerevisiae and demonstrate that transposons are a source of extensive genomic variation. We also apply this method to mapping bacterial transposon insertion sites in a yeast genomic library. This unique whole genome view of transposon location will facilitate our exploration of transposon dynamics, as well as defining bases for individual differences and adaptive potential.

A Two-Component Regulatory System, CsrR-CsrS, Represses Expression of Three Streptococcus pyogenes Virulence Factors, Hyaluronic Acid Capsule, Streptolysin S, and Pyrogenic Exotoxin B

OpenAIRE

Heath, Andrew; DiRita, Victor J.; Barg, Neil L.; Engleberg, N. Cary

1999-01-01

Certain Tn916 insertions in the chromosome of an M1-type, nonmucoid Streptococcus pyogenes isolate (MGAS166) were previously shown to result in stable mucoidy with increased expression of the capsular synthetic genes. The transposon insertions in these strains are directly upstream of an apparent operon encoding a two-component regulatory system, designated csrR-csrS. Compared with MGAS166, these mucoid mutants are more hemolytic and cause significantly more tissue damage in a murine model of...

Genome-wide analysis of Tol2 transposon reintegration in zebrafish.

Science.gov (United States)

Kondrychyn, Igor; Garcia-Lecea, Marta; Emelyanov, Alexander; Parinov, Sergey; Korzh, Vladimir

2009-09-08

Tol2, a member of the hAT family of transposons, has become a useful tool for genetic manipulation of model animals, but information about its interactions with vertebrate genomes is still limited. Furthermore, published reports on Tol2 have mainly been based on random integration of the transposon system after co-injection of a plasmid DNA harboring the transposon and a transposase mRNA. It is important to understand how Tol2 would behave upon activation after integration into the genome. We performed a large-scale enhancer trap (ET) screen and generated 338 insertions of the Tol2 transposon-based ET cassette into the zebrafish genome. These insertions were generated by remobilizing the transposon from two different donor sites in two transgenic lines. We found that 39% of Tol2 insertions occurred in transcription units, mostly into introns. Analysis of the transposon target sites revealed no strict specificity at the DNA sequence level. However, Tol2 was prone to target AT-rich regions with weak palindromic consensus sequences centered at the insertion site. Our systematic analysis of sequential remobilizations of the Tol2 transposon from two independent sites within a vertebrate genome has revealed properties such as a tendency to integrate into transcription units and into AT-rich palindrome-like sequences. This information will influence the development of various applications involving DNA transposons and Tol2 in particular.

Tenascin-C in the extracellular matrix promotes the selection of highly proliferative and tubulogenesis-defective endothelial cells

Energy Technology Data Exchange (ETDEWEB)

Alves, Tercia Rodrigues [Universidade do Estado do Rio de Janeiro (UERJ), Instituto de Biologia Roberto Alcantara Gomes, Departamento de Biologia Celular, Laboratorio de Biologia da Celula Endotelial e da Angiogenese (LabAngio), Rio de Janeiro (Brazil); Universidade Federal do Rio de Janeiro (UFRJ), Programa de Biologia Celular e do Desenvolvimento, Instituto de Ciencias Biomedicas, INNT/INCT/MCT, Rio de Janeiro (Brazil); Carvalho da Fonseca, Anna Carolina [Universidade Federal do Rio de Janeiro (UFRJ), Programa de Biologia Celular e do Desenvolvimento, Instituto de Ciencias Biomedicas, INNT/INCT/MCT, Rio de Janeiro (Brazil); Nunes, Sara Santana; Oliveira da Silva, Aline [Universidade do Estado do Rio de Janeiro (UERJ), Instituto de Biologia Roberto Alcantara Gomes, Departamento de Biologia Celular, Laboratorio de Biologia da Celula Endotelial e da Angiogenese (LabAngio), Rio de Janeiro (Brazil); Dubois, Luiz Gustavo Feijo; Faria, Jane; Kahn, Suzana Assad [Universidade Federal do Rio de Janeiro (UFRJ), Programa de Biologia Celular e do Desenvolvimento, Instituto de Ciencias Biomedicas, INNT/INCT/MCT, Rio de Janeiro (Brazil); Viana, Nathan Bessa [Universidade Federal do Rio de Janeiro, Laboratorio de Pincas Oticas, Coordenacao de Programas de Estudos Avancados, Instituto de Ciencias Biomedicas, Rio de Janeiro (Brazil); Universidade Federal do Rio de Janeiro, Instituto de Fisica, Rio de Janeiro (Brazil); Marcondes, Jorge [Universidade Federal do Rio de Janeiro, Hospital Universitario Clementino Fraga Filho, Servico de Neurocirurgia, Rio de Janeiro (Brazil); Legrand, Chantal [Institut Universitaire d' Hematologie, Universite Paris-Diderot, Paris 7, INSERM U553, Paris (France); Moura-Neto, Vivaldo [Universidade Federal do Rio de Janeiro (UFRJ), Programa de Biologia Celular e do Desenvolvimento, Instituto de Ciencias Biomedicas, INNT/INCT/MCT, Rio de Janeiro (Brazil); and others

2011-09-10

The extracellular matrix (ECM) contains important cues for tissue homeostasis and morphogenesis. The matricellular protein tenascin-C (TN-C) is overexpressed in remodeling tissues and cancer. In the present work, we studied the effect of different ECM-which exhibited a significant diversity in their TN-C content-in endothelial survival, proliferation and tubulogenic differentiation: autologous (endothelial) ECM devoid of TN-C, but bearing large amounts of FN; fibroblast ECM, bearing both high TN-C and FN contents; and finally, glioma-derived matrices, usually poor in FN, but very rich in TN-C. HUVECs initially adhered to the immobilized matrix produced by U373 MG glioma cells, but significantly detached and died by anoikis (50 to 80%) after 24 h, as compared with cells incubated with endothelial and fibroblast matrices. Surviving endothelial cells (20 to 50%) became up to 6-fold more proliferative and formed 74-97% less tube-like structures in vitro than cells grown on non-tumoral matrices. An antibody against the EGF-like repeats of tenascin-C (TN-C) partially rescued cells from the tubulogenic defect, indicating that this molecule is responsible for the selection of highly proliferative and tubulogenic defective endothelial cells. Interestingly, by using defined substrata, in conditions that mimic glioma and normal cell ECM composition, we observed that fibronectin (FN) modulates the TN-C-induced selection of endothelial cells. Our data show that TN-C is able to modulate endothelial branching morphogenesis in vitro and, since it is prevalent in matrices of injured and tumor tissues, also suggest a role for this protein in vascular morphogenesis, in these physiological contexts.

Tenascin-C in the extracellular matrix promotes the selection of highly proliferative and tubulogenesis-defective endothelial cells

International Nuclear Information System (INIS)

Alves, Tercia Rodrigues; Carvalho da Fonseca, Anna Carolina; Nunes, Sara Santana; Oliveira da Silva, Aline; Dubois, Luiz Gustavo Feijo; Faria, Jane; Kahn, Suzana Assad; Viana, Nathan Bessa; Marcondes, Jorge; Legrand, Chantal; Moura-Neto, Vivaldo

2011-01-01

The extracellular matrix (ECM) contains important cues for tissue homeostasis and morphogenesis. The matricellular protein tenascin-C (TN-C) is overexpressed in remodeling tissues and cancer. In the present work, we studied the effect of different ECM-which exhibited a significant diversity in their TN-C content-in endothelial survival, proliferation and tubulogenic differentiation: autologous (endothelial) ECM devoid of TN-C, but bearing large amounts of FN; fibroblast ECM, bearing both high TN-C and FN contents; and finally, glioma-derived matrices, usually poor in FN, but very rich in TN-C. HUVECs initially adhered to the immobilized matrix produced by U373 MG glioma cells, but significantly detached and died by anoikis (50 to 80%) after 24 h, as compared with cells incubated with endothelial and fibroblast matrices. Surviving endothelial cells (20 to 50%) became up to 6-fold more proliferative and formed 74-97% less tube-like structures in vitro than cells grown on non-tumoral matrices. An antibody against the EGF-like repeats of tenascin-C (TN-C) partially rescued cells from the tubulogenic defect, indicating that this molecule is responsible for the selection of highly proliferative and tubulogenic defective endothelial cells. Interestingly, by using defined substrata, in conditions that mimic glioma and normal cell ECM composition, we observed that fibronectin (FN) modulates the TN-C-induced selection of endothelial cells. Our data show that TN-C is able to modulate endothelial branching morphogenesis in vitro and, since it is prevalent in matrices of injured and tumor tissues, also suggest a role for this protein in vascular morphogenesis, in these physiological contexts.

Elevated Plasma Cardiac Troponin T Levels Caused by Skeletal Muscle Damage in Pompe Disease.

Science.gov (United States)

Wens, Stephan C A; Schaaf, Gerben J; Michels, Michelle; Kruijshaar, Michelle E; van Gestel, Tom J M; In 't Groen, Stijn; Pijnenburg, Joon; Dekkers, Dick H W; Demmers, Jeroen A A; Verdijk, Lex B; Brusse, Esther; van Schaik, Ron H N; van der Ploeg, Ans T; van Doorn, Pieter A; Pijnappel, W W M Pim

2016-02-01

Elevated plasma cardiac troponin T (cTnT) levels in patients with neuromuscular disorders may erroneously lead to the diagnosis of acute myocardial infarction or myocardial injury. In 122 patients with Pompe disease, the relationship between cTnT, cardiac troponin I, creatine kinase (CK), CK-myocardial band levels, and skeletal muscle damage was assessed. ECG and echocardiography were used to evaluate possible cardiac disease. Patients were divided into classic infantile, childhood-onset, and adult-onset patients. cTnT levels were elevated in 82% of patients (median 27 ng/L, normal values normal in all patients, whereas CK-myocardial band levels were increased in 59% of patients. cTnT levels correlated with CK levels in all 3 subgroups (Pmass index measured with echocardiography was normal in all the 3 subgroups. cTnT mRNA expression in skeletal muscle was not detectable in controls but was strongly induced in patients with Pompe disease. cTnT protein was identified by mass spectrometry in patient-derived skeletal muscle tissue. Elevated plasma cTnT levels in patients with Pompe disease are associated with skeletal muscle damage, rather than acute myocardial injury. Increased cTnT levels in Pompe disease and likely other neuromuscular disorders should be interpreted with caution to avoid unnecessary cardiac interventions. © 2016 American Heart Association, Inc.

Drug: D00105 [KEGG MEDICUS

Lifescience Database Archive (English)

Full Text Available D00105 Drug Estradiol (JAN/USP/INN); Climara (TN); Divigel (TN); Estrace (TN); Estraderm (TN); Estr...asorb (TN); Estring (TN); Estrogel (TN); Innofem (TN); Vagifem (TN); Vivelle (TN) ... C18H24O2... D00105.gif ... Other ... DG01584 ... Estrogen receptor agonist ... DG01986 ... Estrogen Cyp substrate ... DG01892 ... CYP1A2 s... code: G03CA03 Chemical group: DG00462 ... Estrogen NR3A1 (ESR1) [HSA:2099] [KO:K08550]; NR3A2 (ESR2) [HSA:210

31 CFR 601.4 - Use of paper; interest-bearing securities of the United States.

Science.gov (United States)

2010-07-01

... PAPER FOR UNITED STATES CURRENCY AND OTHER SECURITIES § 601.4 Use of paper; interest-bearing securities of the United States. The existing distinctive papers shall be used for the printing of interest...

Legionella dumoffii Tex-KL Mutated in an Operon Homologous to traC-traD is Defective in Epithelial Cell Invasion.

Science.gov (United States)

Qin, Tian; Ken-Ichiro, Iida; Ren, Hong Yu; Zhou, Hai Jian; Yoshida, Shin-Ichi

2016-06-01

To understand the mechanism of invasion by Legionella dumoffii. The L. dumoffii strain Tex-KL was mutated using the Tn903 derivative, Tn903dIIlacZ. After screening 799 transposon insertion mutants, we isolated one defective mutant. We then constructed the gene-disrupted mutant, KL16, and studied its invasion of and intracellular growth in HeLa and A549 cells, and in A/J mice survival experiments. The structure of traC-traD operon was analyzed by RT-PCR. The transposon insertion was in a gene homologous to Salmonella typhi traC, which is required for the assembly of F pilin into the mature F pilus structure and for conjugal DNA transmission. Results from RT-PCR suggested that the traC-traD region formed an operon. We found that when the traC gene was disrupted, invasion and intracellular growth of L. dumoffii Tex-KL were impaired in human epithelial cells. When mice were infected by intranasal inoculation with a traC deficient mutant, their survival significantly increased when compared to mice infected with the wild-type strain.. Our results indicated that the traC-traD operon is required for the invasion and intracellular growth abilities of L. dumoffii Tex-KL in epithelial cells. Copyright © 2016 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.

Genome-wide analysis of Tol2 transposon reintegration in zebrafish

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Parinov Sergey

2009-09-01

Full Text Available Abstract Background Tol2, a member of the hAT family of transposons, has become a useful tool for genetic manipulation of model animals, but information about its interactions with vertebrate genomes is still limited. Furthermore, published reports on Tol2 have mainly been based on random integration of the transposon system after co-injection of a plasmid DNA harboring the transposon and a transposase mRNA. It is important to understand how Tol2 would behave upon activation after integration into the genome. Results We performed a large-scale enhancer trap (ET screen and generated 338 insertions of the Tol2 transposon-based ET cassette into the zebrafish genome. These insertions were generated by remobilizing the transposon from two different donor sites in two transgenic lines. We found that 39% of Tol2 insertions occurred in transcription units, mostly into introns. Analysis of the transposon target sites revealed no strict specificity at the DNA sequence level. However, Tol2 was prone to target AT-rich regions with weak palindromic consensus sequences centered at the insertion site. Conclusion Our systematic analysis of sequential remobilizations of the Tol2 transposon from two independent sites within a vertebrate genome has revealed properties such as a tendency to integrate into transcription units and into AT-rich palindrome-like sequences. This information will influence the development of various applications involving DNA transposons and Tol2 in particular.

Human and Swine Hosts Share Vancomycin-Resistant Enterococcus faecium CC17 and CC5 and Enterococcus faecalis CC2 Clonal Clusters Harboring Tn1546 on Indistinguishable Plasmids

DEFF Research Database (Denmark)

Freitas, Ana R.; Coque, Teresa M.; Novais, Carla

2011-01-01

clonally related Enterococcus faecium clonal complex 5 (CC5) isolates (17 sequence type 6 [ST6], 6 ST5, 5 ST185, 1 ST147, and 1 ST493) were obtained from feces of swine and healthy humans. This collection included isolates widespread among pigs of European Union (EU) countries since the mid-1990s. Each ST...... comprised isolates showing similar pulsed-field gel electrophoresis (PFGE) patterns (≤6 bands difference; >82% similarity). Some CC5 PFGE subtype strains from swine were indistinguishable from hospital vancomycin-resistant enterococci (VRE) causing infections. A truncated variant of Tn1546 (encoding...... resistance to vancomycin) and tcrB (coding for resistance to copper) were consistently located on 150- to 190-kb plasmids (rep(pLG1)). E. faecium CC17 (ST132) isolates from pig manure and two clinical samples showed identical PFGE profiles and contained a 60-kb mosaic plasmid (rep(Inc18) plus rep...

Complexity of Complement Resistance Factors Expressed by Acinetobacter baumannii Needed for Survival in Human Serum.

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Sanchez-Larrayoz, Amaro F; Elhosseiny, Noha M; Chevrette, Marc G; Fu, Yang; Giunta, Peter; Spallanzani, Raúl G; Ravi, Keerthikka; Pier, Gerald B; Lory, Stephen; Maira-Litrán, Tomás

2017-10-15

Acinetobacter baumannii is a bacterial pathogen with increasing impact in healthcare settings, due in part to this organism's resistance to many antimicrobial agents, with pneumonia and bacteremia as the most common manifestations of disease. A significant proportion of clinically relevant A. baumannii strains are resistant to killing by normal human serum (NHS), an observation supported in this study by showing that 12 out of 15 genetically diverse strains of A. baumannii are resistant to NHS killing. To expand our understanding of the genetic basis of A. baumannii serum resistance, a transposon (Tn) sequencing (Tn-seq) approach was used to identify genes contributing to this trait. An ordered Tn library in strain AB5075 with insertions in every nonessential gene was subjected to selection in NHS. We identified 50 genes essential for the survival of A. baumannii in NHS, including already known serum resistance factors, and many novel genes not previously associated with serum resistance. This latter group included the maintenance of lipid asymmetry genetic pathway as a key determinant in protecting A. baumannii from the bactericidal activity of NHS via the alternative complement pathway. Follow-up studies validated the role of eight additional genes identified by Tn-seq in A. baumannii resistance to killing by NHS but not by normal mouse serum, highlighting the human species specificity of A. baumannii serum resistance. The identification of a large number of genes essential for serum resistance in A. baumannii indicates the degree of complexity needed for this phenotype, which might reflect a general pattern that pathogens rely on to cause serious infections. Copyright © 2017 by The American Association of Immunologists, Inc.

Insertional inactivation of a chromosomal locus that modulates expression of potential virulence determinants in Staphylococcus aureus.

OpenAIRE

Cheung, A L; Wolz, C; Yeaman, M R; Bayer, A S

1995-01-01

A single insertion of transposon Tn551 into a unique chromosomal locus of Staphylococcus aureus ISP479C has resulted in a pleiotropic effect on the expression of both extracellular and cell wall proteins. In particular, the expression of cell wall protein A and clumping activity with fibrinogen were rendered undetectable in the mutant 1E3 compared with the parent. The secretion of alpha-hemolysin in mutant 1E3 was modestly increased. Southern blot and phenotypic analyses indicated that this l...

Tics in TACs: A Step into an Avalanche? Systematic Literature Review and Conclusions.

Science.gov (United States)

Wöber, Christian

2017-11-01

Trigeminal autonomic cephalalgias (TACs) comprise cluster headache, paroxysmal hemicrania, short-lasting unilateral neuralgiform headache attacks, and hemicrania continua. In some cases, trigeminal neuralgia (TN, "tic douloureux") or TN-like pain may co-occur with TACs. This article will review the co-occurrence and overlap of TACs and tics in order to contribute to a better understanding of the issue and an improved management of the patients. For performing a systematic literature review Pubmed was searched using a total of ten terms. The articles identified were screened for further articles of relevance. TACs are related to tics in various ways. TN or TN-like paroxysms may co-occur with CH, PH, and HC, labeled as cluster-tic syndrome, PH-tic syndrome, and HC-tic syndrome. Such co-occurrence was not only found in the primary TACs but also in secondary headaches resembling TACs. The initial onset of TAC and tic may be simultaneous or separated by months or years. In acute attacks, tic and TAC may occur concurrently or much more often independently of each other. The term "cluster-tic syndrome" was also used in patients with a single type of pain in a twilight zone between TACs and TN fulfilling none of the relevant diagnostic criteria. Short-lasting neuralgiform headache attacks overlap with TN in terms of clinical features, imaging findings, and therapy. © 2017 American Headache Society.

Prognostic Value of Triple-Negative Phenotype at the Time of Locally Recurrent, Conservatively Treated Breast Cancer

International Nuclear Information System (INIS)

Parikh, Rahul R.; Housman, Douglas; Yang Qifeng; Toppmeyer, Deborah; Wilson, Lynn D.; Haffty, Bruce G.

2008-01-01

Purpose: To evaluate the prognostic value of triple-negative (TN) ER, PR, Her2/neu basal-like carcinoma of the breast, at the time of ipsilateral breast tumor recurrence (IBTR) after conservative surgery and radiation treatment (RT). Methods and Materials: A tissue microarray was constructed of 47 IBTR specimens of patients who experienced an IBTR after conservative surgery and RT that were processed and stained for ER, PR, and HER2/neu. Results: At a median post-recurrence follow-up of 7.5 years, the 5-year overall survival (OS) and disease metastasis-free survival (DMFS) after IBTR were 91.4% and 83.0%, respectively. Median time to tumor recurrence (TTR) and IBTR was shorter in the TN phenotype (3.88 vs. 5.00 years; p = 0.09). The TN tumors were not associated with size of local recurrence or recurrence elsewhere in the breast. Despite administration of standard chemotherapy at the time of IBTR, the 5-year DMFS and 5-year OS for the TN cohort were 48.6% and 72.7%, respectively. The 5-year DMFS was 48.6% for TN tumors and 90.8% for non-TN tumors (p < 0.01). By univariate analysis, significant factors associated with poor 5-year DMFS and OS after IBTR included: TN phenotype (p < 0.01), TTR 3 years or less (p < 0.01), local recurrence at or near the original tumor site (p = 0.08). In multivariate analysis, TN was a significant independent predictor of poorer 5-year DMFS (relative risk, 5.91; 95% confidence interval, 1.83-19.01; p < 0.01) after IBTR. Conclusions: Although patients experiencing an IBTR have a relatively favorable prognosis, those with IBTR events of the TN phenotype had a rather poor prognosis despite receiving standard chemotherapy. Strategies with novel systemic therapies to improve outcomes in patients experiencing IBTR of the TN phenotype are warranted

Ca2+-induced PRE-NMR changes in the troponin complex reveal the possessive nature of the cardiac isoform for its regulatory switch.

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Nicole M Cordina

Full Text Available The interaction between myosin and actin in cardiac muscle, modulated by the calcium (Ca2+ sensor Troponin complex (Tn, is a complex process which is yet to be fully resolved at the molecular level. Our understanding of how the binding of Ca2+ triggers conformational changes within Tn that are subsequently propagated through the contractile apparatus to initiate muscle activation is hampered by a lack of an atomic structure for the Ca2+-free state of the cardiac isoform. We have used paramagnetic relaxation enhancement (PRE-NMR to obtain a description of the Ca2+-free state of cardiac Tn by describing the movement of key regions of the troponin I (cTnI subunit upon the release of Ca2+ from Troponin C (cTnC. Site-directed spin-labeling was used to position paramagnetic spin labels in cTnI and the changes in the interaction between cTnI and cTnC subunits were then mapped by PRE-NMR. The functionally important regions of cTnI targeted in this study included the cTnC-binding N-region (cTnI57, the inhibitory region (cTnI143, and two sites on the regulatory switch region (cTnI151 and cTnI159. Comparison of 1H-15N-TROSY spectra of Ca2+-bound and free states for the spin labeled cTnC-cTnI binary constructs demonstrated the release and modest movement of the cTnI switch region (∼10 Å away from the hydrophobic N-lobe of troponin C (cTnC upon the removal of Ca2+. Our data supports a model where the non-bound regulatory switch region of cTnI is highly flexible in the absence of Ca2+ but remains in close vicinity to cTnC. We speculate that the close proximity of TnI to TnC in the cardiac complex is favourable for increasing the frequency of collisions between the N-lobe of cTnC and the regulatory switch region, counterbalancing the reduction in collision probability that results from the incomplete opening of the N-lobe of TnC that is unique to the cardiac isoform.

Complete nucleotide sequence of CTX-M-15-plasmids from clinical Escherichia coli isolates: insertional events of transposons and insertion sequences.

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Annemieke Smet

Full Text Available BACKGROUND: CTX-M-producing Escherichia coli strains are regarded as major global pathogens. METHODOLOGY/PRINCIPAL FINDINGS: The nucleotide sequence of three plasmids (pEC_B24: 73801-bp; pEC_L8: 118525-bp and pEC_L46: 144871-bp from Escherichia coli isolates obtained from patients with urinary tract infections and one plasmid (pEC_Bactec: 92970-bp from an Escherichia coli strain isolated from the joint of a horse with arthritis were determined. Plasmid pEC_Bactec belongs to the IncI1 group and carries two resistance genes: bla(TEM-1 and bla(CTX-M-15. It shares more than 90% homology with a previously published bla(CTX-M-plasmid from E. coli of human origin. Plasmid pEC_B24 belongs to the IncFII group whereas plasmids pEC_L8 and pEC_L46 represent a fusion of two replicons of type FII and FIA. On the pEC_B24 backbone, two resistance genes, bla(TEM-1 and bla(CTX-M-15, were found. Six resistance genes, bla(TEM-1, bla(CTX-M-15, bla(OXA-1, aac6'-lb-cr, tetA and catB4, were detected on the pEC_L8 backbone. The same antimicrobial drug resistance genes, with the exception of tetA, were also identified on the pEC_L46 backbone. Genome analysis of all 4 plasmids studied provides evidence of a seemingly frequent transposition event of the bla(CTX-M-15-ISEcp1 element. This element seems to have a preferred insertion site at the tnpA gene of a bla(TEM-carrying Tn3-like transposon, the latter itself being inserted by a transposition event. The IS26-composite transposon, which contains the bla(OXA-1, aac6'-lb-cr and catB4 genes, was inserted into plasmids pEC_L8 and pEC_L46 by homologous recombination rather than a transposition event. Results obtained for pEC_L46 indicated that IS26 also plays an important role in structural rearrangements of the plasmid backbone and seems to facilitate the mobilisation of fragments from other plasmids. CONCLUSIONS: Collectively, these data suggests that IS26 together with ISEcp1 could play a critical role in the evolution of

Bursts and horizontal evolution of DNA transposons in the speciation of pseudotetraploid salmonids

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Davidson William S

2007-11-01

Full Text Available Abstract Background Several genome duplications have occurred in the evolutionary history of teleost fish. In returning to a stable diploid state, the polyploid genome reorganized, and large portions are lost, while the fish lines evolved to numerous species. Large scale transposon movement has been postulated to play an important role in the genome reorganization process. We analyzed the DNA sequence of several large loci in Salmo salar and other species for the presence of DNA transposon families. Results We have identified bursts of activity of 14 families of DNA transposons (12 Tc1-like and 2 piggyBac-like families, including 11 novel ones in genome sequences of Salmo salar. Several of these families have similar sequences in a number of closely and distantly related fish, lamprey, and frog species as well as in the parasite Schistosoma japonicum. Analysis of sequence similarities between copies within the families of these bursts demonstrates several waves of transposition activities coinciding with salmonid species divergence. Tc1-like families show a master gene-like copying process, illustrated by extensive but short burst of copying activity, while the piggyBac-like families show a more random copying pattern. Recent families may include copies with an open reading frame for an active transposase enzyme. Conclusion We have identified defined bursts of transposon activity that make use of master-slave and random mechanisms. The bursts occur well after hypothesized polyploidy events and coincide with speciation events. Parasite-mediated lateral transfer of transposons are implicated.

Stimulated forces demonstrated: Why the trouton-noble experiment failed and how to make it succeed

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Cornille, Patrick; Naudin, Jean-Louis; Szames, Alexandre

1999-01-01

At the turn of the 20th century, Frederick T. Trouton and Henry R. Noble (TN) performed a little known æther drift experiment which came to be known as the electrostatic analogue of Michelson-Morley's famous experiment. If the æther were real, they said, a capacitor charged with ``high'' voltage should exhibit a ``jerk'' and a subsequent ``spontaneous'' torque, thus demonstrating the existence of an æther wind. Trouton and Noble failed to observe the torque and paved the way to the special theory of relativity. We have replicated the TN experiment. We obtained positive results: the principle of relativity is disproved. Other TN-like experiments allowed us to observe the stimulated rotational motions (TN's long-sought effect), the stimulated translational motions (Biefeld-Brown effect), and the sustained rotation of charged, suspended capacitors. Video movies are presented at the conference. TN's failure is analyzed. The concept of stimulated forces is explained in light of its relevance to the future of space locomotion.

Serial High-Sensitivity Troponin T in Post-Primary Angioplasty Exercise Test

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Humberto Andres Vaz

2016-04-01

Full Text Available Abstract Background: The kinetics of high-sensitivity troponin T (hscTnT release should be studied in different situations, including functional tests with transient ischemic abnormalities. Objective: To evaluate the release of hscTnT by serial measurements after exercise testing (ET, and to correlate hscTnT elevations with abnormalities suggestive of ischemia. Methods: Patients with acute ST-segment elevation myocardial infarction (STEMI undergoing primary angioplasty were referred for ET 3 months after infarction. Blood samples were collected to measure basal hscTnT immediately before (TnT0h, 2 (TnT2h, 5 (TnT5h, and 8 hours (TnT8h after ET. The outcomes were peak hscTnT, TnT5h/TnT0h ratio, and the area under the blood concentration-time curve (AUC for hscTnT levels. Log-transformation was performed on hscTnT values, and comparisons were assessed with the geometric mean ratio, along with their 95% confidence intervals. Statistical significance was assessed by analysis of covariance with no adjustment, and then, adjusted for TnT0h, age and sex, followed by additional variables (metabolic equivalents, maximum heart rate achieved, anterior wall STEMI, and creatinine clearance. Results: This study included 95 patients. The highest geometric means were observed at 5 hours (TnT5h. After adjustments, peak hscTnT, TnT5h/TnT0h and AUC were 59% (p = 0.002, 59% (p = 0.003 and 45% (p = 0.003 higher, respectively, in patients with an abnormal ET as compared to those with normal tests. Conclusion: Higher elevations of hscTnT may occur after an abnormal ET as compared to a normal ET in patients with STEMI.

Serial High-Sensitivity Troponin T in Post-Primary Angioplasty Exercise Test

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Vaz, Humberto Andres, E-mail: humbertovaz@cardiol.br; Vanz, Ana Paula; Castro, Iran [Instituto de Cardiologia - Fundação Universitária de Cardiologia, Porto Alegre, RS (Brazil)

2016-04-15

The kinetics of high-sensitivity troponin T (hscTnT) release should be studied in different situations, including functional tests with transient ischemic abnormalities. To evaluate the release of hscTnT by serial measurements after exercise testing (ET), and to correlate hscTnT elevations with abnormalities suggestive of ischemia. Patients with acute ST-segment elevation myocardial infarction (STEMI) undergoing primary angioplasty were referred for ET 3 months after infarction. Blood samples were collected to measure basal hscTnT immediately before (TnT{sub 0h}), 2 (TnT{sub 2h}), 5 (TnT{sub 5h}), and 8 hours (TnT{sub 8h}) after ET. The outcomes were peak hscTnT, TnT{sub 5h}/TnT{sub 0h} ratio, and the area under the blood concentration-time curve (AUC) for hscTnT levels. Log-transformation was performed on hscTnT values, and comparisons were assessed with the geometric mean ratio, along with their 95% confidence intervals. Statistical significance was assessed by analysis of covariance with no adjustment, and then, adjusted for TnT{sub 0h}, age and sex, followed by additional variables (metabolic equivalents, maximum heart rate achieved, anterior wall STEMI, and creatinine clearance). This study included 95 patients. The highest geometric means were observed at 5 hours (TnT{sub 5h}). After adjustments, peak hscTnT, TnT{sub 5h}/TnT{sub 0h} and AUC were 59% (p = 0.002), 59% (p = 0.003) and 45% (p = 0.003) higher, respectively, in patients with an abnormal ET as compared to those with normal tests. Higher elevations of hscTnT may occur after an abnormal ET as compared to a normal ET in patients with STEMI.

Církevní kritika výstřední módy císařského dvora Karla IV. a datování nástěnných maleb karlštejnského schodištního cyklu kolem let 1365 - 1367

Czech Academy of Sciences Publication Activity Database

Hlobil, Ivo

2006-01-01

Roč. 13, příl. (2006), s. 19-22 ISSN 1212-1487. [Schodištní cykly Velké věže hradu Karlštejna. Stav po restaurování. Praha, 05.05.2004-07.05.2004] Institutional research plan: CEZ:AV0Z80330511 Keywords : wall painting * Karlštejn * Charles IV Subject RIV: AL - Art, Architecture, Cultural Heritage

Effect of extended exposure to low-dose radiation on autoimmune diseases of immunologically suppressed MRL/MpTn-gld/gld mice

International Nuclear Information System (INIS)

Ootsuyama, Akira; Okazaki, Ryuji; Norimura, Toshiyuki

2003-01-01

The purpose of this paper is to analyze the relationship between alterations of splenic T-cell subpopulations and the amelioration of autoimmune diseases of MRL/MpTn-gld/gld mice (MRL/gld mice) after extended exposure to low-dose radiation. After the onset of disease, 4-month-old MRL/gld mice were exposed to doses of 0.05, 0.2, and 0.5 Gy/day for 4 weeks (5 days/week), for total doses of 1, 4, and 10 Gy, respectively. The MRL/gld mice that were irradiated with 0.2 and 0.5 Gy/day showed an obvious decrease in the proportion of splenic CD4 - CD8 - T cells and remission of their autoimmune diseases. After the last irradiation, apoptotic cells were found in the white pulp of the spleen of the MRL/gld mice irradiated with 0.2 Gy/day, but not in the MRL/MpJ-+/+ mice (MRL/wild mice), which experienced a similar treatment. Before the onset of disease, 3-month-old MRL/gld mice subjected to 0.2 Gy/day showed a decrease in the proportion of splenic CD4 - CD8 - T cells and less remission of their autoimmune diseases than the 4-month-old mice. These results suggest that the accumulated CD4 - CD8 - T cells are more sensitive to radiation than other T cell subpopulations, and that decreasing CD4 - CD8 - T cells with extended exposure to low-dose radiation leads to the amelioration of autoimmune disease. (author)

A novel low-temperature-active β-glucosidase from symbiotic Serratia sp. TN49 reveals four essential positions for substrate accommodation.

Science.gov (United States)

Zhou, Junpei; Zhang, Rui; Shi, Pengjun; Huang, Huoqing; Meng, Kun; Yuan, Tiezheng; Yang, Peilong; Yao, Bin

2011-10-01

A 2,373-bp full-length gene (bglA49) encoding a 790-residue polypeptide (BglA49) with a calculated mass of 87.8 kDa was cloned from Serratia sp. TN49, a symbiotic bacterium isolated from the gut of longhorned beetle (Batocera horsfieldi) larvae. The deduced amino acid sequence of BglA49 showed the highest identities of 80.1% with a conceptually translated protein from Pantoea sp. At-9b (EEW02556), 38.3% with the identified glycoside hydrolase (GH) family 3 β-glucosidase from Clostridium stercorarium NCBI 11754 (CAB08072), and sp. G5 (ABL09836) and Paenibacillus sp. C7 (AAX35883). The recombinant enzyme (r-BglA49) was expressed in Escherichia coli and displayed the typical characteristics of low-temperature-active enzymes, such as low temperature optimum (showing apparent optimal activity at 35°C), activity at low temperatures (retaining approximately 60% of its maximum activity at 20°C and approximately 25% at 10°C). Compared with the thermophilic GH 3 β-glucosidase, r-BglA49 had fewer hydrogen bonds and salt bridges and less proline residues. These features might relate to the increased structure flexibility and higher catalytic activity at low temperatures of r-BglA49. The molecular docking study of four GH 3 β-glucosidases revealed five conserved positions contributing to substrate accommodation, among which four positions of r-BglA49 (R192, Y228, D260, and E449) were identified to be essential based on site-directed mutagenesis analysis.

Phage transposon mutagenesis.

Science.gov (United States)

Siegrist, M Sloan; Rubin, Eric J

2009-01-01

Phage transduction is an attractive method of genetic manipulation in mycobacteria. PhiMycoMarT7 is well suited for transposon mutagenesis as it is temperature sensitive for replication and contains T7 promoters that promote transcription, a highly active transposase gene, and an Escherichia coli oriR6 K origin of replication. Mycobacterial transposon mutant libraries produced by PhiMycoMarT7 transduction are amenable to both forward and reverse genetic studies. In this protocol, we detail the preparation of PhiMycoMarT7, including a description of the phage, reconstitution of the phage, purification of plaques, preparation of phage stock, and titering of phage stock. We then describe the transduction procedure and finally outline the isolation of individual transposon mutants.

Evaluation of analytical performance of a new high-sensitivity immunoassay for cardiac troponin I.

Science.gov (United States)

Masotti, Silvia; Prontera, Concetta; Musetti, Veronica; Storti, Simona; Ndreu, Rudina; Zucchelli, Gian Carlo; Passino, Claudio; Clerico, Aldo

2018-02-23

The study aim was to evaluate and compare the analytical performance of the new chemiluminescent immunoassay for cardiac troponin I (cTnI), called Access hs-TnI using DxI platform, with those of Access AccuTnI+3 method, and high-sensitivity (hs) cTnI method for ARCHITECT platform. The limits of blank (LoB), detection (LoD) and quantitation (LoQ) at 10% and 20% CV were evaluated according to international standardized protocols. For the evaluation of analytical performance and comparison of cTnI results, both heparinized plasma samples, collected from healthy subjects and patients with cardiac diseases, and quality control samples distributed in external quality assessment programs were used. LoB, LoD and LoQ at 20% and 10% CV values of the Access hs-cTnI method were 0.6, 1.3, 2.1 and 5.3 ng/L, respectively. Access hs-cTnI method showed analytical performance significantly better than that of Access AccuTnI+3 method and similar results to those of hs ARCHITECT cTnI method. Moreover, the cTnI concentrations measured with Access hs-cTnI method showed close linear regressions with both Access AccuTnI+3 and ARCHITECT hs-cTnI methods, although there were systematic differences between these methods. There was no difference between cTnI values measured by Access hs-cTnI in heparinized plasma and serum samples, whereas there was a significant difference between cTnI values, respectively measured in EDTA and heparin plasma samples. Access hs-cTnI has analytical sensitivity parameters significantly improved compared to Access AccuTnI+3 method and is similar to those of the high-sensitivity method using ARCHITECT platform.

Uusehitised Tallinna vanalinnas

Index Scriptorium Estoniae

2003-01-01

Aegade jooksul Tallinna vanalinna ehitatud uued ehitised ( kino "Sõprus" Vana-Posti tn-l, Kirjanike maja Harju tn-l, Väike-Karja tn. lillekauplus, Raeköök Dunkri tn-l, Kaubahall, WW Passaaz Aia tn-l, kino "Bi-Ba-Bo"(ehitatud 1930-ndail), klaaspaviljon ( nn. Jänese vorst) ja De La Gardie kaubamaja Viru tn-l jt.)

Population dynamics of Vibrio fischeri during infection of Euprymna scolopes.

Science.gov (United States)

McCann, Jessica; Stabb, Eric V; Millikan, Deborah S; Ruby, Edward G

2003-10-01

The luminous bacterium Vibrio fischeri colonizes a specialized light-emitting organ within its squid host, Euprymna scolopes. Newly hatched juvenile squid must acquire their symbiont from ambient seawater, where the bacteria are present at low concentrations. To understand the population dynamics of V. fischeri during colonization more fully, we used mini-Tn7 transposons to mark bacteria with antibiotic resistance so that the growth of their progeny could be monitored. When grown in culture, there was no detectable metabolic burden on V. fischeri cells carrying the transposon, which inserts in single copy in a specific intergenic region of the V. fischeri genome. Strains marked with mini-Tn7 also appeared to be equivalent to the wild type in their ability to infect and multiply within the host during coinoculation experiments. Studies of the early stages of colonization suggested that only a few bacteria became associated with symbiotic tissue when animals were exposed for a discrete period (3 h) to an inoculum of V. fischeri cells equivalent to natural population levels; nevertheless, all these hosts became infected. When three differentially marked strains of V. fischeri were coincubated with juvenile squid, the number of strains recovered from an individual symbiotic organ was directly dependent on the size of the inoculum. Further, these results indicated that, when exposed to low numbers of V. fischeri, the host may become colonized by only one or a few bacterial cells, suggesting that symbiotic infection is highly efficient.

Plasmid borne Carbapenem-Hydrolyzing Class D β-Lactamases (CHDLs) and AdeABC efflux pump conferring carbapenem-tigecycline resistance among Acinetobacter baumannii isolates harboring TnAbaRs.

Science.gov (United States)

Savari, Mohammad; Ekrami, Alireza; Shoja, Saeed; Bahador, Abbas

2017-03-01

Here we studied the prevalence and mechanisms of simultaneous resistance to carbapenem and tigecycline and accumulation of resistance determinants reservoirs in genome of Acinetobacter baumannii (A. baumannii) clinical isolates. Susceptibility of the isolates were measured to 18 antimicrobial agents. Genetic diversity of the microbial population was determined using the International Clonal lineage typing (IC typing), multiple locus VNTR analysis (MLVA) and plasmid profiling methods. To detect the AbaRs, Carbapenem-Hydrolyzing Class D β-Lactamases (CHDLs) genes, AdeABC efflux pump genes and resistance determinants, PCR was used. Filter mating experiments were used to prove that if carbapenem resistance genes are located on conjugative plasmids or not. Among the A. baumannii clinical isolates, 40.8% were carbapenem-tigecycline resistant and in this population, 46.9% were belonging to IC I, IC II or IC III and 53.1% were IC variants. These isolates had fallen in 40 MLVA types and were harboring plasmids in multiple numbers and sizes. In this study, bla OXA-23-like was the most prevalent CHDL and conjugation analysis proved that the carbapenem resistance genes are located on conjugative plasmids. All efflux pump genes, except for adeC, were detected in all carbapenem-tigecycline resistant A. baumannii (CTRAb) isolates. Resistance determinants were distributed in both TnAbaRs and R plasmids with a shift toward the R plasmids. Emerging of carbapenem resistant A. baumannii (CRAB) with simultaneous resistance to the last line therapy including tigecycline represent emerging of extensively drug resistance (XDR) and pandrug resistance (PDR) phenotypes that would be a great threat to our public health system. Copyright © 2017 Elsevier Ltd. All rights reserved.

The Effect of Silicon on some Morpho-physiological Characteristics and Grain Yield of Sorghum (Sorghum bicolor L. under Salt Stress

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S Hasibi

2016-12-01

Full Text Available Introduction Nowadays, salinity is one of the limiting factors for crop production in arid and semi-arid regions. On the other hand, sorghum (Sorghum bicolor L. is a self-pollinated and short-day plant, which partly has been adapted to salinity and water stress conditions; also play an important role in humans, livestock and poultry nourishments. All studies have showed the positive effects of Silicon on growth and yield of plants in both normal and stress conditions. The aim of this experiment was to improve salinity tolerance of Sorghum by application of Silicon. Materials and Methods A split plot experiment based on randomized complete block design with three replications in both normal and salt stress conditions was carried out at research farm of Shahid Bahonar University of Kerman in 2013. Silicon treatments (0 and 6 mM were considered as main plot and various sorghum genotypes (payam, sepideh, TN-4-70, TN-04-71, TN-04-39, TN-04-107, TN-04-100, TN-04-37, TN-04-68, TN-04-83, TN-04-62 and TN-04-95 were assigned to sub plots. The sodium silicate was used as silica source. The data were analyzed by SAS software using combine analysis. Means comparisons were accomplished by Duncan multiple range test at 5% probability level. Some of the measured traits were as follow: Relative water content (Ritchie and Nguyen, 1990, Relative permeability (33, leaf area index and chlorophyll index (by SPAD. Results and Discussion According to the results, use of silicon led to increase of RWC under salinity stress, while RWC decreased by 13% when no silicon applied. Salinity significantly decreased 1000-grain weight. Maximum grain yield obtained from TN-04-37 (987.6 g m-2 under normal condition with foliar application of silicon. Application of silicon under stress condition led to 38% increase in grain yield of Sepideh compared to control. Under salt stress, silicon also increased shoot dry weight in TN-04-107, TN-04-70, TN-04-37, Payam and Sepideh genotypes

Use of HLA-B*58:01 genotyping to prevent allopurinol induced severe cutaneous adverse reactions in Taiwan: national prospective cohort study

Science.gov (United States)

Ko, Tai-Ming; Tsai, Chang-Youh; Chen, Shih-Yang; Chen, Kuo-Shu; Yu, Kuang-Hui; Chu, Chih-Sheng; Huang, Chung-Ming; Wang, Chrong-Reen; Weng, Chia-Tse; Yu, Chia-Li; Hsieh, Song-Chou; Tsai, Jer-Chia; Lai, Wen-Ter; Tsai, Wen-Chan; Yin, Guang-Dar; Ou, Tsan-Teng; Cheng, Kai-Hung; Yen, Jeng-Hsien; Liou, Teh-Ling; Lin, Tsung-Hsien; Chen, Der-Yuan; Hsiao, Pi-Jung; Weng, Meng-Yu; Chen, Yi-Ming; Chen, Chen-Hung; Liu, Ming-Fei; Yen, Hsueh-Wei; Lee, Jia-Jung; Kuo, Mei-Chuan; Wu, Chen-Ching; Hung, Shih-Yuan; Luo, Shue-Fen; Yang, Ya-Hui; Chuang, Hui-Ping; Chou, Yi-Chun; Liao, Hung-Ting; Wang, Chia-Wen; Huang, Chun-Lin; Chang, Chia-Shuo; Lee, Ming-Ta Michael; Chen, Pei; Wong, Chih-Shung; Chen, Chien-Hsiun; Wu, Jer-Yuarn; Chen, Yuan-Tsong

2015-01-01

Objective To evaluate the use of prospective screening for the HLA-B*58:01 allele to identify Taiwanese individuals at risk of severe cutaneous adverse reactions (SCARs) induced by allopurinol treatment. Design National prospective cohort study. Setting 15 medical centres in different regions of Taiwan, from July 2009 to August 2014. Participants 2926 people who had an indication for allopurinol treatment but had not taken allopurinol previously. Participants were excluded if they had undergone a bone marrow transplant, were not of Han Chinese descent, and had a history of allopurinol induced hypersensitivity. DNA purified from 2910 participants’ peripheral blood was used to assess the presence of HLA-B*58:01. Main outcome measures Incidence of allopurinol induced SCARs with and without screening. Results Participants who tested positive for HLA-B*58:01 (19.6%, n=571) were advised to avoid allopurinol, and were referred to an alternate drug treatment or advised to continue with their prestudy treatment. Participants who tested negative (80.4%, n=2339) were given allopurinol. Participants were interviewed once a week for two months to monitor symptoms. The historical incidence of allopurinol induced SCARs, estimated by the National Health Insurance research database of Taiwan, was used for comparison. Mild, transient rash without blisters developed in 97 (3%) participants during follow-up. None of the participants was admitted to hospital owing to adverse drug reactions. SCARs did not develop in any of the participants receiving allopurinol who screened negative for HLA-B*58:01. By contrast, seven cases of SCARs were expected, based on the estimated historical incidence of allopurinol induced SCARs nationwide (0.30% per year, 95% confidence interval 0.28% to 0.31%; P=0.0026; two side one sample binomial test). Conclusions Prospective screening of the HLA-B*58:01 allele, coupled with an alternative drug treatment for carriers, significantly decreased the incidence

Genetic Control of Plant Root Colonization by the Biocontrol agent, Pseudomonas fluorescens

Energy Technology Data Exchange (ETDEWEB)

Cole, Benjamin J.; Fletcher, Meghan; Waters, Jordan; Wetmore, Kelly; Blow, Matthew J.; Deutschbauer, Adam M.; Dangl, Jeffry L.; Visel, Axel

2015-03-19

Plant growth promoting rhizobacteria (PGPR) are a critical component of plant root ecosystems. PGPR promote plant growth by solubilizing inaccessible minerals, suppressing pathogenic microorganisms in the soil, and directly stimulating growth through hormone synthesis. Pseudomonas fluorescens is a well-established PGPR isolated from wheat roots that can also colonize the root system of the model plant, Arabidopsis thaliana. We have created barcoded transposon insertion mutant libraries suitable for genome-wide transposon-mediated mutagenesis followed by sequencing (TnSeq). These libraries consist of over 105 independent insertions, collectively providing loss-of-function mutants for nearly all genes in the P.fluorescens genome. Each insertion mutant can be unambiguously identified by a randomized 20 nucleotide sequence (barcode) engineered into the transposon sequence. We used these libraries in a gnotobiotic assay to examine the colonization ability of P.fluorescens on A.thaliana roots. Taking advantage of the ability to distinguish individual colonization events using barcode sequences, we assessed the timing and microbial concentration dependence of colonization of the rhizoplane niche. These data provide direct insight into the dynamics of plant root colonization in an in vivo system and define baseline parameters for the systematic identification of the bacterial genes and molecular pathways using TnSeq assays. Having determined parameters that facilitate potential colonization of roots by thousands of independent insertion mutants in a single assay, we are currently establishing a genome-wide functional map of genes required for root colonization in P.fluorescens. Importantly, the approach developed and optimized here for P.fluorescens>A.thaliana colonization will be applicable to a wide range of plant-microbe interactions, including biofuel feedstock plants and microbes known or hypothesized to impact on biofuel-relevant traits including biomass productivity

Pim-1 Kinase Phosphorylates Cardiac Troponin I and Regulates Cardiac Myofilament Function

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Ni Zhu

2018-03-01

Full Text Available Background/Aims: Pim-1 is a serine/threonine kinase that is highly expressed in the heart, and exerts potent cardiac protective effects through enhancing survival, proliferation, and regeneration of cardiomyocytes. Its myocardial specific substrates, however, remain unknown. In the present study, we aim to investigate whether Pim-1 modulates myofilament activity through phosphorylation of cardiac troponin I (cTnI, a key component in regulating myofilament function in the heart. Methods: Coimmunoprecipitation and immunofluorescent assays were employed to investigate the interaction of Pim-1 with cTnI in cardiomyocytes. Biochemical, site directed mutagenesis, and mass spectrometric analyses were utilized to identify the phosphorylation sites of Pim1 in cTnI. Myofilament functional assay using skinned cardiac fiber was used to assess the effect of Pim1-mediated phosphorylation on cardiac myofilament activity. Lastly, the functional significance of Pim1-mediated cTnI in heart disease was determined in diabetic mice. Results: We found that Pim-1 specifically interacts with cTnI in cardiomyocytes and this interaction leads to Pim1-mediated cTnI phosphorylation, predominantly at Ser23/24 and Ser150. Furthermore, our functional assay demonstrated that Pim-1 induces a robust phosphorylation of cTnI within the troponin complex, thus leading to a decreased Ca2+ sensitivity. Insulin-like growth factor 1 (IGF-1, a peptide growth factor that has been shown to stimulate myocardial contractility, markedly induces cTnI phosphorylation at Ser23/24 and Ser150 through increasing Pim-1 expression in cardiomyocytes. In a high-fat diabetic mice model, the expression of Pim1 in the heart is significantly decreased, which is accompanied by a decreased phosphorylation of cTnI at Ser23/24 and Ser150, further implicating the pathological significance of the Pim1/cTnI axis in the development of diabetic cardiomyopathy. Conclusion: Our results demonstrate that Pim-1 is a

Atmospheric processes affecting the separation of volcanic ash and SO2 in volcanic eruptions: inferences from the May 2011 Grímsvötn eruption

Science.gov (United States)

Prata, Fred; Woodhouse, Mark; Huppert, Herbert E.; Prata, Andrew; Thordarson, Thor; Carn, Simon

2017-09-01

The separation of volcanic ash and sulfur dioxide (SO2) gas is sometimes observed during volcanic eruptions. The exact conditions under which separation occurs are not fully understood but the phenomenon is of importance because of the effects volcanic emissions have on aviation, on the environment, and on the earth's radiation balance. The eruption of Grímsvötn, a subglacial volcano under the Vatnajökull glacier in Iceland during 21-28 May 2011 produced one of the most spectacular examples of ash and SO2 separation, which led to errors in the forecasting of ash in the atmosphere over northern Europe. Satellite data from several sources coupled with meteorological wind data and photographic evidence suggest that the eruption column was unable to sustain itself, resulting in a large deposition of ash, which left a low-level ash-rich atmospheric plume moving southwards and then eastwards towards the southern Scandinavian coast and a high-level predominantly SO2 plume travelling northwards and then spreading eastwards and westwards. Here we provide observational and modelling perspectives on the separation of ash and SO2 and present quantitative estimates of the masses of ash and SO2 that erupted, the directions of transport, and the likely impacts. We hypothesise that a partial column collapse or sloughing fed with ash from pyroclastic density currents (PDCs) occurred during the early stage of the eruption, leading to an ash-laden gravity intrusion that was swept southwards, separated from the main column. Our model suggests that water-mediated aggregation caused enhanced ash removal because of the plentiful supply of source water from melted glacial ice and from entrained atmospheric water. The analysis also suggests that ash and SO2 should be treated with separate source terms, leading to improvements in forecasting the movement of both types of emissions.

Genome-Wide Mutagenesis in Borrelia burgdorferi.

Science.gov (United States)

Lin, Tao; Gao, Lihui

2018-01-01

Signature-tagged mutagenesis (STM) is a functional genomics approach to identify bacterial virulence determinants and virulence factors by simultaneously screening multiple mutants in a single host animal, and has been utilized extensively for the study of bacterial pathogenesis, host-pathogen interactions, and spirochete and tick biology. The signature-tagged transposon mutagenesis has been developed to investigate virulence determinants and pathogenesis of Borrelia burgdorferi. Mutants in genes important in virulence are identified by negative selection in which the mutants fail to colonize or disseminate in the animal host and tick vector. STM procedure combined with Luminex Flex ® Map™ technology and next-generation sequencing (e.g., Tn-seq) are the powerful high-throughput tools for the determination of Borrelia burgdorferi virulence determinants. The assessment of multiple tissue sites and two DNA resources at two different time points using Luminex Flex ® Map™ technology provides a robust data set. B. burgdorferi transposon mutant screening indicates that a high proportion of genes are the novel virulence determinants that are required for mouse and tick infection. In this protocol, an effective signature-tagged Himar1-based transposon suicide vector was developed and used to generate a sequence-defined library of nearly 4800 mutants in the infectious B. burgdorferi B31 clone. In STM, signature-tagged suicide vectors are constructed by inserting unique DNA sequences (tags) into the transposable elements. The signature-tagged transposon mutants are generated when transposon suicide vectors are transformed into an infectious B. burgdorferi clone, and the transposable element is transposed into the 5'-TA-3' sequence in the B. burgdorferi genome with the signature tag. The transposon library is created and consists of many sub-libraries, each sub-library has several hundreds of mutants with same tags. A group of mice or ticks are infected with a mixed

Time dependence of volcano inflation: mass influx or viscoelastic relaxation? Insights from Grímsvötn volcano, Iceland

Science.gov (United States)

Segall, P.

2017-12-01

Distinguishing magma chamber pressurization from relaxation of a viscoelastic aureole surrounding the chamber based on geodetic measurements has remained challenging. Elastic models with mass inflow proportional to the pressure difference between the chamber and a deep reservoir predict exponentially decaying flux. For a spherical chamber surrounded by a Maxwell viscoelastic shell with pressure dependent recharge, the surface deformation is the sum of two exponentials (Segall, 2016). GPS displacements following eruptions of Grímsvötn, Iceland in 2004 and 2011 exhibit rapid post-eruptive inflation (time scale of 0.1 yr), followed by inflation with a much longer time constant. Markov Chain Monte Carlo inversion with the viscoelastic model shows the GPS time series can be fit with viscosity of 2e16 Pa-s, and a relatively incompressible magma, B = beta_c/ (beta_m + beta_c) > 0.6, where beta_m and beta_c are chamber and magma compressibility. The latter appears to conflict with the ratio of erupted volume to geodetically inferred source volume change, rv 10, obtained for the best fitting spherical (Mogi ) source (Hreinsdóttir, 2014). Since rv = 1/B, this implies a relatively compressible melt, B 0.1. Reexamination of the co-eruptive GPS and tilt data with the more general ellipsoidal model of Cervelli (2013), reveals that the best fitting sources are oblate (b/a 3), deeper, and with larger volume changes, rv 3, relative to spherical models. Oblate magma chambers are consistent with seismic tomography. FEM calculations including free surface effects lead to even larger co-eruptive volume changes, smaller rv and hence larger B. I conclude that the data are consistent with rapid post-eruptive inflation driven by viscoelastic relaxation with a relatively incompressible magma, although other interpretations will be discussed.

Early colonization pattern of maize (Zea mays L. Poales, Poaceae roots by Herbaspirillum seropedicae (Burkholderiales, Oxalobacteraceae

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Rose A. Monteiro

2008-12-01

Full Text Available The bacterium Herbaspirillum seropedicae is an endophytic diazotroph found in several plants, including economically important poaceous species. However, the mechanisms involved in the interaction between H. seropedicae and these plants are not completely characterized. We investigated the attachment of Herbaspirillum to maize roots and the invasion of the roots by this bacterium using H. seropedicae strain SMR1 transformed with the suicide plasmid pUTKandsRed, which carries a mini-Tn5 transposon containing the gene for the Discosoma red fluorescent protein (Dsred constitutively expressed together with the kanamycin resistance gene. Integration of the mini-Tn5 into the bacterial chromosome yielded the mutant H. seropedicae strain RAM4 which was capable of expressing Dsred and could be observed on and inside fresh maize root samples. Confocal microscopy of maize roots inoculated with H. seropedicae three days after germination showed that H. seropedicae cell were attached to the root surface 30 min after inoculation, were visible in the internal tissues after twenty-four hours and in the endodermis, the central cylinder and xylem after three days.

Simple mucin-type carbohydrates in normal and malignant human endometrium

DEFF Research Database (Denmark)

Ravn, V; Mandel, U; Svenstrup, B

1995-01-01

The simple mucin-type carbohydrate antigens, Tn, sialosyl-Tn, and T, are tumor-associated antigens of adenocarcinomas. We evaluated by immunohistochemistry the expression of Tn, sialosyl-Tn (s-Tn), T, and sialosyl-T (s-T) antigens in normal nonsecretory, early gestational, and malignant human...... and malignant endometrium, and the expression of s-T antigen was positively correlated with E2 levels in serum. Our findings suggest a hormonal influence on expression of simple mucin-type carbohydrates in human endometrium. However, the accumulation of Tn and s-Tn antigens in malignant endometrial cells seem...

Combined exposure of ELF magnetic fields and X-rays increased mutant yields compared with X-rays alone in pTN89 plasmids

International Nuclear Information System (INIS)

Koyama, Shin; Nakahara, Takehisa; Sakurai, Tomonori; Komatsubara, Yoshiki; Miyakoshi, Junji; Isozumi, Yasuhito

2005-01-01

We have examined mutations in the supF gene carried by pTN89 plasmids in Escherichia coli (E. coli) to examine the effects of extremely low frequency magnetic fields (ELFMFs) and/or X-rays to the plasmids. The plasmids were subjected to sham exposure or exposed to an ELFMF (5 mT), with or without X-ray irradiation (10 Gy). For the combined treatments, exposure to the ELFMF was immediately before or after X-ray irradiation. The mutant fractions were 0.94 x 10 -5 for X-rays alone, 1.58 x 10 -5 for an ELFMF followed by X-rays, and 3.64 x 10 -5 for X-rays followed by an ELFMF. Increased mutant fraction was not detected following exposure to a magnetic field alone, or after sham exposure. The mutant fraction for X-rays followed by an ELFMF was significantly higher than those of other treatments. Sequence analysis of the supF mutant plasmids revealed that base substitutions were dominant on exposure to X-rays alone and X-rays plus an ELFMF. Several types of deletions were detected in only the combined treatments, but not with X-rays alone. We could not find any mutant colonies in sham irradiated and an ELFMF alone treatment, but exposure to ELFMFs immediately before or after X-ray irradiation may enhance the mutations. Our results indicate that an ELFMF increases mutation and alters the spectrum of mutations. (author)

Integrones: los coleccionistas de genes Integrons: gene collectors

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J. A. Di Conza

2010-02-01

Full Text Available Los integrones son estructuras genéticas que han despertado gran interés, debido a que algunos de ellos vehiculizan genes de resistencia a los antimicrobianos. Están formados por un fragmento que codifica una integrasa (intI y, a continuación, una secuencia attI a la que se unen los genes en casetes que codifican diferentes mecanismos de resistencia. Dentro de intI, en su extremo 3´, hay una secuencia promotora Pc a partir de la cual se transcriben los casetes de resistencia integrados, ya que estos genes carecen de promotor. Sin embargo, estos casetes presentan una secuencia específica denominada attC, la cual es reconocida por la integrasa que se une, por recombinación, a la secuencia attI del integrón en la orientación adecuada para su expresión. Los integrones se han clasificado según la secuencia de su integrasa, pero en la actualidad se prefiere clasificarlos según su localización. Se habla, en general, de "integrones móviles" para referirse a aquellos asociados a secuencias de inserción, transposones y/o plásmidos conjugativos, los que en su mayoría median mecanismos de resistencia, y de "superintegrones", de localización cromosómica y con grandes arreglos de genes en casetes. Los integrones móviles de clase 1 son los más abundantes en aislamientos clínicos y suelen estar asociados a transposones del subgrupo Tn21, seguidos por los de clase 2, derivados principalmente de Tn7. Estos elementos no son móviles por sí mismos, pero su asociación con elementos que sí lo son facilita su transferencia horizontal, lo que explica su amplia difusión entre las bacterias. Esta revisión intenta recopilar la información disponible acerca de los integrones móviles descritos en Argentina hasta la fecha.Integrons gained great interest due to their participation in resistance gene recruitment and expression. Their basic structure includes a fragment that encodes an integrase (intI followed by a recognition sequence (attI into

Certifying the TN-BRP and TN-REG transportable storage demonstration casks

International Nuclear Information System (INIS)

Abbott, D.G.; Nolan, D.J.; Yoshimura, H.R.

1991-01-01

The US DOE has obtained US NRC certification to transport two transportable storage casks for a demonstration project. Because the casks had been built before the decision was made to obtain NRC certification, only limited modifications could be made to the casks. NRC's review resulted in several technical concerns that were subsequently resolved by design modifications, testing, and further analysis. Certification activities included qualifying the ferritic steel body material, modifying the borated stainless steel basket design, and extensive impact limiter testing. Recommendations for certifying future casks are presented based on experience with these casks

Carbapenem-resistance and pathogenicity of bovine Acinetobacter indicus-like isolates.

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Peter Klotz

Full Text Available The objective of this study was to characterize blaOXA-23 harbouring Acinetobacter indicus-like strains from cattle including genomic and phylogenetic analyses, antimicrobial susceptibility testing and evaluation of pathogenicity in vitro and in vivo. Nasal and rectal swabs (n = 45 from cattle in Germany were screened for carbapenem-non-susceptible Acinetobacter spp. Thereby, two carbapenem resistant Acinetobacter spp. from the nasal cavities of two calves could be isolated. MALDI-TOF mass spectrometry and 16S rDNA sequencing identified these isolates as A. indicus-like. A phylogenetic tree based on partial rpoB sequences indicated closest relation of the two bovine isolates to the A. indicus type strain A648T and human clinical A. indicus isolates, while whole genome comparison revealed considerable intraspecies diversity. High mimimum inhibitory concentrations were observed for carbapenems and other antibiotics including fluoroquinolones and gentamicin. Whole genome sequencing and PCR mapping revealed that both isolates harboured blaOXA-23 localized on the chromosome and surrounded by interrupted Tn2008 transposon structures. Since the pathogenic potential of A. indicus is unknown, pathogenicity was assessed employing the Galleria (G. mellonella infection model and an in vitro cytotoxicity assay using A549 human lung epithelial cells. Pathogenicity in vivo (G. mellonella killing assay and in vitro (cytotoxicity assay of the two A. indicus-like isolates was lower compared to A. baumannii ATCC 17978 and similar to A. lwoffii ATCC 15309. The reduced pathogenicity of A. indicus compared to A. baumannii correlated with the absence of important virulence genes encoding like phospholipase C1+C2, acinetobactin outer membrane protein BauA, RND-type efflux system proteins AdeRS and AdeAB or the trimeric autotransporter adhesin Ata. The emergence of carbapenem-resistant A. indicus-like strains from cattle carrying blaOXA-23 on transposable elements and

Atmospheric processes affecting the separation of volcanic ash and SO2 in volcanic eruptions: inferences from the May 2011 Grímsvötn eruption

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F. Prata

2017-09-01

Full Text Available The separation of volcanic ash and sulfur dioxide (SO2 gas is sometimes observed during volcanic eruptions. The exact conditions under which separation occurs are not fully understood but the phenomenon is of importance because of the effects volcanic emissions have on aviation, on the environment, and on the earth's radiation balance. The eruption of Grímsvötn, a subglacial volcano under the Vatnajökull glacier in Iceland during 21–28 May 2011 produced one of the most spectacular examples of ash and SO2 separation, which led to errors in the forecasting of ash in the atmosphere over northern Europe. Satellite data from several sources coupled with meteorological wind data and photographic evidence suggest that the eruption column was unable to sustain itself, resulting in a large deposition of ash, which left a low-level ash-rich atmospheric plume moving southwards and then eastwards towards the southern Scandinavian coast and a high-level predominantly SO2 plume travelling northwards and then spreading eastwards and westwards. Here we provide observational and modelling perspectives on the separation of ash and SO2 and present quantitative estimates of the masses of ash and SO2 that erupted, the directions of transport, and the likely impacts. We hypothesise that a partial column collapse or sloughing fed with ash from pyroclastic density currents (PDCs occurred during the early stage of the eruption, leading to an ash-laden gravity intrusion that was swept southwards, separated from the main column. Our model suggests that water-mediated aggregation caused enhanced ash removal because of the plentiful supply of source water from melted glacial ice and from entrained atmospheric water. The analysis also suggests that ash and SO2 should be treated with separate source terms, leading to improvements in forecasting the movement of both types of emissions.

Andisols from Tondano Area, North Sulawesi: Properties and Classification

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Hikmatullah

2008-01-01

Full Text Available Three pedons of Andisol (TN-1, TN-2 and TN-3 developed from young volcanic materials of the Lokon, Soputan, and Lengkoan volcanoes respectively in the Tondano area, North Sulawesi, were studied in the field, and 18 soil samples were analysed in the laboratory for physical, chemical, and mineralogical properties, and they were classified according to Keys to Soil Taxonomy 2003. The results indicated that all the pedons meet the requirements of the andic soil properties, and thus classified into Andisol order. Pedon TN-1 meets bulk density 85%, and (Alo + 0.5Feo content extracted by ammonium oxalate > 2.0%, while pedons TN-2 and TN-3 meet the requirements of P retention > 25%, (Alo + 0.5Feo content > 0.4%, volcanic glass content > 5%, and value of [%(Alo+0.5Feo x 15.625 + (% volcanic glass] is > 36.25. Composition of sand mineral fraction indicate that pedon TN-1 and TN-3 show andesitic to basaltic volcanic materials, whereas pedon TN-2 with high olivin content belongs to basaltic volcanic materials. Clay minerals of all the pedons was dominated by hydrated-halloysite with few of disordered-kaolinite, which indicated a little weathering of the pedons. The pedons were classified at family level as Typic Hapludand, medial, amorphic, isothermic (TN-1, Humic Udivitrand, ashy, amorphic, isothermic (TN-2, and Alfic Hapludand, medial, glassy, isothermic (TN-3.

Immunohistochemical expression of tenascin in melanocytic tumours of dogs.

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Sevastre, B; van Ederen, A M; Terlou, M; Gruys, E; Nederbragt, H

2007-01-01

The aim of this study was to investigate tenascin-C (TN) immunolabelling and labelling for endothelium by von Willebrand Factor (vWF) in melanocytic tumours of dogs as compared with normal tissues, to evaluate the TN distribution in these types of tumours and to investigate whether a relation could be established between TN and angiogenesis in different types of tumour. Samples of normal dog skin (n=8), benign skin melanocytomas (n=10), malignant oral melanomas (n=9) and malignant toe melanomas (n=5) were studied. The percentages of TN and vWF immunolabelling per total microscopical area were analysed by morphometric methods. In normal skin, TN was found at dermo-epidermal junctions, around hair follicles, in the smooth muscles of hair follicles, and in the walls of blood vessels. TN immunolabelling (distribution and intensity) in melanocytomas was comparable with that found in normal skin. In melanomas, TN expression was considerably increased, its intensity in toe melanomas being twice that observed in oral melanomas. The degree of TN immunolabelling was not related to the histological malignancy of the melanomas. In melanomas, TN was found in the connective tissue surrounding the tumour cell nests and in narrow stromal strands inside the tumour. Regions infiltrated with lymphocytes were devoid of TN. The presence of TN around capillaries in melanocytomas and melanomas was investigated by double-immunolabelling (for TN and vWF). The intensity of vWF and TN immunolabelling was higher in melanomas than in melanocytomas, and higher in toe melanomas than in oral melanomas; however, no clear relation between TN expression and immunolabelling for vWF was found.

Mariner and the ITm Superfamily of Transposons.

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Tellier, Michael; Bouuaert, Corentin Claeys; Chalmers, Ronald

2015-04-01

The IS630-Tc1-mariner (ITm) family of transposons is one of the most widespread in nature. The phylogenetic distribution of its members shows that they do not persist for long in a given lineage, but rely on frequent horizontal transfer to new hosts. Although they are primarily selfish genomic-parasites, ITm transposons contribute to the evolution of their hosts because they generate variation and contribute protein domains and regulatory regions. Here we review the molecular mechanism of ITm transposition and its regulation. We focus mostly on the mariner elements, which are understood in the greatest detail owing to in vitro reconstitution and structural analysis. Nevertheless, the most important characteristics are probably shared across the grouping. Members of the ITm family are mobilized by a cut-and-paste mechanism and integrate at 5'-TA dinucleotide target sites. The elements encode a single transposase protein with an N-terminal DNA-binding domain and a C-terminal catalytic domain. The phosphoryl-transferase reactions during the DNA-strand breaking and joining reactions are performed by the two metal-ion mechanism. The metal ions are coordinated by three or four acidic amino acid residues located within an RNase H-like structural fold. Although all of the strand breaking and joining events at a given transposon end are performed by a single molecule of transposase, the reaction is coordinated by close communication between transpososome components. During transpososome assembly, transposase dimers compete for free transposon ends. This helps to protect the host by dampening an otherwise exponential increase in the rate of transposition as the copy number increases.

Self-reported cocaine use is not associated with elevations in high-sensitivity troponin I.

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Jordan, Candice D; Korley, Frederick K; Stolbach, Andrew I

2017-06-01

High-sensitivity troponin (hsTn) assays detect 10 times lower concentrations of cardiac troponin than conventional assays. We examined the effects of self-reported cocaine use to determine whether those with acute cocaine use being evaluated for ACS are more likely to have elevated hsTnI than those nonusers being evaluated for ACS. We conducted a sub-analysis of a prospective cohort of ED patients evaluated for acute coronary syndrome. Recent cocaine use was determined by structured patient interviews. High-sensitivity troponin (Abbott) and conventional troponin I (Abbott, cTnI) were measured on samples drawn at presentation. Urine toxicology screen for cocaine metabolite was obtained at the discretion of treating clinicians. Of 1862 patients enrolled, 444 reported prior cocaine use and 99 reported cocaine use within the preceding month. Median hsTn in patients with last cocaine use within 24 h, 2-7 days, 1 week-1 month, >1 month, and no prior cocaine use were: 9 (IQR: 3-17) ng/L, 6 (IQR: 3-24.3) ng/L, 6 (IQR: 3-89.5) ng/L, 3 (IQR: 3-18.5) ng/L and 3 (IQR: 3-17) ng/L, respectively. Urine toxicology assays (UTox) for cocaine were performed in 640 (34.4%) patients. The median hsTn for those who were UTox+, UTox - and those without a UTox were: 9 ng/L (IQR: 3-48.5), 9 ng/L (IQR: 3-40) and 3 ng/L (IQR: 3-12), respectively. There were no differences in the prevalence of new troponin elevations (hsTn >99th percentile but cTnI cocaine use compared to those without recent cocaine use. In this first investigation of hsTn in patients with self-reported recent cocaine use, we have determined that hsTn does not lead to an increase in the prevalence of troponin elevation in cocaine users.

Obstructive sleep apnea: no independent association to troponins.

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Hall, Trygve Sørdahl; Herrscher, Tobias; Jarolim, Petr; Fagerland, Morten W; Jensen, Torstein; Hallén, Jonas; Agewall, Stefan; Atar, Dan

2014-05-01

Cardiac troponins (cTn) are to date the most sensitive and specific biochemical markers of myocardial injury. Abnormal breathing patterns in patients with obstructive sleep apnea (OSA) may cause myocardial cell stress detectable by novel cTn assays. The objectives of this study were to investigate whether a new single-molecule cTnI (S-cTnI) assay and a commercially available high-sensitivity cTnT (hs-cTnT) assay would detect myocyte injury in individuals evaluated for possible OSA, and to explore their relation to variables of disordered breathing during sleep. Consecutive individuals referred to Lovisenberg Diakonale Hospital's sleep laboratory between 1 October 2009 and 1 March 2010 were included. We measured cTn in specimens collected the morning after sleep and studied these in relation to variables recorded during polygraphy or polysomnography. All 222 (100 %) individuals had measurable cTn levels using either assay. Stratified into categories according to the apnea-hypopnea index (AHI), patients with OSA (AHI ≥5) had a different distribution of S-cTnI (P = 0.036) and hs-cTnT (P = 0.002) compared to those without (AHI <5). The median (quartiles 1-3) were 3.0 (1.9-6.0) versus 2.3 (1.6-3.8) ng/l for S-cTnI, and 7.0 (5.5-8.7) versus 6.2 (4.9-7.2) ng/l for hs-cTnT. However, in multiple median regression analyses adjusted for conventional predictors, neither S-cTnI (P = 0.57) nor hs-cTnT (P = 0.80) were significantly associated with AHI. This study reveals no association independent of conventional predictors between OSA and myocardial cell injury measured by S-cTnI and hs-cTnT assays. Our findings support a search for novel biomarkers for prognostication of OSA.

Macrophage migration inhibitory factor stimulated by Helicobacter pylori increases proliferation of gastric epithelial cells

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Xia, Harry Hua-Xiang; Lam, Shiu Kum; Chan, Annie O.O.; Lin, Marie Chia Mi; Kung, Hsiang Fu; Ogura, Keiji; Berg, Douglas E.; Wong, Benjamin C. Y.

2005-01-01

AIM: Helicobacter pylori (H pylori) is associated with increased gastric inflammatory and epithelial expression of macrophage migration inhibitory factor (MIF) and gastric epithelial cell proliferation. This study aimed at determining whether H pylori directly stimulates release of MIF in monocytes, whether the cag pathogenicity island (PAI) is involved for this function, and whether MIF stimulated by H pylori increases gastric epithelial cell proliferation in vitro. METHODS: A cytotoxic wild-type H pylori strain (TN2)and its three isogenic mutants (TN2△cag, TN2△cagA and TN2△cagE) were co-cultured with cells of a human monocyte cell line, THP-1, for 24 h at different organism/cell ratios. MIF in the supernatants was measured by an ELISA. Cells of a human gastric cancer cell line, MKN45, were then co-cultured with the supernatants, with and without monoclonal anti-MIF antibody for 24 h. The cells were further incubated for 12 h after addition of 3H-thymidine, and the levels of incorporation of 3H-thymidine were measured with a liquid scintillation counter. RESULTS: The wild-type strain and the isogenic mutants, TN2△cagA and TN2△cagE, increased MIF release at organism/cell ratios of 200/1 and 400/1, but not at the ratios of 50/1 and 100/1. However, the mutant TN2△cag did not increase the release of MIF at any of the four ratios. 3H-thymidine readings for MKN-45 cells were significantly increased with supernatants derived from the wild-type strain and the mutants TN2△cagA and TN2△cagE, but not from the mutant TN2△cag. Moreover, in the presence of monoclonal anti-MIF antibody, the stimulatory effects of the wild-type strain on cell proliferation disappeared. CONCLUSION: H pylori stimulates MIF release in monocytes, likely through its cag PAI, but not related to cagA or cagE. H pylori-stimulated monocyte culture supernatant increases gastric cell proliferation, which is blocked by anti-MIF antibody, suggesting that MIF plays an important role in H

Effects of Diet and Genetics on Growth Performance of Pigs in Response to Repeated Exposure to Heat Stress

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Wendy M. Rauw

2017-10-01

Full Text Available Heat stress (HS is one of the costliest issues in the U.S. pork industry. Aims of the present study were to determine the consequences of repeated exposure to HS on growth performance, and the effects of a high fiber diet, the genetic potential for high lean tissue accretion, and the genetic potential for residual feed intake (RFI on resilience to HS. Barrows (n = 97 from three genetic lines (commercial, high RFI, low RFI where subjected three times to a 4-day HS treatment (HS1, HS2, and HS3 which was preceded by a 9-day neutral (TN adaptation period (TN1 and alternated by 7-day periods of neutral temperatures (TN2, TN3, and TN4. Body weight gain (BWG, feed intake (FI, feed conversion efficiency (FCE, RFI, and the drop in BWG and FI between TN and HS were estimated for each period, and slaughter traits were measured at the end of TN4. Commercial pigs had lower FI when fed a high fiber diet compared to a regular diet (2.70 ± 0.08 vs. 2.96 ± 0.08 kg/d; P < 0.05, while no differences were found for BWG, RFI or FCE. HS reduced FI, BWG, and FCE, increased RFI, and resulted in leaner pigs that generate smaller carcasses at slaughter. In TN, commercial pigs grew faster than the low and high RFI pigs (1.22 ± 0.06 vs. 0.720 ± 0.05 and 0.657 ± 0.07; P < 0.001 but growth rates were not significantly different between the lines during HS. Growth rates for the low RFI and high RFI pigs were similar both during TN and during HS. Pigs of interest for genetic improvement are those that are able to maintain growth rates during HS. Our results show that response in growth to HS was repeatable over subsequent 4-d HS cycles, which suggests the potential for including this response in the breeding index. The best performing animals during HS are likely those that are not highly superior for growth in TN.

A landscape-scale study of land use and parent material effects on soil organic carbon and total nitrogen in the Konya Basin, Turkey

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Mayes, M. T.; Marin-Spiotta, E.; Ozdogan, M.; Erdogan, M. A.

2011-12-01

shown by previous modeling or soil survey efforts. We provide baseline information on SOC and TN that can inform benchmarks for future soil monitoring and land use planning in an arid region that is likely to be highly impacted by future climatic changes, agricultural intensification and urban development. Our results suggest the importance of accounting for soil physical properties, and land use effects that are dependent on soil parent materials in future efforts to model or account for SOC and TN in similar ancient agricultural landscapes.

S-glutathionylation of troponin I (fast) increases contractile apparatus Ca2+ sensitivity in fast-twitch muscle fibres of rats and humans.

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Mollica, J P; Dutka, T L; Merry, T L; Lamboley, C R; McConell, G K; McKenna, M J; Murphy, R M; Lamb, G D

2012-03-15

-glutathionylation of TnI(f) (latter examined only in toad). Following 40 min of cycling exercise in human subjects (at ∼60% peak oxygen consumption), TnI(f) in vastus lateralis muscle displayed a marked increase in S-glutathionylation (∼4-fold). These findings show that S-glutathionylation of TnI(f), most probably at Cys133, increases the Ca2+ sensitivity of the contractile apparatus, and that this occurs in exercising humans, with likely beneficial effects on performance.

Multi-centre evaluation of recent troponin assays for the diagnosis of NSTEMI

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Camille Chenevier-Gobeaux

2018-07-01

Full Text Available Objectives: We aimed to compare the use of nine different cardiac troponin (cTn assays (2 cTnT and 7 cTnI for the diagnosis of NSTEMI in a single multi-centre population. Design and methods: One hundred and fifty-eight patients were included (mean age 60 years, SD 17 years, including 23 patients (14% with NSTEMI. Results: The analytical comparison highlighted a large heterogeneity of cTn assays, as reflected by percentages of patients with detectable cTn, correlation coefficients, Passing-Bablok comparisons and concordance coefficients. Correlations within cTnI assays were good and correlation within cTnT assays was excellent. Diagnostic performances demonstrated that each cTn assay has specific threshold values. Furthermore, some assays (HS-cTnI and T, cTnI-Pathfast and cTnI-Centaur indicated high sensitivity and negative predictive value using the limit of detection (LoD diagnostic strategy. For the latter assays, a significant increase in specificity was found when using the 99th percentile or the H0-H3 strategies, in comparison to the LoD strategy. When applying the European Society of Cardiology H0-H3 algorithm, comparable diagnostic performances were obtained. Conclusion: All 9 cTn assays indicated overall good diagnostic performances for the diagnosis of NSTEMI in emergency departments when the recommended algorithm based on the variation of cTn value between two measurements at admission and 3 h later was used. Keywords: Cardiac troponin, High-sensitivity assay, Chest pain, Emergency department, NSTEMI, Analytical evaluation

Fluorescent Protein-Based Ca2+ Sensor Reveals Global, Divalent Cation-Dependent Conformational Changes in Cardiac Troponin C.

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Myriam A Badr

Full Text Available Cardiac troponin C (cTnC is a key effector in cardiac muscle excitation-contraction coupling as the Ca2+ sensing subunit responsible for controlling contraction. In this study, we generated several FRET sensors for divalent cations based on cTnC flanked by a donor fluorescent protein (CFP and an acceptor fluorescent protein (YFP. The sensors report Ca2+ and Mg2+ binding, and relay global structural information about the structural relationship between cTnC's N- and C-domains. The sensors were first characterized using end point titrations to decipher the response to Ca2+ binding in the presence or absence of Mg2+. The sensor that exhibited the largest responses in end point titrations, CTV-TnC, (Cerulean, TnC, and Venus was characterized more extensively. Most of the divalent cation-dependent FRET signal originates from the high affinity C-terminal EF hands. CTV-TnC reconstitutes into skinned fiber preparations indicating proper assembly of troponin complex, with only ~0.2 pCa unit rightward shift of Ca2+-sensitive force development compared to WT-cTnC. Affinity of CTV-TnC for divalent cations is in agreement with known values for WT-cTnC. Analytical ultracentrifugation indicates that CTV-TnC undergoes compaction as divalent cations bind. C-terminal sites induce ion-specific (Ca2+ versus Mg2+ conformational changes in cTnC. Our data also provide support for the presence of additional, non-EF-hand sites on cTnC for Mg2+ binding. In conclusion, we successfully generated a novel FRET-Ca2+ sensor based on full length cTnC with a variety of cellular applications. Our sensor reveals global structural information about cTnC upon divalent cation binding.

Yam storage protein dioscorins from Dioscorea alata and Dioscorea japonica exhibit distinct immunomodulatory activities in mice.

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Lin, Pei-Lan; Lin, Kuo-Wei; Weng, Ching-Feng; Lin, Kuo-Chih

2009-06-10

The aim of this study was to elucidate the effect of the major storage protein dioscorin isolated from two different yam species, Tainong No. 1 (TN1-dioscorins) and Japanese yam (Dj-dioscorins), on the immune activities of mice. Dj-dioscorins, like TN1-dioscorins, could induce expression of the pro-inflammatory cytokines and stimulate phagocytosis of RAW 264.7. Intraperitoneal injection of the TN1-dioscorins into mice stimulated phagocytosis of bone marrow, spleen, and thymic cells. In contrast, the T and B cells in bone marrow, spleen, and thymus isolated from mice injected with Dj-dioscorins had higher proliferative responses to mitogens. Furthermore, Dj-dioscorins enhanced proliferation of CD4(+), CD8(+), and Tim3(+) (Th1) cells in spleen and CD19(+) cells in both spleen and thymus. Supplement of Dj-dioscorins in the lymphoid cells isolated from Dj-dioscorins primed mice induced cell proliferation of both spleen and thymic cells. These findings indicated that TN1-dioscorins have a higher ability to stimulate the phagocytic activity of the lymphoid cells than Dj-dioscorins, whereas Dj-dioscorins possess more abilities than TN1-dioscorins to enhance the proliferation of the lymphoid cells.

Prognostic implications of high-sensitivity cardiac troponin T assay in a real-world population with non-ST-elevation acute coronary syndrome.

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Magnoni, Marco; Gallone, Guglielmo; Ceriotti, Ferruccio; Vergani, Vittoria; Giorgio, Daniela; Angeloni, Giulia; Maseri, Attilio; Cianflone, Domenico

2018-09-01

High-sensitivity cardiac troponin T (hsTnT) was recently approved for clinical use by the Food and Drug Administration. The transition from contemporary to hsTnT assays requires a thorough understanding of the clinical differences between these assays. HsTnT may provide a more accurate prognostic stratification than contemporary cardiac troponin I (cTnI) in patients with non-ST-segment elevation acute coronary syndromes (NSTE-ACS). HsTnT and cTnI were measured in 644 patients with CK-MB negative NSTE-ACS who were enrolled in the prospective multicenter SPAI (Stratificazione Prognostica dell'Angina Instabile) study. Patients were stratified at the 99th percentile reference limit for each assay. The primary endpoint was cardiovascular death (CVD) or non-fatal myocardial infarction (MI); the secondary endpoint was the occurrence of unstable angina (UA). Follow-up lasted 180 days. Patients with hsTnT ≥99th percentile were at higher risk of CVD/MI (30-day: 5.9% vs 0.8%, p  = 0.001; 180-day: 11.1% vs 4.7%, p  = 0.004), also after adjusting for TIMI Risk Score. No significant difference in CVD/MI at 180-day was found between hsTnT-positive/cTnI-negative and hsTnT-negative/cTnI-negative patients (adjHR 1.61, 95% CI 0.74-3.49, p  = 0.232). Occurrence of UA was not differently distributed between hsTnT groups dichotomized at the 99th percentile (12.4% vs 12.5% p  = 0.54). Our investigation on a real-world NSTE-ACS population showed good prognostic performance of hsTnT in the risk stratification of the hard endpoint, but did not demonstrate the improved prognostic ability of hsTnT over contemporary cTn. Neither troponin assay predicted the recurrence of UA, suggesting the acute rise of cardiac troponin as a marker of severity, but not the occurrence of future coronary instability.

Early diagnosis of myocardial infarction using absolute and relative changes in cardiac troponin concentrations.

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Irfan, Affan; Reichlin, Tobias; Twerenbold, Raphael; Meister, Marc; Moehring, Berit; Wildi, Karin; Bassetti, Stefano; Zellweger, Christa; Gimenez, Maria Rubini; Hoeller, Rebeca; Murray, Karsten; Sou, Seoung Mann; Mueller, Mira; Mosimann, Tamina; Reiter, Miriam; Haaf, Philip; Ziller, Ronny; Freidank, Heike; Osswald, Stefan; Mueller, Christian

2013-09-01

Absolute changes in high-sensitivity cardiac troponin T (hs-cTnT) seem to have higher diagnostic accuracy in the early diagnosis of acute myocardial infarction compared with relative changes. It is unknown whether the same applies to high-sensitivity cardiac troponin I (hs-cTnI) assays and whether the combination of absolute and relative change might further increase accuracy. In a prospective, international multicenter study, high-sensitivity cardiac troponin (hs-cTn) was measured with 3 novel assays (hs-cTnT, Roche Diagnostics Corp, Indianapolis, Ind; hs-cTnI, Beckman Coulter Inc, Brea, Calif; hs-cTnI, Siemens, Munich, Germany) in a blinded fashion at presentation and after 1 and 2 hours in a blinded fashion in 830 unselected patients with suspected acute myocardial infarction. The final diagnosis was adjudicated by 2 independent cardiologists. The area under the receiver operating characteristic curve for diagnosing acute myocardial infarction was significantly higher for 1- and 2-hour absolute versus relative hs-cTn changes for all 3 assays (P Siemens, 0.96 [95% CI, 0.93-0.99]) were high and provided some benefit compared with the use of absolute change alone for hs-cTnT, but not for the hs-cTnI assays. Reclassification analysis confirmed the superiority of absolute changes versus relative changes. Absolute changes seem to be the preferred metrics for both hs-cTnT and hs-cTnI in the early diagnosis of acute myocardial infarction. The combination of absolute and relative changes provides a small added value for hs-cTnT, but not for hs-cTnI. Copyright © 2013 Elsevier Inc. All rights reserved.

The Foldback-like element Galileo belongs to the P superfamily of DNA transposons and is widespread within the Drosophila genus.

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Marzo, Mar; Puig, Marta; Ruiz, Alfredo

2008-02-26

Galileo is the only transposable element (TE) known to have generated natural chromosomal inversions in the genus Drosophila. It was discovered in Drosophila buzzatii and classified as a Foldback-like element because of its long, internally repetitive, terminal inverted repeats (TIRs) and lack of coding capacity. Here, we characterized a seemingly complete copy of Galileo from the D. buzzatii genome. It is 5,406 bp long, possesses 1,229-bp TIRs, and encodes a 912-aa transposase similar to those of the Drosophila melanogaster 1360 (Hoppel) and P elements. We also searched the recently available genome sequences of 12 Drosophila species for elements similar to Dbuz\\Galileo by using bioinformatic tools. Galileo was found in six species (ananassae, willistoni, peudoobscura, persimilis, virilis, and mojavensis) from the two main lineages within the Drosophila genus. Our observations place Galileo within the P superfamily of cut-and-paste transposons and extend considerably its phylogenetic distribution. The interspecific distribution of Galileo indicates an ancient presence in the genus, but the phylogenetic tree built with the transposase amino acid sequences contrasts significantly with that of the species, indicating lineage sorting and/or horizontal transfer events. Our results also suggest that Foldback-like elements such as Galileo may evolve from DNA-based transposon ancestors by loss of the transposase gene and disproportionate elongation of TIRs.

Selectivity verification of cardiac troponin monoclonal antibodies for cardiac troponin detection by using conventional ELISA

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Fathil, M. F. M.; Arshad, M. K. Md; Gopinath, Subash C. B.; Adzhri, R.; Ruslinda, A. R.; Hashim, U.

2017-03-01

This paper presents preparation and characterization of conventional enzyme-linked immunosorbent assay (ELISA) for cardiac troponin detection to determine the selectivity of the cardiac troponin monoclonal antibodies. Monoclonal antibodies, used to capture and bind the targets in this experiment, are cTnI monoclonal antibody (MAb-cTnI) and cTnT monoclonal antibody (MAb-cTnT), while both cardiac troponin I (cTnI) and T (cTnT) are used as targets. ELISA is performed inside two microtiter plates for MAb-cTnI and MAb-cTnT. For each plate, monoclonal antibodies are tested by various concentrations of cTnI and cTnT ranging from 0-6400 µg/l. The binding selectivity and level of detection between monoclonal antibodies and antigen are determined through visual observation based on the color change inside each well on the plate. ELISA reader is further used to quantitatively measured the optical density of the color changes, thus produced more accurate reading. The results from this experiment are utilized to justify the use of these monoclonal antibodies as bio-receptors for cardiac troponin detection by using field-effect transistor (FET)-based biosensors coupled with substrate-gate in the future.

Cost effectiveness analysis of HLA-B*58:01 genotyping prior to initiation of allopurinol for gout.

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Plumpton, Catrin O; Alfirevic, Ana; Pirmohamed, Munir; Hughes, Dyfrig A

2017-10-01

To determine whether prospective testing for HLA-B*58:01, as a strategy to prevent serious adverse reactions to allopurinol in patients with gout, is cost-effective from the perspective of the National Health Service in the UK. A systematic review and meta-analysis for the association of HLA-B*58:01 with cutaneous and hypersensitivity adverse drug reactions informed a decision analytic and Markov model to estimate lifetime costs and outcomes associated with testing vs standard care (with febuxostat prescribed for patients who test positive). Scenario analyses assessed alternative treatment assumptions and patient populations. The number of patients needed to test to prevent one case of adverse drug reaction was 11 286 (95% central range (CR): 2573, 53 594). Cost and quality-adjusted life-year (QALY) gains were small, £103 (95% CR: £98, £106) and 0.0023 (95% CR: -0.0006, 0.0055), respectively, resulting in an incremental cost-effectiveness ratio (ICER) of £44 954 per QALY gained. The probability of testing being cost-effective at a threshold of £30 000 per QALY was 0.25. Reduced costs of testing or febuxostat resulted in an ICER below £30 000 per QALY gained. The ICER for patients with chronic renal insufficiency was £38 478 per QALY gained. Routine testing for HLA-B*58:01 in order to reduce the incidence of adverse drug reactions in patients being prescribed allopurinol for gout is unlikely to be cost-effective in the UK; however testing is expected to become cost-effective with reductions in the cost of genotyping, and with the future availability of cheaper, generic febuxostat. © The Author 2017. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Unusual genome complexity in Lactobacillus salivarius JCM1046.

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Raftis, Emma J; Forde, Brian M; Claesson, Marcus J; O'Toole, Paul W

2014-09-08

Lactobacillus salivarius strains are increasingly being exploited for their probiotic properties in humans and animals. Dissemination of antibiotic resistance genes among species with food or probiotic-association is undesirable and is often mediated by plasmids or integrative and conjugative elements. L. salivarius strains typically have multireplicon genomes including circular megaplasmids that encode strain-specific traits for intestinal survival and probiotic activity. Linear plasmids are less common in lactobacilli and show a very limited distribution in L. salivarius. Here we present experimental evidence that supports an unusually complex multireplicon genome structure in the porcine isolate L. salivarius JCM1046. JCM1046 harbours a 1.83 Mb chromosome, and four plasmids which constitute 20% of the genome. In addition to the known 219 kb repA-type megaplasmid pMP1046A, we identified and experimentally validated the topology of three additional replicons, the circular pMP1046B (129 kb), a linear plasmid pLMP1046 (101 kb) and pCTN1046 (33 kb) harbouring a conjugative transposon. pMP1046B harbours both plasmid-associated replication genes and paralogues of chromosomally encoded housekeeping and information-processing related genes, thus qualifying it as a putative chromid. pLMP1046 shares limited sequence homology or gene synteny with other L. salivarius plasmids, and its putative replication-associated protein is homologous to the RepA/E proteins found in the large circular megaplasmids of L. salivarius. Plasmid pCTN1046 harbours a single copy of an integrated conjugative transposon (Tn6224) which appears to be functionally intact and includes the tetracycline resistance gene tetM. Experimental validation of sequence assemblies and plasmid topology resolved the complex genome architecture of L. salivarius JCM1046. A high-coverage draft genome sequence would not have elucidated the genome complexity in this strain. Given the expanding use of L. salivarius

Cardiac Troponin Elevation Predicts Mortality in Patients Undergoing Orthotopic Liver Transplantation

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David Snipelisky

2013-01-01

Full Text Available Introduction. While patients undergoing orthotopic liver transplantation (OLT have high cardiovascular event rates, preoperative risk stratification may not necessarily predict those susceptible patients. Troponin T (TnT may help predict patients at risk for cardiovascular complications. Methods. Consecutive patients undergoing OLT at Mayo Clinic in Florida between 1998 and 2010 who had TnT obtained within 10 days following surgery were included. Three groups were compared based on TnT level: (1 normal (TnT ≤0.01 ng/mL, (2 intermediate (TnT 0.02–0.11 ng/mL, and (3 elevated (TnT >0.11 ng/mL. Overall and cardiovascular mortality was assessed. Results. Of the 78 patients included, there was no difference in age, gender, severity of liver disease, and echocardiographic findings. Patients in the normal and intermediate TnT groups had a lower overall mortality rate (14.3% and 0%, resp. when compared with those with elevated TnT (50%; P=0.001. Patients in the elevated TnT group had a cardiovascular mortality rate of 37.5% compared with 1.4% in the other groups combined (P<0.01. The elevated TnT group had a much higher mortality rate when compared with those in the intermediate group (P<0.0001. Conclusion. TnT may accurately help risk stratify patients in the early postoperative setting to better predict cardiovascular complications.

24-Hour Kinetics of Cardiac Troponin-T Using a "High-Sensitivity" Assay in Thoroughbred Chuckwagon Racing Geldings after Race and Associated Clinical Sampling Guidelines.

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Shields, E; Seiden-Long, I; Massie, S; Leguillette, R

2018-01-01

A "high-sensitivity" cardiac troponin-T (hscTnT) assay recently has been validated for use in horses and is a specific biomarker of myocardial damage. Postexercise release kinetics of cTnT utilizing the hscTnT assay have yet to be established in horses. To determine: (1) cTnT release kinetics in racing Thoroughbreds after a high-intensity 5/8th mile Chuckwagon race; (2) the effects of age on pre- and postrace cTnT concentrations; and (3) sampling guidelines for clinicians evaluating horses presenting after exercise. Samples were obtained from 38 Thoroughbred geldings aged 5-16 years before racing and immediately, 2, 3, 4, 6, 12, and 24 hour postrace. Prospective, observational study with convenience sampling. A fifth-generation hscTnT assay was used for plasma sample analysis, and concentrations were compared at all time-points. Correlations were determined between cTnT concentrations and age. Biochemistry analysis was performed to assess rhabdomyolysis, renal failure, and exercise-induced dehydration. All horses with measureable cTnT concentrations had significant postexercise increases in cTnT with a median peak (8.0 ng/L) at 3-hour postrace. All horses had peak postexercise cTnT concentrations 2- to 6-hour postrace ≤ the 99th percentile upper reference limit of 23.2 ng/L, after which all cTnT concentrations decreased until returning to baseline by 12-24 hours. There was no correlation over time between cTnT concentrations and age. In racing Thoroughbreds completing short-duration, high-intensity Chuckwagon races, cTnT concentrations are expected to be increased 2- to 6-hour postrace and to decrease by 12-24 hours while remaining ≤23.2 ng/L throughout. This study contributes to establishing guidelines for clinical use of the hscTnT assay in exercising horses. Copyright © 2017 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

Formulation and in vitro evaluation of theophylline matrix tablets prepared by direct compression: Effect of polymer blends

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El-Bagory, Ibrahim; Barakat, Nahla; Ibrahim, Mohamed A.; El-Enazi, Fouza

2011-01-01

The deformation mechanism of pharmaceutical powders, used in formulating directly compressed matrix tablets, affects the characteristics of the formed tablets. Three polymers of different deformation mechanisms were tested for their impact on theophylline directly compressed tablets namely Kollidon SR (KL SR, plastic deformation), Ethylcellulose (EC, elastic deformation) and Carnauba wax (CW, brittle deformation) at different compression forces. However, tablets based mainly on KL SR, the plastically deformed polymer (TN1) exhibited the highest hardness values compared to the other formulae which are based on either blends of KL SR with CW, the very brittle deformed polymer. The upper detected force for TN formulae and the lower punch force were found to dependent mainly on the powder deformation. This difference is attributed to the work done during the compression phase as well as the work lost during the decompression phase. Furthermore, the release profiles of TN from formulae TN2 and TN4 that are based on the composition (2KL SR:1EC) and (1KL SR:2EC), respectively, were consistent with different deformation mechanisms of KL SR and EC and on the physicochemical properties like the water absorptive capacity of EC. Upon increasing the weight ratio of KL SR (TN2), the release rate was greatly retarded (39.4%, 37.1%, 35.0% and 33.6% released after 8 h at 5, 10, 15 and 20 kN. PMID:24115902

Comparison of cardiac troponins I and T measured with high-sensitivity methods for evaluation of prognosis in atrial fibrillation: an ARISTOTLE substudy.

Science.gov (United States)

Hijazi, Ziad; Siegbahn, Agneta; Andersson, Ulrika; Lindahl, Bertil; Granger, Christopher B; Alexander, John H; Atar, Dan; Gersh, Bernard J; Hanna, Michael; Harjola, Veli-Pekka; Horowitz, John; Husted, Steen; Hylek, Elaine M; Lopes, Renato D; McMurray, John J V; Wallentin, Lars

2015-02-01

Although cardiac troponin is associated with outcomes in atrial fibrillation (AF), the complementary prognostic information provided by cardiac troponin I (cTnI) and cTnT is unknown. This study investigated the distribution, determinants, and prognostic value of cTnI and cTnT concentrations in patients with AF. Samples were collected. At the time of randomization, we analyzed cTnI and cTnT concentrations of 14806 AF patients in the Apixaban for Reduction in Stroke and Other Thromboembolic Events in Atrial Fibrillation (ARISTOTLE) trial using high-sensitivity assays. Correlations (Spearman), determinants (multivariable linear regression), and outcomes (adjusted Cox models and c-statistics) were investigated. Concentrations of cTnI and cTnT were correlated (r = 0.70) and measurable in most participants [cTnI 98.5% (median 5.4 ng/L, ≥99th percentile in 9.2%) and cTnT 93.5% (median 10.9 ng/L, ≥99th percentile in 34.4%)]. Renal impairment was the most important factor affecting the concentrations of both troponins. cTnI increase was more associated with heart failure, vascular disease, and persistent/permanent AF, and cTnT with age, male sex, and diabetes. Over a median 1.9 years of follow-up, patients with both troponins above the median had significantly higher risk for stroke/systemic embolism [hazard ratio (HR) 1.72 (95% CI 1.31-2.27)], cardiac death [3.14 (2.35-4.20)], and myocardial infarction [2.99 (1.78-5.03)] than those with both troponins below median (all P Chemistry.

Mobile Genetic Elements and Evolution of CRISPR-Cas Systems: All the Way There and Back

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Makarova, Kira S.

2017-01-01

Abstract The Clustered Regularly Interspaced Palindromic Repeats (CRISPR)-CRISPR-associated proteins (Cas) systems of bacterial and archaeal adaptive immunity show multifaceted evolutionary relationships with at least five classes of mobile genetic elements (MGE). First, the adaptation module of CRISPR-Cas that is responsible for the formation of the immune memory apparently evolved from a Casposon, a self-synthesizing transposon that employs the Cas1 protein as the integrase and might have brought additional cas genes to the emerging immunity loci. Second, a large subset of type III CRISPR-Cas systems recruited a reverse transcriptase from a Group II intron, providing for spacer acquisition from RNA. Third, effector nucleases of Class 2 CRISPR-Cas systems that are responsible for the recognition and cleavage of the target DNA were derived from transposon-encoded TnpB nucleases, most likely, on several independent occasions. Fourth, accessory nucleases in some variants of types I and III toxin and type VI effectors RNases appear to be ultimately derived from toxin nucleases of microbial toxin–antitoxin modules. Fifth, the opposite direction of evolution is manifested in the recruitment of CRISPR-Cas systems by a distinct family of Tn7-like transposons that probably exploit the capacity of CRISPR-Cas to recognize unique DNA sites to facilitate transposition as well as by bacteriophages that employ them to cope with host defense. Additionally, individual Cas proteins, such as the Cas4 nuclease, were recruited by bacteriophages and transposons. The two-sided evolutionary connection between CRISPR-Cas and MGE fits the “guns for hire” paradigm whereby homologous enzymatic machineries, in particular nucleases, are shuttled between MGE and defense systems and are used alternately as means of offense or defense. PMID:28985291

Thermodynamic properties of Heusler Fe2VSi

Science.gov (United States)

Ito, Masakazu; Kai, Keita; Furuta, Tatsuya; Manaka, Hirotaka; Terada, Norio; Hiroi, Masahiko; Kondo, Akihiro; Kindo, Koichi

2018-05-01

We investigated temperature, T, dependence of magnetization, M(T), electrical resistivity, ρ(T), and specific heat, Cp(T), for the Heusler compound Fe2VSi. M(T) shows anomalies at TN1 ˜ 115 K and at TN2 ˜ 35 K. The anomaly at TN1 is caused by the magnetic transition with a crystal structural change. On the other hand, ρ(T) and Cp(T) show only anomaly at TN1, and no trace of anomaly at TN2 is observed. Because of the irreversibility of M(T), which is the characteristic of spin-glass freezing, appears below TN2, a spin-glass freezing may occur at TN2. From the analogy of the Heusler compound (Fe1-xVx ) 3Si with the cubic D03 crystal structure, (0 ≤ x ≤ 0.2), we suggested that the atomic disorder of V site by the Fe atoms gives rise to the magnetic frustration. This could be cause for the spin-glass freezing. By the Clausius-Clapeyron relation, pressure, P, derivative of TN1, (d/TN 1 d P ), is estimated to be ˜-10 K/Gpa.

Fabrication of Inverted Bulk-Heterojunction Organic Solar Cell with Ultrathin Titanium Oxide Nanosheet as an Electron-Extracting Buffer Layer

Science.gov (United States)

Itoh, Eiji; Maruyama, Yasutake; Fukuda, Katsutoshi

2012-02-01

The contributions and deposition conditions of ultrathin titania nanosheet (TN) crystallites were studied in an inverted bulk-heterojunction (BHJ) cell in indium tin oxide (ITO)/titania nanosheet/poly(3-hexylthiophene) (P3HT):phenyl-C61-butyric acid methylester (PCBM) active layer/MoOx/Ag multilayered photovoltaic devices. Only one or two layers of poly(diallyldimethylammonium chloride) (PDDA) and TN multilayered film deposited by the layer-by-layer deposition technique effectively decreased the leakage current and increased both open circuit voltage (VOC) and fill factor (FF), and power conversion efficiency (η) was increased nearly twofold by the insertion of two TN layers. The deposition of additional TN layers caused the reduction in FF, and the abnormal S-shaped curves above VOC for the devices with three and four TN layers were ascribed to the interfacial potential barrier at the ITO/TN interface and the series resistance across the multilayers of TN and PDDA. The performance of the BHJ cell with TN was markedly improved, and the S-shaped curves were eliminated following the the insertion of anatase-phase titanium dioxide between the ITO and TN layers owing to the decrease in the interfacial potential barrier.

Type IV Pili are required for virulence, twitching motility, and biofilm formation of acidovorax avenae subsp. Citrulli.

Science.gov (United States)

Bahar, Ofir; Goffer, Tal; Burdman, Saul

2009-08-01

Acidovorax avenae subsp. citrulli is the causal agent of bacterial fruit blotch (BFB), a threatening disease of watermelon, melon, and other cucurbits. Despite the economic importance of BFB, relatively little is known about basic aspects of the pathogen's biology and the molecular basis of its interaction with host plants. To identify A. avenae subsp. citrulli genes associated with pathogenicity, we generated a transposon (Tn5) mutant library on the background of strain M6, a group I strain of A. avenae subsp. citrulli, and screened it for reduced virulence by seed-transmission assays with melon. Here, we report the identification of a Tn5 mutant with reduced virulence that is impaired in pilM, which encodes a protein involved in assembly of type IV pili (TFP). Further characterization of this mutant revealed that A. avenae subsp. citrulli requires TFP for twitching motility and wild-type levels of biofilm formation. Significant reductions in virulence and biofilm formation as well as abolishment of twitching were also observed in insertional mutants affected in other TFP genes. We also provide the first evidence that group I strains of A. avenae subsp. citrulli can colonize and move through host xylem vessels.

Modulation of glacier ablation by tephra coverage from Eyjafjallajökull and Grímsvötn volcanoes, Iceland: an automated field experiment

Science.gov (United States)

Möller, Rebecca; Möller, Marco; Kukla, Peter A.; Schneider, Christoph

2018-01-01

We report results from a field experiment investigating the influence of volcanic tephra coverage on glacier ablation. These influences are known to be significantly different from those of moraine debris on glaciers due to the contrasting grain size distribution and thermal conductivity. Thus far, the influences of tephra deposits on glacier ablation have rarely been studied. For the experiment, artificial plots of two different tephra types from Eyjafjallajökull and Grímsvötn volcanoes were installed on a snow-covered glacier surface of Vatnajökull ice cap, Iceland. Snow-surface lowering and atmospheric conditions were monitored in summer 2015 and compared to a tephra-free reference site. For each of the two volcanic tephra types, three plots of variable thickness ( ˜ 1.5, ˜ 8.5 and ˜ 80 mm) were monitored. After limiting the records to a period of reliable measurements, a 50-day data set of hourly records was obtained, which can be downloaded from the Pangaea data repository (pangaea.de" target="_blank">https://www.pangaea.de; doi:10.1594/PANGAEA.876656). The experiment shows a substantial increase in snow-surface lowering rates under the ˜ 1.5 and ˜ 8.5 mm tephra plots when compared to uncovered conditions. Under the thick tephra cover some insulating effects could be observed. These results are in contrast to other studies which depicted insulating effects for much thinner tephra coverage on bare-ice glacier surfaces. Differences between the influences of the two different petrological types of tephra exist but are negligible compared to the effect of tephra coverage overall.

Tunicamycin promotes apoptosis in leukemia cells through ROS generation and downregulation of survivin expression.

Science.gov (United States)

Lim, Eun Jin; Heo, Jeonghoon; Kim, Young-Ho

2015-08-01

Tunicamycin (TN), one of the endoplasmic reticulum stress inducers, has been reported to inhibit tumor cell growth and exhibit anticarcinogenic activity. However, the mechanism by which TN initiates apoptosis remains poorly understood. In the present study, we investigated the effect of TN on the apoptotic pathway in U937 cells. We show that TN induces apoptosis in association with caspase-3 activation, generation of reactive oxygen species (ROS), and downregulation of survivin expression. P38 MAPK (mitogen-activated protein kinase) and the generation of ROS signaling pathway play crucial roles in TN-induced apoptosis in U937 cells. We hypothesized that TN-induced activation of p38 MAPK signaling pathway is responsible for cell death. To test this hypothesis, we selectively inhibited MAPK during treatment with TN. Our data demonstrated that inhibitor of p38 (SB), but not ERK (PD) or JNK (SP), partially maintained apoptosis during treatment with TN. Pre-treatment with NAC and GSH markedly prevented cell death, suggesting a role for ROS in this process. Ectopic expression of survivin in U937 cells attenuated TN-induced apoptosis by suppression of caspase-3 cleavage, mitochondrial membrane potential, and cytochrome c release in U937 cells. Taken together, our results show that TN modulates multiple components of the apoptotic response of human leukemia cells and raise the possibility of a novel therapeutic strategy for hematological malignancies.

Impact of Water Management on Rice Varieties, Yield, and Water Productivity under the System of Rice Intensification in Southern Taiwan

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Victoriano Joseph Pascual

2016-12-01

Full Text Available The system of rice intensification (SRI uses less water and enhances rice yield through synergy among several agronomic management practices. This claim was investigated to determine the effects of crop growth, yield and irrigation water use, using two thirds of the recommended SRI practices and two rice varieties, namely Tainan11 (TN11 and Tidung30 (TD30. Irrigation regimes were (a intermittent irrigation with three-day intervals (TD303 and TN113; (b intermittent irrigation with seven-day intervals (TD307 and TN117 and (c continuous flooding (TD30F and TN11F. Results showed that intermittent irrigation of three- and seven-day intervals produced water savings of 55% and 74% compared with continuous flooding. Total water productivity was greater with intermittent irrigation at seven-day intervals producing 0.35 kg·grain/m3 (TN117 and 0.46 kg·grain/m3 (TD307. Average daily headed panicle reduced by 166% and 196% for TN113 and TN117 compared with TN11F, with similar reduction recorded for TD303 (150% and TD307 (156% compared with TD30F. Grain yield of TD30 was comparable among irrigation regimes; however, it reduced by 30.29% in TN117 compared to TN11F. Plant height and leaf area were greater in plants exposed to intermittent irrigation of three-day intervals.

A mutation in the aroE gene affects pigment production, virulence, and chemotaxis in Xanthomonas oryzae pv. oryzae.

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Kim, Hong-Il; Noh, Tae-Hwan; Lee, Chang-Soo; Park, Young-Jin

2015-01-01

Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight (BB) in rice. To study its function, a random insertion mutation library of Xoo was constructed using the Tn5 transposon. A mutant strain with decreased virulence against the susceptible rice cultivar IR24 was isolated from the library (aroE mutant), which also had extremely low pigment production. Thermal asymmetric interlaced-polymerase chain reaction (TAIL-PCR) and sequence analysis of the mutant revealed that the transposon was inserted into the aroE gene (encoding shikimate dehydrogenase). To investigate gene expression changes in the pigment- and virulence-deficient mutant, DNA microarray analysis was performed, which showed downregulation of 20 genes involved in the chemotaxis of Xoo. Our findings reveal that mutation of the aroE gene affects virulence and pigment production, as well as expression of genes involved in Xoo chemotaxis. Copyright © 2014 Elsevier GmbH. All rights reserved.

Head-to-head comparison of cardiac troponin T and troponin I in patients without acute coronary syndrome

DEFF Research Database (Denmark)

Árnadóttir, Ásthildur; Falk Klein, Christine; Iversen, Kasper

2017-01-01

BACKGROUND: Cardiac-specific troponin T (cTnT) and troponin I (cTnI) are considered diagnostically equal in patients with acute coronary syndrome (ACS). The aim of this systematic review was to compare the prevalence and prognostic strength of elevations of cTnT and cTnI in patients with other...

Sex differences of troponin test performance in chest pain patients.

Science.gov (United States)

Slagman, Anna; Searle, Julia; Vollert, Jörn O; Storchmann, Harald; Büschenfelde, Dirk Meyer Zum; von Recum, Johannes; Vlasny, Daniela; Ale-Abaei, Angela; Koch, Matthias; Müller, Christian; Müller, Reinhold; Somasundaram, Rajan; Möckel, Martin

2015-01-01

Current guidelines recommend troponin as the preferred biomarker to diagnose acute myocardial infarction (AMI) irrespective of the patient's sex. Recent reports have shown that sex-specific cut-offs should be considered but studies investigating sex-differences in the diagnostic accuracy of cardiac troponins are sparse. To evaluate whether the diagnostic performance of cardiac troponin at admission (cTn) under routine conditions is influenced by patient's sex. Between 15th of February 2009 and 15th of February 2010, women (n=1648) and men (n=2305) who presented to the emergency department with chest pain (n=3954) were enrolled. The diagnostic performance of the routine, contemporary sensitive cTn assays (TnI; Stratus® CS, Siemens and TnT; Roche Diagnostics) at baseline for the diagnosis of non-ST-elevation myocardial infarction (NSTEMI) was analyzed. NSTEMI was diagnosed in 7.3% (n=287) of all patients. Men were more likely to be diagnosed with NSTEMI (8.8%; n=202) as compared to women (5.2%; n=85; psex, with a lower sensitivity and NPV in women. The definition and implementation of sex-specific cut-off values for cTn into clinical routine seems to be highly recommendable. Copyright © 2015. Published by Elsevier Ireland Ltd.

Baixa temperatura noturna e deficiência hídrica na fotossíntese de cana‑de‑açúcar

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Daniela Favero São Pedro Machado

2013-05-01

Full Text Available O objetivo deste trabalho foi avaliar as respostas fotossintéticas da cana‑de‑açúcar aos efeitos simultâneos e isolados de baixa temperatura noturna (TN e deficiência hídrica (DH. Após 128 dias do plantio, as plantas da cultivar IACSP94-2094 foram submetidas aos tratamentos: controle, sem DH e com TN de 20°C (TN20; com DH e TN de 20°C (DH/TN20; sem DH e com TN de 12°C (TN12; e com DH e TN de 12°C (DH/TN12 por cinco dias. Após o período de tratamento, as plantas foram irrigadas e submetidas à TN de 20°C por mais quatro dias, para recuperação. Houve decréscimos na assimilação de CO2 em todos os tratamentos. A recuperação total da assimilação de CO2 foi observada apenas nas plantas do tratamento TN12. A ocorrência simultânea da baixa temperatura noturna e da deficiência hídrica causou dano acentuado e persistente na condutância estomática, na capacidade máxima da ribulose‑1,5‑bisfosfato carboxilase, no transporte aparente de elétrons, no fator de eficiência e na eficiência operacional do fotossistema II, o que resultou em limitações difusivas, bioquímicas e fotoquímicas da fotossíntese das plantas de cana‑de‑açúcar.

Characterization of extended-spectrum β-lactamases (ESBLs)-producing Salmonella in retail raw chicken carcasses.

Science.gov (United States)

Qiao, Jing; Zhang, Qiang; Alali, Walid Q; Wang, Jiawei; Meng, Lingyuan; Xiao, Yingping; Yang, Hua; Chen, Sheng; Cui, Shenghui; Yang, Baowei

2017-05-02

Extended-spectrum β-lactamases (ESBLs)-producing Salmonella is considered a serious concern to public health worldwide. However, limited information is available on ESBLs-producing Salmonella in retail chicken products in China. The objective of this study was to characterize ESBLs-producing Salmonella isolates from retail chickens in China. A total of 890 Salmonella isolates from retail chicken carcasses collected from 4 provinces were firstly screened for ESBLs-production phenotype via the double-disk synergy test method. A total of 96 (10.8%, n=890) ESBLs-producing Salmonella were identified and subjected to PFGE analysis, characterization for the presence of ESBLs encoding genes, transposons, carbapenemase and virulence genes. A total of 59 PFGE profiles were detected in these 96 isolates, among which 57.3% were found to harbor bla TEM-1 , whereas 30.2%, 24.0%, 18.8% and 7.3% were carrying bla OXA-1 , bla CTX-M-15 , bla CTX-M-3 and bla PSE-1 genes, respectively. Moreover, 42 (43.8%) isolates co-carried 2 ESBLs-producing genes, and two (2.1%) isolates co-carried 3 genes. Furthermore, 24 (25.0%) ESBLs-producing isolates carried VIM and 10 (10.4%) carried KPC encoding genes that closely associated with carbapenems resistance. Eighty-eight isolates harbored transposons ranging from 4.2% for Tn903 to 76.0% for Tn21. Out of the 88 Salmonella that harbored transposons, 25%, 22.7%, 23.9%, 10.2% and 1.1% of isolates were found to carry 2, 3, 4, 5 and 6 transposons, respectively. The minimum inhibitory concentration (MIC) values for cephalosporins (ceftriaxone, cefoperazone and cefoxitin) to ESBLs-producing isolates were from 4 to 1024μg/mL, for nalidixic acid were from 64 to 512μg/mL, for fluoroquinolones (ciprofloxacin, levofloxacin and gatifloxacin) were from 4 to 256μg/mL. Twenty-nine virulence genes were detected in the 96 ESBLs-producing isolates with 2.1% harbored spvR (lowest) and 90.6% harbored marT and steB (highest). All isolates carried at least one

In vitro characterization and inhibition of the CXCR4/CXCL12 chemokine axis in human uveal melanoma cell lines

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Antecka Emilia

2007-11-01

Full Text Available Abstract Purpose The CXCR4/CXCL12 chemokine axis may play a critical role in guiding CXCR4+ circulating malignant cells to organ specific locations that actively secrete its ligand CXCL12 (SDF-1 such as bone, brain, liver, and lungs. We sought to characterize the presence of the CXCR4/CXCL12 axis in five uveal melanoma (UM cell lines in vitro. The ability of TN14003, a synthetic peptide inhibitor that targets the CXCR4 receptor complex, to inhibit this axis was also assessed. Methods Immunocytochemistry was performed against CXCR4 to confirm expression of this chemokine receptor in all five UM cell lines. Flow cytometry was preformed to evaluate CXCR4 cell surface expression on all five UM cell lines. A proliferation assay was also used to test effects TN14003 would have on cellular proliferation. Inhibition of cellular migration by specifically inhibiting the CXCR4/CXCL12 axis with TN14003 was also investigated. The binding efficacy of TN14003 to the CXCR4 receptor was assessed through flow cytometric methods. Results The CXCR4 receptor was present on all five UM cell lines. All five cell lines expressed different relative levels of surface CXCR4. TN14003 did not affect the proliferation of the five cell lines (p > 0.05. All cell lines migrated towards the chemokine CXCL12 at a level greater than the negative control (p Conclusion Interfering with the CXCR4/CXCL12 axis, using TN14003 was shown to effectively down regulate UM cell migration in vitro. Knowing that UM expresses the CXCR4 receptor, these CXCR4+ cells may be less likely to colonize distant organs that secrete the CXCL12 ligand, if treated with an inhibitor that binds CXCR4. Further studies should be pursued in order to test TN14003 efficacy in vivo.

Tumor exosomes induce tunneling nanotubes in lipid raft-enriched regions of human mesothelioma cells

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Thayanithy, Venugopal [Department of Medicine, Division of Hematology, Oncology and Transplantation, University of Minnesota, Minneapolis, MN 55455 (United States); Babatunde, Victor [Moore Laboratory, Department of Cell Biology, Sloan-Kettering Institute, Memorial Sloan-Kettering Cancer Center, New York, NY 10021 (United States); Dickson, Elizabeth L. [Department of Obstetrics and Gynecology, Division of Gynecologic Oncology, University of Minnesota, Minneapolis, MN 55455 (United States); Wong, Phillip [Department of Medicine, Division of Hematology, Oncology and Transplantation, University of Minnesota, Minneapolis, MN 55455 (United States); Oh, Sanghoon; Ke, Xu; Barlas, Afsar; Fujisawa, Sho; Romin, Yevgeniy [Molecular Cytology, Memorial Sloan-Kettering Cancer Center, New York, NY 10021 (United States); Moreira, André L. [Department of Pathology, Memorial Sloan-Kettering Cancer Center, New York, NY 10021 (United States); Downey, Robert J. [Department of Surgery, Memorial Sloan-Kettering Cancer Center, New York, NY 10021 (United States); Steer, Clifford J. [Departments of Medicine and Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455 (United States); Subramanian, Subbaya [Department of Surgery, University of Minnesota, Minneapolis, MN 55455 (United States); Manova-Todorova, Katia [Molecular Cytology, Memorial Sloan-Kettering Cancer Center, New York, NY 10021 (United States); Moore, Malcolm A.S. [Moore Laboratory, Department of Cell Biology, Sloan-Kettering Institute, Memorial Sloan-Kettering Cancer Center, New York, NY 10021 (United States); Lou, Emil, E-mail: emil-lou@umn.edu [Department of Medicine, Division of Hematology, Oncology and Transplantation, University of Minnesota, Minneapolis, MN 55455 (United States)

2014-04-15

Tunneling nanotubes (TnTs) are long, non-adherent, actin-based cellular extensions that act as conduits for transport of cellular cargo between connected cells. The mechanisms of nanotube formation and the effects of the tumor microenvironment and cellular signals on TnT formation are unknown. In the present study, we explored exosomes as potential mediators of TnT formation in mesothelioma and the potential relationship of lipid rafts to TnT formation. Mesothelioma cells co-cultured with exogenous mesothelioma-derived exosomes formed more TnTs than cells cultured without exosomes within 24–48 h; and this effect was most prominent in media conditions (low-serum, hyperglycemic medium) that support TnT formation (1.3–1.9-fold difference). Fluorescence and electron microscopy confirmed the purity of isolated exosomes and revealed that they localized predominantly at the base of and within TnTs, in addition to the extracellular environment. Time-lapse microscopic imaging demonstrated uptake of tumor exosomes by TnTs, which facilitated intercellular transfer of these exosomes between connected cells. Mesothelioma cells connected via TnTs were also significantly enriched for lipid rafts at nearly a 2-fold higher number compared with cells not connected by TnTs. Our findings provide supportive evidence of exosomes as potential chemotactic stimuli for TnT formation, and also lipid raft formation as a potential biomarker for TnT-forming cells. - Highlights: • Exosomes derived from malignant cells can stimulate an increased rate in the formation of tunneling nanotubes. • Tunneling nanotubes can serve as conduits for intercellular transfer of these exosomes. • Most notably, exosomes derived from benign mesothelial cells had no effect on nanotube formation. • Cells forming nanotubes were enriched in lipid rafts at a greater number compared with cells not forming nanotubes. • Our findings suggest causal and potentially synergistic association of exosomes and

The clinicopathologic characteristics and prognostic significance of triple-negativity in node-negative breast cancer

International Nuclear Information System (INIS)

Rhee, Jiyoung; Kim, Tae-You; Han, Sae-Won; Oh, Do-Youn; Kim, Jee Hyun; Im, Seock-Ah; Han, Wonshik; Ae Park, In; Noh, Dong-Young; Bang, Yung-Jue

2008-01-01

Triple-negative (TN) breast cancer, which is defined as being negative for the estrogen receptor (ER), the progesterone receptor (PR), and the human epidermal growth factor receptor 2 (HER-2), represents a subset of breast cancer with different biologic behaviour. We investigated the clinicopathologic characteristics and prognostic indicators of lymph node-negative TN breast cancer. Medical records were reviewed from patients with node-negative breast cancer who underwent curative surgery at Seoul National University Hospital between Jan. 2000 and Jun. 2003. Clinicopathologic variables and clinical outcomes were evaluated. Among 683 patients included, 136 had TN breast cancer and 529 had non-TN breast cancer. TN breast cancer correlated with younger age (< 35 y, p = 0.003), and higher histologic and nuclear grade (p < 0.001). It also correlated with a molecular profile associated with biological aggressiveness: negative for bcl-2 expression (p < 0.001), positive for the epidermal growth factor receptor (p = 0.003), and a high level of p53 (p < 0.001) and Ki67 expression (p < 0.00). The relapse rates during the follow-up period (median, 56.8 months) were 14.7% for TN breast cancer and 6.6% for non-TN breast cancer (p = 0.004). Relapse free survival (RFS) was significantly shorter among patients with TN breast cancer compared with those with non-TN breast cancer (4-year RFS rate 85.5% vs. 94.2%, respectively; p = 0.001). On multivariate analysis, young age, close resection margin, and triple-negativity were independent predictors of shorter RFS. TN breast cancer had higher relapse rate and more aggressive clinicopathologic characteristics than non-TN in node-negative breast cancer. Thus, TN breast cancer should be integrated into the risk factor analysis for node-negative breast cancer

Tumor exosomes induce tunneling nanotubes in lipid raft-enriched regions of human mesothelioma cells

International Nuclear Information System (INIS)

Thayanithy, Venugopal; Babatunde, Victor; Dickson, Elizabeth L.; Wong, Phillip; Oh, Sanghoon; Ke, Xu; Barlas, Afsar; Fujisawa, Sho; Romin, Yevgeniy; Moreira, André L.; Downey, Robert J.; Steer, Clifford J.; Subramanian, Subbaya; Manova-Todorova, Katia; Moore, Malcolm A.S.; Lou, Emil

2014-01-01

Tunneling nanotubes (TnTs) are long, non-adherent, actin-based cellular extensions that act as conduits for transport of cellular cargo between connected cells. The mechanisms of nanotube formation and the effects of the tumor microenvironment and cellular signals on TnT formation are unknown. In the present study, we explored exosomes as potential mediators of TnT formation in mesothelioma and the potential relationship of lipid rafts to TnT formation. Mesothelioma cells co-cultured with exogenous mesothelioma-derived exosomes formed more TnTs than cells cultured without exosomes within 24–48 h; and this effect was most prominent in media conditions (low-serum, hyperglycemic medium) that support TnT formation (1.3–1.9-fold difference). Fluorescence and electron microscopy confirmed the purity of isolated exosomes and revealed that they localized predominantly at the base of and within TnTs, in addition to the extracellular environment. Time-lapse microscopic imaging demonstrated uptake of tumor exosomes by TnTs, which facilitated intercellular transfer of these exosomes between connected cells. Mesothelioma cells connected via TnTs were also significantly enriched for lipid rafts at nearly a 2-fold higher number compared with cells not connected by TnTs. Our findings provide supportive evidence of exosomes as potential chemotactic stimuli for TnT formation, and also lipid raft formation as a potential biomarker for TnT-forming cells. - Highlights: • Exosomes derived from malignant cells can stimulate an increased rate in the formation of tunneling nanotubes. • Tunneling nanotubes can serve as conduits for intercellular transfer of these exosomes. • Most notably, exosomes derived from benign mesothelial cells had no effect on nanotube formation. • Cells forming nanotubes were enriched in lipid rafts at a greater number compared with cells not forming nanotubes. • Our findings suggest causal and potentially synergistic association of exosomes and

Human plasminogen binding protein tetranectin

DEFF Research Database (Denmark)

Kastrup, J S; Rasmussen, H; Nielsen, B B

1997-01-01

The recombinant human plasminogen binding protein tetranectin (TN) and the C-type lectin CRD of this protein (TN3) have been crystallized. TN3 crystallizes in the tetragonal space group P4(2)2(1)2 with cell dimensions a = b = 64.0, c = 75.7 A and with one molecule per asymmetric unit. The crystals...... to at least 2.5 A. A full data set has been collected to 3.0 A. The asymmetric unit contains one monomer of TN. Molecular replacement solutions for TN3 and TN have been obtained using the structure of the C-type lectin CRD of rat mannose-binding protein as search model. The rhombohedral space group indicates...

Complete nucleotide sequence and analysis of two conjugative broad host range plasmids from a marine microbial biofilm.

Directory of Open Access Journals (Sweden)

Peter Norberg

Full Text Available The complete nucleotide sequence of plasmids pMCBF1 and pMCBF6 was determined and analyzed. pMCBF1 and pMCBF6 form a novel clade within the IncP-1 plasmid family designated IncP-1 ς. The plasmids were exogenously isolated earlier from a marine biofilm. pMCBF1 (62 689 base pairs; bp and pMCBF6 (66 729 bp have identical backbones, but differ in their mercury resistance transposons. pMCBF1 carries Tn5053 and pMCBF6 carries Tn5058. Both are flanked by 5 bp direct repeats, typical of replicative transposition. Both insertions are in the vicinity of a resolvase gene in the backbone, supporting the idea that both transposons are "res-site hunters" that preferably insert close to and use external resolvase functions. The similarity of the backbones indicates recent insertion of the two transposons and the ongoing dynamics of plasmid evolution in marine biofilms. Both plasmids also carry the insertion sequence ISPst1, albeit without flanking repeats. ISPs1is located in an unusual site within the control region of the plasmid. In contrast to most known IncP-1 plasmids the pMCBF1/pMCBF6 backbone has no insert between the replication initiation gene (trfA and the vegetative replication origin (oriV. One pMCBF1/pMCBF6 block of about 2.5 kilo bases (kb has no similarity with known sequences in the databases. Furthermore, insertion of three genes with similarity to the multidrug efflux pump operon mexEF and a gene from the NodT family of the tripartite multi-drug resistance-nodulation-division (RND system in Pseudomonas aeruginosa was found. They do not seem to confer antibiotic resistance to the hosts of pMCBF1/pMCBF6, but the presence of RND on promiscuous plasmids may have serious implications for the spread of antibiotic multi-resistance.

Organic matter geochemical signatures (TOC, TN, C/N ratio, δ13C and δ15N) of surface sediment from lakes distributed along a climatological gradient on the western side of the southern Andes.

Science.gov (United States)

Contreras, Sergio; Werne, Josef P; Araneda, A; Urrutia, R; Conejero, C A

2018-07-15

Paleolimnological studies in western South America, where meteorological stations are scarce, are critical to obtain more realistic and reliable regional reconstructions of past climate and environmental changes, including vegetation and water budget variability. However, climate and environmental geochemical indicators must be tested before they can be applied with confidence. Here we present a survey of lacustrine surface sediment (core top, 0 to ~1cm) biogeochemical proxies (total organic carbon [TOC], total nitrogen [TN], carbon/nitrogen ratio [C/N ratio] and bulk organic δ 13 C and total δ 15 N) from a suite of 72 lakes spanning the transition from a Mediterranean climate with a patchwork of cultivated vegetation, pastureland, and conifers in central Chile to a rainy temperate climate dominated by broadleaf deciduous and evergreen forest further south. Sedimentary data are compared to the latitudinal and orographic climatic trends of the region based on the climatology (precipitation and temperature) produced with Climate Forecast System Reanalysis (CFSR) data and the modern Southern Hemisphere Westerly Winds (SWW) location. The geochemical data show inflection points at ~42°S latitude and ~1500m elevation that are likely related to the northern limit of influence of the SWW and elevation of the snow line, respectively. Overall the organic proxies were able to mimic climatic trends (Mean Annual Precipitation [MAP] and temperature [MAT]), indicating that they are a useful tool to be included in paleoclimatological reconstruction of the region. Copyright © 2018 Elsevier B.V. All rights reserved.

The bandit, a new DNA transposon from a hookworm-possible horizontal genetic transfer between host and parasite.

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Thewarach Laha

2007-09-01

Full Text Available An enhanced understanding of the hookworm genome and its resident mobile genetic elements should facilitate understanding of the genome evolution, genome organization, possibly host-parasite co-evolution and horizontal gene transfer, and from a practical perspective, development of transposon-based transgenesis for hookworms and other parasitic nematodes.A novel mariner-like element (MLE was characterized from the genome of the dog hookworm, Ancylostoma caninum, and termed bandit. The consensus sequence of the bandit transposon was 1,285 base pairs (bp in length. The new transposon was flanked by perfect terminal inverted repeats of 32 nucleotides in length with a common target site duplication TA, and it encoded an open reading frame (ORF of 342 deduced amino acid residues. Phylogenetic comparisons confirmed that the ORF encoded a mariner-like transposase, which included conserved catalytic domains, and that the bandit transposon belonged to the cecropia subfamily of MLEs. The phylogenetic analysis also indicated that the Hsmar1 transposon from humans was the closest known relative of bandit, and that bandit and Hsmar1 constituted a clade discrete from the Tc1 subfamily of MLEs from the nematode Caenorhabditis elegans. Moreover, homology models based on the crystal structure of Mos1 from Drosophila mauritiana revealed closer identity in active site residues of the catalytic domain including Ser281, Lys289 and Asp293 between bandit and Hsmar1 than between Mos1 and either bandit or Hsmar1. The entire bandit ORF was amplified from genomic DNA and a fragment of the bandit ORF was amplified from RNA, indicating that this transposon is actively transcribed in hookworms.A mariner-like transposon termed bandit has colonized the genome of the hookworm A. caninum. Although MLEs exhibit a broad host range, and are identified in other nematodes, the closest phylogenetic relative of bandit is the Hsmar1 element of humans. This surprising finding suggests

Structural magnetic resonance imaging can identify trigeminal system abnormalities in classical trigeminal neuralgia

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Danielle DeSouza

2016-10-01

Full Text Available Classical trigeminal neuralgia (TN is a chronic pain disorder that has been described as one ofthe most severe pains one can suffer. The most prevalent theory of TN etiology is that the trigeminal nerve is compressed at the root entry zone (REZ by blood vessels. However, there is significant evidence showing a lack of neurovascular compression (NVC for many cases of classical TN. Furthermore, a considerable number of patients who are asymptomatic have MR evidence of NVC. Since there is no validated animal model that reproduces the clinical features of TN, our understanding of TN pathology mainly comes from biopsy studies that have limitations. Sophisticated structural MRI techniques including diffusion tensor imaging provide new opportunities to assess the trigeminal nerves and CNS to provide insight into TN etiology and pathogenesis. Specifically, studies have used high-resolution structural MRI methods to visualize patterns of trigeminal nerve-vessel relationships and to detect subtle pathological features at the trigeminal REZ. Structural MRI has also identified CNS abnormalities in cortical and subcortical gray matter and white matter and demonstrated that effective neurosurgical treatment for TN is associated with a reversal of specific nerve and brain abnormalities. In conclusion, this review highlights the advanced structural neuroimaging methods that are valuable tools to assess the trigeminal system in TN and may inform our current understanding of TN pathology. These methods may in the future have clinical utility for the development of neuroimaging-based biomarkers of TN.

A Noise Trimming and Positional Significance of Transposon Insertion System to Identify Essential Genes in Yersinia pestis

Science.gov (United States)

Yang, Zheng Rong; Bullifent, Helen L.; Moore, Karen; Paszkiewicz, Konrad; Saint, Richard J.; Southern, Stephanie J.; Champion, Olivia L.; Senior, Nicola J.; Sarkar-Tyson, Mitali; Oyston, Petra C. F.; Atkins, Timothy P.; Titball, Richard W.

2017-02-01

Massively parallel sequencing technology coupled with saturation mutagenesis has provided new and global insights into gene functions and roles. At a simplistic level, the frequency of mutations within genes can indicate the degree of essentiality. However, this approach neglects to take account of the positional significance of mutations - the function of a gene is less likely to be disrupted by a mutation close to the distal ends. Therefore, a systematic bioinformatics approach to improve the reliability of essential gene identification is desirable. We report here a parametric model which introduces a novel mutation feature together with a noise trimming approach to predict the biological significance of Tn5 mutations. We show improved performance of essential gene prediction in the bacterium Yersinia pestis, the causative agent of plague. This method would have broad applicability to other organisms and to the identification of genes which are essential for competitiveness or survival under a broad range of stresses.

Design of convolutional tornado code

Science.gov (United States)

Zhou, Hui; Yang, Yao; Gao, Hongmin; Tan, Lu

2017-09-01

As a linear block code, the traditional tornado (tTN) code is inefficient in burst-erasure environment and its multi-level structure may lead to high encoding/decoding complexity. This paper presents a convolutional tornado (cTN) code which is able to improve the burst-erasure protection capability by applying the convolution property to the tTN code, and reduce computational complexity by abrogating the multi-level structure. The simulation results show that cTN code can provide a better packet loss protection performance with lower computation complexity than tTN code.

Genome-wide discovery of novel M1T1 group A streptococcal determinants important for fitness and virulence during soft-tissue infection.

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Yoann Le Breton

2017-08-01

Full Text Available The Group A Streptococcus remains a significant human pathogen causing a wide array of disease ranging from self-limiting to life-threatening invasive infections. Epithelium (skin or throat colonization with progression to the subepithelial tissues is the common step in all GAS infections. Here, we used transposon-sequencing (Tn-seq to define the GAS 5448 genetic requirements for in vivo fitness in subepithelial tissue. A near-saturation transposon library of the M1T1 GAS 5448 strain was injected subcutaneously into mice, producing suppurative inflammation at 24 h that progressed to prominent abscesses with tissue necrosis at 48 h. The library composition was monitored en masse by Tn-seq and ratios of mutant abundance comparing the output (12, 24 and 48 h versus input (T0 mutant pools were calculated for each gene. We identified a total of 273 subcutaneous fitness (scf genes with 147 genes (55 of unknown function critical for the M1T1 GAS 5448 fitness in vivo; and 126 genes (53 of unknown function potentially linked to in vivo fitness advantage. Selected scf genes were validated in competitive subcutaneous infection with parental 5448. Two uncharacterized genes, scfA and scfB, encoding putative membrane-associated proteins and conserved among Gram-positive pathogens, were further characterized. Defined scfAB mutants in GAS were outcompeted by wild type 5448 in vivo, attenuated for lesion formation in the soft tissue infection model and dissemination to the bloodstream. We hypothesize that scfAB play an integral role in enhancing adaptation and fitness of GAS during localized skin infection, and potentially in propagation to other deeper host environments.

Identification and characterization of the multidrug resistance gene cfr in a Panton-Valentine leukocidin-positive sequence type 8 methicillin-resistant Staphylococcus aureus IVa (USA300) isolate.

LENUS (Irish Health Repository)

Shore, Anna C

2010-12-01

The staphylococcal cfr gene mediates resistance to phenicols, lincosamides, oxazolidinones, pleuromutilins, and streptogramin A, a phenotype that has been termed PhLOPS(A). The cfr gene has mainly been associated with coagulase-negative staphylococcal isolates from animals, and only a few cfr-positive methicillin-resistant Staphylococcus aureus (MRSA) isolates have been described so far. This study reports the first description of a cfr-positive MRSA isolate (M05\\/0060) belonging to the pandemic Panton-Valentine leukocidin (PVL)-positive sequence type 8 MRSA IVa\\/USA300 (ST8-MRSA-IVa\\/USA300) clone. The cfr gene was detected in M05\\/0060 using a DNA microarray which was used to screen PVL-positive MRSA isolates for the presence of virulence genes, typing markers, and antimicrobial resistance genes. Antimicrobial susceptibility testing revealed that M05\\/0060 exhibited the cfr-associated resistance phenotype. Molecular analysis identified the presence of cfr and a second phenicol resistance gene, fexA, on a novel 45-kb conjugative plasmid, which was designated pSCFS7. Within pSCFS7, a DNA segment consisting of cfr, a truncated copy of insertion sequence IS21-558, and a region with homology to the DNA invertase gene bin3 of transposon Tn552 from Bacillus mycoides was integrated into the transposase gene tnpB of the fexA-carrying transposon Tn558. The emergence of a multidrug-resistant cfr-positive variant of ST8-MRSA-IVa\\/USA300 is alarming and requires ongoing surveillance. Moreover, the identification of a novel conjugative plasmid carrying the cfr gene indicates the ability of cfr to spread to other MRSA strains.

Genome-wide discovery of novel M1T1 group A streptococcal determinants important for fitness and virulence during soft-tissue infection.

Science.gov (United States)

Le Breton, Yoann; Belew, Ashton T; Freiberg, Jeffrey A; Sundar, Ganesh S; Islam, Emrul; Lieberman, Joshua; Shirtliff, Mark E; Tettelin, Hervé; El-Sayed, Najib M; McIver, Kevin S

2017-08-01

The Group A Streptococcus remains a significant human pathogen causing a wide array of disease ranging from self-limiting to life-threatening invasive infections. Epithelium (skin or throat) colonization with progression to the subepithelial tissues is the common step in all GAS infections. Here, we used transposon-sequencing (Tn-seq) to define the GAS 5448 genetic requirements for in vivo fitness in subepithelial tissue. A near-saturation transposon library of the M1T1 GAS 5448 strain was injected subcutaneously into mice, producing suppurative inflammation at 24 h that progressed to prominent abscesses with tissue necrosis at 48 h. The library composition was monitored en masse by Tn-seq and ratios of mutant abundance comparing the output (12, 24 and 48 h) versus input (T0) mutant pools were calculated for each gene. We identified a total of 273 subcutaneous fitness (scf) genes with 147 genes (55 of unknown function) critical for the M1T1 GAS 5448 fitness in vivo; and 126 genes (53 of unknown function) potentially linked to in vivo fitness advantage. Selected scf genes were validated in competitive subcutaneous infection with parental 5448. Two uncharacterized genes, scfA and scfB, encoding putative membrane-associated proteins and conserved among Gram-positive pathogens, were further characterized. Defined scfAB mutants in GAS were outcompeted by wild type 5448 in vivo, attenuated for lesion formation in the soft tissue infection model and dissemination to the bloodstream. We hypothesize that scfAB play an integral role in enhancing adaptation and fitness of GAS during localized skin infection, and potentially in propagation to other deeper host environments.

Curcumin targets breast cancer stem-like cells with microtentacles that persist in mammospheres and promote reattachment.

Science.gov (United States)

Charpentier, Monica S; Whipple, Rebecca A; Vitolo, Michele I; Boggs, Amanda E; Slovic, Jana; Thompson, Keyata N; Bhandary, Lekhana; Martin, Stuart S

2014-02-15

Cancer stem-like cells (CSC) and circulating tumor cells (CTC) have related properties associated with distant metastasis, but the mechanisms through which CSCs promote metastasis are unclear. In this study, we report that breast cancer cell lines with more stem-like properties display higher levels of microtentacles (McTN), a type of tubulin-based protrusion of the plasma cell membrane that forms on detached or suspended cells and aid in cell reattachment. We hypothesized that CSCs with large numbers of McTNs would more efficiently attach to distant tissues, promoting metastatic efficiency. The naturally occurring stem-like subpopulation of the human mammary epithelial (HMLE) cell line presents increased McTNs compared with its isogenic non-stem-like subpopulation. This increase was supported by elevated α-tubulin detyrosination and vimentin protein levels and organization. Increased McTNs in stem-like HMLEs promoted a faster initial reattachment of suspended cells that was inhibited by the tubulin-directed drug, colchicine, confirming a functional role for McTNs in stem cell reattachment. Moreover, live-cell confocal microscopy showed that McTNs persist in breast stem cell mammospheres as flexible, motile protrusions on the surface of the mammosphere. Although exposed to the environment, they also function as extensions between adjacent cells along cell-cell junctions. We found that treatment with the breast CSC-targeting compound curcumin rapidly extinguished McTN in breast CSC, preventing reattachment from suspension. Together, our results support a model in which breast CSCs with cytoskeletal alterations that promote McTNs can mediate attachment and metastasis but might be targeted by curcumin as an antimetastatic strategy. ©2013 AACR.

The default mode network and recurrent depression: a neurobiological model of cognitive risk factors.

Science.gov (United States)

Marchetti, Igor; Koster, Ernst H W; Sonuga-Barke, Edmund J; De Raedt, Rudi

2012-09-01

A neurobiological account of cognitive vulnerability for recurrent depression is presented based on recent developments of resting state neural networks. We propose that alterations in the interplay between task positive (TP) and task negative (TN) elements of the Default Mode Network (DMN) act as a neurobiological risk factor for recurrent depression mediated by cognitive mechanisms. In the framework, depression is characterized by an imbalance between TN-TP components leading to an overpowering of TP by TN activity. The TN-TP imbalance is associated with a dysfunctional internally-focused cognitive style as well as a failure to attenuate TN activity in the transition from rest to task. Thus we propose the TN-TP imbalance as overarching neural mechanism involved in crucial cognitive risk factors for recurrent depression, namely rumination, impaired attentional control, and cognitive reactivity. During remission the TN-TP imbalance persists predisposing to vulnerability of recurrent depression. Empirical data to support this model is reviewed. Finally, we specify how this framework can guide future research efforts.

New transposon tools tailored for metabolic engineering of Gram-negative microbial cell factories

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Esteban eMartínez-García

2014-10-01

Full Text Available Re-programming microorganisms to modify their existing functions and/or to bestow bacteria with entirely new-to-Nature tasks have largely relied so far on specialized molecular biology tools. Such endeavors are not only relevant in the burgeoning metabolic engineering arena, but also instrumental to explore the functioning of complex regulatory networks from a fundamental point of view. À la carte modification of bacterial genomes thus calls for novel tools to make genetic manipulations easier. We propose the use of a series of new broad-host-range mini-Tn5 vectors, termed pBAMDs, for the delivery of gene(s into the chromosome of Gram-negative bacteria and for generating saturated mutagenesis libraries in gene function studies. These delivery vectors endow the user with the possibility of easy cloning and subsequent insertion of functional cargoes with three different antibiotic resistance markers (kanamycin, streptomycin, and gentamicin. After validating the pBAMD vectors in the environmental bacterium Pseudomonas putida KT2440, their use was also illustrated by inserting the entire poly(3-hydroxybutyrate (PHB synthesis pathway from Cupriavidus necator in the chromosome of a phosphotransacetylase mutant of Escherichia coli. PHB is a completely biodegradable polyester with a number of industrial applications that make it attractive as a potential replacement of oil-based plastics. The non-selective nature of chromosomal insertions of the biosynthetic genes was evidenced by a large landscape of PHB synthesis levels in independent clones. One clone was selected and further characterized as a microbial cell factory for PHB accumulation, and it achieved polymer accumulation levels comparable to those of a plasmid-bearing recombinant. Taken together, our results demonstrate that the new mini-Tn5 vectors can be used to confer interesting phenotypes in Gram-negative bacteria that would be very difficult to engineer through direct manipulation of the

New Transposon Tools Tailored for Metabolic Engineering of Gram-Negative Microbial Cell Factories

Energy Technology Data Exchange (ETDEWEB)

Martínez-García, Esteban; Aparicio, Tomás; Lorenzo, Víctor de; Nikel, Pablo I., E-mail: pablo.nikel@cnb.csic.es [Systems and Synthetic Biology Program, Centro Nacional de Biotecnología (CNB-CSIC), Madrid (Spain)

2014-10-28

Re-programming microorganisms to modify their existing functions and/or to bestow bacteria with entirely new-to-Nature tasks have largely relied so far on specialized molecular biology tools. Such endeavors are not only relevant in the burgeoning metabolic engineering arena but also instrumental to explore the functioning of complex regulatory networks from a fundamental point of view. À la carte modification of bacterial genomes thus calls for novel tools to make genetic manipulations easier. We propose the use of a series of new broad-host-range mini-Tn5-vectors, termed pBAMDs, for the delivery of gene(s) into the chromosome of Gram-negative bacteria and for generating saturated mutagenesis libraries in gene function studies. These delivery vectors endow the user with the possibility of easy cloning and subsequent insertion of functional cargoes with three different antibiotic-resistance markers (kanamycin, streptomycin, and gentamicin). After validating the pBAMD vectors in the environmental bacterium Pseudomonas putida KT2440, their use was also illustrated by inserting the entire poly(3-hydroxybutyrate) (PHB) synthesis pathway from Cupriavidus necator in the chromosome of a phosphotransacetylase mutant of Escherichia coli. PHB is a completely biodegradable polyester with a number of industrial applications that make it attractive as a potential replacement of oil-based plastics. The non-selective nature of chromosomal insertions of the biosynthetic genes was evidenced by a large landscape of PHB synthesis levels in independent clones. One clone was selected and further characterized as a microbial cell factory for PHB accumulation, and it achieved polymer accumulation levels comparable to those of a plasmid-bearing recombinant. Taken together, our results demonstrate that the new mini-Tn5-vectors can be used to confer interesting phenotypes in Gram-negative bacteria that would be very difficult to engineer through direct manipulation of the structural genes.

New Transposon Tools Tailored for Metabolic Engineering of Gram-Negative Microbial Cell Factories

International Nuclear Information System (INIS)

Martínez-García, Esteban; Aparicio, Tomás; Lorenzo, Víctor de; Nikel, Pablo I.

2014-01-01

Re-programming microorganisms to modify their existing functions and/or to bestow bacteria with entirely new-to-Nature tasks have largely relied so far on specialized molecular biology tools. Such endeavors are not only relevant in the burgeoning metabolic engineering arena but also instrumental to explore the functioning of complex regulatory networks from a fundamental point of view. À la carte modification of bacterial genomes thus calls for novel tools to make genetic manipulations easier. We propose the use of a series of new broad-host-range mini-Tn5-vectors, termed pBAMDs, for the delivery of gene(s) into the chromosome of Gram-negative bacteria and for generating saturated mutagenesis libraries in gene function studies. These delivery vectors endow the user with the possibility of easy cloning and subsequent insertion of functional cargoes with three different antibiotic-resistance markers (kanamycin, streptomycin, and gentamicin). After validating the pBAMD vectors in the environmental bacterium Pseudomonas putida KT2440, their use was also illustrated by inserting the entire poly(3-hydroxybutyrate) (PHB) synthesis pathway from Cupriavidus necator in the chromosome of a phosphotransacetylase mutant of Escherichia coli. PHB is a completely biodegradable polyester with a number of industrial applications that make it attractive as a potential replacement of oil-based plastics. The non-selective nature of chromosomal insertions of the biosynthetic genes was evidenced by a large landscape of PHB synthesis levels in independent clones. One clone was selected and further characterized as a microbial cell factory for PHB accumulation, and it achieved polymer accumulation levels comparable to those of a plasmid-bearing recombinant. Taken together, our results demonstrate that the new mini-Tn5-vectors can be used to confer interesting phenotypes in Gram-negative bacteria that would be very difficult to engineer through direct manipulation of the structural genes.

Characterization and validation of new tools for measuring site-specific cardiac troponin I phosphorylation.

Science.gov (United States)

Thoemmes, Stephen F; Stutzke, Crystal A; Du, Yanmei; Browning, Michael D; Buttrick, Peter M; Walker, Lori A

2014-01-31

Phosphorylation of cardiac troponin I is a well established mechanism by which cardiac contractility is modulated. However, there are a number of phosphorylation sites on TnI which contribute singly or in combination to influence cardiac function. Accordingly, methods for accurately measuring site-specific TnI phosphorylation are needed. Currently, two strategies are employed: mass spectrometry, which is costly, difficult and has a low throughput; and Western blotting using phospho-specific antibodies, which is limited by the availability of reagents. In this report, we describe a cohort of new site-specific TnI phosphoantibodies, generated against physiologically relevant phosphorylation sites, that are superior to the current commercially available antibodies: to phospho-serine 22/23 which shows a >5-fold phospho-specificity for phosphorylated TnI; to phospho-serine 43, which has >3-fold phospho-specificity for phosphorylated TnI; and phospho-serine 150 which has >2-fold phospho-specificity for phosphorylated TnI. These new antibodies demonstrated greater sensitivity and specificity for the phosphorylated TnI than the most widely used commercially available reagents. For example, at a protein load of 20 μg of total cardiac extract, a commercially available antibody recognized both phosphorylated and dephosphorylated TnI to the same degree. At the same protein load our phospho-serine 22/23 antibody exhibited no cross-reactivity with dephosphorylated TnI. These new tools should allow a more accurate assessment and a better understanding of the role of TnI phosphorylation in the response of the heart to pathologic stress. Copyright © 2013 Elsevier B.V. All rights reserved.

Enplanement and All Cargo Activity

Science.gov (United States)

1994-04-01

277,614 6.5% 255.910 146 STX ALEXANDR HAMILTON cHRISTIAxv=EDVI 276,060 -9.1% 303,657 147 ORS AUSTIN STRAUBEL INTE GREWI DAY ,In 270,871 5.0% 256.033 148...MOCI4I COUNTY ll 35 "TN TRENTON GIBSON COUNTY TGC 22 TN HUMBOLDT HUMBOLDT MUNI MS3 16 TN SET ER ROBERT SIBLEY SZY 14 TN DYERSBURG DYERSBURG MINI DYR

The impact of hepatitis B carrier on cardiac troponin I in 100-km ultramarathon runners.

Science.gov (United States)

Li, Li-Hua; How, Chorng-Kuang; Kao, Wei-Fong; Chiu, Yu-Hui; Meng, Chen; Hsu, Cheng-Chin; Tsay, Yeou-Guang

2017-06-01

Prolonged endurance exercise is known to cause elevation of cardiac troponin I (cTnI). Previous studies have reported the correlation of several factors with exercise-induced cTnI release. However, the investigation of the predictors for elevated cTnI and postrace kinetics of cTnI after ultramarathon running is lacking, especially in an Oriental population. Twenty-six participants, including eight hepatitis B virus carrier (HBVc) runners, who finished a 100-km ultramarathon in Taiwan were enrolled. For each participant, blood samples were collected 1 week before the race, as well as immediately and 24 hours after the finish. The results showed that 19 runners (73.1%) had postrace elevated cTnI levels and eight (30.8%) had elevated cTnI values lasting more than 24 hours after the run. A multiple linear regression analysis demonstrated that the HBV status was a factor related to the high level of cTnI after 24 hours of running (β=0.03, p=0.08). The recovery of plasma cTnI levels was delayed in ultramarathon runners with latent HBV infection. Among HBVc runners, multiple linear regression analyses showed age (β=-0.01), previous running experience (β=-0.06), training distance (β=0.37), and 4 hours of running distance (β=-0.04) as significant predictors of higher postrace cTnI levels. For most athletes, cTnI values significantly increased immediately following the race in the absence of adverse clinical sequelae, and HBVc runners had higher and prolonged cTnI levels. While several factors are identified for such HBV effects, the specific causes need further elucidation. Copyright © 2017. Published by Elsevier Taiwan LLC.

Association of elevated serum cardiac troponin-I level and complications in acute heart failurecases

Directory of Open Access Journals (Sweden)

Farjana Akhter

2013-07-01

Full Text Available Acute heart failure is one of the major causes of morbidity and mortality all over the world. Available published data has suggested that patients of acute heart failure with elevated level of serum cardiac troponin-I (cTn-I have more adverse outcomes than that of acute heart failure with normal cTn-I level. Elevated level of serum cTn-I is a potential risk factor for acute heart failure. This study was carried out in the department of Biochemistry, Dhaka Medical College and National Institute of Cardiovascular Disease (NICVD during the period from January 2010 to December 2010. In this study, 100 patients with acute heart failure were enrolled. Out of 100 cases, 50 had elevated serum cTn-I (cTn-I ³ 1.0 ng/ml and 50 had normal serum cTn-I (cTn-I < 1.0 ng/ml. The adverse outcome of the two groups were recorded and compared. Patients with high and normal serum cTn-I had mean age of 52.40 ± 8.10 years and 54.64 ± 7.26 years respectively while male and female cases were equally distributed. Left ventricular systolic dysfunction (lower ejection fraction was significantly (p<0.05 higher among cases with elevated cTn-I level compared to those with normal level. The rate of renal failure (raised serum creatinine, impaired liver functions (raised ALT and AST and abnormal serum electrolytes were significantly higher among the patients with elevated cTn-I compared to those with normal level. The study showed that elevated serum cTn-I level was a good biomarker to indicate adverse complications in acute heart failure cases. Ibrahim Med. Coll. J. 2013; 7(2: 32-34

Changes in the dynamics of the cardiac troponin C molecule explain the effects of Ca2+-sensitizing mutations.

Science.gov (United States)

Stevens, Charles M; Rayani, Kaveh; Singh, Gurpreet; Lotfalisalmasi, Bairam; Tieleman, D Peter; Tibbits, Glen F

2017-07-14

Cardiac troponin C (cTnC) is the regulatory protein that initiates cardiac contraction in response to Ca 2+ TnC binding Ca 2+ initiates a cascade of protein-protein interactions that begins with the opening of the N-terminal domain of cTnC, followed by cTnC binding the troponin I switch peptide (TnI SW ). We have evaluated, through isothermal titration calorimetry and molecular-dynamics simulation, the effect of several clinically relevant mutations (A8V, L29Q, A31S, L48Q, Q50R, and C84Y) on the Ca 2+ affinity, structural dynamics, and calculated interaction strengths between cTnC and each of Ca 2+ and TnI SW Surprisingly the Ca 2+ affinity measured by isothermal titration calorimetry was only significantly affected by half of these mutations including L48Q, which had a 10-fold higher affinity than WT, and the Q50R and C84Y mutants, each of which had affinities 3-fold higher than wild type. This suggests that Ca 2+ affinity of the N-terminal domain of cTnC in isolation is insufficient to explain the pathogenicity of these mutations. Molecular-dynamics simulation was used to evaluate the effects of these mutations on Ca 2+ binding, structural dynamics, and TnI interaction independently. Many of the mutations had a pronounced effect on the balance between the open and closed conformations of the TnC molecule, which provides an indirect mechanism for their pathogenic properties. Our data demonstrate that the structural dynamics of the cTnC molecule are key in determining myofilament Ca 2+ sensitivity. Our data further suggest that modulation of the structural dynamics is the underlying molecular mechanism for many disease mutations that are far from the regulatory Ca 2+ -binding site of cTnC. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Triple-negative (ER, PgR, HER-2/neu breast cancer in Indian women

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Vinayak W Patil

2011-03-01

Full Text Available Vinayak W Patil1, Rajeev Singhai1, Amit V Patil2, Prakash D Gurav21Department of Biochemistry, Grant Medical College and Sir JJ Group of Hospitals, Mumbai, India; 2Department of Surgery, Government Medical College, Miraj, IndiaAbstract: The aim of our study was to analyze triple-negative (TN breast cancer, which is defined as being negative for the estrogen receptor (ER, the progesterone receptor (PgR, and the human epidermal growth factor receptor 2 (HER-2/neu and which represents a subset of breast cancer with different biologic behavior. We investigated the clinicopathological characteristics and prognostic indicators of lymph node-negative TN breast cancer. Medical records were reviewed from patients with node-negative breast cancer who underwent curative surgery at Grant Medical College and Sir JJ Group of Hospitals, Mumbai, India, from May 2007 to October 2010. Clinicopathological variables and clinical outcomes were evaluated. Among 683 patients included, 136 had TN breast cancer and 529 had non-TN breast cancer. TN breast cancer correlated with younger age (<35 years, P = 0.003 and a higher histopathologic and nuclear grade (P < 0.001. It also correlated with a molecular profile associated with biological aggressiveness: negative for Bcl-2 expression (P < 0.001, positive for the epidermal growth factor receptor (P = 0.003, and a high level of p53 (P < 0.001 and Ki-67 expression (P < 0.00. The relapse rates during the follow-up period (median 56.8 months were 14.7% for TN breast cancer and 6.6% for non-TN breast cancer (P = 0.004. Relapse-free survival (RFS was significantly shorter among patients with TN breast cancer compared with those with non-TN breast cancer: 3.5-year RFS rate 85.5% versus 94.2%, respectively; P = 0.001. On multivariate analysis, young age, close resection margin, and triple negativity were independent predictors of shorter RFS. TN breast cancer had a higher relapse rate and more aggressive clinicopathological

Equivalent Markov-Renewal Processes.

Science.gov (United States)

1979-12-01

a Fa P(X0 - JON Joml(t I’Q m-lim (tm 0 J 0 A0 jA rm A- Let td be the semi algebra of sets of the form (X0 cA 0 ,-*.,Xnc An, T1 < t l,..*,Tn < t n... IHQ (tl)Q(t 2 )"Q(tn)U - Y(tI)Y(t 2). "’Y(tn). If 0 - wy then Bl - wjjl - w, so (tl)Q(t2)... Q(tn)U - WnQ(tl)Q(t 2) ... Q(tn)U - 1O¥(t )¥(t,) ..•.y

Cardiac troponin I levels in canine pyometra

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Hagman Ragnvi

2007-02-01

Full Text Available Abstract Background Myocardial injury may contribute to unexpected deaths due to pyometra. To detect myocardial damage, measurement of cardiac troponin I (cTnI is currently the most sensitive and specific method. The aims of the present study were to evaluate presence of myocardial damage in canine pyometra by analysis of cTnI, to explore whether myocardial injury was associated with systemic inflammatory response syndrome (SIRS and to evaluate whether other clinical or laboratory parameters were associated with cTnI increase. Methods Preoperative plasma levels of cTnI were investigated in 58 female dogs with pyometra and 9 controls. The value of physical examination findings, haematological, serum biochemical and pro-inflammatory (CRP and TNF-α parameters as possible predictors of increased cTnI levels was also evaluated. Results Seven dogs with pyometra (12% and one control dog (11% had increased levels of cTnI. In the pyometra group, the levels ranged between 0.3–0.9 μg l-1 and in the control dog the level was 0.3 μg l-1. The cTnI levels did not differ significantly between the two groups. No cardiac abnormalities were evident on preoperative physical examinations. Four of the pyometra patients died within two weeks of surgery, of which two were examined post mortem. In one of these cases (later diagnosed with myocarditis and disseminated bacterial infection the cTnI levels increased from 0.9 μg l-1 preoperatively to 180 μg l-1 the following day when also heart arrhythmia was also detected. The other patient had cTnI levels of 0.7 μg l-1 with no detectable heart pathology post mortem. CTnI increase was not associated with presence of SIRS. There was a trend for the association of cTnI increase with increased mortality. No preoperative physical examination findings and few but unspecific laboratory parameters were associated with increased cTnI levels. Conclusion Increased cTnI levels were observed in 12% of the dogs with pyometra. The

The nervus terminalis in amphibians: anatomy, chemistry and relationship with the hypothalamic gonadotropin-releasing hormone system.

Science.gov (United States)

Muske, L E; Moore, F L

1988-01-01

The nervus terminalis (TN), a component of the olfactory system, is found in most vertebrates. The TN of some fishes and mammals contains neurons immunoreactive (ir) to gonadotropin-releasing hormone (LHRH), and to several other neuropeptides and neurotransmitter systems, but there is little information on TN chemistry in other vertebrate taxa. Using immunocytochemical techniques, we found LHRH-ir neurons in amphibian TNs. In anurans, but not in a urodele, the TN was also found to contain Phe-Met-Arg-Phe-NH2 (FMRFamide) immunoreactivity. LHRH-ir neurons of the TN and those of the septal-hypothalamic system are morphologically homogeneous and form a distinct anatomical continuum in amphibians. Based upon topographical and cytological criteria, we hypothesize that LHRH-ir systems in vertebrates might derive embryonically from the TN.

Two-electron oxidation of cobalt phthalocyanines by thionyl chloride: Implications for lithium/thionyl chloride batteries. Technical report

Energy Technology Data Exchange (ETDEWEB)

Bernstein, P.A.; Lever, A.B.

1989-10-20

Cyclic voltammetry, DPV and electronic spectroscopy are used to study the reaction between thionyl chloride and cobalt phthalocyanine. SOCl2 reacts with (Co(I)Tn Pc(2-)) and Co(II)Tn Pc(2-) to give two-electron oxidized species. Implications for Li/SOCl2 batteries are discussed. Thionyl chloride also forms a mono SOCl2 adduct with Co(II)TnPc(2-). Driving forces (Delta E values) were calculated for CoTnPc comproportionation and CoTnPc + SOCl2 reactions. Rest potential measurements of a Li/SOCl2 cells show that addition of AlCl3 stabilizes the LiCl product as LiAlCl4. A catalytic two-electron mechanism is indicated for the reduction of thionyl chloride in a Li/SOCl2/(CoTnPc,C) battery.

Comparação entre troponina I cardíaca e CK-MB massa em síndrome coronariana aguda sem supra de ST Comparación entre troponina i cardíaca y ck-mb masa en síndrome coronario agudo sin supradesnivel de ST Comparison between cardiac troponin I and CK-MB mass in acute coronary syndrome without st elevation

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Elizabete Silva dos Santos

2011-03-01

Full Text Available FUNDAMENTO: Há incertezas do valor prognóstico comparativo entre troponina I cardíaca (cTnI e CK-MB em síndrome coronariana aguda (SCA. OBJETIVO: Comparar o valor prognóstico entre a cTnI e a CK-MB massa em pacientes com SCA sem supradesnível do segmento ST. MÉTODOS: Foram analisados 1.027 pacientes, de modo prospectivo, em um centro terciário de cardiologia. Combinações dos biomarcadores foram examinadas: cTnI normal, CK-MB massa normal (65,5%; cTnI normal, CK-MB massa elevada (3,9%; cTnI elevada, CK-MB massa normal (8,8%; cTnI elevada, CK-MB massa elevada (20,7%. Análise multivariada de variáveis clínicas, eletrocardiográficas e laboratoriais determinou o valor prognóstico independente dos biomarcadores para o evento de morte ou (reinfarto em 30 dias. RESULTADOS: Pacientes com pelo menos um biomarcador elevado foram mais idosos (p = 0,02 e do sexo masculino (p FUNDAMENTO: Hay dudas sobre el valor pronóstico comparativo entre troponina I cardíaca (cTnI y CK-MB en síndrome coronario agudo (SCA. OBJETIVO: Comparar el valor pronóstico entre la cTnI y la CK-MB masa en pacientes con SCA sin supradesnivel del segmento ST. MÉTODOS: Fueron analizados 1.027 pacientes, de modo prospectivo, en un centro terciario de cardiología. Combinaciones de los biomarcadores fueron examinadas: cTnI normal, CK-MB masa normal (65,5%; cTnI normal, CK-MB masa elevada (3,9%; cTnI elevada, CK-MB masa normal (8,8%; cTnI elevada, CK-MB masa elevada (20,7%. Análisis multivariado de variables clínicas, electrocardiográficas y de laboratorio determinó el valor pronóstico independiente de los biomarcadores para el evento de muerte o (reinfarto en 30 días. RESULTADOS: Pacientes con por lo menos un biomarcador elevado eron más añosos (p = 0,02 y del sexo masculino (p BACKGROUND: There is uncertainty as to the comparative prognostic value between cardiac troponin I (cTnI and CK-MB in acute coronary syndrome (ACS. OBJECTIVE: To compare the prognostic

Projekt VPN pro přístup klientů do do korporátní sítě obchodního řetězce v regionu střední Evropa

OpenAIRE

Ondrák, Richard

2008-01-01

Tato práce se zabývá problematikou vzdáleného přístupu klientů do korporátní sítě nadnárodního obchodního řetězce Tesco Stores v regionu střední Evropy. Na základě zhodnocení současných možností vzdáleného přístupu bude cílem práce navrhnout nové vhodné regionální řešení, které jednak nahradí všechny dosud používané systémy, a jednak splní všechny požadavky a zvyšující se nároky firemních uživatelů i dodavatelů. Řešení bude projektováno s ohledem zejména na bezpečnost, snadné použití, co nejm...

Enhancement of the potential to utilize octopine in the nonfluorescent Pseudomonas sp. strain 92

International Nuclear Information System (INIS)

Gill, S.S.; Boivin, R.; Dion, P.

1991-01-01

The nonfluorescent Pseudomonas sp. strain 92 requires the presence of a supplementary carbon source for growth on octopine, whereas the spontaneous mutant RB100 has acquired the capacity to utilize this opine as the sole carbon and nitrogen source. Insertional mutagenesis of RB100 with transposon Tn5 generated mutants which were unable to grow on octopine and others which grew slowly on this substrate. Both types of mutants yielded revertants that had regained the ability to utilize octopine. Some of the revertants had lost the transposon, whereas in others the transposon was retained but with rearrangements of the insertion site. Genes of octopine catabolism from strain 92 were cloned on a cosmid vector to generate pK3. The clone pK3 conferred the ability to utilize octopine as the sole carbon and nitrogen source on the host Pseudomonas putida KT2440. Although they conferred an equivalent growth phenotype, the mutant genes carried by RB100 and the cloned genes on pK3 differed in their regulation. Utilization of [ 14 C]octopine was inducible by octopine in RB100 and was constitutive in KT2440(pK3)

Remobilization of Sleeping Beauty transposons in the germline of Xenopus tropicalis

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Yergeau Donald A

2011-11-01

Full Text Available Abstract Background The Sleeping Beauty (SB transposon system has been used for germline transgenesis of the diploid frog, Xenopus tropicalis. Injecting one-cell embryos with plasmid DNA harboring an SB transposon substrate together with mRNA encoding the SB transposase enzyme resulted in non-canonical integration of small-order concatemers of the transposon. Here, we demonstrate that SB transposons stably integrated into the frog genome are effective substrates for remobilization. Results Transgenic frogs that express the SB10 transposase were bred with SB transposon-harboring animals to yield double-transgenic 'hopper' frogs. Remobilization events were observed in the progeny of the hopper frogs and were verified by Southern blot analysis and cloning of the novel integrations sites. Unlike the co-injection method used to generate founder lines, transgenic remobilization resulted in canonical transposition of the SB transposons. The remobilized SB transposons frequently integrated near the site of the donor locus; approximately 80% re-integrated with 3 Mb of the donor locus, a phenomenon known as 'local hopping'. Conclusions In this study, we demonstrate that SB transposons integrated into the X. tropicalis genome are effective substrates for excision and re-integration, and that the remobilized transposons are transmitted through the germline. This is an important step in the development of large-scale transposon-mediated gene- and enhancer-trap strategies in this highly tractable developmental model system.

Serum cardiac troponin I and cardiac troponin T concentrations in dogs with gastric dilatation-volvulus.

Science.gov (United States)

Schober, Karsten E; Cornand, Corinna; Kirbach, Babett; Aupperle, Heike; Oechtering, Gerhard

2002-08-01

To determine whether serum concentrations of cardiac troponin I (cTnI) and cardiac troponin T (cTnT) are increased in dogs with gastric dilatationvolvulus (GDV) and whether concentrations correlate with severity of ECG abnormalities or outcome. Prospective case series. 85 dogs with GDV. Serum cTnl and cTnT concentrations were measured 12 to 24, 48, 72, and 96 hours after surgery. Dogs were grouped on the basis of severity of ECG abnormalities and outcome. cTnl and cTnT were detected in serum from 74 (87%) and 43 (51%) dogs, respectively. Concentrations were significantly different among groups when dogs were grouped on the basis of severity of ECG abnormalities (none or mild vs moderate vs severe). Dogs that died (n = 16) had significantly higher serum cTnI (24.9 ng/ml) and cTnT (0.18 ng/ml) concentrations than did dogs that survived (2.05 and dogs with high serum cardiac troponin concentrations. Results indicate that concentrations of cTnI and cTnT suggestive of myocardial cell injury can commonly be found in serum from dogs with GDV and that serum cardiac troponin concentrations are associated with severity of ECG abnormalities and outcome.

Characterization of a volcanic ash episode in southern Finland caused by the Grimsvötn eruption in Iceland in May 2011

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V.-M. Kerminen

2011-12-01

Full Text Available The volcanic eruption of Grimsvötn in Iceland in May 2011 affected surface-layer air quality at several locations in Northern Europe. In Helsinki, Finland, the main pollution episode lasted for more than 8 h around the noon of 25 May. We characterized this episode by relying on detailed physical, chemical and optical aerosol measurements. The analysis was aided by air mass trajectory calculations, satellite measurements, and dispersion model simulations. During the episode, volcanic ash particles were present at sizes from less than 0.5 μm up to sizes >10 μm. The mass mean diameter of ash particles was a few μm in the Helsinki area, and the ash enhanced PM₁₀ mass concentrations up to several tens of μg m⁻³. Individual particle analysis showed that some ash particles appeared almost non-reacted during the atmospheric transportation, while most of them were mixed with sea salt or other type of particulate matter. Also sulfate of volcanic origin appeared to have been transported to our measurement site, but its contribution to the aerosol mass was minor due the separation of ash-particle and sulfur dioxide plumes shortly after the eruption. The volcanic material had very little effect on PM₁ mass concentrations or sub-micron particle number size distributions in the Helsinki area. The aerosol scattering coefficient was increased and visibility was slightly decreased during the episode, but in general changes in aerosol optical properties due to volcanic aerosols seem to be difficult to be distinguished from those induced by other pollutants present in a continental boundary layer. The case investigated here demonstrates clearly the power of combining surface aerosol measurements, dispersion model simulations and satellite measurements in analyzing surface air pollution episodes caused by volcanic eruptions. None of these three approaches alone would be sufficient to forecast, or even to unambiguously identify

Enzyme immunoassay and proteomic characterization of troponin I as a marker of mammalian muscle compounds in raw meat and some meat products.

Science.gov (United States)

Zvereva, Elena A; Kovalev, Leonid I; Ivanov, Alexei V; Kovaleva, Marina A; Zherdev, Anatoly V; Shishkin, Sergey S; Lisitsyn, Andrey B; Chernukha, Irina M; Dzantiev, Boris B

2015-07-01

The skeletal muscle protein troponin I (TnI) has been characterized as a potential thermally stable and species-specific biomarker of mammalian muscle tissues in raw meat and meat products. This study proposed a technique for the quantification of TnI comprising protein extraction and sandwich enzyme-linked immunosorbent assay (ELISA). The technique is characterized by a TnI detection limit of 4.8 ng/ml with quantifiable concentrations ranging from 8.7 to 52 ng/ml. The method was shown to be suitable for detection of TnI in mammalian (beef, pork, lamb, and horse) meat but not in poultry (chicken, turkey, and duck) meat. In particular, the TnI content in beef was 0.40 3 ± 0.058 mg/g of wet tissue. The TnI estimations obtained for the pork and beef samples using ELISA were comparable to the proteomic analysis results. Thus, the quantitative study of TnI can be a convenient way to assess the mammalian muscle tissue content of various meat products. Copyright © 2015. Published by Elsevier Ltd.

Dominance of multidrug resistant CC271 clones in macrolide-resistant streptococcus pneumoniae in Arizona

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Bowers Jolene R

2012-01-01

Full Text Available Abstract Background Rates of resistance to macrolide antibiotics in Streptococcus pneumoniae are rising around the world due to the spread of mobile genetic elements harboring mef(E and erm(B genes and post-vaccine clonal expansion of strains that carry them. Results Characterization of 592 clinical isolates collected in Arizona over a 10 year period shows 23.6% are macrolide resistant. The largest portion of the macrolide-resistant population, 52%, is dual mef(E/erm(B-positive. All dual-positive isolates are multidrug-resistant clonal lineages of Taiwan19F-14, mostly multilocus sequence type 320, carrying the recently described transposon Tn2010. The remainder of the macrolide resistant S. pneumoniae collection includes 31% mef(E-positive, and 9% erm(B-positive strains. Conclusions The dual-positive, multidrug-resistant S. pneumoniae clones have likely expanded by switching to non-vaccine serotypes after the heptavalent pneumococcal conjugate vaccine release, and their success limits therapy options. This upsurge could have a considerable clinical impact in Arizona.

Characterization of an immunodominant cancer-specific O-glycopeptide epitope in murine podoplanin (OTS8)

DEFF Research Database (Denmark)

Steentoft, Catharina; Schjoldager, Katrine T; Cló, Emiliano

2010-01-01

antibody 237, developed to a spontaneous murine fibrosarcoma, was shown to be directed to murine podoplanin (OTS8) with truncated Tn O-glycans. Our understanding of such cancer-specific auto-antibodies to truncated glycoforms of glycoproteins is limited. Here we have investigated immunogenicity...... of a chemoenzymatically produced Tn-glycopeptide derived from the putative murine podoplanin O-glycopeptide epitope. We found that the Tn O-glycopeptide was highly immunogenic in mice and produced a Tn-glycoform specific response with no reactivity against unglycosylated peptides or the O-glycopeptide with extended O......-glycan (STn and T glycoforms). The immunodominant epitope was strictly dependent on the peptide sequence, required Tn at a specific single Thr residue (Thr(77)), and antibodies to the epitope were not found in naive mice. We further tested a Tn O-glycopeptide library derived from human podoplanin...

Value of serum tenascin-C in patients with acute myocardial infarction

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Rania Gaber

2016-09-01

Conclusions: Serum TN-C might be a novel marker reflecting active structural remodeling in the myocardium following infarction, with high TN-C levels at acute stages possibly predicting progression of LV remodeling. Also, the incorporation of a combination of serum TN-C and plasma BNP levels may improve risk stratification for congestive heart failure after AMI. Further studies on large scale are needed for more evaluation of TN-C role in HF.

The nervus terminalis in the chick: a FMRFamide-immunoreactive and AChE-positive nerve.

Science.gov (United States)

Wirsig-Wiechmann, C R

1990-07-16

The chick terminal nerve (TN) was examined by immunocytochemical and histochemical methods. Molluscan cardioexcitatory peptide-immunoreactive (FMRFamide-ir) and acetylcholinesterase (AChE)-positive TN perikarya and fibers were distributed along olfactory and trigeminal nerves. FMRFamide-ir TN fibers terminated in the olfactory lamina propria and epithelium and in ganglia along the rostroventral nasal septum. This initial description of several populations of avian TN neurons should provide the foundation for future developmental studies of this system.

Index of NACA Technical Publications, 1915 - 1949

Science.gov (United States)

1950-03-31

Feb. 1947. Weller, R.; Middlehurst, D. J. and Steiner , R.: The Photoviscous Properties of Allen, H. Julian; Heaslet, Max A. and Fluids. TN 841, Feb...TM 1216, March Taylor, C. Fayette: A Suggested Method 1949. for Measuring Turbulence. TN 380, Iglisch, Rudolf : Exact Calculation of June 1931. Laminar...TN 472, Sept. 1933. TN�, Aug. 1928. Wallace, Rudolf N.: The Effect of Split Knight,-Montgomery and Wenzinger, Carl Trailing-Edge Wing Flaps on the

The mecillinam resistome reveals a role for peptidoglycan endopeptidases in stimulating cell wall synthesis in Escherichia coli.

Science.gov (United States)

Lai, Ghee Chuan; Cho, Hongbaek; Bernhardt, Thomas G

2017-07-01

Bacterial cells are typically surrounded by an net-like macromolecule called the cell wall constructed from the heteropolymer peptidoglycan (PG). Biogenesis of this matrix is the target of penicillin and related beta-lactams. These drugs inhibit the transpeptidase activity of PG synthases called penicillin-binding proteins (PBPs), preventing the crosslinking of nascent wall material into the existing network. The beta-lactam mecillinam specifically targets the PBP2 enzyme in the cell elongation machinery of Escherichia coli. Low-throughput selections for mecillinam resistance have historically been useful in defining mechanisms involved in cell wall biogenesis and the killing activity of beta-lactam antibiotics. Here, we used transposon-sequencing (Tn-Seq) as a high-throughput method to identify nearly all mecillinam resistance loci in the E. coli genome, providing a comprehensive resource for uncovering new mechanisms underlying PG assembly and drug resistance. Induction of the stringent response or the Rcs envelope stress response has been previously implicated in mecillinam resistance. We therefore also performed the Tn-Seq analysis in mutants defective for these responses in addition to wild-type cells. Thus, the utility of the dataset was greatly enhanced by determining the stress response dependence of each resistance locus in the resistome. Reasoning that stress response-independent resistance loci are those most likely to identify direct modulators of cell wall biogenesis, we focused our downstream analysis on this subset of the resistome. Characterization of one of these alleles led to the surprising discovery that the overproduction of endopeptidase enzymes that cleave crosslinks in the cell wall promotes mecillinam resistance by stimulating PG synthesis by a subset of PBPs. Our analysis of this activation mechanism suggests that, contrary to the prevailing view in the field, PG synthases and PG cleaving enzymes need not function in multi-enzyme complexes

The mecillinam resistome reveals a role for peptidoglycan endopeptidases in stimulating cell wall synthesis in Escherichia coli.

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Ghee Chuan Lai

2017-07-01

Full Text Available Bacterial cells are typically surrounded by an net-like macromolecule called the cell wall constructed from the heteropolymer peptidoglycan (PG. Biogenesis of this matrix is the target of penicillin and related beta-lactams. These drugs inhibit the transpeptidase activity of PG synthases called penicillin-binding proteins (PBPs, preventing the crosslinking of nascent wall material into the existing network. The beta-lactam mecillinam specifically targets the PBP2 enzyme in the cell elongation machinery of Escherichia coli. Low-throughput selections for mecillinam resistance have historically been useful in defining mechanisms involved in cell wall biogenesis and the killing activity of beta-lactam antibiotics. Here, we used transposon-sequencing (Tn-Seq as a high-throughput method to identify nearly all mecillinam resistance loci in the E. coli genome, providing a comprehensive resource for uncovering new mechanisms underlying PG assembly and drug resistance. Induction of the stringent response or the Rcs envelope stress response has been previously implicated in mecillinam resistance. We therefore also performed the Tn-Seq analysis in mutants defective for these responses in addition to wild-type cells. Thus, the utility of the dataset was greatly enhanced by determining the stress response dependence of each resistance locus in the resistome. Reasoning that stress response-independent resistance loci are those most likely to identify direct modulators of cell wall biogenesis, we focused our downstream analysis on this subset of the resistome. Characterization of one of these alleles led to the surprising discovery that the overproduction of endopeptidase enzymes that cleave crosslinks in the cell wall promotes mecillinam resistance by stimulating PG synthesis by a subset of PBPs. Our analysis of this activation mechanism suggests that, contrary to the prevailing view in the field, PG synthases and PG cleaving enzymes need not function in multi

Cardiospecific troponins in non-ischemic cardiological pathologies

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Giuseppe Lippi

2006-10-01

Full Text Available Cardiospecific troponin T (TnT and I (TnI are low-molecularweight proteins that form part of the troponin complex and are integral components of the myofibrillar contractile apparatus of the heart. Loss of integrity of cardiac myocyte membranes causes release of cardiac troponins into the circulation, which can be detected by highly sensitive assays for cTnT and cTnI developed to the stat diagnosis of acute coronary syndrome. Regardless of preanalytical and analytical sources (incorrect collection or handling of the specimen, malfunctioning of the analyzer, heterophilic antibodies, a diagnostic troponin value is expression of myocardial damage, though it does not provide definitive clue on the nature of such an increment. In fact, increases in serum cardiac troponins also occur in the absence of cardiac ischemia and may sometimes led to an inappropriate or unjustified clinical decision making. Abnormal troponin values in plasma are frequently observed in various clinical contexts independent from the acute coronary disease, like myocarditis, pulmonary embolism, acute heart failure, septic shock, and as a result of cardiotoxic drugs as well as after therapeutic procedures like coronary angioplasty, electrophysiological ablations, or electrical cardioversion. Awareness of this issue is essential to either prevent unjustified alarmism or underestimation of clinical situations that may finally compromise the patient’s health.

Clinicopathological Characteristics of Triple-negative Breast Cancers in the Northeast Region of Turkey

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Bülent Yıldız

2014-06-01

Full Text Available Background: Triple-negative (TN breast cancer is a subtype of breast cancer characterised by a loss of estrogen receptor (ER, progesterone receptor (PR expression, and the absence of human epidermal growth factor (HER2 overexpression. Aims: To identify the relationships between clinicopathological characteristics of TN breast cancers in the northeast region of Turkey and disease free survival (DFS and overall survival (OS. Study Design: Retrospective clinical study. Methods: Seven hundred and eighty non-metastatic breast cancer patients were enrolled in this study. The relationships between TN breast cancer and other breast cancers with respect to clinicopathological characteristics, as well as DFS and OS, were studied. Results: The triple-negative phenotype was detected in 204 patients (27.1%. Patients with triple-negative breast cancer had more grade 2-3 tumours compared to those with other types of breast cancer (92.5% versus 84.3%, p=0.004. Invasive ductal carcinoma histology, on the other hand, was less prevalent in patients with TN breast cancer (77% versus 84.5%, p=0.016. No significant differences were identified between the groups in other clinicopathological variables. Relapse and mortality rates were higher in the TN group during the follow-up of both groups [57 (27.9% versus 89 (16.2%, p<0.001 for relapse; 27 (13.2% versus 37 (6.8%, p=0.005 for mortality]. The univariate analysis demonstrated shorter DFS and OS for patients with TN breast cancer compared to those with other types of breast cancer. In the multivariate analysis, patients with TN breast cancer were 2.21 times more likely to develop relapse, while the likelihood of death increased 3.21-fold (p<0.001 and p<0.001. Conclusion: Triple-negative breast cancers demonstrate a more aggressive clinical course compared to other breast cancers. More effective strategies should be developed for the treatment of this subgroup of breast cancer.

Combining Shigella Tn-seq data with gold-standard E. coli gene deletion data suggests rare transitions between essential and non-essential gene functionality.

Science.gov (United States)

Freed, Nikki E; Bumann, Dirk; Silander, Olin K

2016-09-06

Gene essentiality - whether or not a gene is necessary for cell growth - is a fundamental component of gene function. It is not well established how quickly gene essentiality can change, as few studies have compared empirical measures of essentiality between closely related organisms. Here we present the results of a Tn-seq experiment designed to detect essential protein coding genes in the bacterial pathogen Shigella flexneri 2a 2457T on a genome-wide scale. Superficial analysis of this data suggested that 481 protein-coding genes in this Shigella strain are critical for robust cellular growth on rich media. Comparison of this set of genes with a gold-standard data set of essential genes in the closely related Escherichia coli K12 BW25113 revealed that an excessive number of genes appeared essential in Shigella but non-essential in E. coli. Importantly, and in converse to this comparison, we found no genes that were essential in E. coli and non-essential in Shigella, implying that many genes were artefactually inferred as essential in Shigella. Controlling for such artefacts resulted in a much smaller set of discrepant genes. Among these, we identified three sets of functionally related genes, two of which have previously been implicated as critical for Shigella growth, but which are dispensable for E. coli growth. The data presented here highlight the small number of protein coding genes for which we have strong evidence that their essentiality status differs between the closely related bacterial taxa E. coli and Shigella. A set of genes involved in acetate utilization provides a canonical example. These results leave open the possibility of developing strain-specific antibiotic treatments targeting such differentially essential genes, but suggest that such opportunities may be rare in closely related bacteria.

Identification and evolutionary dynamics of cacta DNA transposons in brassica

International Nuclear Information System (INIS)

Nouroz, F.; Noreen, S.; Harrison, J.S.H.

2017-01-01

Transposable elements are the major drivers of genome evolution and plasticity. Due to their transposition mode, they are classified into two major classes as Retrotransposons and DNA transposons. The En/Spm or CACTA elements are diverse group of DNA transposons proliferating in plant genomes. Various bioinformatics and molecular approaches were used for identification and distribution of CACTA transposons in Brassica genome. A combination of dot plot analysis and BLASTN searches yielded 35 autonomous and 7 non-autonomous CACTA elements in Brassica. The elements ranged in sizes from 1.2 kb non-autonomous elements to 11kb autonomous elements, terminated by 3 bp Target Site Duplication (TSD) and ~15 bp conserved Terminal Inverted Repeat (TIR) motifs (5'-CACTACAAGAAAACA-3'), with heterogeneous internal regions. The transposase (TNP) was identified from autonomous CACTA elements, while other protein domains from Brassica and other plants CACTA revealed similar organizations with minor differences. Both transposases (TNPD, TNPA) are present in most CACTA, while a few CACTA harboured an additional ATHILA ORF1-like domain. The PCR analysis amplified the CACTA transposases from 40 Brassica accessions (A, B, and C-genome) suggesting their distribution among various Brassica crops. A detailed characterization and evolutionary analysis of the identified CACTA elements allowed some to be placed in genome-specific groups, while most of them (Brassica-Arabidopsis elements) have followed the same evolutionary line. The distribution of CACTA in Brassica concluded that 3 bp TSDs generating CACTA transposons contributed significantly to genome size and evolution of Brassica genome. (author)

Impact of high-intensity interval training and moderate-intensity continuous training on resting and postexercise cardiac troponin T concentration.

Science.gov (United States)

Nie, Jinlei; Zhang, Haifeng; Kong, Zhaowei; George, Keith; Little, Jonathan P; Tong, Tomas K; Li, Feifei; Shi, Qingde

2018-03-01

What is the central question of this study? Does exercise training impact resting and postexercise cardiac troponin T (cTnT) concentration? What is the main finding and its importance? This randomized controlled intervention study demonstrated that 12 weeks of either high-intensity interval training or moderate-intensity continuous training largely abolished the exercise-induced elevation in cTnT when exercise was performed at the same absolute intensity. There was no impact of training on resting cTnT or postexercise appearance of cTnT when exercise was performed at the same relative intensity. These findings provide new information that might help clinicians with decision-making in relationship to basal and postexercise values of cTnT in individuals with different training status. We evaluated the influence of 12 weeks of high-intensity interval training [HIIT; repeated 4 min cycling at 90% of maximal oxygen uptake (V̇O2max) interspersed with 3 min rest, 200-300 kJ per session, 3 or 4 days each week] and work-equivalent moderate-intensity continuous training (MICT; continuous cycling at 60% V̇O2max) on resting cardiac troponin T (cTnT) and the appearance of exercise-induced cTnT. Forty-eight sedentary obese young women were randomly assigned to HIIT, MICT or a control group. The V̇O2max and body composition were measured before and after training. At baseline, cTnT was assessed using a high-sensitivity assay at rest and immediately, 2 and 4 h after 45 min cycling at 60% V̇O2max. After a 12 week training period, cTnT was assessed before and after 45 min cycling at the same relative and absolute intensities as before training. Training led to higher V̇O2max and lower fat mass in both HIIT and MICT groups (all P training, cTnT was significantly elevated in all three groups (by 35-118%, all P training, both resting and postexercise cTnT concentrations (same relative intensity) were similar to pretraining values. In contrast, postexercise cTnT (same

Chemical composition and antimicrobial activity of the essential oils of Pinus peuce (Pinaceae) growing wild in R. Macedonia.

Science.gov (United States)

Karapandzova, Marija; Stefkova, Gjose; Cvetkovikj, Ivana; Trajkovska-Dokik, Elena; Kaftandzieva, Ana; Kulevanova, Svetlana

2014-11-01

The chemical composition and antimicrobial activity of the essential oils isolated from twigs with needles (T+N) and from twigs without needles (T-N) from wild Pinus peuce Griseb. (Pinaceae), from three different locations in R. Macedonia, were investigated. Essential oil yields of T+N ranged from 7.5 mL/kg to 12.5 mL/kg and for T-N from 13.8 mL/kg to 17.3 mL/kg. GC/FID/MS analysis of the essential oils revealed eighty-four components, representing 93.7-95.7% and 91.2-92.0% of the T+N and T-N oils, respectively. The major components in T+N and T-N oils were monoterpenes: α-pinene (23.8-39.9%, 21.2-23.3%), camphene (2.2-5.5%, 0.7-2.0%), β-pinene (10.1-17.1%, 8.2-16.4%), myrcene (1.2-1.41%, 1.6-2.5%), limonene+β-phellandrene (6.8-14.0%, 8.8-23.6%) and bornyl acetate (2.3-6.9%, 1.1-3.4%), followed by the sesquiterpenes: trans-(E)-caryophyllene (3.6-4.3%, 3.2-7.3%), germacrene D (7.1-9.5%, 5.0-10.3%) and δ-cadinene (2.1-3.1%, 3.3-4.2%, respectively). Antimicrobial screening of the essential oils was made by disk diffusion and broth dilution methods against 13 bacterial isolates of Gram-positive and Gram-negative bacteria and one strain of Candida albicans. T-N essential oils showed antimicrobial activity toward Streptococcus pneumoniae, Staphylococcus aureus, S. epidermidis and Candida albicans as well as Streptococcus agalactiae, Acinetobacter spp. and Haemophilus influenzae. The antimicrobial activity of T+N essential oils was greater, especially against Streptococcus agalactiae, S. pyogenes, Enterococcus and Candida albicans, followed by Haemophilus influenzae, Acinetobacter spp., Escherichia coli, Salmonella enteritidis, Staphylococcus aureus and S. epidermidis. Minimal inhibitory concentrations (MICs) of all tested essential oils ranged from 15-125 μL/mL. Summarizing the obtained results, the antimicrobial activity of Pinus peuce T+N and T-N essential oils collected from different localities in R. Macedonia varied considerably. These alterations in the

D-arabinose metabolism in Escherichia coli B: induction and cotransductional mapping of the L-fucose-D-arabinose pathway enzymes.

Science.gov (United States)

Elsinghorst, E A; Mortlock, R P

1988-12-01

D-Arabinose is degraded by Escherichia coli B via some of the L-fucose pathway enzymes and a D-ribulokinase which is distinct from the L-fuculokinase of the L-fucose pathway. We found that L-fucose and D-arabinose acted as the apparent inducers of the enzymes needed for their degradation. These enzymes, including D-ribulokinase, appeared to be coordinately regulated, and mutants which constitutively synthesized the L-fucose enzymes also constitutively synthesized D-ribulokinase. In contrast to D-arabinose-positive mutants of E. coli K-12, in which L-fuculose-1-phosphate and D-ribulose-1-phosphate act as inducers of the L-fucose pathway, we found that these intermediates did not act as inducers in E. coli B. To further characterize the E. coli B system, some of the L-fucose-D-arabinose genes were mapped by using bacteriophage P1 transduction. A transposon Tn10 insertion near the E. coli B L-fucose regulon was used in two- and three-factor reciprocal crosses. The gene encoding D-ribulokinase, designated darK, was found to map within the L-fucose regulon, and the partial gene order was found to be Tn10-fucA-darK-fucI-fucK-thyA.

Diagnosis of type 1 and type 2 myocardial infarction using a high-sensitivity cardiac troponin I assay with sex-specific 99th percentiles based on the third universal definition of myocardial infarction classification system.

Science.gov (United States)

Sandoval, Yader; Smith, Stephen W; Schulz, Karen M; Murakami, MaryAnn M; Love, Sara A; Nicholson, Jennifer; Apple, Fred S

2015-04-01

The frequency and characteristics of myocardial infarction (MI) subtypes per the Third Universal Definition of MI (TUDMI) classification system using high-sensitivity (hs) cardiac troponin assays with sex-specific cutoffs is not well known. We sought to describe the diagnostic characteristics of type 1 (T1MI) and type 2 (T2MI) MI using an hs-cardiac troponin I (hs-cTnI) assay with sex-specific cutoffs. A total of 310 consecutive patients with serial cTnI measurements obtained on clinical indication were studied with contemporary and hs-cTnI assays. Ninety-ninth percentile sex-specific upper reference limits (URLs) for the hs-cTnI assay were 16 ng/L for females and 34 ng/L for males. The TUDMI consensus recommendations were used to define and adjudicate MI based on each URL. A total of 127 (41%) patients had at least 1 hs-cTnI exceeding the sex-specific 99th percentiles, whereas 183 (59%) had hs-cTnI within the reference interval. Females had more myocardial injury related to supply/demand ischemia than males (39% vs 18%, P = 0.01), whereas males had more multifactorial or indeterminate injury (52% vs 33%, P = 0.05). By hs-cTnI, there were 32 (10%) acute MIs, among which 10 (3%) were T1MI and 22 (7%) were T2MI. T2MI represented 69% (22 out of 32) of all acute MIs, whereas T1MI represented 31% (10 out of 32). Ninety-five patients (31%) had an increased hs-cTnI above the 99th percentile but did not meet criteria for acute MI. The most common triggers for T2MI were tachyarrhythmias, hypotension/shock, and hypertension. By contemporary cTnI, more MIs (14 T1MI and 29 T2MI) were diagnosed. By contemporary cTnI, there were 43 MIs, 14 T1MI, and 29 T2MI. Fewer MI diagnoses were found with the hs-cTnI assay, contrary to the commonly accepted idea that hs-cTnI will lead to excessive false-positive diagnoses. © 2015 American Association for Clinical Chemistry.

External validation of heart-type fatty acid binding protein, high-sensitivity cardiac troponin, and electrocardiography as rule-out for acute myocardial infarction.

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Van Hise, Christopher B; Greenslade, Jaimi H; Parsonage, William; Than, Martin; Young, Joanna; Cullen, Louise

2018-02-01

To externally validate a clinical decision rule incorporating heart fatty acid binding protein (h-FABP), high-sensitivity troponin (hs-cTn) and electrocardiogram (ECG) for the detection of acute myocardial infarction (AMI) on presentation to the Emergency Department. We also investigated whether this clinical decision rule improved identification of AMI over algorithms incorporating hs-cTn and ECG only. This study included data from 789 patients from the Brisbane ADAPT cohort and 441 patients from the Christchurch TIMI RCT cohort. The primary outcome was index AMI. Sensitivity, specificity, positive predictive value and negative predictive value were used to assess the diagnostic accuracy of the algorithms. 1230 patients were recruited, including 112 (9.1%) with AMI. The algorithm including h-FABP and hs-cTnT had 100% sensitivity and 32.4% specificity. The algorithm utilising h-FABP and hs-cTnI had similar sensitivity (99.1%) and higher specificity (43.4%). The hs-cTnI and hs-cTnT algorithms without h-FABP both had a sensitivity of 98.2%; a result that was not significantly different from either algorithm incorporating h-FABP. Specificity was higher for the hs-cTnI algorithm (68.1%) compared to the hs-cTnT algorithm (33.0%). The specificity of the algorithm incorporating hs-cTnI alone was also significantly higher than both of the algorithms incorporating h-FABP (p<0.01). For patients presenting to the Emergency Department with chest pain, an algorithm incorporating h-FABP, hs-cTn and ECG has high accuracy and can rule out up to 40% of patients. An algorithm incorporating only hs-cTn and ECG has similar sensitivity and may rule out a higher proportion of patients. Each of the algorithms can be used to safely identify patients as low risk for AMI on presentation to the Emergency Department. Copyright © 2017 The Canadian Society of Clinical Chemists. All rights reserved.

Measurement of cardiac troponin I utilizing a point of care analyzer in healthy alpacas.

Science.gov (United States)

Blass, Keith A; Kraus, Marc S; Rishniw, Mark; Mann, Sabine; Mitchell, Lisa M; Divers, Thomas J

2011-12-01

Myocardial disease in camelids is poorly characterized. Nutritional (selenium deficiency) and toxic (ionophore toxicity) myocardial disease have been reported in camelids. Diagnosis and management of these and other myocardial diseases might be enhanced by evaluating cardiac troponin I (cTnI) concentrations. No information about cTnI reference intervals in camelids is currently available. (A) To determine cTnI concentrations obtained using a point of care i-STAT(®)1 analyzer (Heska Corporation) in healthy alpacas; (B) to compare alpaca cTnI concentrations between heparinized whole blood and plasma samples and between 2 different storage conditions (4 °C for 24 h or -80 °C for 30 days); (C) to examine assay reproducibility using the i-STAT(®)1. 23 healthy alpacas were evaluated. Blood and plasma samples were analyzed by the i-STAT(®)1 within 1 h of collection. Aliquots of plasma were stored at either 4 °C for 24 h or -80 °C for 30 days, and then analyzed. Assay reproducibility was determined by comparing 2 plasma or whole blood cTnI concentrations measured on the same sample over a 10 min period. Analyzer-specific plasma cTnI concentrations in clinically normal alpacas had a median of blood concentrations showed good agreement. Storage did not affect cTnI concentrations (p > 0.75). Plasma cTnI concentrations had coefficient of repeatability of 0.02 ng/mL. The i-STAT(®)1 can measure cTnI in alpacas on both plasma and whole blood and provides similar values for both samples. Storage at 4 °C for 24 h or -80 °C for 30 days does not affect estimates of plasma cTnI. Evaluation of cTnI might be of value in assessing cardiac disease in this species. Copyright © 2011 Elsevier B.V. All rights reserved.

Preprocedural High-Sensitivity Cardiac Troponin T and Clinical Outcomes in Patients With Stable Coronary Artery Disease Undergoing Elective Percutaneous Coronary Intervention.

Science.gov (United States)

Zanchin, Thomas; Räber, Lorenz; Koskinas, Konstantinos C; Piccolo, Raffaele; Jüni, Peter; Pilgrim, Thomas; Stortecky, Stefan; Khattab, Ahmed A; Wenaweser, Peter; Bloechlinger, Stefan; Moschovitis, Aris; Frenk, Andre; Moro, Christina; Meier, Bernhard; Fiedler, Georg M; Heg, Dik; Windecker, Stephan

2016-06-01

Cardiac troponin detected by new-generation, highly sensitive assays predicts clinical outcomes among patients with stable coronary artery disease (SCAD) treated medically. The prognostic value of baseline high-sensitivity cardiac troponin T (hs-cTnT) elevation in SCAD patients undergoing elective percutaneous coronary interventions is not well established. This study assessed the association of preprocedural levels of hs-cTnT with 1-year clinical outcomes among SCAD patients undergoing percutaneous coronary intervention. Between 2010 and 2014, 6974 consecutive patients were prospectively enrolled in the Bern Percutaneous Coronary Interventions Registry. Among patients with SCAD (n=2029), 527 (26%) had elevated preprocedural hs-cTnT above the upper reference limit of 14 ng/L. The primary end point, mortality within 1 year, occurred in 20 patients (1.4%) with normal hs-cTnT versus 39 patients (7.7%) with elevated baseline hs-cTnT (P<0.001). Patients with elevated hs-cTnT had increased risks of all-cause (hazard ratio 5.73; 95% confidence intervals 3.34-9.83; P<0.001) and cardiac mortality (hazard ratio 4.68; 95% confidence interval 2.12-10.31; P<0.001). Preprocedural hs-TnT elevation remained an independent predictor of 1-year mortality after adjustment for relevant risk factors, including age, sex, and renal failure (adjusted hazard ratio 2.08; 95% confidence interval 1.10-3.92; P=0.024). A graded mortality risk was observed across higher tertiles of elevated preprocedural hs-cTnT, but not among patients with hs-cTnT below the upper reference limit. Preprocedural elevation of hs-cTnT is observed in one fourth of SCAD patients undergoing elective percutaneous coronary intervention. Increased levels of preprocedural hs-cTnT are proportionally related to the risk of death and emerged as independent predictors of all-cause mortality within 1 year. URL: http://www.clinicaltrials.gov. Unique identifier: NCT02241291. © 2016 American Heart Association, Inc.

Structure of the C-type lectin carbohydrate recognition domain of human Tetranectin

DEFF Research Database (Denmark)

Kastrup, Jette Sandholm Jensen; Nielsen, Bettina Bryde; Rasmussen, Hanne B.

1998-01-01

Tetranectin (TN) is a C-type lectin involved in fibrinolysis, being the only endogenous ligand known to bind specifically to the kringle 4 domain of plasminogen. TN was originally isolated from plasma, but shows a wide tissue distribution. Furthermore, TN has been found in the extracellular matri...... molecules. One sulfate ion has been located at the surface of TN3, forming contacts to Glu120, Lys148, Asn106 of a symmetry-related molecule, and to an ethanol molecule....

Interdependence of the kinetics of NTP hydrolysis and the stability of the RecA-ssDNA complex.

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Katz, F S; Bryant, F R

2001-09-18

The ssDNA-dependent NTP hydrolysis activity of the RecA protein was examined using a series of dTn oligomers ranging in size from dT10 to dT2000 as the ssDNA effector. There were three distinct manifestations of the dTn-dependent NTP hydrolysis reaction, depending on the length of the dTn effector that was used. With longer dTn oligomers, NTP hydrolysis occurred with a turnover number of 20-25 min(-1) and the observed S0.5 value for the NTP was independent of the concentration of the dTn oligomer (DNA concentration-independent hydrolysis). With dTn oligomers of intermediate length, NTP hydrolysis still occurred with a turnover number of 20-25 min(-1), but the observed S0.5 for the NTP decreased with increasing dTn concentration until reaching a value similar to that obtained with the longer dTn oligomers (DNA concentration-dependent hydrolysis). With shorter dTn oligomers, the NTP hydrolysis activity was effectively eliminated. Although this general progression of kinetic behavior was observed for the three structurally related NTPs (dATP, ATP, and GTP), the dTn oligomer length at which DNA concentration-independent, DNA concentration-dependent, and no NTP hydrolysis was observed depended on the NTP being considered. For example, dATP (S0.5 = 35 microM) was hydrolyzed in the presence of dT20, whereas ATP (S0.5 = 70 microM) and GTP (S0.5 = 1200 microM) required at least dT50 and dT200 for hydrolysis, respectively. These results are discussed in terms of a kinetic model in which the stability of the RecA-ssDNA-NTP complex is dependent on the intrinsic S0.5 value of the NTP being hydrolyzed.

European multicenter analytical evaluation of the Abbott ARCHITECT STAT high sensitive troponin I immunoassay.

Science.gov (United States)

Krintus, Magdalena; Kozinski, Marek; Boudry, Pascal; Capell, Nuria Estañ; Köller, Ursula; Lackner, Karl; Lefèvre, Guillaume; Lennartz, Lieselotte; Lotz, Johannes; Herranz, Antonio Mora; Nybo, Mads; Plebani, Mario; Sandberg, Maria B; Schratzberger, Wolfgang; Shih, Jessie; Skadberg, Øyvind; Chargui, Ahmed Taoufik; Zaninotto, Martina; Sypniewska, Grazyna

2014-11-01

International recommendations highlight the superior value of cardiac troponins (cTns) for early diagnosis of myocardial infarction along with analytical requirements of improved precision and detectability. In this multicenter study, we investigated the analytical performance of a new high sensitive cardiac troponin I (hs-cTnI) assay and its 99th percentile upper reference limit (URL). Laboratories from nine European countries evaluated the ARCHITECT STAT high sensitive troponin I (hs-TnI) immunoassay on the ARCHITECT i2000SR/i1000SR immunoanalyzers. Imprecision, limit of blank (LoB), limit of detection (LoD), limit of quantitation (LoQ) linearity of dilution, interferences, sample type, method comparisons, and 99th percentile URLs were evaluated in this study. Total imprecision of 3.3%-8.9%, 2.0%-3.5% and 1.5%-5.2% was determined for the low, medium and high controls, respectively. The lowest cTnI concentration corresponding to a total CV of 10% was 5.6 ng/L. Common interferences, sample dilution and carryover did not affect the hs-cTnI results. Slight, but statistically significant, differences with sample type were found. Concordance between the investigated hs-cTnI assay and contemporary cTnI assay at 99th percentile cut-off was found to be 95%. TnI was detectable in 75% and 57% of the apparently healthy population using the lower (1.1 ng/L) and upper (1.9 ng/L) limit of the LoD range provided by the ARCHITECT STAT hs-TnI package insert, respectively. The 99th percentile values were gender dependent. The new ARCHITECT STAT hs-TnI assay with improved analytical features meets the criteria of high sensitive Tn test and will be a valuable diagnostic tool.

Total Nitrogen Sources of the Three Gorges Reservoir--A Spatio-Temporal Approach.

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Chunping Ren

Full Text Available Understanding the spatial and temporal variation of nutrient concentrations, loads, and their distribution from upstream tributaries is important for the management of large lakes and reservoirs. The Three Gorges Dam was built on the Yangtze River in China, the world's third longest river, and impounded the famous Three Gorges Reservoir (TGR. In this study, we analyzed total nitrogen (TN concentrations and inflow data from 2003 till 2010 for the main upstream tributaries of the TGR that contribute about 82% of the TGR's total inflow. We used time series analysis for seasonal decomposition of TN concentrations and used non-parametric statistical tests (Kruskal-Walli H, Mann-Whitney U as well as base flow segmentation to analyze significant spatial and temporal patterns of TN pollution input into the TGR. Our results show that TN concentrations had significant spatial heterogeneity across the study area (Tuo River> Yangtze River> Wu River> Min River> Jialing River>Jinsha River. Furthermore, we derived apparent seasonal changes in three out of five upstream tributaries of the TGR rivers (Kruskal-Walli H ρ = 0.009, 0.030 and 0.029 for Tuo River, Jinsha River and Min River in sequence. TN pollution from non-point sources in the upstream tributaries accounted for 68.9% of the total TN input into the TGR. Non-point source pollution of TN revealed increasing trends for 4 out of five upstream tributaries of the TGR. Land use/cover and soil type were identified as the dominant driving factors for the spatial distribution of TN. Intensifying agriculture and increasing urbanization in the upstream catchments of the TGR were the main driving factors for non-point source pollution of TN increase from 2003 till 2010. Land use and land cover management as well as chemical fertilizer use restriction were needed to overcome the threats of increasing TN pollution.

Gene-specific cell labeling using MiMIC transposons.

Science.gov (United States)

Gnerer, Joshua P; Venken, Koen J T; Dierick, Herman A

2015-04-30

Binary expression systems such as GAL4/UAS, LexA/LexAop and QF/QUAS have greatly enhanced the power of Drosophila as a model organism by allowing spatio-temporal manipulation of gene function as well as cell and neural circuit function. Tissue-specific expression of these heterologous transcription factors relies on random transposon integration near enhancers or promoters that drive the binary transcription factor embedded in the transposon. Alternatively, gene-specific promoter elements are directly fused to the binary factor within the transposon followed by random or site-specific integration. However, such insertions do not consistently recapitulate endogenous expression. We used Minos-Mediated Integration Cassette (MiMIC) transposons to convert host loci into reliable gene-specific binary effectors. MiMIC transposons allow recombinase-mediated cassette exchange to modify the transposon content. We developed novel exchange cassettes to convert coding intronic MiMIC insertions into gene-specific binary factor protein-traps. In addition, we expanded the set of binary factor exchange cassettes available for non-coding intronic MiMIC insertions. We show that binary factor conversions of different insertions in the same locus have indistinguishable expression patterns, suggesting that they reliably reflect endogenous gene expression. We show the efficacy and broad applicability of these new tools by dissecting the cellular expression patterns of the Drosophila serotonin receptor gene family. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

Plasma Levels of High Sensitivity Cardiac Troponin T in Adults with Repaired Tetralogy of Fallot

Science.gov (United States)

Lai, Clare T. M.; Wong, Sophia J.; Ip, Janice J. K.; Wong, Wai-keung; Tsang, Kwong-cheong; Lam, Wendy W. M.; Cheung, Yiu-fai

2015-01-01

Detectable low circulating level of cardiac troponin T (cTnT) may reflect subclinical myocardial injury. We tested the hypothesis that circulating levels of hs-cTnT are altered in adults with repaired tetralogy of Fallot (TOF) and associated with ventricular volume load and function. Eighty-eight TOF patients and 48 controls were studied. Plasma hs-cTnT levels were determined using a highly sensitive assay (hs-cTnT). The right (RV) and left ventricular (LV) volumes and ejection fraction (EF) were measured using 3D echocardiography and, in 52 patients, cardiac magnetic resonance (CMR). The median (interquartile range) for male and female patients were 4.87 (3.83–6.62) ng/L and 3.11 (1.00–3.87) ng/L, respectively. Thirty percent of female but none of the male patients had increased hs-cTnT levels. Female patients with elevated hs-cTnT levels, compared to those without, had greater RV end-diastolic and end-systolic volumes and LV systolic dyssynchrony index (all p < 0.05). For patient cohort only, hs-cTnT levels correlated positively with CMR-derived RV end-diastolic volume and negatively with echocardiography-derived LV and RV EF (all p < 0.05). Multiple linear regression identified sex and RV EF as significant correlates of log-transformed hs-cTnT levels. Increased hs-cTnT levels occur in 30% of female patients after TOF repair, and are associated with greater RV volumes and worse RV EF. PMID:26360613

BWR - Spent Fuel Transport and Storage with the TNTM9/4 and TNTM24BH Casks

International Nuclear Information System (INIS)

Wattez, L.; Marguerat, Y.; Hoesli, C.

2006-01-01

The Swiss Nuclear Utilities have started in 2001 to store spent fuel in dry metallic dual-purpose casks at ZWILAG, the Swiss interim storage facility. BKW FMB Energy Ltd., the Muehleberg Nuclear Power Plant owner, is involved in this process and has elected to store its BWR spent fuel in a new high capacity dual-purpose cask, the TNeTeM24BH from the COGEMA Logistics/TRANSNUCLEAR TN TM 24 family. The Muehleberg BWR spent fuels are transported by road in a medium size shuttle transport cask and then transferred to a heavy transport/storage cask (dry transfer) in the hot cell of ZWILAG site. For that purpose, COGEMA Logistics designed and supplied: - Two shuttle casks, TN TM 9/4, mainly devoted to transport of spent fuel from Muehleberg NPP to ZWILAG. Licensed according to IAEA 1996, the TN TM 9/4 is a 40 ton transport cask, for 7 BWR high bum-up spent fuel assemblies. - A series of new high capacity dual-purpose casks, TN TM 24BH, holding 69 BWR spent fuels. Two transport campaigns took place in 2003 and 2004. For each campaign, ten TN TM 9/4 round trips are performed, and one TN TM 24BH is loaded. 5 additional TN TM 24BH are being manufactured for BKW, and the next transport campaigns are scheduled from 2006. The TN TM 24BH high capacity dual purpose cask and the TN TM 9/4 transport cask characteristics and capabilities will then be detailed. (authors)

Modelling Ca2+ bound Troponin in Excitation Contraction Coupling

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Henry G. Zot

2016-09-01

Full Text Available To explain disparate decay rates of cytosolic Ca2+ and structural changes in the thin filaments during a twitch, we model the time course of Ca2+ bound troponin (Tn resulting from the free Ca2+ transient of fast skeletal muscle. In fibers stretched beyond overlap, the decay of Ca2+ as measured by a change in fluo 3 fluorescence is significantly slower than the intensity decay of the meridional 1/38.5 nm-1 reflection of Tn; this is not simply explained by considering only the Ca2+ binding properties of Tn alone (Matsuo, T., Iwamoto, H., and Yagi, N. (2010. Biophys. J. 99, 193-200. We apply a comprehensive model that includes the known Ca2+ binding properties of Tn in the context of the thin filament with and without cycling crossbridges. Calculations based on the model predict that the transient of Ca2+ bound Tn correlates with either the fluo 3 time course in muscle with overlapping thin and thick filaments or the intensity of the meridional 1/38.5 nm-1 reflection in overstretched muscle. Hence, cycling crossbridges delay the dissociation of Ca2+ from Tn. Correlation with the fluo 3 fluorescence change is not causal given that the transient of Ca2+ bound Tn depends on sarcomere length, whereas the fluo-3 fluorescence change does not. Transient positions of tropomyosin calculated from the time course of Ca2+ bound Tn are in reasonable agreement with the transient of measured perturbations of the Tn repeat in overlap and non-overlap muscle preparations.

TNNI3K is a novel mediator of myofilament function and phosphorylates cardiac troponin I

International Nuclear Information System (INIS)

Wang, Hui; Wang, Lin; Song, Li; Zhang, Yan-Wan; Ye, Jue; Xu, Rui-Xia; Shi, Na; Meng, Xian-Min

2013-01-01

The phosphorylation of cardiac troponin I (cTnI) plays an important role in the contractile dysfunction associated with heart failure. Human cardiac troponin I-interacting kinase (TNNI3K) is a novel cardiac-specific functional kinase that can bind to cTnI in a yeast two-hybrid screen. The purpose of this study was to investigate whether TNNI3K can phosphorylate cTnI at specific sites and to examine whether the phosphorylation of cTnI caused by TNNI3K can regulate cardiac myofilament contractile function. Co-immunoprecipitation was performed to confirm that TNNI3K could interact with cTnI. Kinase assays further indicated that TNNI3K did not phosphorylate cTnI at Ser23/24 and Ser44, but directly phosphorylated Ser43 and Thr143 in vitro. The results obtained for adult rat cardiomyocytes also indicated that enhanced phosphorylation of cTnI at Ser43 and Thr143 correlated with rTNNI3K (rat TNNI3K) overexpression, and phosphorylation was reduced when rTNNI3K was knocked down. To determine the contractile function modulated by TNNI3K-mediated phosphorylation of cTnI, cardiomyocyte contraction was studied in adult rat ventricular myocytes. The contraction of cardiomyocytes increased with rTNNI3K overexpression and decreased with rTNNI3K knockdown. We conclude that TNNI3K may be a novel mediator of cTnI phosphorylation and contribute to the regulation of cardiac myofilament contraction function

Tetranectin Knockout Mice Develop Features of Parkinson Disease

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Er-song Wang

2014-07-01

Full Text Available Background/Aims: Aggregation of insoluble α-synuclein to form Lewy bodies (LBs may contribute to the selective loss of midbrain dopaminergic neurons in Parkinson disease (PD. Lack of robust animal models has impeded elucidation of the molecular mechanisms of LB formation and other critical aspects of PD pathogenesis. Methods: We established a mouse model with targeted deletion of the plasminogen-binding protein tetranectin (TN gene (TN-/- and measured the behavioral and histopathological features of PD. Results: Aged (15-to 20-month-old TN-/- mice displayed motor deficits resembling PD symptoms, including limb rigidity and both slower ambulation (bradykinesia and reduced rearing activity in the open field. In addition, these mice exhibited more numerous α-synuclein-positive LB-like inclusions within the substantia nigra pars compacta (SNc and reduced numbers of SNc dopaminergic neurons than age-matched wild type (WT mice. These pathological changes were also accompanied by loss of dopamine terminals in the dorsal striatum. Conclusion: The TN-/- mouse exhibits several key features of PD and so may be a valuable model for studying LB formation and testing candidate neuroprotective therapies for PD and other synucleinopathies.

The ( and ( Contrast as a Case of Fossilized Pronunciation Error for Turkish English Teachers

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Mehmet Demirezen

2007-04-01

Full Text Available The ƒËƒÁƒÍ, called ¡§ash¡¨ and ƒË„ƒÍ, called ¡§schwa¡¨ are very frequent phonemes inEnglish language that confuse the Turkish teachers of English. The main reason for thisconfusion is that both of them do not exist in modern standard Turkish. Another reasonis the reduction of ƒËƒÁƒÍ into ƒË„ƒÍ in word formation process with the accompaniment ofthe change of primary stress to another syllable. For example, milk maƒ°n, post maƒ°n, andgate maƒ°n are phrases by nature, but in a process of compounding they take themorphological form of milkmaƒtn, poƒ°stmaƒtn, and gaƒ°temaƒtn through a phonological decayof the /ƒÁ/ phoneme into /„/ phoneme with the reduction of the primary stress of the freeword maƒ°n into the weak stress form like -maƒtn, relegating it into the position of a boundmorpheme. This article, through a model lesson plan, will explore such sound changesthat confuse the Turkish English teachers.

Factors independently associated with cardiac troponin I levels in young and healthy adults from the general population.

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Bossard, Matthias; Thériault, Sébastien; Aeschbacher, Stefanie; Schoen, Tobias; Kunz, Seraina; von Rotz, Mirco; Estis, Joel; Todd, John; Risch, Martin; Mueller, Christian; Risch, Lorenz; Paré, Guillaume; Conen, David

2017-02-01

Determinants of cardiomyocyte injury as quantified by high-sensitivity cardiac troponin I (cTnI) in young and healthy individuals, and sex-specific 99th percentiles are largely unknown. Our study included 2077 adults from the general population aged 25-41 years without cardiovascular disease. cTnI was measured using a high-sensitivity assay. We performed stepwise backward linear regression analyses to identify variables independently associated with hs-cTnI levels, and calculated narrow-sense heritability from 1638-genotyped participants. Median age was 37 years. cTnI was quantifiable in all but 11 participants (99.5 %). Median (interquartile range) cTnI was significantly higher in men than in women [0.99 (0.71; 1.65) versus 0.47 (0.33; 0.71) ng/L, p age, systolic blood pressure, heart rate, left ventricular mass, N-terminal pro B-type natriuretic peptide, and creatine kinase (all p age, and systolic blood pressure belong to the strongest determinants of hs-cTnI in healthy adults. The 99th percentile was three times higher in men compared to women. Hence, sex-specific cut-off values may be preferable when applying hs-cTnI for screening purposes. Our results may also improve the interpretation of cTn levels in daily clinical practice.

Moving the largest capacity PWR dual-purpose cask in the world from Goesgen NPP to the Zwilag interim storage site

International Nuclear Information System (INIS)

Delannay, M.; Dudragne, S.

2002-01-01

The Swiss Goesgen nuclear power plant (NPP) has decided to use two different methods for the disposal of its spent fuel. (1) To reprocess some of its spent fuel in dedicated facilities. Some of the vitrified waste from the reprocessing will be shipped back to Switzerland using the new COGEMA Logistics, TN81 cask. (2) To ship the other part of its spent fuel to the central interim storage facility of Zwilag (Switzerland) using a COGEMA Logistics dual-purpose TN24G cask. The TN24G is the heaviest and largest dual-purpose cask manufactured so far by COGEMA Logistics in Europe. It is intended for the transport and storage of 37 pressurised water-reactor (PWR) spent fuel assemblies. Four casks were delivered by COGEMA Logistics to Goesgen NPP. Three transports of loaded TN24G casks between Goesgen and Zwilag were successfully performed at the beginning of 2002 with the new COGEMA Logistics Q76 wagon specifically designed to transport heavy casks. This article describes the procedure of operations and shipments for the first TN24G casks up to storage at Zwilag. The fourth transport of loaded TN24G was due to happen in October 2002. The TN24G cask, as part of the TN24 casks family, proved to be a very efficient solution for the KKG spent fuel management. (author)

High-Throughput Genetic Screens Identify a Large and Diverse Collection of New Sporulation Genes in Bacillus subtilis.

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Meeske, Alexander J; Rodrigues, Christopher D A; Brady, Jacqueline; Lim, Hoong Chuin; Bernhardt, Thomas G; Rudner, David Z

2016-01-01

The differentiation of the bacterium Bacillus subtilis into a dormant spore is among the most well-characterized developmental pathways in biology. Classical genetic screens performed over the past half century identified scores of factors involved in every step of this morphological process. More recently, transcriptional profiling uncovered additional sporulation-induced genes required for successful spore development. Here, we used transposon-sequencing (Tn-seq) to assess whether there were any sporulation genes left to be discovered. Our screen identified 133 out of the 148 genes with known sporulation defects. Surprisingly, we discovered 24 additional genes that had not been previously implicated in spore formation. To investigate their functions, we used fluorescence microscopy to survey early, middle, and late stages of differentiation of null mutants from the B. subtilis ordered knockout collection. This analysis identified mutants that are delayed in the initiation of sporulation, defective in membrane remodeling, and impaired in spore maturation. Several mutants had novel sporulation phenotypes. We performed in-depth characterization of two new factors that participate in cell-cell signaling pathways during sporulation. One (SpoIIT) functions in the activation of σE in the mother cell; the other (SpoIIIL) is required for σG activity in the forespore. Our analysis also revealed that as many as 36 sporulation-induced genes with no previously reported mutant phenotypes are required for timely spore maturation. Finally, we discovered a large set of transposon insertions that trigger premature initiation of sporulation. Our results highlight the power of Tn-seq for the discovery of new genes and novel pathways in sporulation and, combined with the recently completed null mutant collection, open the door for similar screens in other, less well-characterized processes.

High-Throughput Genetic Screens Identify a Large and Diverse Collection of New Sporulation Genes in Bacillus subtilis

Science.gov (United States)

Brady, Jacqueline; Lim, Hoong Chuin; Bernhardt, Thomas G.; Rudner, David Z.

2016-01-01

The differentiation of the bacterium Bacillus subtilis into a dormant spore is among the most well-characterized developmental pathways in biology. Classical genetic screens performed over the past half century identified scores of factors involved in every step of this morphological process. More recently, transcriptional profiling uncovered additional sporulation-induced genes required for successful spore development. Here, we used transposon-sequencing (Tn-seq) to assess whether there were any sporulation genes left to be discovered. Our screen identified 133 out of the 148 genes with known sporulation defects. Surprisingly, we discovered 24 additional genes that had not been previously implicated in spore formation. To investigate their functions, we used fluorescence microscopy to survey early, middle, and late stages of differentiation of null mutants from the B. subtilis ordered knockout collection. This analysis identified mutants that are delayed in the initiation of sporulation, defective in membrane remodeling, and impaired in spore maturation. Several mutants had novel sporulation phenotypes. We performed in-depth characterization of two new factors that participate in cell–cell signaling pathways during sporulation. One (SpoIIT) functions in the activation of σE in the mother cell; the other (SpoIIIL) is required for σG activity in the forespore. Our analysis also revealed that as many as 36 sporulation-induced genes with no previously reported mutant phenotypes are required for timely spore maturation. Finally, we discovered a large set of transposon insertions that trigger premature initiation of sporulation. Our results highlight the power of Tn-seq for the discovery of new genes and novel pathways in sporulation and, combined with the recently completed null mutant collection, open the door for similar screens in other, less well-characterized processes. PMID:26735940

ESTIMATIVA DE TEMPERATURAS ABSOLUTAS USANDO DADOS CLIMÁTICOS E ORBITAIS SOBRE O TERRITÓRIO BRASILEIRO

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PABRÍCIO MARCOS OLIVEIRA LOPES

2011-01-01

Full Text Available This work aimed to determine over the Brazilian territory extreme air temperatures: maximum (Tx and minimum (Tn absolute, using multiple regression techniques, orbital data and climatological. The data of Tx and Tn were observed between the years of 1961 and 1990 in 204 meteorological stations by the National Institute of Meteorology (INMET, and topography information was obtained from the program Shuttle Radar Topography Mission (SRTM. The multiple regression models were validated with the observational data of 30 meteorological stations in different locations than those generated. The statistical analysis was performed using bootstrap techniques. The results showed that the multiple regression model for Tn presented good agreement with the observed data (R² = 0.86 showing no tendencies to overestimate or underestimate Tn. The Tn varied between -12 to 25 °C and the Tx between 31 and 42 °C. For the Tx the results were not as satisfactory. In a first approach, the limits of the Tn can be used for data quality control of the meteorological station. The map of Tn can be used to subsidize research in agroclimatic zoning in the region where these data are not available.

Radically altered T cell receptor signaling in glycopeptide-specific T cell hybridoma induced by antigen with minimal differences in the glycan group

DEFF Research Database (Denmark)

Jensen, T; Nielsen, M; Gad, Monika

2001-01-01

A T cell hybridoma raised against the synthetic glycopeptide T(72)(Tn) was used to study whether the initial TCR signaling events are markedly different when the hybridoma is stimulated with glycopeptides closely related to the cognate glycopeptide antigen. T(72)(Tn) has an alpha-D-GalNAc group O......)(alpha-D-GlcNAc), which differs from T(72)(Tn) solely by the orientation of a hydroxy group in the carbohydrate structure, completely failed to induce detectable tyrosine phosphorylation and IL-2 secretion. APC pulsed with S(72)(Tn), which differs from T(72)(Tn) by not having a methyl group in the serine......-linked to the central threonine in the decapeptide VITAFTEGLK, and the hybridoma is known to be highly specific for this carbohydrate group. T(72)(Tn)-pulsed APC induced tyrosine phosphorylation of the TCR-zeta 21- and 23-kDa proteins and the downstream p42/44 MAP kinase and strong IL-2 secretion. APC pulsed with T(72...

Combining High Sensitivity Cardiac Troponin I and Cardiac Troponin T in the Early Diagnosis of Acute Myocardial Infarction.

Science.gov (United States)

van der Linden, Noreen; Wildi, Karin; Twerenbold, Raphael; Pickering, John W; Than, Martin; Cullen, Louise; Greenslade, Jaimi; Parsonage, William; Nestelberger, Thomas; Boeddinghaus, Jasper; Badertscher, Patrick; Rubini Giménez, Maria; Klinkenberg, Lieke J J; Bekers, Otto; Schöni, Aline; Keller, Dagmar I; Sabti, Zaid; Puelacher, Christian; Cupa, Janosch; Schumacher, Lukas; Kozhuharov, Nikola; Grimm, Karin; Shrestha, Samyut; Flores, Dayana; Freese, Michael; Stelzig, Claudia; Strebel, Ivo; Miró, Òscar; Rentsch, Katharina; Morawiec, Beata; Kawecki, Damian; Kloos, Wanda; Lohrmann, Jens; Richards, A Mark; Troughton, Richard; Pemberton, Christopher; Osswald, Stefan; van Dieijen-Visser, Marja P; Mingels, Alma M; Reichlin, Tobias; Meex, Steven J R; Mueller, Christian

2018-04-24

Background -Combining two signals of cardiomyocyte injury, cardiac troponin I (cTnI) and T (cTnT), might overcome some individual pathophysiological and analytical limitations and thereby increase diagnostic accuracy for acute myocardial infarction (AMI) with a single blood draw. We aimed to evaluate the diagnostic performance of combinations of high sensitivity (hs) cTnI and hs-cTnT for the early diagnosis of AMI. Methods -The diagnostic performance of combining hs-cTnI (Architect, Abbott) and hs-cTnT (Elecsys, Roche) concentrations (sum, product, ratio and a combination algorithm) obtained at the time of presentation was evaluated in a large multicenter diagnostic study of patients with suspected AMI. The optimal rule out and rule in thresholds were externally validated in a second large multicenter diagnostic study. The proportion of patients eligible for early rule out was compared with the ESC 0/1 and 0/3 hour algorithms. Results -Combining hs-cTnI and hs-cTnT concentrations did not consistently increase overall diagnostic accuracy as compared with the individual isoforms. However, the combination improved the proportion of patients meeting criteria for very early rule-out. With the ESC 2015 guideline recommended algorithms and cut-offs, the proportion meeting rule out criteria after the baseline blood sampling was limited (6-24%) and assay dependent. Application of optimized cut-off values using the sum (9 ng/L) and product (18 ng2/L2) of hs-cTnI and hs-cTnT concentrations led to an increase in the proportion ruled-out after a single blood draw to 34-41% in the original (sum: negative predictive value (NPV) 100% (95%CI: 99.5-100%); product: NPV 100% (95%CI: 99.5-100%) and in the validation cohort (sum: NPV 99.6% (95%CI: 99.0-99.9%); product: NPV 99.4% (95%CI: 98.8-99.8%). The use of a combination algorithm (hs-cTnI <4 ng/L and hs-cTnT <9 ng/L) showed comparable results for rule out (40-43% ruled out; NPV original cohort 99.9% (95%CI: 99.2-100%); NPV

Equivalence of the O( n) vector ferromagnetic and antiferromagnetic models

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Sousa, J. Ricardo de

The effective-field renormalization group (EFRG) approach is used to find the Néel temperature ( TN) of the O( n) vector model with antiferromagnetic (AF) interaction. The EFRG method is illustrated by employing approximations in which clusters with one ( N‧=1) and two ( N=2) spins are used. The critical temperature TN is obtained as a function of component ( n) and coordination ( z) numbers. For all values of n and z we show that TN= Tc, where Tc is the Curie temperature for the ferromagnetic (F) case. As a comparison, the results of the quantum Heisenberg model ( n=3) with F and AF interactions are also presented, and we find that TN> Tc, which is different from the classical result Tc= TN.

Genome-Wide Sensitivity Analysis of the Microsymbiont Sinorhizobium meliloti to Symbiotically Important, Defensin-Like Host Peptides

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Markus F. F. Arnold

2017-08-01

Full Text Available The model legume species Medicago truncatula expresses more than 700 nodule-specific cysteine-rich (NCR signaling peptides that mediate the differentiation of Sinorhizobium meliloti bacteria into nitrogen-fixing bacteroids. NCR peptides are essential for a successful symbiosis in legume plants of the inverted-repeat-lacking clade (IRLC and show similarity to mammalian defensins. In addition to signaling functions, many NCR peptides exhibit antimicrobial activity in vitro and in vivo. Bacterial resistance to these antimicrobial activities is likely to be important for symbiosis. However, the mechanisms used by S. meliloti to resist antimicrobial activity of plant peptides are poorly understood. To address this, we applied a global genetic approach using transposon mutagenesis followed by high-throughput sequencing (Tn-seq to identify S. meliloti genes and pathways that increase or decrease bacterial competitiveness during exposure to the well-studied cationic NCR247 peptide and also to the unrelated model antimicrobial peptide polymyxin B. We identified 78 genes and several diverse pathways whose interruption alters S. meliloti resistance to NCR247. These genes encode the following: (i cell envelope polysaccharide biosynthesis and modification proteins, (ii inner and outer membrane proteins, (iii peptidoglycan (PG effector proteins, and (iv non-membrane-associated factors such as transcriptional regulators and ribosome-associated factors. We describe a previously uncharacterized yet highly conserved peptidase, which protects S. meliloti from NCR247 and increases competitiveness during symbiosis. Additionally, we highlight a considerable number of uncharacterized genes that provide the basis for future studies to investigate the molecular basis of symbiotic development as well as chronic pathogenic interactions.

Hypertrophic cardiomyopathy-linked mutation in troponin T causes myofibrillar disarray and pro-arrhythmic action potential changes in human iPSC cardiomyocytes.

Science.gov (United States)

Wang, Lili; Kim, Kyungsoo; Parikh, Shan; Cadar, Adrian Gabriel; Bersell, Kevin R; He, Huan; Pinto, Jose R; Kryshtal, Dmytro O; Knollmann, Bjorn C

2018-01-01

Mutations in cardiac troponin T (TnT) are linked to increased risk of ventricular arrhythmia and sudden death despite causing little to no cardiac hypertrophy. Studies in mice suggest that the hypertrophic cardiomyopathy (HCM)-associated TnT-I79N mutation increases myofilament Ca sensitivity and is arrhythmogenic, but whether findings from mice translate to human cardiomyocyte electrophysiology is not known. To study the effects of the TnT-I79N mutation in human cardiomyocytes. Using CRISPR/Cas9, the TnT-I79N mutation was introduced into human induced pluripotent stem cells (hiPSCs). We then used the matrigel mattress method to generate single rod-shaped cardiomyocytes (CMs) and studied contractility, Ca handling and electrophysiology. Compared to isogenic control hiPSC-CMs, TnT-I79N hiPSC-CMs exhibited sarcomere disorganization, increased systolic function and impaired relaxation. The Ca-dependence of contractility was leftward shifted in mutation containing cardiomyocytes, demonstrating increased myofilament Ca sensitivity. In voltage-clamped hiPSC-CMs, TnT-I79N reduced intracellular Ca transients by enhancing cytosolic Ca buffering. These changes in Ca handling resulted in beat-to-beat instability and triangulation of the cardiac action potential, which are predictors of arrhythmia risk. The myofilament Ca sensitizer EMD57033 produced similar action potential triangulation in control hiPSC-CMs. The TnT-I79N hiPSC-CM model not only reproduces key cellular features of TnT-linked HCM such as myofilament disarray, hypercontractility and diastolic dysfunction, but also suggests that this TnT mutation causes pro-arrhythmic changes of the human ventricular action potential. Copyright © 2017 Elsevier Ltd. All rights reserved.

[Comparison analysis of muscle enzymes in children with myocarditis and Duchene/Becker muscular dystrophy].

Science.gov (United States)

Zhang, Yali; Wang, Hong; Yu, Xuexin; Xing, Yanlin; Wang, Ce; He, Rong

2016-09-28

To compare the changes in muscle enzyme between children with myocarditis and Duchene/Becker muscular dystrophy (DMD/BMD), and to seek the explanations for variation.
 The retrospective analysis for 83 myocarditis children (myocarditis group) and 69 DMD/BMD children (DMD/BMD group), who were collected from Department of Pediatric of Shengjing Hospital affiliated to China Medical University since January 2008 to May 2015, was carried out. At the same time, 24 healthy children from the Department of Pediatric Development served as a control group. The examination indexes included creatine kinase (CK), creatine kinase-isoenzyme MB (CK-MB), creatine kinase isoenzyme MB mass (CK-MB mass), cardiac troponin I (cTnI) and high-sensitive-cTnT (hs-cTnT).
 1) In the myocarditis group, the CK increased from 100 to 1 000 U/L, reached a peak after 5 days, which lasted for a week and then dropped to the normal; the CK-MB reached a peak after 5 to 7 days and dropped to the normal a month later; the CK-MB mass reached a peak on the first day and dropped to the normal after 3 weeks; the cTn reached to a peak after 5 days and dropped to the normal after about 17 days; hs-cTnT reached to a peak on the first day and dropped to the normal after about 19 days. 2) In the DMD/BMD group, the CK increased significantly and 27 cases had a CK value of more than 10 000 U/L. After the treatment for 1 to 2 weeks, their enzyme rose again after a slight drop. In terms of cTnI, 6 cases showed a moderate increase, 5 of them couldn't drop to the normal level until more than 3 weeks later; the hs-cTnT increased in the 45 cases, which lasted for more than 3 weeks in the 31 cases of them and showed a tendency of persisting increase.
 The cTnI and hs-cTnT rise significantly and possess wider observation window than CK and CK-MB mass in myocarditis children, with more sensitive and specific changes. The myocardial damage can occur before myasthenia and keep this trend for a long time in the DMD

Temporal changes in nitrogen and phosphorus concentrations with comparisons to conservation practices and agricultural activities in the Lower Grand River, Missouri and Iowa, and selected watersheds, 1969–2015

Science.gov (United States)

Krempa, Heather M.; Flickinger, Allison K.

2017-08-01

This report presents the results of a cooperative study by the U.S. Geological Survey and Missouri Department of Natural Resources to estimate total nitrogen (TN) and total phosphorus (TP) concentrations at monitoring sites within and near the Lower Grand River hydrological unit. The primary objectives of the study were to quantify temporal changes in TN and TP concentrations and compare those concentrations to conservation practices and agricultural activities. Despite increases in funding during 2011–15 for conservation practices in the Lower Grand River from the Mississippi River Basin Healthy Watersheds Initiative, decreases in flow-normalized TN and TP concentrations during this time at the long-term Grand River site were less than at other long-term sites, which did not receive funding from the Mississippi River Basin Healthy Watersheds Initiative. The relative differences in the magnitude of flow-normalized TN and TP concentrations among long-term sites are directly related to the amount of agricultural land use within the watershed. Significant relations were determined between nitrogen from cattle manure and flow-normalized TN concentrations at selected long-term sites, indicating livestock manure may be a substantial source of nitrogen within the selected long-term site watersheds. Relations between flow-normalized TN and TP concentrations with Conservation Reserve Program acres and with nitrogen and phosphorus from commercial fertilizer indicate that changes in these factors alone did not have a substantial effect on stream TN and TP concentrations; other landscape activities, runoff, within-bank nutrients that are suspended during higher streamflows, or a combination of these have had a greater effect on stream TN and TP concentrations; or there is a lag time that is obscuring relations. Temporal changes in flow-adjusted TN and TP concentrations were not substantial at Lower Grand River Mississippi River Basin Healthy Watersheds Initiative sites

Differences in elongation of very long chain fatty acids and fatty acid metabolism between triple-negative and hormone receptor-positive breast cancer.

Science.gov (United States)

Yamashita, Yuji; Nishiumi, Shin; Kono, Seishi; Takao, Shintaro; Azuma, Takeshi; Yoshida, Masaru

2017-08-29

Triple-negative breast cancer (TN) is more aggressive than other subtypes of breast cancer and has a lower survival rate. Furthermore, detailed biological information about the disease is lacking. This study investigated characteristics of metabolic pathways in TN. We performed the metabolome analysis of 74 breast cancer tissues and the corresponding normal breast tissues using LC/MS. Furthermore, we classified the breast cancer tissues into ER-positive, PgR-positive, HER2-negative breast cancer (EP+H-) and TN, and then the differences in their metabolic pathways were investigated. The RT-PCR and immunostaining were carried out to examine the expression of ELOVL1, 2, 3, 4, 5, 6, and 7. We identified 142 of hydrophilic metabolites and 278 of hydrophobic lipid metabolites in breast tissues. We found the differences between breast cancer and normal breast tissues in choline metabolism, glutamine metabolism, lipid metabolism, and so on. Most characteristic of comparison between EP+H- and TN were differences in fatty acid metabolism was which were related to the elongation of very long chain fatty acids were detected between TN and EP+H-. Real-time RT-PCR showed that the mRNA expression levels of ELOVL1, 5, and 6 were significantly upregulated by 8.5-, 4.6- and 7.0-fold, respectively, in the TN tumors compared with their levels in the corresponding normal breast tissue samples. Similarly, the mRNA expression levels of ELOVL1, 5, and 6 were also significantly higher in the EP+H- tissues than in the corresponding normal breast tissues (by 4.9-, 3.4-, and 2.1-fold, respectively). The mRNA expression level of ELOVL6 was 2.6-fold higher in the TN tumors than in the EP+H- tumors. During immunostaining, the TN and EP+H- tumors demonstrated stronger ELOVL1 and 6 staining than the corresponding normal breast tissues, but ELOVL5 was not stained strongly in the TN or EP+H- tumors. Furthermore, the TN tumors exhibited stronger ELOVL1 and 6 staining than the EP+H- tumors. Marked

Cross-sectional study of high-sensitivity cardiac troponins T and I in a hospital and community outpatient setting.

Science.gov (United States)

Potter, Julia M; Simpson, Aaron J; Kerrigan, Jennifer; Southcott, Emma; Salib, Marie M; Koerbin, Gus; Hickman, Peter E

2017-02-01

Cardiac troponins are specific for the heart, but not for the acute coronary syndrome. We wanted to assess how common elevated cardiac troponin concentrations were, in a population with significant non-cardiac disease. We measured both hs-cTnT and hs-cTnI on all samples submitted to the laboratory during one 24h period, and assessed the magnitude of the cTn concentration with the location and severity of disease of the patient. Community patients and patients from the maternity ward had the lowest cTn concentrations with results above the 99th percentile being only 0-2% of the total. As expected, the highest proportion of results >99th percentile came from Coronary Care and Intensive Care. However, substantial numbers of persons on Medical and Surgical wards, without a primary diagnosis of cardiac disease, also had cTn >99th percentile. Particularly for cTnT, there was a highly significant odds ratio predicting mortality when results above and below the 99th percentile were compared. Significant illnesses apart from the acute coronary syndrome are important causes of a rise in cTn to above the 99th percentile, and appear to reflect the total body burden of disease. Even when the high hs-cTn concentration is not due to the acute coronary syndrome, there is a significant association with all-cause mortality. Copyright © 2016 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Hypoxic Living and Exercise Training Alter Adipose Tissue Leptin/Leptin Receptor in Rats

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Yingli Lu

2016-11-01

Full Text Available Background: Hypobaric hypoxia results in weight loss in obese individuals, and exercise training is advocated for the treatment of obesity and its related metabolic dysfunctions. The purpose of this study was to investigate the effects of hypoxic living and exercise training on obesity and adipose tissue leptin/leptin receptor in dietary-induced obese rats. Methods: One hundred and thirty high-fat diet fed Sprague-Dawley rats were assigned into one of the following groups (n=10 each: control, sedentary hypoxic living for 1 to 4 weeks (SH1, SH2, SH3, and SH4, living and exercise training in normoxic conditions for 1 to 4 weeks (TN1, TN2, TN3, and TN4, and living and exercise training in hypoxic conditions for 1 to 4 weeks (TN1, TN2, TN3, and TN4. Epididymal adipose tissue expression levels of leptin and leptin receptor were determined. Results: Compared to hypoxic living and living and exercise training in normoxic conditions, living and exercise training in hypoxic conditions for 3-4 weeks resulted in lower Lee index (P<0.05 to P<0.01, and higher expression of leptin and leptin receptor (P<0.05 to P<0.01 in adipose tissue. Conclusion: In a rodent model of altitude training, living and exercise training in hypoxic conditions resulted in greater alterations in obesity and adipose tissue leptin/leptin receptor than hypoxic living alone and living and exercise training in normoxic conditions.

European experience in transport/storage cask for vitrified residues

International Nuclear Information System (INIS)

Otton, Camille; Sicard, Damien

2007-01-01

Available in abstract form only. Full text of publication follows: Because of the evolution of burnup of spent fuel to be reprocessed, the high activity vitrified residues would not be transported in the existing cask designs. Therefore, TN International has decided in the late nineties to develop a brand new design of casks with optimized capacity able to store and transport the most active and hottest canisters: the TN TM 81 casks currently in use in Switzerland and the TN TM 85 cask which shall permit in the near future in Germany the storage and the transport of the most active vitrified residues defining a thermal power of 56 kW (kilowatts). The challenges for the TN TM 81 and TN TM 85 cask designs were that the geometry entry data were very restrictive and were combined with a fairly wide range set by the AREVA NC Specification relative to vitrified residue canister. The TN TM 81 and the TN TM 85 casks have been designed to fully anticipate shipment constraints of the present vitrified residue production. It also used the feedback of current shipments and the operational constraints and experience of receiving and shipping facilities. The casks had to fit as much as possible in the existing procedures for the already existing flasks such as the TN TM 28 cask and TS 28 V cask, all along the logistics chain of loading, unloading, transport and maintenance. (authors)

Intercellular Communication in Malignant Pleural Mesothelioma: Properties of Tunneling Nanotubes

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Justin William Ady

2014-10-01

Full Text Available Malignant pleural mesothelioma is a particularly aggressive and locally invasive malignancy with a poor prognosis despite advances in understanding of cancer cell biology and development of new therapies. At the cellular level, cultured mesothelioma cells present a mesenchymal appearance and a strong capacity for local cellular invasion. One important but underexplored area of mesothelioma cell biology is intercellular communication. Our group has previously characterized in multiple histological subtypes of mesothelioma a unique cellular protrusion known as tunneling nanotubes (TnTs. TnTs are long, actin filament-based, narrow cytoplasmic extensions that are non-adherent when cultured in vitro and are capable of shuttling cellular cargo between connected cells. Our prior work confirmed the presence of nanotube structures in tumors resected from patients with human mesothelioma. In our current study, we quantified the number of TnTs/cell among various mesothelioma subtypes and normal mesothelial cells using confocal microscopic techniques. We also examined TnT length among adherent cells and cells in suspension. We further examined potential approaches to the in vivo study of TnTs in animal models of cancer. We have developed novel approaches to study TnTs in aggressive solid tumor malignancies and define fundamental characteristics of TnTs in malignant mesothelioma. There is mounting evidence that TnTs play an important role in intercellular communication in mesothelioma and thus merit further investigation of their role in vivo.

Using adrenaline during neonatal resuscitation may have an impact on serum cardiac troponin-T levels.

Science.gov (United States)

Helmer, Caroline; Skranes, Janne H; Liestøl, Knut; Fugelseth, Drude

2015-09-01

It has been suggested that serum cardiac troponin-T (cTnT) can predict the severity of neonatal hypoxic-ischaemic encephalopathy. We evaluated whether cTnT was better correlated with adrenaline during cardiopulmonary resuscitation (CPR) than with the severity of the insult itself, based on the Apgar scores. Serum cTnT was analysed in 47 asphyxiated newborn infants treated with hypothermia. Blood samples and resuscitation data were collected from medical records, and multiple linear regressions were used to evaluate the effect of the treatment and the Apgar scores on cTnT levels. The infants were divided into three groups: the no CPR group (n = 29) just received stimulation and ventilation, the CPR minus adrenaline group (n = 9) received cardiac compression and ventilation and the CPR plus adrenaline group (n = 9) received complete CPR, including adrenaline. In the univariate analysis, the five and ten-minute Apgar scores were significantly lower in the CPR plus adrenaline group and the cTnT was significantly higher. Multiple regression analysis showed significantly higher cTnT values in the CPR plus adrenaline group, but no significant relationship between cTnT and the Apgar scores. Although cTnT correlated with the severity of the insult in neonatal hypoxic-ischaemic encephalopathy, the levels may have been affected by adrenaline administered during CPR. ©2015 Foundation Acta Paediatrica. Published by John Wiley & Sons Ltd.

Expressão de sulfomucinas e de carboidratos de mucinas simples no esôfago de Barrett Expression of sulphomucins and simple-mucin type carbohydrate in Barrett's esophagus

Directory of Open Access Journals (Sweden)

Frederico Henrique C. Melo

2007-04-01

Full Text Available INTRODUÇÃO: A expressão de mucinas sulfatadas e de antígenos de carboidratos simples de mucinas tem sido relacionada com o risco de transformação maligna no trato gastrintestinal. OBJETIVOS: Avaliar a expressão de mucinas sulfatadas e dos antígenos Tn, sTn, T e sT no esôfago de Barrett (EB. MATERIAL E MÉTODO: Estudaram-se 50 biópsias de EB. As amostras foram processadas rotineiramente para exame histológico (hematoxilina e eosina [HE] e ácido periódico de Schiff [PAS]/azul de alcião. Cortes adicionais foram corados para detecção de mucinas sulfatadas (diamina férrica e dos antígenos Tn, sTn, T e sT por imuno-histoquímica. O EB foi classificado quanto aos subtipos de metaplasia intestinal e sua extensão nas amostras. A expressão de Tn, sTn, T e sT no EB foi discriminada quanto à positividade em células caliciformes e colunares. RESULTADOS: Detectou-se metaplasia intestinal incompleta com secreção de sulfomucinas em 47 casos; em três não se identificou a lesão nos cortes realizados para a histoquímica. Observou-se metaplasia intestinal tipo III em 44 dos 47 casos analisados (93,6% e do tipo II isoladamente em três casos (6,4%. Não se detectou metaplasia intestinal do tipo I. Tn expressou-se fortemente em células colunares em 94% dos casos e foi focal/negativo nas caliciformes; sTn exibiu forte marcação nas células caliciformes em 88% dos casos. Os demais marcadores foram negativos na maior parte dos casos (T negativo em 82% e sT em 87,8%. CONCLUSÕES: A expressão de sulfomucinas e de antígenos de carboidratos simples de mucinas foi homogênea no EB. A lesão caracterizou-se por metaplasia intestinal incompleta com extenso componente de tipo III e expressão de Tn e sTn. Esses dados sugerem que os marcadores testados não permitem discriminar lesões com diferentes potenciais de transformação maligna.BACKGROUND: The expression of sulphomucins and simple-mucin type carbohydrate antigens has been used as marker

Blood troponin levels in acute cardiac events depends on space weather activity components (a correlative study).

Science.gov (United States)

Stoupel, Eliiyahu; Radishauskas, Richardas; Bernotiene, Gailute; Tamoshiunas, Abdonas; Virvichiute, Daiva

2018-02-05

Many biological processes are influenced by space weather activity components such as solar activity (SA), geomagnetic activity (GMA) and cosmic ray activity (CRA). Examples are total mortality, acute myocardial infarction (AMI), stroke (cerebrovascular accident), sudden cardiac death, some congenital maladies (congenital heart disease and Down syndrome), many events in neonatology, ophtalmology, blood pressure regulation, blood coagulation, inflammation, etc. The aim of this study was to check if the level of blood troponins (Tns) - markers of myocardial damage and recognized components of modern description of AMI - is connected with the mentioned space weather parameters. Patients admitted to a 3000-bed tertiary university hospital in Kaunas, Lithuania, with suspected AMI were the object of the study. Data for the time between 2008 and 2013 - 72 consecutive months - were studied. Of the patients, 1896 (1398 male, 498 female) had elevated troponin I (Tn I) or troponin T (Tn T, sensitive Tn) levels. Normal values were 0.00-0.03 ng/mL for Tn I and 0.00-14.00 ng/mL for Tn T. Monthly means and standard deviation of Tn I and Tn T were compared with monthly markers of SA, GMA and CRA. Pearson correlation coefficients and their probabilities were established (in addition to the consecutive graphs of both comparing physical and biological data). The cosmophysical data came from space service institutions in the United States, Russia and Finland. AMI was diagnosed in 1188 patients (62.66%), and intermediate coronary syndrome in 698 patients (36.81%). There were significant links of the Tn blood levels with four SA indices and CRA (neutron activity in imp/min); there was no significant correlation with GMA indices Ap and Cp (p=0.27 and p=0.235). Tn T levels significantly correlated with the GMA indices and not with the SA and CRA levels (Ap: r=0.77, p=0.0021; Cp: r=0.729, p=0.0047). First, the monthly level of blood Tn I in ACS is significantly correlated with the indices

Origins of Phosphorus and Nitrogen in the Seto Inland Sea, Japan

OpenAIRE

Yanagi, Tetsuo; Tanaka, Tsuyoshi; 柳, 哲雄; 田中, 剛

2013-01-01

Origins of total phosphorus (TP) and total nitrogen (TN) were investigated in the Seto Inland Sea, Japan, using the unit response function method. About 58 % of TP and TN in the Seto Inland Sea were found to have originated from the open ocean, 9 % of TP and 14 % of TN originate from rivers, 33 % of TP and 29 % of TN originate from bottom sediments. We consequently suggest that it would be very difficult to prevent the occurrence of red tides and oxygen deficient water masses in the Seto Inla...

Technical Network

CERN Multimedia

2007-01-01

In order to optimize the management of the Technical Network (TN), to ease the understanding and purpose of devices connected to the TN, and to improve security incident handling, the Technical Network Administrators and the CNIC WG have asked IT/CS to verify the "description" and "tag" fields of devices connected to the TN. Therefore, persons responsible for systems connected to the TN will receive email notifications from IT/CS asking them to add the corresponding information in the network database. Thank you very much for your cooperation. The Technical Network Administrators & the CNIC WG

Correlation of Optic Nerve Microcirculation with Papillomacular Bundle Structure in Treatment Naive Normal Tension Glaucoma

Directory of Open Access Journals (Sweden)

Wataru Kobayashi

2014-01-01

Full Text Available Purpose. To assess the association between optic nerve head (ONH microcirculation, central papillomacular bundle (CPB structure, and visual function in eyes with treatment naive normal tension glaucoma (NTG. Methods. This study included 40 eyes of 40 patients with NTG and 20 eyes of 20 normal patients. We used laser speckle flowgraphy (LSFG to measure mean blur rate (MBR in all eyes and calculated the ratio of MBR in the horizontal quadrants of tissue area ONH (temporal/nasal ratio of MBR in the tissue area: T/N MT. Clinical findings also included retinal nerve fiber layer thickness (RNFLT and ganglion cell complex thickness (GCCT in the CPB and macular areas, best-corrected visual acuity (BCVA, mean deviation (MD, and refractive error. Results. T/N MT was correlated with both BCVA and MD. The OCT parameters most highly correlated with T/N MT were macular RNFLT and mid-CPB RNFLT. Furthermore, T/N MT, mid-CPB RNFLT, and macular RNFLT were higher in NTG than in normal eyes. A discrimination analysis revealed that T/N MT and refractive error were independent factors indicating NTG. Conclusions. Our results suggest that T/N MT is a candidate biomarker of NTG. Furthermore, T/N MT reflects visual function, including acuity and sensitivity, and CPB structure.

Reducing human nitrogen use for food production.

Science.gov (United States)

Liu, Junguo; Ma, Kun; Ciais, Philippe; Polasky, Stephen

2016-07-22

Reactive nitrogen (N) is created in order to sustain food production, but only a small fraction of this N ends up being consumed as food, the rest being lost to the environment. We calculated that the total N input (TN) of global food production was 171 Tg N yr(-1) in 2000. The production of animal products accounted for over 50% of the TN, against 17% for global calories production. Under current TN per unit of food production and assuming no change in agricultural practices and waste-to-food ratios, we estimate that an additional TN of 100 Tg N yr(-1) will be needed by 2030 for a baseline scenario that would meet hunger alleviation targets for over 9 billion people. Increased animal production will have the largest impact on increasing TN, which calls for new food production systems with better N-recycling, such as cooperation between crop and livestock producing farms. Increased N-use efficiency, healthier diet and decreased food waste could mitigate this increase and even reduce TN in 2030 by 8% relative to the 2000 level. Achieving a worldwide reduction of TN is a major challenge that requires sustained actions to improve nitrogen management practices and reduce nitrogen losses into the environment.

Dynamical sampling and frame representations with bounded operators

DEFF Research Database (Denmark)

Christensen, Ole; Hasannasab, Marzieh; Rashidi, Ehsan

2017-01-01

The purpose of this paper is to study frames for a Hilbert space H, having the form {Tnφ}n=0∞ for some φ∈H and an operator T:H→H. We characterize the frames that have such a representation for a bounded operator T, and discuss the properties of this operator. In particular, we prove that the image...... chain of T has finite length N in the overcomplete case; furthermore {Tnφ}n=0∞ has the very particular property that {Tnφ}n=0N−1∪{Tnφ}n=N+ℓ∞ is a frame for H for all ℓ∈N0. We also prove that frames of the form {Tnφ}n=0∞ are sensitive to the ordering of the elements and to norm...

Novel effect of spin dynamics with suppression of charge and orbital ordering in Nd0.5Ca0.5MnO3 under the influence of ac electric field

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Sarwar, T.; Qamar, A.; Nadeem, M.

2017-07-01

Dynamics of spin ordering in the manganite Nd0.5Ca0.5MnO3 have been investigated in this paper. It was observed that the complex mixed magnetic ordering in pellets is comprised of antiferromagnetic ordering at 160 K (TN) and complete charge ordering at 250 K (TCO). Under ac field, appearance of unstable ferromagnetic correlations is observed above TCO, which is badly frustrated due to strong spin disorder induced by Jahn Teller distortions. Impedance measurements reveal the spin glass like scenario, suppressing the strong antiferromagnetic and charge ordering states below TN.

Diagnostic and prognostic value of high-sensitivity cardiac troponin T in patients with syncope.

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Christ, Michael; Geier, Felicitas; Popp, Steffen; Singler, Katrin; Smolarsky, Alexander; Bertsch, Thomas; Müller, Christian; Greve, Yvonne

2015-02-01

We examined the diagnostic and predictive value of high-sensitivity cardiac troponin T (cTnThs) in patients with syncope. We performed an analysis of consecutive patients with syncope presenting to the emergency department. The primary end point was the accuracy to diagnose a cardiac syncope. In addition, the study explored the prognostic relevance of cTnThs in patients with cardiac and noncardiac syncope. A total of 360 patients were enrolled (median age, 70.5 years; male, 55.8%; 23.9% aged >80 years). Cardiac syncope was present in 22% of patients, reflex syncope was present in 40% of patients, syncope due to orthostatic hypotension was present in 20% of patients, and unexplained syncope was present in 17.5% of patients. A total of 148 patients (41%) had cTnThs levels above the 99% confidence interval (CI) (cutoff point). The diagnostic accuracy for cTnThs levels to determine the diagnosis of cardiac syncope was quantified by the area under the curve (0.77; CI, 0.72-0.83; P value of cTnThs levels within 30 days: Patients with increased cTnThs levels had a 52% likelihood for adverse events, patients with cTnThs levels below the cutoff point had a low risk (negative predictive value, 83.5%). Increased cTnThs levels indicate adverse prognosis in patients with noncardiac causes of syncope, but not in patients with cardiac syncope being a risk factor for adverse outcome by itself. Patients with syncope presenting to the emergency department have a high proportion of life-threatening conditions. cTnThs levels show a limited diagnostic and predictive accuracy for the identification of patients with syncope at high risk. Copyright © 2015 Elsevier Inc. All rights reserved.

Consequences of implementing a cardiac troponin assay with improved sensitivity at Swedish coronary care units: an analysis from the SWEDEHEART registry.

Science.gov (United States)

Eggers, Kai M; Lindahl, Bertil; Melki, Dina; Jernberg, Tomas

2016-08-07

Cardiac troponin (cTn) assays with improved sensitivity are increasingly utilized for the assessment of patients admitted because of suspected acute coronary syndrome (ACS). However, data on the clinical consequences of the implementation of such assays are limited. In a retrospective register-based study (37 710 coronary care unit admissions; SWEDEHEART registry), we compared the case mix, the use of diagnostic procedures, treatments, and 1-year all-cause mortality 1 year before the implementation of a cTn assay with improved sensitivity (study period 1) and 1 year thereafter (study period 2). During study period 2, more at-risk patients were admitted and more patients had cTn levels above the myocardial infarction cut-off (ACS patients +13.1%; non-ACS patients +160.1%). cTn levels above this cut-off exhibited stronger associations with mortality risk in study period 2 (adjusted HR 4.45 [95% confidence interval, CI, 3.36-5.89]) compared with period 1 (adjusted HR 2.43 [95% CI 2.11-2.80]), similar as for the cTn ratio relative to the respective 99th percentile. While there was no multivariable-adjusted increase in the use of diagnostic procedures, significant trends towards more differentiated treatment depending on the cause of cTn elevation, i.e. ACS or non-ACS, were noted. The implementation of a cTn assay with improved sensitivity was associated with an increase in the number of patients who due to their cTn-status were identified as suitable for beneficial therapies. There was no inappropriate increase in hospital resource utilization. As such, cTn assays with improved sensitivity provide an opportunity to improve the clinical management of patients with suspected ACS. Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2016. For permissions please email: journals.permissions@oup.com.

[Accumulation of soil organic carbon and total nitrogen in Pinus yunnanensis forests at different age stages].

Science.gov (United States)

Miao, Juan; Zhou, Chuan-Yan; Li, Shi-Jie; Yan, Jun-Hua

2014-03-01

Taking three Pinus yunnanensis forests at different ages (19, 28 and 45 a) in Panxian County of Guizhou Province as test objects, we investigated vertical distributions and accumulation rates of soil organic carbon (SOC) and total nitrogen (TN), as well as their relationships with soil bulk density. For the three forests at different age stages, SOC and TN changed consistently along the soil profile, declining with the soil depth. Both SOC and TN storage increased with the forest age. The SOC and TN storage amounts were 96.24, 121.65 and 148.13 t x hm(-2), and 10.76, 12.96 and 13.08 t x hm(-2) for the forest stands with 19 a, 28 a and 45 a, respectively. SOC had a significant positive correlation with soil TN, while both of them had a significant negative relationship with the soil bulk density. The accumulation rates of both SOC and TN storage at different growth periods were different, and the rate in the period from age 19 to 28 was higher than in the period from age 28 to 45.

Spatial Regression and Prediction of Water Quality in a Watershed with Complex Pollution Sources.

Science.gov (United States)

Yang, Xiaoying; Liu, Qun; Luo, Xingzhang; Zheng, Zheng

2017-08-16

Fast economic development, burgeoning population growth, and rapid urbanization have led to complex pollution sources contributing to water quality deterioration simultaneously in many developing countries including China. This paper explored the use of spatial regression to evaluate the impacts of watershed characteristics on ambient total nitrogen (TN) concentration in a heavily polluted watershed and make predictions across the region. Regression results have confirmed the substantial impact on TN concentration by a variety of point and non-point pollution sources. In addition, spatial regression has yielded better performance than ordinary regression in predicting TN concentrations. Due to its best performance in cross-validation, the river distance based spatial regression model was used to predict TN concentrations across the watershed. The prediction results have revealed a distinct pattern in the spatial distribution of TN concentrations and identified three critical sub-regions in priority for reducing TN loads. Our study results have indicated that spatial regression could potentially serve as an effective tool to facilitate water pollution control in watersheds under diverse physical and socio-economical conditions.

The value of determination of blood cardiac troponin T, IL-8 and TNF-α contents for the diagnosis and prognosis assessment in patients with viral myocarditis

International Nuclear Information System (INIS)

Tao Qian; Zhou Xuanyan; Li Enjie

2006-01-01

Objective: To investigate the value of determination of blood cardiac troponin T (cTnT), IL-8 and TNF-α contents for the diagnosis and prognosis assessment in patients with viral myocarditis (VMC). Methods: Plasma cTnT (with immuno-infiltration method) and serum IL - 8, TNF ( with ELISA) contents were measured in 86 patients with VMC at different stages ( 1 months, 6 months), and 30 controls. Results: The positive rates of blood cTnT, IL-8 and TNF-α in acute VMC patients were significantly higher than those in patients with history less than 6 months as well as with history more than 6 months and the controls group (P 0.05). The cure rate in patients with positive cTnT was significantly lower than that in patients with negative cTnT both at 1 months and 6 months (P<0.005). Conclusion: Blood cTnT, IL-8 and TNF-α are sensitive and specific predictors for diagnosis and prognosis assessment in patients with VMC. (authors)

The relationship of plasma creatinine (as eGFR) and high-sensitivity cardiac troponin and NT-proBNP concentrations in a hospital and community outpatient population.

Science.gov (United States)

Potter, Julia M; Simpson, Aaron J; Kerrigan, Jennifer; Southcott, Emma; Salib, Marie M; Koerbin, Gus; Hickman, Peter E

2017-10-01

While persons with overt renal failure have a well-described rise in troponin and NT-proBNP, it is less well described what the relationship is between cardiac markers and persons with impaired renal function, not requiring dialysis. We have collected ALL samples referred to our pathology practice over a 24h period and measured hs-cTnI, hs-cTnT, NT-proBNP, calculated the eGFR, and related our measurements to clinical outcomes. For both men and women, for all of hs-cTnI, hs-cTnT and NT-proBNP, there was a graded response, as renal function worsened, the concentration of the cardiac marker increased. There is a graded inverse relationship between eGFR and the concentrations of hs-cTnI, hs-cTnT and NT-proBNP. For women only there appeared to be an increase in mortality at lowest eGFR. Copyright © 2017 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Radon and thoron concentrations in traditional cave dwellings and soil beds

International Nuclear Information System (INIS)

Shang Bing; Cui Hongxing; Li Hongzhou

2003-01-01

This paper describes the measured results of 222 Rn and 220 Tn concentrations in six cave dwellings and adobe Kang in Ning county of Gansu Province by using Rn-Tn solid-state nuclear track detectors. 222 Rn and 220 Tn concentrations are 161 and 307 Bq·m -3 (10 cm from soil wall), 99 and 158 Bq·m -3 (middle of the room) in cave dwellings, and 133 and 317 Bq·m -3 (5 cm above adobe Kang), 179 and 145 Bq·m -3 (20 cm above adobe Kang) on soil beds, respectively. 222 Rn and 220 Tn concentrations near the wall are higher than those obtained other place in the room. 222 Rn and 220 Tn concentrations above the adobe Kang are related to the distance from the surface of adobe Kang and heating situation. The residents sleep directly on the beds, especially for the children, the time staying on the adobe Kang are longer than other position. The dose of 222 Rn and 220 Tn from the traditional adobe Kang should be continuously studied

Novel biomarkers for risk stratification in pulmonary arterial hypertension

Directory of Open Access Journals (Sweden)

Thomas Zelniker

2015-10-01

Full Text Available Risk stratification in pulmonary arterial hypertension (PAH is paramount to identifying individuals at highest risk of death. So far, there are only limited parameters for prognostication in patients with PAH. 95 patients with confirmed PAH were included in the present analysis and followed for a total of 4 years. Blood samples were analysed for serum levels of N-terminal pro-brain natriuretic peptide, high-sensitivity troponin T (hsTnT, pro-atrial natriuretic peptide (proANP, growth differentiation factor 15, soluble fms-like tyrosine kinase 1 and placental growth factor. 27 (28.4% patients died during a follow-up of 4 years. Levels of all tested biomarkers, except for placental growth factor, were significantly elevated in nonsurvivors compared with survivors. Receiver operating characteristic analyses demonstrated that cardiac biomarkers had the highest power in predicting mortality. In particular, proANP exhibited the highest area under the curve, followed by N-terminal pro-brain natriuretic peptide and hsTnT. Furthermore, proANP and hsTnT added significant additive prognostic value to the established markers in categorical and continuous net reclassification index. Moreover, after Cox regression, proANP (hazard ratio (HR 1.91, hsTnT (HR 1.41, echocardiographic right ventricular impairment (HR 1.30 and 6-min walk test (HR 0.97 per 10 m remained the only significant parameters in prognostication of mortality. Our data suggest benefits of the implementation of proANP and hsTnT as additive biomarkers for risk stratification in patients with PAH.

A practical approach for the validation and clinical implementation of a high-sensitivity cardiac troponin I assay across a North American city

Directory of Open Access Journals (Sweden)

Peter A. Kavsak

2015-04-01

Full Text Available Objectives: Despite several publications on the analytical performance of high-sensitivity cardiac troponin (hs-cTn assays, there has been little information on how laboratories should validate and implement these assays into clinical service. Our study provides a practical approach for the validation and implementation of a hs-cTn assay across a large North American City. Design and methods: Validation for the Abbott ARCHITECT hs-cTnI assay (across 5 analyzers consisted of verification of limit of blank (LoB, precision (i.e., coefficient of variation; CV testing at the reported limit of detection (LoD and within and outside the 99th percentile, linearity testing, cTnI versus hs-cTnI patient comparison within and between analyzers (Passing and Bablok and non-parametric analyses. Education, clinical communications, and memorandums were issued in advance to inform all staff across the city as well as a selected reminder the day before live-date to important users. All hospitals switched to the hs-cTnI assay concurrently (the contemporary cTnI assay removed with laboratory staff instructed to repeat samples previously measured with the contemporary cTnI assay with the hs-cTnI assay only by physician request. Results: Across the 5 analyzers and 6 reagent packs the overall LoB was 0.6 ng/L (n=60 with a CV of 33% at an overall mean of 1.2 ng/L (n=60; reported LoD=1.0 ng/L, with linearity demonstrated from 45,005 ng/L to 1.1 ng/L. Precision testing with a normal patient-pool QC material (mean range across 5 analyzers was 3.9–4.4 ng/L yielded a range of CVs from 7% to 10% (within-run and CVs from 7% to 18% (between-run with the high patient-pool QC material (mean range across 5 analyzers was 29.6–36.3 ng/L yielding a range of CVs from 2% to 5% (within-run and CVs from 4% to 8% (between-run. There was agreement between hs-cTnI versus cTnI with the patient samples (slope ranges: 0.89–1.03; intercept ranges: 1.9–3

Transport of Spent Nuclear Fuels, High and Intermediate Level Wastes: A Continuous Challenge

International Nuclear Information System (INIS)

Otton, C.; Blachet, L.

2009-01-01

For more than 45 years TN International has been involved in the radioactive materials transportation field. Since the beginning the used nuclear fuel transportation has been its core business. During all these years TN International, now part of AREVA, has been able to anticipate and fulfil the needs for new transport or storage casks design to fit the nuclear industry evolutions. A whole fleet of casks able to transport all the materials of the nuclear fuel cycle has been developed. In this presentation we will focus on the casks for the spent fuel, high level waste and intermediate level waste transportation. Answering to the constant evolution of the nuclear industry transport needs is a challenge that TN International faces routinely. Concerning the spent nuclear fuel transportation, TN International has developed in the early 80's a fleet of TN12 type casks fitted with several types of baskets able to safely transport all the spent fuel from the nuclear power plant or the research laboratories to AREVA La Hague plant. The current challenge is the design of a new transport cask generation taking into account the needs of the industry for the next 30 years. The replacement of the TN12 cask generation is to be scheduled as the regulations have changed and the fuel characteristics have evolved. The new generation of casks will take into account all the technical evolutions made during the TN12 thirty years of use. MOX spent fuel has now its dedicated cask: the TN112 which certificate of approval has been obtained in July 2008. This cask is able to transport 12 MOX spent fuel elements with a short cooling time. The first loading of the cask has been performed in 2008 in the EDF nuclear power plant of Saint-Laurent-des-Eaux. Concerning the high level waste such as the La Hague vitrified residues a whole fleet of casks has been developed such as the TN 28 VT dedicated to transport, the TN81 and TN85 dedicated to transport and storage. These casks have permitted the

Identifying transposon insertions and their effects from RNA-sequencing data.

Science.gov (United States)

de Ruiter, Julian R; Kas, Sjors M; Schut, Eva; Adams, David J; Koudijs, Marco J; Wessels, Lodewyk F A; Jonkers, Jos

2017-07-07

Insertional mutagenesis using engineered transposons is a potent forward genetic screening technique used to identify cancer genes in mouse model systems. In the analysis of these screens, transposon insertion sites are typically identified by targeted DNA-sequencing and subsequently assigned to predicted target genes using heuristics. As such, these approaches provide no direct evidence that insertions actually affect their predicted targets or how transcripts of these genes are affected. To address this, we developed IM-Fusion, an approach that identifies insertion sites from gene-transposon fusions in standard single- and paired-end RNA-sequencing data. We demonstrate IM-Fusion on two separate transposon screens of 123 mammary tumors and 20 B-cell acute lymphoblastic leukemias, respectively. We show that IM-Fusion accurately identifies transposon insertions and their true target genes. Furthermore, by combining the identified insertion sites with expression quantification, we show that we can determine the effect of a transposon insertion on its target gene(s) and prioritize insertions that have a significant effect on expression. We expect that IM-Fusion will significantly enhance the accuracy of cancer gene discovery in forward genetic screens and provide initial insight into the biological effects of insertions on candidate cancer genes. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

The evaluation of B-type Natriuretic Peptide and Troponin I in acute myocardial infarction and unstable angina

Directory of Open Access Journals (Sweden)

Nafija Serdarevic

2014-09-01

Full Text Available Introduction: The diagnostic utility of B-type natriuretic peptide (BNP has prompted interest in its use as an aid in the detection of early heart failure and assessment of diseases. The first objective of this study was measurement of BNP and troponin I (TnI blood levels in patients with acute myocardial infarction (AMI and unstable angina. The second objective of this study was to find a correlation between TnI and BNP in blood.Methods: The concentrations of BNP and TnI in 150 blood levels were determined using CMIA (chemiluminescent microparticle immunoassay Architect and 2000 (Abbott diagnostics. The retrospective study included 100 patients who were hospitalized at the Department of Internal Medicine of the University Clinical Center Sarajevo and 50 healthy control. The reference blood range of BNP is 0-100 pg/mL and TnI is 0.00-0.4 ng/mL.Results: In the patients with AMI the mean value of BNP is 764.48 ± 639.52 pg/mL and TnI is 2.50 ± 2.28ng/mL. The patients with unstable angina have BNP 287.18 ± 593.20 pg/mL and TnI 0.10 ± 0.23 ng/mL. Our studies have shown that the correlation between BNP and TnI was statistically significant for p< 0.05 using Student t test with correlation coefficient r = 0.36. Conclusions: BNP and TnI levels can help to identify the patients with a high risk for cardiovascular diseases.

Prospective comparison of long-term pain relief rates after first-time microvascular decompression and stereotactic radiosurgery for trigeminal neuralgia.

Science.gov (United States)

Wang, Doris D; Raygor, Kunal P; Cage, Tene A; Ward, Mariann M; Westcott, Sarah; Barbaro, Nicholas M; Chang, Edward F

2018-01-01

OBJECTIVE Common surgical treatments for trigeminal neuralgia (TN) include microvascular decompression (MVD), stereotactic radiosurgery (SRS), and radiofrequency ablation (RFA). Although the efficacy of each procedure has been described, few studies have directly compared these treatment modalities on pain control for TN. Using a large prospective longitudinal database, the authors aimed to 1) directly compare long-term pain control rates for first-time surgical treatments for idiopathic TN, and 2) identify predictors of pain control. METHODS The authors reviewed a prospectively collected database for all patients who underwent treatment for TN between 1997 and 2014 at the University of California, San Francisco. Standardized collection of data on preoperative clinical characteristics, surgical procedure, and postoperative outcomes was performed. Data analyses were limited to those patients who received a first-time procedure for treatment of idiopathic TN with > 1 year of follow-up. RESULTS Of 764 surgical procedures performed at the University of California, San Francisco, for TN (364 SRS, 316 MVD, and 84 RFA), 340 patients underwent first-time treatment for idiopathic TN (164 MVD, 168 SRS, and 8 RFA) and had > 1 year of follow-up. The analysis was restricted to patients who underwent MVD or SRS. Patients who received MVD were younger than those who underwent SRS (median age 63 vs 72 years, respectively; p 5 years of follow-up (60 of 164 and 64 of 168 patients, respectively). Immediate or short-term (making depends upon many factors. This information can help physicians counsel patients with idiopathic TN on treatment selection.

Restricted N-terminal truncation of cardiac troponin T: a novel mechanism for functional adaptation to energetic crisis.

Science.gov (United States)

Feng, Han-Zhong; Biesiadecki, Brandon J; Yu, Zhi-Bin; Hossain, M Moazzem; Jin, J-P

2008-07-15

The N-terminal variable region of cardiac troponin T (TnT) is a regulatory structure that can be selectively removed during myocardial ischaemia reperfusion by mu-calpain proteolysis. Here we investigated the pathophysiological significance of this post-translational modification that removes amino acids 1-71 of cardiac TnT. Working heart preparations were employed to study rat acute myocardial infarction and transgenic mouse hearts over-expressing the N-terminal truncated cardiac TnT (cTnT-ND). Ex vivo myocardial infarction by ligation of the left anterior descending coronary artery induced heart failure and produced cTnT-ND not only in the infarct but also in remote zones, including the right ventricular free wall, indicating a whole organ response in the absence of systemic neurohumoral mechanisms. Left ventricular pressure overload in mouse working hearts produced increased cTnT-ND in both ventricles, suggesting a role of haemodynamic stress in triggering an acute whole organ proteolytic regulation. Transgenic mouse hearts in which the endogenous intact cardiac TnT was partially replaced by cTnT-ND showed lowered contractile velocity. When afterload increased from 55 mmHg to 90 mmHg, stroke volume decreased in the wild type but not in the transgenic mouse hearts. Correspondingly, the left ventricular rapid-ejection time of the transgenic mouse hearts was significantly longer than that of wild type hearts, especially at high afterload. The restricted deletion of the N-terminal variable region of cardiac troponin T demonstrates a novel mechanism by which the thin filament regulation adapts to sustain cardiac function under stress conditions.

Characteristics of Nitrogen Loss through Surface-Subsurface Flow on Red Soil Slopes of Southeast China

Science.gov (United States)

Zheng, Haijin; Liu, Zhao; Zuo, Jichao; Wang, Lingyun; Nie, Xiaofei

2017-12-01

Soil nitrogen (N) loss related to surface flow and subsurface flow (including interflow and groundwater flow) from slope lands is a global issue. A lysimetric experiment with three types of land cover (grass cover, GC; litter cover, LC; and bare land, BL) were carried out on a red soil slope land in southeast China. Total Nitrogen (TN) loss through surface flow, interflow and groundwater flow was observed under 28 natural precipitation events from 2015 to 2016. TN concentrations from subsurface flow on BL and LC plots were, on average, 2.7-8.2 and 1.5-4.4 times greater than TN concentrations from surface flow, respectively; the average concentration of TN from subsurface flow on GC was about 36-56% of that recorded from surface flow. Surface flow, interflow and groundwater flow contributed 0-15, 2-9 and 76-96%, respectively, of loss load of TN. Compared with BL, GC and LC intercepted 83-86% of TN loss through surface runoff; GC intercepted 95% of TN loss through subsurface flow while TN loss through subsurface flow on LC is 2.3 times larger than that on BL. In conclusion, subsurface flow especially groundwater flow is the dominant hydrological rout for N loss that is usually underestimated. Grass cover has the high retention of N runoff loss while litter mulch will increase N leaching loss. These findings provide scientific support to control N runoff loss from the red soil slope lands by using suitable vegetation cover and mulching techniques.

Technical Network

CERN Multimedia

2007-01-01

In order to optimise the management of the Technical Network (TN), to facilitate understanding of the purpose of devices connected to the TN and to improve security incident handling, the Technical Network Administrators and the CNIC WG have asked IT/CS to verify the "description" and "tag" fields of devices connected to the TN. Therefore, persons responsible for systems connected to the TN will receive e-mails from IT/CS asking them to add the corresponding information in the network database at "network-cern-ch". Thank you very much for your cooperation. The Technical Network Administrators & the CNIC WG

Gigantic magnetoelectric effect caused by magnetic-field-induced canted antiferromagnetic-paramagnetic transition in quasi-two-dimensional Ca2CoSi2O7 crystal

Science.gov (United States)

Akaki, M.; Tozawa, J.; Akahoshi, D.; Kuwahara, H.

2009-05-01

We have investigated the magnetic and dielectric properties of Ca2CoSi2O7 crystal. The dielectricity and magnetism of Ca2CoSi2O7 are strongly coupled below a canted antiferromagnetic transition temperature (TN). Magnetic fields induce electric polarization below TN. Interestingly, the magnetic-field-induced electric polarization is detected even without poling electric fields. Below TN, a canted antiferromagnetic-paramagnetic transition is induced by magnetic fields. The large magnetocapacitance is observed around TN. The origin of the large magnetocapacitance is due to the magnetic-field-induced the canted antiferromagnetic-paramagnetic transition.

Atrophic changes in the trigeminal nerves of patients with trigeminal neuralgia due to neurovascular compression and their association with the severity of compression and clinical outcomes.

Science.gov (United States)

Leal, Paulo Roberto Lacerda; Barbier, Charlotte; Hermier, Marc; Souza, Miguel Angelo; Cristino-Filho, Gerardo; Sindou, Marc

2014-06-01

The aim of this study was to prospectively evaluate atrophic changes in trigeminal nerves (TGNs) using measurements of volume (V) and cross-sectional area (CSA) from high-resolution 3-T MR images obtained in patients with unilateral trigeminal neuralgia (TN), and to correlate these data with patient and neurovascular compression (NVC) characteristics and with clinical outcomes. Anatomical TGN parameters (V and CSA) were obtained in 50 patients (30 women and 20 men; mean age 56.42 years, range 22-79 years) with classic TN before treatment with microvascular decompression (MVD). Parameters were compared between the symptomatic (ipsilateralTN) and asymptomatic (contralateralTN) sides of the face. Twenty normal control subjects were also included. Two independent observers blinded to the side of pain separately analyzed the images. Measurements of V (from the pons to the entrance of the nerve into Meckel's cave) and CSA (at 5 mm from the entry of the TGN into the pons) for each TGN were performed using imaging software and axial and coronal projections, respectively. These data were correlated with patient characteristics (age, duration of symptoms before MVD, side of pain, sex, and area of pain distribution), NVC characteristics (type of vessel involved in NVC, location of compression along the nerve, site of compression around the circumference of the root, and degree of compression), and clinical outcomes at the 2-year follow-up after surgery. Comparisons were made using Bonferroni's test. Interobserver variability was assessed using the Pearson correlation coefficient. The mean V of the TGN on the ipsilateralTN (60.35 ± 21.74 mm(3)) was significantly smaller (p controls (78.62 ± 24.62 mm(3) and 89.09 ± 14.72 mm(3), respectively). The mean CSA of the TGN on the ipsilateralTN (4.17 ± 1.74 mm(2)) was significantly smaller than those for the contralateralTN and controls (5.41 ± 1.89 mm(2) and 5.64 ± 0.85 mm(2), respectively). The ipsilateralTN with NVC Grade III

Suicidal autointegration of sleeping beauty and piggyBac transposons in eukaryotic cells.

Directory of Open Access Journals (Sweden)

Yongming Wang

2014-03-01

Full Text Available Transposons are discrete segments of DNA that have the distinctive ability to move and replicate within genomes across the tree of life. 'Cut and paste' DNA transposition involves excision from a donor locus and reintegration into a new locus in the genome. We studied molecular events following the excision steps of two eukaryotic DNA transposons, Sleeping Beauty (SB and piggyBac (PB that are widely used for genome manipulation in vertebrate species. SB originates from fish and PB from insects; thus, by introducing these transposons to human cells we aimed to monitor the process of establishing a transposon-host relationship in a naïve cellular environment. Similarly to retroviruses, neither SB nor PB is capable of self-avoidance because a significant portion of the excised transposons integrated back into its own genome in a suicidal process called autointegration. Barrier-to-autointegration factor (BANF1, a cellular co-factor of certain retroviruses, inhibited transposon autointegration, and was detected in higher-order protein complexes containing the SB transposase. Increasing size sensitized transposition for autointegration, consistent with elevated vulnerability of larger transposons. Both SB and PB were affected similarly by the size of the transposon in three different assays: excision, autointegration and productive transposition. Prior to reintegration, SB is completely separated from the donor molecule and followed an unbiased autointegration pattern, not associated with local hopping. Self-disruptive autointegration occurred at similar frequency for both transposons, while aberrant, pseudo-transposition events were more frequently observed for PB.

Molecular characterization and diversity of a novel non-autonomous mutator-like transposon family in brassica

International Nuclear Information System (INIS)

Nouroz, F.

2015-01-01

Transposable elements (TEs) are capable of mobilizing from one genomic location to other, with changes in their copy numbers. Mutator-like elements (MULEs) are DNA transposons characterized by 9 bp target site duplications (TSDs), with high variability in sequence and length, and include non-conserved terminal inverted repeats (TIRs). We identified and characterized a family of Mutator-like elements designated as Shahroz. The structural and molecular analyses revealed that family had a small number of mostly defective non-autonomous MULEs and has shown limited activity in the evolutionary history of the Brassica A-genome. The Shahroz elements range in size from 2734 to 3160 bp including 76 bp imperfect TIRs and 9 bp variable TSDs. The individual copies have shown high homology (52-99%) in their entire lengths. The study revealed that the elements are less in numbers but active in Brassica rapa genomes and PCR amplification revealed their specificity and amplification in A-genome containing diploid and polyploids Brassica. The phylogenetic analysis of Brassica MULEs with other plant Mutator elements revealed that no correlation exists between Brassica MULEs and other elements suggesting a separate line of evolution. Analyzing the regions flanking the insertions revealed that the insertions have showed a preference for AT rich regions. The detailed study of these insertions revealed that although less in number and small sizes, they have played a role in Brassica genome evolution by their mobilization. (author)

Elevated cardiac troponin I predicts cardiovascular or cerebrovascular events in hypertensive patients

International Nuclear Information System (INIS)

Zhang Li'na; Cao Yanfei; Qiu Yifan

2011-01-01

Objective: Whether elevated cTnI is associated with cardiovascular or cerebrovascular events in patients with hypertension (HT) without left ventricular(LV) systolic dysfunction is not clear. Method: We measured cTnI serum level in 170 patients with essential HT without LV systolic dysfunction (LVEF 55%),renal failure,and prior cardiovascular or cerebrovascular diseases. Besides, control group of 40 normal presons was established and following up (45±38)months. Results: Level of cTnI was elevated (≥0.04 ng/ml) in 15 (8.8%) of the 170 patients and in 0 (0%) of the 40 normal controls. The rate of diabetes mellitus(DM), the cardiothoracic ratio, serum NT-proBNP value, and LV mass index were significantly higher in patients with than without elevated cTnI (DM, 9/15 versus 25/155, P 2 , P=0.0001). Kaplan-Meier analysis demonstrated that significantly fewer (P<0.000001) patients with, than without elevated cTnI remained free of events (hospitalization due to cardiovascular or cerebrovascular disease). Stepwise Cox multivariate analysis revealed that elevated cTnT (hazard ratio, 6.59, P=0.000001) and smoking (hazard ratio, 2.26, P=0.04) were independent predictors of events. Conclusion: The present findings indicate that cTnI is a biomarker and useful predictor of future cardiovascular or cerebrovascular events in hypertensive patients. (authors)

Chronic defensiveness and neuroendocrine dysfunction reflect a novel cardiac troponin T cut point: The SABPA study.

Science.gov (United States)

Malan, Leoné; Hamer, Mark; von Känel, Roland; Lambert, Gavin W; Delport, Rhena; Steyn, Hendrik S; Malan, Nicolaas T

2017-11-01

Sympatho-adrenal responses are activated as an innate defense coping (DefS) mechanism during emotional stress. Whether these sympatho-adrenal responses drive cardiac troponin T (cTnT) increases are unknown. Therefore, associations between cTnT and sympatho-adrenal responses were assessed. A prospective bi-ethnic cohort, excluding atrial fibrillation, myocardial infarction and stroke cases, was followed for 3 years (N=342; 45.6±9.0years). We obtained serum high-sensitive cTnT and exposure measures [Coping-Strategy-Indicator, depression/Patient-Health-Questionnarie-9, 24h BP, 24h heart-rate-variability (HRV) and 24h urinary catecholamines]. Blacks showed moderate depression (45% vs. 16%) and 24h hypertension (67% vs. 42%) prevalence compared to Whites. A receiver-operating-characteristics cTnT cut-point 4.2ng/L predicting hypertension in Blacks was used as binary outcome measure in relation to exposure measures [AUC 0.68 (95% CI 0.60-0.76); sensitivity/specificity 63/70%; P≤0.001]. Bi-ethnic cTnT-incidence was similar (Blacks=27%, Whites=25%) with cTnT-recovery better in Blacks (9%) compared to Whites (5%), P=0.001. In cross-sectional analyses, elevated cTnT was related to DefS [OR 1.08 (95% CI 0.99-1.16); P=0.06]; 24h BP [OR 1.03-1.04 (95% CI 1.01-1.08); P≤0.02] and depressed HRV [OR 2.19 (95% CI 1.09-4.41); P=0.03] in Blacks, but not in Whites. At 3year follow-up, elevated cTnT was related to attenuated urine norepinephrine:creatinine ratio in Blacks [OR 1.46 (95% CI 1.01-2.10); P=0.04]. In Whites, a cut point of 5.6ng/L cTnT predicting hypertension was not associated with exposure measures. Central neural control systems exemplified a brain-heart stress pathway. Desensitization of sympatho-adrenal responses occurred with initial neural- (HRV) followed by neuroendocrine dysfunction (norepinephrine:creatinine) in relation to elevated cTnT. Chronic defensiveness may thus drive the desensitization or physiological depression, reflecting ischemic heart disease

Comparação entre troponina I cardíaca e CK-MB massa em síndrome coronariana aguda sem supra de ST

Directory of Open Access Journals (Sweden)

Elizabete Silva dos Santos