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Sample records for erm helgi arst

  1. ERM Ideas and Innovations

    ERIC Educational Resources Information Center

    England, Lenore

    2012-01-01

    Welcome to the new "Journal of Electronic Resources Librarianship" column entitled "ERM Ideas and Innovations," which will appear in each issue of JERL for the next 2 years, and possibly more, beginning with this issue. The overall focus of each column will be to introduce and expand ideas, discuss innovations, and ultimately encourage and foster…

  2. ERM proteins at a glance.

    PubMed

    McClatchey, Andrea I

    2014-08-01

    The cell cortex is a dynamic and heterogeneous structure that governs cell identity and behavior. The ERM proteins (ezrin, radixin and moesin) are major architects of the cell cortex, and they link plasma membrane phospholipids and proteins to the underlying cortical actin cytoskeleton. Recent studies in several model systems have uncovered surprisingly dynamic and complex molecular activities of the ERM proteins and have provided new mechanistic insight into how they build and maintain cortical domains. Among many well-established and essential functions of ERM proteins, this Cell Science at a Glance article and accompanying poster will focus on the role of ERMs in organizing the cell cortex during cell division and apical morphogenesis. These examples highlight an emerging appreciation that the ERM proteins both locally alter the mechanical properties of the cell cortex, and control the spatial distribution and activity of key membrane complexes, establishing the ERM proteins as a nexus for the physical and functional organization of the cell cortex and making it clear that they are much more than scaffolds. This article is part of a Minifocus on Establishing polarity. PMID:24951115

  3. ERM proteins at a glance

    PubMed Central

    McClatchey, Andrea I.

    2014-01-01

    ABSTRACT The cell cortex is a dynamic and heterogeneous structure that governs cell identity and behavior. The ERM proteins (ezrin, radixin and moesin) are major architects of the cell cortex, and they link plasma membrane phospholipids and proteins to the underlying cortical actin cytoskeleton. Recent studies in several model systems have uncovered surprisingly dynamic and complex molecular activities of the ERM proteins and have provided new mechanistic insight into how they build and maintain cortical domains. Among many well-established and essential functions of ERM proteins, this Cell Science at a Glance article and accompanying poster will focus on the role of ERMs in organizing the cell cortex during cell division and apical morphogenesis. These examples highlight an emerging appreciation that the ERM proteins both locally alter the mechanical properties of the cell cortex, and control the spatial distribution and activity of key membrane complexes, establishing the ERM proteins as a nexus for the physical and functional organization of the cell cortex and making it clear that they are much more than scaffolds. This article is part of a Minifocus on Establishing polarity. For further reading, please see related articles: ‘Establishment of epithelial polarity – GEF who's minding the GAP?’ by Siu Ngok et al. (J. Cell Sci. 127, 3205–3215). ‘Integrins and epithelial cell polarity’ by Jessica Lee and Charles Streuli (J. Cell Sci. 127, 3217–3225). PMID:24951115

  4. ERM Ideas and Innovations: Digital Repository Management as ERM

    ERIC Educational Resources Information Center

    Pinkas, María M.; Lin, Na

    2014-01-01

    This article describes the application of electronic resources management (ERM) to digital repository management at the Health Sciences and Human Services Library at the University of Maryland, Baltimore. The authors discuss electronic resources management techniques, through the application of "Techniques for Electronic Management,"…

  5. ERMes: Open Source Simplicity for Your E-Resource Management

    ERIC Educational Resources Information Center

    Doering, William; Chilton, Galadriel

    2009-01-01

    ERMes, the latest version of electronic resource management system (ERM), is a relational database; content in different tables connects to, and works with, content in other tables. ERMes requires Access 2007 (Windows) or Access 2008 (Mac) to operate as the database utilizes functionality not available in previous versions of Microsoft Access. The…

  6. ERM proteins: from cellular architecture to cell signaling.

    PubMed

    Louvet-Vallée, S

    2000-08-01

    ERM (ezrin/radixin/moesin) proteins, concentrated in actin rich cell-surface structures, cross-link actin filaments with the plasma membrane. They are involved in the formation of microvilli, cell-cell adhesion, maintenance of cell shape, cell motility and membrane trafficking. Recent analyses reveal that they are not only involved in cytoskeleton organization but also in signaling pathway. They play an important role in the activation of members of the Rho family by recruiting their regulators. The functions of ERM proteins are regulated by their conformational charges: the intramolecular interaction between the N- and C-terminal domains of ERM proteins charges masks several binding sites, leading to a dormant protein. Different activation signals regulate ERM proteins functions by modulating these intramolecular interactions. The involvement of ERM proteins in many signaling pathways has led to study their role during development of different species. PMID:11071040

  7. New MLSB Resistance Gene erm(43) in Staphylococcus lentus

    PubMed Central

    Schwendener, Sybille

    2012-01-01

    The search for a specific rRNA methylase motif led to the identification of the new macrolide, lincosamide, and streptogramin B resistance gene erm(43) in Staphylococcus lentus. An inducible resistance phenotype was demonstrated by cloning and expressing erm(43) and its regulatory region in Staphylococcus aureus. The erm(43) gene was detected in two different DNA fragments, of 6,230 bp and 1,559 bp, that were each integrated at the same location in the chromosome in several S. lentus isolates of human, dog, and chicken origin. PMID:22733067

  8. ERM proteins regulate growth cone responses to Sema3A

    PubMed Central

    Mintz, C. David; Carcea, Ioana; McNickle, Daniel G.; Dickson, Tracey C.; Ge, Yongchao; Salton, Stephen R.J.; Benson, Deanna L.

    2008-01-01

    Axonal growth cones initiate and sustain directed growth in response to cues in their environment. A variety of events such as receptor internalization, kinase activation, and actin rearrangement can be stimulated by guidance cues and are essential for mediating targeted growth cone behavior. Surprisingly little is known about how such disparate actions are coordinated. Our data suggest that ezrin, radixin, and moesin (ERMs), a family of highly homologous, multifunctional proteins may be able to coordinate growth cone responses to the guidance cue, Sema3A. We show that active ERMs concentrate asymmetrically in neocortical growth cones, are rapidly and transiently inactivated by Sema3A, and are required for Sema3A-mediated growth cone collapse and guidance. The FERM domain of active ERMs regulates internalization of the Sema3A receptor, Npn1 and its co-receptor, L1CAM, while the ERM C-terminal domain binds and caps F-actin. Our data support a model in which ERMs can coordinate membrane and actin dynamics in response to Sema3A. PMID:18651636

  9. ERMS guidebook: Cleaner air, reduced cost, Illinois EPA

    SciTech Connect

    1998-04-01

    The Illinois EPA has prepared this guidebook to help participants in the Emissions Reduction Market System and other interested persons understand how the system is designed and going to be operated. This guidebook serves as a supplement to other important reference materials such as the ERMS regulations (35 Ill. Admin. Code 205) and the Environmental Protection Act (Sec. 98.).

  10. Ecoremediation (ERM) and Saprobiology - is there a link?

    NASA Astrophysics Data System (ADS)

    Svirčev, Zorica; Marković, Slobodan; Krstić, Svetislav; Krstić, Kosta; Obreht, Igor

    2010-05-01

    Healthy environment is rapidly becoming very important value in human life and a criterion of the quality of living. To achieve this target, many of us consider the 'returning to nature' and respecting the ancient laws of nature as the possible solution. Accelerated or fast eutrophization is detected destiny in majority of fresh water ecosystems today, mainly due to global climate changes and adverse human impact. However, this process can be tackled by different activities, ecoremedation (ERM) having an especially important place. Ecoremediation is a system of utilizing the natural processes for revival and protection of the environment. ERM methods may reduce and avert the consequences of agricultural pollution, tourism, transport, industry, land fills and (over)population. In its essence, ERM represents the 'returning to nature' approach aiming to preserve or re-establish the natural balance of the ecosystems, but also a human endevour that enables new jobs and by-side activities important for economic and social (sustainable) development of the human society. In this study we propose the model of eutrophication control achieved on four important levels in the functional dynamics of the ecosystems. ERM is placed as key element in prevention, increasing of decomposition processes through different modes of aeration, decreasing of the total capacity of the ecosystem usually via constructed wetlands, planting and removing of the riparian vegetation.

  11. Core sequence in the RNA motif recognized by the ErmE methyltransferase revealed by relaxing the fidelity of the enzyme for its target.

    PubMed Central

    Hansen, L H; Vester, B; Douthwaite, S

    1999-01-01

    Under physiological conditions, the ErmE methyltransferase specifically modifies a single adenosine within ribosomal RNA (rRNA), and thereby confers resistance to multiple antibiotics. The adenosine (A2058 in Escherichia coli 23S rRNA) lies within a highly conserved structure, and is methylated efficiently, and with equally high fidelity, in rRNAs from phylogenetically diverse bacteria. However, the fidelity of ErmE is reduced when magnesium is removed, and over twenty new sites of ErmE methylation appear in E. coli 16S and 23S rRNAs. These sites show widely different degrees of reactivity to ErmE. The canonical A2058 site is largely unaffected by magnesium depletion and remains the most reactive site in the rRNA. This suggests that methylation at the new sites results from changes in the RNA substrate rather than the methyltransferase. Chemical probing confirms that the rRNA structure opens upon magnesium depletion, exposing potential new interaction sites to the enzyme. The new ErmE sites show homology with the canonical A2058 site, and have the consensus sequence aNNNcgGAHAg (ErmE methylation occurs exclusively at adenosines (underlined); these are preceded by a guanosine, equivalent to G2057; there is a high preference for the adenosine equivalent to A2060; H is any nucleotide except G; N is any nucleotide; and there are slight preferences for the nucleotides shown in lower case). This consensus is believed to represent the core of the motif that Erm methyltransferases recognize at their canonical A2058 site. The data also reveal constraints on the higher order structure of the motif that affect methyltransferase recognition. PMID:9917069

  12. Erythromycin-induced ribosome stall in the ermA leader: a barricade to 5'-to-3' nucleolytic cleavage of the ermA transcript.

    PubMed Central

    Sandler, P; Weisblum, B

    1989-01-01

    The Staphylococcus aureus ermA gene, whose product confers resistance to the macrolide-lincosamide-streptogramin B family of antibiotics, is induced at the level of translation by nanomolar concentrations of erythromycin. Erythromycin also specifically stabilizes ermA transcripts, and the induced stabilization requires in-phase translation of at least one of two small leader peptides in the 5' leader region of the transcript. Erythromycin-induced mRNA stabilization was tested in three constructions in which the ermA transcript was elongated by making insertions at the ermA transcription start. Whereas mRNA downstream of the leader peptide is stabilized by erythromycin, mRNA upstream is not. In the presence of erythromycin, specific mRNA decay intermediates in both the extended ermA genes and the wild-type ermA gene were detected by both Northern blotting and S1 nuclease mapping. The 5' ends of the intermediates map to the sequences that encode each of the two ermA leader peptides, suggesting that the intermediates are produced by stalled erythromycin-bound ribosomes acting as barricades to degradation by 5'-to-3' RNases. In addition, whereas erythromycin was found previously to stabilize ermA transcripts only physically, an ermC-cat-86 hybrid transcript was stabilized both physically and functionally by erythromycin. Images PMID:2592348

  13. Redundant control of migration and adhesion by ERM proteins in vascular smooth muscle cells

    SciTech Connect

    Baeyens, Nicolas; Latrache, Iman; Yerna, Xavier; Noppe, Gauthier; Horman, Sandrine; Morel, Nicole

    2013-11-22

    Highlights: •The three ERM proteins are expressed in vascular smooth muscle cell. •ERM depletion inhibited PDGF-evoked migration redundantly. •ERM depletion increased cell adhesion redundantly. •ERM depletion did not affect PDGF-evoked Ca signal, Rac1 activation, proliferation. •ERM proteins control PDGF-induced migration by regulating adhesion. -- Abstract: Ezrin, radixin, and moesin possess a very similar structure with a C-terminal actin-binding domain and a N-terminal FERM interacting domain. They are known to be involved in cytoskeleton organization in several cell types but their function in vascular smooth muscle cells (VSMC) is still unknown. The aim of this study was to investigate the role of ERM proteins in cell migration induced by PDGF, a growth factor involved in pathophysiological processes like angiogenesis or atherosclerosis. We used primary cultured VSMC obtained from rat aorta, which express the three ERM proteins. Simultaneous depletion of the three ERM proteins with specific siRNAs abolished the effects of PDGF on cell architecture and migration and markedly increased cell adhesion and focal adhesion size, while these parameters were only slightly affected by depletion of ezrin, radixin or moesin alone. Rac1 activation, cell proliferation, and Ca{sup 2+} signal in response to PDGF were unaffected by ERM depletion. These results indicate that ERM proteins exert a redundant control on PDGF-induced VSMC migration by regulating focal adhesion turn-over and cell adhesion to substrate.

  14. NHERF2 is crucial in ERM phosphorylation in pulmonary endothelial cells

    PubMed Central

    2013-01-01

    Background EBP50 and NHERF2 adaptor proteins are incriminated in various signaling pathways of the cell. They can bind ERM proteins and mediate ERM-membrane protein interactions. Results Binding of ERM to EBP50 and NHERF2 was compared in pulmonary artery endothelial cells by immunoprecipitation. NHERF2 associates with all three ERM, but EBP50 appeared to be a weak binding partner if at all. Furthermore, we detected co-localization of NHERF2 and phospho-ERM at the cell membrane and in the filopodia of dividing cells. Silencing of NHERF2 prevented agonist or angiogenesis induced phosphorylation of ERM, while overexpression of the adaptor elevated the phosphorylation level of ERM, likely catalyzed by Rho kinase 2, which co-immunoprecipitated with NHERF2/ERM in control EC, but did not bind to ERM in NHERF2 depleted cells. Dependence of ERM phosphorylation on NHERF2 was also shown in Matrigel tube formation assay, and NHERF2 was proved to be important in angiogenesis as well. Furthermore, when NHERF2 was depleted or cells were overexpressing a mutant form of NHERF2 unable to bind ERM, we found attenuated cell attachment with ECIS measurements, while it was supported by overexpression of wild type NHERF2. Conclusions Pivotal role of NHERF2 in the phosphorylation process of ERM in pulmonary artery endothelial cells is shown. We propose that NHERF2 provides a common anchoring surface for ERM and Rho kinase 2. Our results demonstrate the essential role of NHERF2 in endothelial cell adhesion/migration and angiogenesis. PMID:24364877

  15. Our ASHRM journey continues: ERM for our patients' safety.

    PubMed

    Oppenberg, Andrew A

    2013-01-01

    First and foremost, as your 2013 ASHRM president, I'd like to thank you for all of the significant accomplishments you've helped ASHRM attain this year. And I'll be forever grateful for your support and making my personal and professional dream come true-to serve as your president. Advocating on behalf of all healthcare risk managers and furthering the quest of Getting to Zero for our patients' safety through Enterprise Risk Management, or ERM, has been an honor, because everyone is a risk manager! PMID:24078201

  16. Detection of the macrolide-lincosamide-streptogramin B resistance gene erm(44) and a novel erm(44) variant in staphylococci from aquatic environments.

    PubMed

    Wendlandt, Sarah; Heß, Stefanie; Li, Jun; Feßler, Andrea T; Wang, Yang; Kadlec, Kristina; Gallert, Claudia; Schwarz, Stefan

    2015-08-01

    Resistance to macrolides, lincosamides and streptogramin B antibiotics (MLSB) is not restricted to staphylococci from clinical samples but can also be present in staphylococci from the aquatic environment. Two coagulase-negative staphylococci-Staphylococcus xylosus and S. saprophyticus were obtained from sewage and receiving river water samples and were investigated for the genetic basis of inducible MLSB resistance by whole-genome sequencing. Two rRNA methylases encoded by erm(44) and a novel erm(44) variant were identified, which had only 84% amino acid identity. While fragments of phage DNA were found in the vicinity of the erm(44) gene of S. xylosus, no relics of mobile genetic elements were detected in the sequences flanking the erm(44) variant gene in the S. saprophyticus strain. The functionality of the erm genes was confirmed by cloning and transformation experiments. Based on the obtained sequences, specific PCR assays for both erm genes were developed and used to identify erm(44) in another 7 S. xylosus and 17 S. saprophyticus isolates from aquatic environments. PMID:26207047

  17. Rapid detection of ermB gene in Clostridium difficile by loop-mediated isothermal amplification.

    PubMed

    Lin, Minyi; Liu, Wei; Wang, Pu; Tan, Jiasheng; Zhou, Youlian; Wu, Peiqun; Zhang, Ting; Yuan, Jing; Chen, Ye

    2015-08-01

    Macrolide-lincosamide-streptogramin B resistance in Clostridium difficile is mostly due to the ermB resistance determinant. Here, we describe a sensitive and rapid molecular method to detect ermB in C. difficile to contribute to the wider epidemiological study. Five sets of loop-mediated isothermal amplification (LAMP) primers were designed and optimized for rapid detection of ermB. The specificity and sensitivity of the primers for ermB were detected, and the ermB LAMP assay was compared to conventional PCR with 80 clinical isolates of C. difficile. Real-time monitoring of turbidity and chromogenic reaction were used to determine negative and positive results. A total of 26 pathogenic bacterial strains of different species were found to be negative for ermB, which indicated the high specificity of the primers. ermB was detected in 78.8 % (63/80) of the clinical isolates by both LAMP and conventional PCR. The detection limit of LAMP was 36.1  pg DNA μl- 1 and its sensitivity was 10-fold greater than that of conventional PCR. This study is the first report regarding the development and application of the LAMP assay for detection of the ermB gene in C. difficile strains. The developed LAMP method is sensitive, specific and provides a user-friendly visual approach for the rapid detection of ermB-bearing C. difficile. PMID:26272634

  18. Constitutive and Inducible Expression of the rRNA Methylase Gene erm(B) in Campylobacter

    PubMed Central

    Deng, Fengru; Shen, Jianzhong; Zhang, Maojun; Wu, Congming

    2015-01-01

    Macrolides are the antimicrobials of choice for treating human campylobacteriosis. The recent emergence of erm(B) in Campylobacter bacteria threatens the utility of this class of antibiotics. Here we report the constitutive and inducible expression of erm(B) in Campylobacter isolates derived from diarrheal patients and food-producing animals. Constitutive expression of erm(B) was associated with insertion and deletion in the regulatory region of the gene, providing the first documentation of the differential expression of erm(B) in Campylobacter bacteria. PMID:26259800

  19. FGF2 stimulates SDF-1 expression through the Erm transcription factor in Sertoli cells.

    PubMed

    Yoon, Kyung-Ae; Chae, Young-Mi; Cho, Je-Yoel

    2009-07-01

    Ets-related molecule (Erm) is a member of the Ets transcription factor family. Erm is known to be an important factor for the self-renewal of Spermatogonial stem cells (SSCs) and the maintenance of spermatogenesis. We investigated the molecular mechanism of Erm regulation on SDF-1 in TM4 Sertoli cells. Erm and Sdf-1 levels were up-regulated after FGF2 treatment in TM4 cells, whereas these levels were significantly decreased by FGF2 in ST2 bone marrow stromal cells. Knockdown of Erm by siRNA in the presence of FGF2 decreased the Sdf-1 levels in TM4 cells. The expression levels of Erm were similar and Erm overexpression increased the Sdf-1 in both TM4 and ST2 cells. FGFR subtype analysis revealed that FGFR4 was expressed in TM4 cells but not in ST2 cells. A blocking experiment also confirmed that FGFR4 is partly responsible for the up-regulation of Erm and SDF-1 induced by FGF2 stimulation in TM4 cells. FGF2 and ERM increased the activity of Sdf-1 gene promoter region in a dose-dependent manner. EMSA revealed that ERM strongly binds to the -846 to -851 nucleotide region of the potential Ets binding site (EBS) in the Sdf-1 promoter. In addition, CXCR4, the SDF-1 receptor, was expressed in spermatogonia and Sertoli cells in the seminiferous tubules of the mouse testis. Our results indicate that ERM directly regulates Sdf-1 gene expression by interacting with its cis-acting element in response to FGF2 stimulation in TM4 cells. PMID:19301256

  20. ErmF and ereD Are Responsible for Erythromycin Resistance in Riemerella anatipestifer

    PubMed Central

    Xing, Linlin; Yu, Hui; Qi, Jingjing; Jiang, Pan; Sun, Bingqing; Cui, Junsheng; Ou, Changcan; Chang, Weishan; Hu, Qinghai

    2015-01-01

    To investigate the genetic basis of erythromycin resistance in Riemerella anatipestifer, the MIC to erythromycin of 79 R. anatipestifer isolates from China and one typed strain, ATCC11845, were evaluated. The results showed that 43 of 80 (53.8%) of the tested R. anatipestifer strains showed resistance to erythromycin, and 30 of 43 erythromycin-resistant R. anatipestifer strains carried ermF or ermFU with an MIC in the range of 32–2048 μg/ml, while the other 13 strains carrying the ereD gene exhibited an MIC of 4–16 μg/ml. Of 30 ermF + R. anatipestifer strains, 27 (90.0%) carried the ermFU gene which may have been derived from the CTnDOT-like element, while three other strains carried ermF from transposon Tn4351. Moreover, sequence analysis revealed that ermF, ermFU, and ereD were located within the multiresistance region of the R. anatipestifer genome. PMID:26107936

  1. Identification of ribosomal RNA methyltransferase gene ermF in Riemerella anatipestifer.

    PubMed

    Luo, Hongyan; Liu, Mafeng; Wang, Lanying; Zhou, Wangshu; Wang, Mingshu; Cheng, Anchun; Jia, Renyong; Chen, Shun; Sun, Kunfeng; Yang, Qiao; Chen, Xiaoyue; Zhu, Dekang

    2015-01-01

    Riemerella anatipestifer is a major bacterial pathogen of waterfowl, globally responsible for avian septicaemic disease. As chemotherapy is the predominant method for the prevention and treatment of R. anatipestifer infection in poultry, the widespread use of antibiotics has favoured the emergence of antibiotic-resistant strains. However, little is known about R. anatipestifer susceptibility to macrolide antibiotics and its resistance mechanism. We report for the first time the identification of a macrolide resistance mechanism in R. anatipestifer that is mediated by the ribosomal RNA methyltransferase ermF. We identified the presence of the ermF gene in 64/206 (31%) R. anatipestifer isolates from different regions in China. An ermF deletion strain was constructed to investigate the function of the ermF gene on the resistance to high levels of macrolides. The ermF mutant strain showed significantly decreased resistance to macrolide and lincosamide, exhibiting 1024-, 1024-, 4- and >2048-fold reduction in the minimum inhibitory concentrations for erythromycin, azithromycin, tylosin and lincomycin, respectively. Furthermore, functional analysis of ermF expression in E. coli XL1-blue showed that the R. anatipestifer ermF gene was functional in E. coli XL1-blue and conferred resistance to high levels of erythromycin (100 µg/ml), supporting the hypothesis that the ermF gene is associated with high-level macrolide resistance. Our work suggests that ribosomal RNA modification mediated by the ermF methyltransferase is the predominant mechanism of resistance to erythromycin in R. anatipestifer isolates. PMID:25690020

  2. Characterization of a novel interaction between ELMO1 and ERM proteins.

    PubMed

    Grimsley, Cynthia M; Lu, Mingjian; Haney, Lisa B; Kinchen, Jason M; Ravichandran, Kodi S

    2006-03-01

    ERMs are closely related proteins involved in cell migration, cell adhesion, maintenance of cell shape, and formation of microvilli through their ability to cross-link the plasma membrane with the actin cytoskeleton. ELMO proteins are also known to regulate actin cytoskeleton reorganization through activation of the small GTPbinding protein Rac via the ELMO-Dock180 complex. Here we showed that ERM proteins associate directly with ELMO1 as purified recombinant proteins in vitro and at endogenous levels in intact cells. We mapped ERM binding on ELMO1 to the N-terminal 280 amino acids, which overlaps with the region required for binding to the GTPase RhoG, but is distinct from the C-terminal Dock180 binding region. Consistent with this, ELMO1 could simultaneously bind both radixin and Dock180, although radixin did not alter Rac activation via the Dock180-ELMO complex. Most interestingly, radixin binding did not affect ELMO binding to active RhoG and a trimeric complex of active RhoG-ELMO-radixin could be detected. Moreover, the three proteins colocalized at the plasma membrane. Finally, in contrast to most other ERM-binding proteins, ELMO1 binding occurred independently of the state of radixin C-terminal phosphorylation, suggesting an ELMO1 interaction with both the active and inactive forms of ERM proteins and implying a possible role of ELMO in localizing or retaining ERM proteins in certain cellular sites. Together these data suggest that ELMO1-mediated cytoskeletal changes may be coordinated with ERM protein crosslinking activity during dynamic cellular functions. PMID:16377631

  3. Are all ERMs and ERLs created equal: Is an exceedance by an individual PAH a potential risk?

    SciTech Connect

    Anderson, P.; Pavlou, S.; Titus, E.

    1995-12-31

    One available method to evaluate the potential for concentrations of chemicals in sediments to pose a potential risk to the benthic community, at least on a screening basis, is comparison to ERLs and ERMs. Historically, ERLs and ERMs have been available for mixtures of total, light and heavy PAH. In the past year, ERLs and ERMs have been derived for several individual PAH. This paper identifies some of the possible unintended consequences and inappropriate conclusions that can result when the new ERLs and ERMs are used without regard to their limitations. The limitations arise for several reasons including: the ubiquitous nature of PAH; the varied mixtures from which PAH in sediments may arise; uncertainty in analytical techniques; and, confounding effects of co-occurring chemicals in the samples used to derive ERLs and ERMs for PAH. This paper discusses these limitations and provides guidelines for use of ERLs and ERMs for individual PAH.

  4. SLK-dependent activation of ERMs controls LGN–NuMA localization and spindle orientation

    PubMed Central

    Machicoane, Mickael; de Frutos, Cristina A.; Fink, Jenny; Rocancourt, Murielle; Lombardi, Yannis; Garel, Sonia; Piel, Matthieu

    2014-01-01

    Mitotic spindle orientation relies on a complex dialog between the spindle microtubules and the cell cortex, in which F-actin has been recently implicated. Here, we report that the membrane–actin linkers ezrin/radixin/moesin (ERMs) are strongly and directly activated by the Ste20-like kinase at mitotic entry in mammalian cells. Using microfabricated adhesive substrates to control the axis of cell division, we found that the activation of ERMs plays a key role in guiding the orientation of the mitotic spindle. Accordingly, impairing ERM activation in apical progenitors of the mouse embryonic neocortex severely disturbed spindle orientation in vivo. At the molecular level, ERM activation promotes the polarized association at the mitotic cortex of leucine-glycine-asparagine repeat protein (LGN) and nuclear mitotic apparatus (NuMA) protein, two essential factors for spindle orientation. We propose that activated ERMs, together with Gαi, are critical for the correct localization of LGN–NuMA force generator complexes and hence for proper spindle orientation. PMID:24958772

  5. Ran GTPase promotes oocyte polarization by regulating ERM (Ezrin/Radixin/Moesin) inactivation

    PubMed Central

    Dehapiot, Benoit; Halet, Guillaume

    2013-01-01

    Asymmetric meiotic divisions in mammalian oocytes are driven by the eccentric positioning of the spindle, along with a dramatic reorganization of the overlying cortex, including a loss of microvilli and formation of a thick actin cap. Actin polarization relies on a Ran-GTP gradient centered on metaphase chromosomes; however, the downstream signaling cascade is not completely understood. In a recent study, we have shown that Ran promotes actin cap formation via the polarized activation of Cdc42. The related GTPase Rac is also activated in a polarized fashion in the oocyte cortex and co-localizes with active Cdc42. In other cells, microvilli collapse can be triggered by inactivation of the ERM (Ezrin/Radixin/Moesin) family of actin-membrane crosslinkers under the control of Rac. Accordingly, we show here that Ran-GTP promotes a substantial loss of phosphorylated ERMs in the cortex overlying the spindle in mouse oocytes. However, this polarized phospho-ERM exclusion zone was unaffected by Rac or Cdc42 inhibition. Therefore, we suggest that Ran activates two distinct pathways to regulate actin cap formation and microvilli disassembly in the polarized cortex of mouse oocytes. The possibility of a crosstalk between Rho GTPase and ERM signaling and a role for ERM inactivation in promoting cortical actin dynamics are also discussed. PMID:23656777

  6. Gem1 and ERMES Do Not Directly Affect Phosphatidylserine Transport from ER to Mitochondria or Mitochondrial Inheritance

    PubMed Central

    Nguyen, Tammy T; Lewandowska, Agnieszka; Choi, Jae-Yeon; Markgraf, Daniel F; Junker, Mirco; Bilgin, Mesut; Ejsing, Christer S; Voelker, Dennis R; Rapoport, Tom A; Shaw, Janet M

    2012-01-01

    In yeast, a protein complex termed the ER-Mitochondria Encounter Structure (ERMES) tethers mitochondria to the endoplasmic reticulum. ERMES proteins are implicated in a variety of cellular functions including phospholipid synthesis, mitochondrial protein import, mitochondrial attachment to actin, polarized mitochondrial movement into daughter cells during division, and maintenance of mitochondrial DNA (mtDNA). The mitochondrial-anchored Gem1 GTPase has been proposed to regulate ERMES functions. Here, we show that ERMES and Gem1 have no direct role in the transport of phosphatidylserine (PS) from the ER to mitochondria during the synthesis of phosphatidylethanolamine (PE), as PS to PE conversion is not affected in ERMES or gem1 mutants. In addition, we report that mitochondrial inheritance defects in ERMES mutants are a secondary consequence of mitochondrial morphology defects, arguing against a primary role for ERMES in mitochondrial association with actin and mitochondrial movement. Finally, we show that ERMES complexes are long-lived, and do not depend on the presence of Gem1. Our findings suggest that the ERMES complex may have primarily a structural role in maintaining mitochondrial morphology. PMID:22409400

  7. Small is beautiful: what flies tell us about ERM protein function in development.

    PubMed

    Polesello, Cédric; Payre, François

    2004-06-01

    Actin dynamics is recognized as being a determinant in many developmental processes and pathologies, such as cell polarity, morphogenesis and tumour metastasis. However, how actin interacts with the plasma membrane is poorly understood. Although numerous studies in cell culture point to the crucial role of Ezrin, Radixin and Moesin (ERM) proteins in the actin-membrane link, genetic approaches in mice have not yet revealed their activity during development. Drosophila has recently become an alternative and promising system for the genetic study of ERM protein function. This article focuses on advances made in flies, providing evidence for the evolutionary conservation of functional properties of ERM proteins, in addition to shedding new light on their importance for development. PMID:15183186

  8. Tylosin resistance in Arcanobacterium pyogenes is encoded by an erm X determinant.

    PubMed

    Jost, B Helen; Field, Adam C; Trinh, Hien T; Songer, J Glenn; Billington, Stephen J

    2003-11-01

    Arcanobacterium pyogenes, a commensal on the mucous membranes of many economically important animal species, is also a pathogen, causing abscesses of the skin, joints, and visceral organs as well as mastitis and abortion. In food animals, A. pyogenes is exposed to antimicrobial agents used for growth promotion, prophylaxis, and therapy, notably tylosin, a macrolide antibiotic used extensively for the prevention of liver abscessation in feedlot cattle in the United States. Of 48 A. pyogenes isolates, 11 (22.9%) exhibited inducible or constitutive resistance to tylosin (MIC of > or = 128 microg/ml). These isolates also exhibited resistance to other macrolide and lincosamide antibiotics, suggesting a macrolide-lincosamide resistance phenotype. Of the 11 resistant isolates, genomic DNA from nine hybridized to an erm(X)-specific probe. Cloning and nucleotide sequencing of the A. pyogenes erm(X) gene indicated that it was >95% similar to erm(X) genes from Corynebacterium and Propionibacterium spp. Eight of the erm(X)-containing A. pyogenes isolates exhibited inducible tylosin resistance, which was consistent with the presence of a putative leader peptide upstream of the erm(X) open reading frame. For at least one A. pyogenes isolate, 98-4277-2, erm(X) was present on a plasmid, pAP2, and was associated with the insertion sequence IS6100. pAP2 also carried genes encoding the repressor-regulated tetracycline efflux system determinant Tet 33. The repA gene from pAP2 was nonfunctional in Escherichia coli and at least one A. pyogenes isolate, suggesting that there may be host-encoded factors required for replication of this plasmid. PMID:14576111

  9. Selecting and Implementing an ERMS at Wayne State University: A Case Study

    ERIC Educational Resources Information Center

    Beals, Nancy

    2008-01-01

    The successful development and implementation of an electronic resources management (ERM) system depends on an undertaking that includes many key factors. Some of these key factors include determining user groups and their needs, evaluating implementation and technical issues, testing the system, exploring how the system will be used, setting…

  10. The Brm-HDAC3-Erm repressor complex suppresses dedifferentiation in Drosophila type II neuroblast lineages

    PubMed Central

    Koe, Chwee Tat; Li, Song; Rossi, Fabrizio; Wong, Jack Jing Lin; Wang, Yan; Zhang, Zhizhuo; Chen, Keng; Aw, Sherry Shiying; Richardson, Helena E; Robson, Paul; Sung, Wing-Kin; Yu, Fengwei; Gonzalez, Cayetano; Wang, Hongyan

    2014-01-01

    The control of self-renewal and differentiation of neural stem and progenitor cells is a crucial issue in stem cell and cancer biology. Drosophila type II neuroblast lineages are prone to developing impaired neuroblast homeostasis if the limited self-renewing potential of intermediate neural progenitors (INPs) is unrestrained. Here, we demonstrate that Drosophila SWI/SNF chromatin remodeling Brahma (Brm) complex functions cooperatively with another chromatin remodeling factor, Histone deacetylase 3 (HDAC3) to suppress the formation of ectopic type II neuroblasts. We show that multiple components of the Brm complex and HDAC3 physically associate with Earmuff (Erm), a type II-specific transcription factor that prevents dedifferentiation of INPs into neuroblasts. Consistently, the predicted Erm-binding motif is present in most of known binding loci of Brm. Furthermore, brm and hdac3 genetically interact with erm to prevent type II neuroblast overgrowth. Thus, the Brm-HDAC3-Erm repressor complex suppresses dedifferentiation of INPs back into type II neuroblasts. DOI: http://dx.doi.org/10.7554/eLife.01906.001 PMID:24618901

  11. Characterization of ERM transactivation domain binding to the ACID/PTOV domain of the Mediator subunit MED25

    PubMed Central

    Landrieu, Isabelle; Verger, Alexis; Baert, Jean-Luc; Rucktooa, Prakash; Cantrelle, François-Xavier; Dewitte, Frédérique; Ferreira, Elisabeth; Lens, Zoé; Villeret, Vincent; Monté, Didier

    2015-01-01

    The N-terminal acidic transactivation domain (TAD) of ERM/ETV5 (ERM38–68), a PEA3 group member of Ets-related transcription factors, directly interacts with the ACID/PTOV domain of the Mediator complex subunit MED25. Molecular details of this interaction were investigated using nuclear magnetic resonance (NMR) spectroscopy. The TAD is disordered in solution but has a propensity to adopt local transient secondary structure. We show that it folds upon binding to MED25 and that the resulting ERM–MED25 complex displays characteristics of a fuzzy complex. Mutational analysis further reveals that two aromatic residues in the ERM TAD (F47 and W57) are involved in the binding to MED25 and participate in the ability of ERM TAD to activate transcription. Mutation of a key residue Q451 in the VP16 H1 binding pocket of MED25 affects the binding of ERM. Furthermore, competition experiments show that ERM and VP16 H1 share a common binding interface on MED25. NMR data confirms the occupancy of this binding pocket by ERM TAD. Based on these experimental data, a structural model of a functional interaction is proposed. This study provides mechanistic insights into the Mediator–transactivator interactions. PMID:26130716

  12. First Report of Macrolide Resistance Gene erm(T) Harbored by a Novel Small Plasmid from Erysipelothrix rhusiopathiae

    PubMed Central

    Xu, Chang-Wen; Zhang, An-Yun; Yang, Chun-Mei; Pan, Yun; Guan, Zhong-Bin; Lei, Chang-Wei; Peng, Lin-Yao; Li, Qing-Zhou

    2015-01-01

    The macrolide resistance gene erm(T) was identified for the first time in a porcine Erysipelothrix rhusiopathiae isolate from swine in China. The novel 3,749-bp small plasmid pER29, which carries erm(T), had a G+C content of 31% and four distinct open reading frames. The presence of pER29 increased by at least 128-fold the MICs of clindamycin and erythromycin for E. rhusiopathiae. The fitness cost of pER29 could be responsible for the low frequency of erm(T) in E. rhusiopathiae. PMID:25666150

  13. Conserved SMP domains of the ERMES complex bind phospholipids and mediate tether assembly

    PubMed Central

    AhYoung, Andrew P.; Jiang, Jiansen; Zhang, Jiang; Khoi Dang, Xuan; Loo, Joseph A.; Zhou, Z. Hong; Egea, Pascal F.

    2015-01-01

    Membrane contact sites (MCS) between organelles are proposed as nexuses for the exchange of lipids, small molecules, and other signals crucial to cellular function and homeostasis. Various protein complexes, such as the endoplasmic reticulum-mitochondrial encounter structure (ERMES), function as dynamic molecular tethers between organelles. Here, we report the reconstitution and characterization of subcomplexes formed by the cytoplasm-exposed synaptotagmin-like mitochondrial lipid-binding protein (SMP) domains present in three of the five ERMES subunits—the soluble protein Mdm12, the endoplasmic reticulum (ER)-resident membrane protein Mmm1, and the mitochondrial membrane protein Mdm34. SMP domains are conserved lipid-binding domains found exclusively in proteins at MCS. We show that the SMP domains of Mdm12 and Mmm1 associate into a tight heterotetramer with equimolecular stoichiometry. Our 17-Å-resolution EM structure of the complex reveals an elongated crescent-shaped particle in which two Mdm12 subunits occupy symmetric but distal positions at the opposite ends of a central ER-anchored Mmm1 homodimer. Rigid body fitting of homology models of these SMP domains in the density maps reveals a distinctive extended tubular structure likely traversed by a hydrophobic tunnel. Furthermore, these two SMP domains bind phospholipids and display a strong preference for phosphatidylcholines, a class of phospholipids whose exchange between the ER and mitochondria is essential. Last, we show that the three SMP-containing ERMES subunits form a ternary complex in which Mdm12 bridges Mmm1 to Mdm34. Our findings highlight roles for SMP domains in ERMES assembly and phospholipid binding and suggest a structure-based mechanism for the facilitated transport of phospholipids between organelles. PMID:26056272

  14. Ezrin, Radixin, and Moesin (ERM) proteins function as pleiotropic regulators of human immunodeficiency virus type 1 infection

    SciTech Connect

    Kubo, Yoshinao Yoshii, Hiroaki; Kamiyama, Haruka; Tominaga, Chika; Tanaka, Yuetsu; Sato, Hironori; Yamamoto, Naoki

    2008-05-25

    Ezrin, radixin, and moesin (ERM) proteins supply functional linkage between integral membrane proteins and cytoskeleton in mammalian cells to regulate membrane protein dynamisms and cytoskeleton rearrangement. To assess potential role of the ERM proteins in HIV-1 lifecycle, we examined if suppression of ERM function in human cells expressing HIV-1 infection receptors influences HIV-1 envelope (Env)-mediated HIV-1-vector transduction and cell-cell fusion. Expression of an ezrin dominant negative mutant or knockdown of ezrin, radixin, or moesin with siRNA uniformly decreased transduction titers of HIV-1 vectors having X4-tropic Env. In contrast, transduction titers of R5-tropic Env HIV-1 vectors were decreased only by radixin knockdown: ezrin knockdown had no detectable effects and moesin knockdown rather increased transduction titer. Each of the ERM suppressions had no detectable effects on cell surface expression of CD4, CCR5, and CXCR4 or VSV-Env-mediated HIV-1 vector transductions. Finally, the individual knockdown of ERM mRNAs uniformly decreased efficiency of cell-cell fusion mediated by X4- or R5-tropic Env and HIV-1 infection receptors. These results suggest that (i) the ERM proteins function as positive regulators of infection by X4-tropic HIV-1, (ii) moesin additionally functions as a negative regulator of R5-tropic HIV-1 virus infection at the early step(s) after the membrane fusion, and (iii) receptor protein dynamisms are regulated differently in R5- and X4-tropic HIV-1 infections.

  15. Noise-induced cochlear F-actin depolymerization is mediated via ROCK2/p-ERM signaling

    PubMed Central

    Han, Yu; Wang, Xianren; Chen, Jun; Sha, Su-Hua

    2015-01-01

    Our previous work has suggested that traumatic noise activates Rho-GTPase pathways in cochlear outer hair cells (OHCs), resulting in cell death and noise-induced hearing loss (NIHL). In this study, we investigated Rho effectors, Rho-associated kinases (ROCKs), and the targets of ROCKs, the ezrin-radixin-moesin (ERM) proteins, in the regulation of the cochlear actin cytoskeleton using adult CBA/J mice under conditions of noise-induced temporary threshold shift (TTS) and permanent threshold shift (PTS) hearing loss, which result in changes to the F/G-actin ratio. The levels of cochlear ROCK2 and p-ERM decreased 1 h after either TTS- or PTS-noise exposure. In contrast, ROCK2 and p-ERM in OHCs decreased only after PTS-, not after TTS-noise exposure. Treatment with lysophosphatidic acid, an activator of the Rho pathway, resulted in significant reversal of the F/G-actin ratio changes caused by noise exposure and attenuated OHC death and NIHL. Conversely, the down-regulation of ROCK2 by pretreatment with ROCK2 siRNA reduced the expression of ROCK2 and p-ERM in OHCs, exacerbated TTS to PTS, and worsened OHC loss. Additionally, pretreatment with siRNA against radixin, an ERM protein, aggravated TTS to PTS. Our results indicate that a ROCK2-mediated ERM-phosphorylation signaling cascade modulates noise-induced hair cell loss and NIHL by targeting the cytoskeleton. PMID:25683353

  16. A piRNA-like small RNA interacts with and modulates p-ERM proteins in human somatic cells

    PubMed Central

    Mei, Yuping; Wang, Yuyan; Kumari, Priti; Shetty, Amol Carl; Clark, David; Gable, Tyler; MacKerell, Alexander D.; Ma, Mark Z.; Weber, David J.; Yang, Austin J.; Edelman, Martin J.; Mao, Li

    2015-01-01

    PIWI-interacting RNAs (piRNAs) are thought to silence transposon and gene expression during development. However, the roles of piRNAs in somatic tissues are largely unknown. Here we report the identification of 555 piRNAs in human lung bronchial epithelial (HBE) and non-small cell lung cancer (NSCLC) cell lines, including 295 that do not exist in databases termed as piRNA-like sncRNAs or piRNA-Ls. Distinctive piRNA/piRNA-L expression patterns are observed between HBE and NSCLC cells. piRNA-like-163 (piR-L-163), the top downregulated piRNA-L in NSCLC cells, binds directly to phosphorylated ERM proteins (p-ERM), which is dependent on the central part of UUNNUUUNNUU motif in piR-L-163 and the RRRKPDT element in ERM. The piR-L-163/p-ERM interaction is critical for p-ERM's binding capability to filamentous actin (F-actin) and ERM-binding phosphoprotein 50 (EBP50). Thus, piRNA/piRNA-L may play a regulatory role through direct interaction with proteins in physiological and pathophysiological conditions. PMID:26095918

  17. TcdC does not significantly repress toxin expression in Clostridium difficile 630ΔErm.

    PubMed

    Bakker, Dennis; Smits, Wiep Klaas; Kuijper, Ed J; Corver, Jeroen

    2012-01-01

    In the past decade, Clostridium difficile has emerged as an important gut pathogen. Symptoms of C. difficile infection range from mild diarrhea to pseudomembranous colitis, sometimes resulting in colectomy or death. The main virulence factors of C. difficile are toxin A and toxin B. Besides the genes encoding these toxins (tcdA and tcdB), the pathogenicity locus (PaLoc) also contains genes encoding a sigma factor (tcdR) and a putative anti-sigma factor (tcdC). The important role of TcdR as a sigma factor for toxin expression is undisputed, whereas the role of TcdC as an anti-sigma factor, inhibiting toxin expression, is currently the subject of debate. To clarify the role of TcdC in toxin expression, we generated an isogenic ClosTron-based mutant of tcdC in Clostridium difficile strain 630Δ Erm (CT::tcdC) and determined the transcription levels of the PaLoc genes and the expression levels of the toxins in the wild type strain and the tcdC mutant strain. We found only minor differences in transcription levels of the PaLoc genes between the wild type and CT::tcdC strains and total toxin levels did not significantly differ either. These results suggest that in C. difficile 630Δerm TcdC is not a major regulator of toxin expression under the conditions tested. PMID:22912837

  18. The New Macrolide-Lincosamide-Streptogramin B Resistance Gene erm(45) Is Located within a Genomic Island in Staphylococcus fleurettii

    PubMed Central

    Wipf, Juliette R. K.; Schwendener, Sybille; Nielsen, Jesper Boye; Westh, Henrik

    2015-01-01

    Genome alignment of a macrolide, lincosamide, and streptogramin B (MLSB)-resistant Staphylococcus fleurettii strain with an MLSB-susceptible S. fleurettii strain revealed a novel 11,513-bp genomic island carrying the new erythromycin resistance methylase gene erm(45). This gene was shown to confer inducible MLSB resistance when cloned into Staphylococcus aureus. The erm(45)-containing island was integrated into the housekeeping gene guaA in S. fleurettii and was able to form a circular intermediate but was not transmissible to S. aureus. PMID:25779586

  19. Transport and persistence of tylosin-resistant enterococci, erm genes, and tylosin in soil and drainage water from fields receiving swine manure

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Land application of manure from tylosin-treated swine introduces tylosin-resistant enterococci, erm genes, which confer resistance to tylosin, and tylosin. This study documents the occurrence and transport of tylosin-resistant enterococci, erm genes, and tylosin in tile-drained chisel plow and no-ti...

  20. The development of an on-line ERM system for the research reactors in Korea

    NASA Astrophysics Data System (ADS)

    Kim, Hee Reyoung; Lee, Wanno; Kim, Eun Han; Choi, Geun Sik; Lee, Chang Woo

    2007-08-01

    A real-time on-line environmental radiation monitoring (ERM) system for the research reactor sites of Daejeon and Seoul is established. In the Daejeon site, a radio communication method with a radiofrequency of 468.8 MHz is used between the main computer and the six posts inside the Daejeon research reactor site. A general telephone communication method by a dial modem is applied between the main computer and a comparison point with one post outside the Daejeon research reactor site. In the Seoul site, a null modem communication method is employed between a sub-computer and the three posts inside the Seoul research reactor site, and a high-speed communication network such as ADSL is used between the sub-computer in the Seoul site and the main computer in the Daejeon site. Consequently, the real-time data from a total of 10 places is displayed on-line on a screen and it is statistically treated.

  1. Science applications of the multi-FOV lidar for ATMOS-B1/ERM

    NASA Astrophysics Data System (ADS)

    Sugimoto, Nobuo; Liu, Zhaoyan; Voelger, Peter; Shimizu, Atsushi; Sasano, Yasuhiro; Asai, Kazuhiro; Ishizu, Mitsuo; Itabe, Toshikasu; Imai, Tadashi

    2001-02-01

    A new multi-FOV space-borne lidar named "A-lidar" is being studied by the National Space Development Agency of Japan (NASDA) for the earth radiation mission proposed as a joint program with the European Space Agency (ESA). The mission is named "EarthCARE". It was formerly called ATMOS-B1 or ERM. The lidar has a two-wavelength transmitter (1064 nm and 532 nm), a dual polarization receiver at 1064 nm, and a multi-field-of-view (multi-FOV) receiver at 532 nm. The multi-FOV feature of A-lidar will enable us to solve the multiple scattering problems with space lidar measurements of profiles of clouds and aerosols. The multi-FOV feature can also be used for characterization of aerosols.

  2. Integration of erm(B)-containing elements through large chromosome fragment exchange in Clostridium difficile

    PubMed Central

    Wasels, François; Spigaglia, Patrizia; Barbanti, Fabrizio; Monot, Marc; Villa, Laura; Dupuy, Bruno; Carattoli, Alessandra; Mastrantonio, Paola

    2015-01-01

    In Clostridium difficile, erm(B) genes are located on mobile elements like Tn5398 and Tn6215. In previous studies, some of these elements were transferred by conjugation-like mechanisms, mobilized in trans by helper conjugative systems. In this study, we analyzed the genomes of several recipient strains that acquired either Tn5398 or Tn6215-like elements. We demonstrated that the integration of the transposons in the genome of the recipient cell was always due to homologous recombination events, involving exchange of large chromosomal segments. We did not observed transposon transfer to a C. difficile strain in presence of DNAse, suggesting that a possible transformation-like mechanism occurred in this recipient.

  3. Opening a Can of wERMS: Texas A&M University's Experiences in Implementing Two Electronic Resource Management Systems

    ERIC Educational Resources Information Center

    Hartnett, Eric; Price, Apryl; Smith, Jane; Barrett, Michael

    2010-01-01

    Over the past few years, Texas A&M University (TAMU) has searched for a way to administer its electronic subscriptions as well as the electronic subscriptions shared among the TAMU System. In this article, we address our attempts to implement an effective electronic resource management system (ERMS), both for subscriptions on the main campus and…

  4. The Novel Macrolide-Lincosamide-Streptogramin B Resistance Gene erm(44) Is Associated with a Prophage in Staphylococcus xylosus

    PubMed Central

    Wipf, Juliette R. K.; Schwendener, Sybille

    2014-01-01

    A novel erythromycin ribosome methylase gene, erm(44), that confers resistance to macrolide, lincosamide, and streptogramin B (MLSB) antibiotics was identified by whole-genome sequencing of the chromosome of Staphylococcus xylosus isolated from bovine mastitis milk. The erm(44) gene is preceded by a regulatory sequence that encodes two leader peptides responsible for the inducible expression of the methylase gene, as demonstrated by cloning in Staphylococcus aureus. The erm(44) gene is located on a 53-kb putative prophage designated ΦJW4341-pro. The 56 predicted open reading frames of ΦJW4341-pro are structurally organized into the five functional modules found in members of the family Siphoviridae. ΦJW4341-pro is site-specifically integrated into the S. xylosus chromosome, where it is flanked by two perfect 19-bp direct repeats, and exhibits the ability to circularize. The presence of erm(44) in three additional S. xylosus strains suggests that this putative prophage has the potential to disseminate MLSB resistance. PMID:25092709

  5. The ETS family member ERM contains an alpha-helical acidic activation domain that contacts TAFII60.

    PubMed Central

    Defossez, P A; Baert, J L; Monnot, M; de Launoit, Y

    1997-01-01

    Transcription factors are modular entities built up of discrete domains, some devoted to DNA binding and others permitting transcriptional modulation. The structure of DNA binding domains has been thoroughly investigated and structural classes clearly defined. In sharp contrast, the structural constraints put on transactivating regions, if any, are mostly unknown. Our investigations focus on ERM, a eukaryotic transcription factor of the ETS family. We have previously shown that ERM harbours two transactivating domains (TADs) with distinct functional features: AD1 lies in the first 72 amino acids of ERM, while AD2 sits in the last 62. Here we show that AD1 is a bona fide acidic TAD, for it activated transcription in yeast cells, while AD2 did not. AD1 contains a 20 amino acid stretch predicted to form an alpha-helix that is found unchanged in the related PEA3 and ER81 transcription factors. Circular dichroism analysis revealed that a 32 amino acid peptide encompassing this region is unstructured in water but folds into a helix when the hydrophobic solvent trifluoroethanol is added. The isolated helix was sufficient to activate transcription and mutations predicted to disrupt it dramatically affected AD1-driven transactivation, whereas mutations decreasing its acidity had more gentle effects. A phenylalanine residue within the helix was particularly sensitive to mutations. Finally, we observed that ERM bound TAFII60 via AD1 and bound TBP and TAFII40, presumably via other activation domains. PMID:9358152

  6. Characterization of a new erm-related macrolide resistance gene present in probiotic strains of Bacillus clausii.

    PubMed

    Bozdogan, Bülent; Galopin, Sébastien; Leclercq, Roland

    2004-01-01

    The mechanism of resistance to macrolides, lincosamides, and streptogramins B was studied in four Bacillus clausii strains that are mixed in a probiotic administered to humans for prevention of gastrointestinal side effects due to oral antibiotic chemotherapy and in three reference strains of B. clausii, DSM8716, ATCC 21536, and ATCC 21537. An 846-bp gene called erm(34), which is related to the erm genes conferring resistance to these antibiotics by ribosomal methylation, was cloned from total DNA of B. clausii DSM8716 into Escherichia coli. The deduced amino acid sequence presented 61% identity with that of Erm(D) from B. licheniformis, B. halodurans, and B. anthracis. Pulsed-field gel electrophoresis of total DNA digested by I-CeuI, followed by hybridization with an erm(34)-specific probe, indicated a chromosomal location of the gene in all B. clausii strains. Repeated attempts to transfer resistance to macrolides by conjugation from B. clausii strains to Enterococcus faecalis JH2-2, E. faecium HM1070, and B. subtilis UCN19 were unsuccessful. PMID:14711653

  7. The Endoplasmic Reticulum-Mitochondrion Tether ERMES Orchestrates Fungal Immune Evasion, Illuminating Inflammasome Responses to Hyphal Signals

    PubMed Central

    Tucey, Timothy M.; Verma-Gaur, Jiyoti; Nguyen, Julie; Hewitt, Victoria L.; Lo, Tricia L.; Shingu-Vazquez, Miguel; Robertson, Avril A. B.; Hill, James R.; Pettolino, Filomena A.; Beddoe, Travis; Cooper, Matthew A.; Naderer, Thomas

    2016-01-01

    ABSTRACT The pathogenic yeast Candida albicans escapes macrophages by triggering NLRP3 inflammasome-dependent host cell death (pyroptosis). Pyroptosis is inflammatory and must be tightly regulated by host and microbe, but the mechanism is incompletely defined. We characterized the C. albicans endoplasmic reticulum (ER)-mitochondrion tether ERMES and show that the ERMES mmm1 mutant is severely crippled in killing macrophages despite hyphal formation and normal phagocytosis and survival. To understand dynamic inflammasome responses to Candida with high spatiotemporal resolution, we established live-cell imaging for parallel detection of inflammasome activation and pyroptosis at the single-cell level. This showed that the inflammasome response to mmm1 mutant hyphae is delayed by 10 h, after which an exacerbated activation occurs. The NLRP3 inhibitor MCC950 inhibited inflammasome activation and pyroptosis by C. albicans, including exacerbated inflammasome activation by the mmm1 mutant. At the cell biology level, inactivation of ERMES led to a rapid collapse of mitochondrial tubular morphology, slow growth and hyphal elongation at host temperature, and reduced exposed 1,3-β-glucan in hyphal populations. Our data suggest that inflammasome activation by C. albicans requires a signal threshold dependent on hyphal elongation and cell wall remodeling, which could fine-tune the response relative to the level of danger posed by C. albicans. The phenotypes of the ERMES mutant and the lack of conservation in animals suggest that ERMES is a promising antifungal drug target. Our data further indicate that NLRP3 inhibition by MCC950 could modulate C. albicans-induced inflammation. IMPORTANCE The yeast Candida albicans causes human infections that have mortality rates approaching 50%. The key to developing improved therapeutics is to understand the host-pathogen interface. A critical interaction is that with macrophages: intracellular Candida triggers the NLRP3/caspase-1

  8. The Endoplasmic Reticulum-Mitochondrion Tether ERMES Orchestrates Fungal Immune Evasion, Illuminating Inflammasome Responses to Hyphal Signals.

    PubMed

    Tucey, Timothy M; Verma-Gaur, Jiyoti; Nguyen, Julie; Hewitt, Victoria L; Lo, Tricia L; Shingu-Vazquez, Miguel; Robertson, Avril A B; Hill, James R; Pettolino, Filomena A; Beddoe, Travis; Cooper, Matthew A; Naderer, Thomas; Traven, Ana

    2016-01-01

    The pathogenic yeast Candida albicans escapes macrophages by triggering NLRP3 inflammasome-dependent host cell death (pyroptosis). Pyroptosis is inflammatory and must be tightly regulated by host and microbe, but the mechanism is incompletely defined. We characterized the C. albicans endoplasmic reticulum (ER)-mitochondrion tether ERMES and show that the ERMES mmm1 mutant is severely crippled in killing macrophages despite hyphal formation and normal phagocytosis and survival. To understand dynamic inflammasome responses to Candida with high spatiotemporal resolution, we established live-cell imaging for parallel detection of inflammasome activation and pyroptosis at the single-cell level. This showed that the inflammasome response to mmm1 mutant hyphae is delayed by 10 h, after which an exacerbated activation occurs. The NLRP3 inhibitor MCC950 inhibited inflammasome activation and pyroptosis by C. albicans, including exacerbated inflammasome activation by the mmm1 mutant. At the cell biology level, inactivation of ERMES led to a rapid collapse of mitochondrial tubular morphology, slow growth and hyphal elongation at host temperature, and reduced exposed 1,3-β-glucan in hyphal populations. Our data suggest that inflammasome activation by C. albicans requires a signal threshold dependent on hyphal elongation and cell wall remodeling, which could fine-tune the response relative to the level of danger posed by C. albicans. The phenotypes of the ERMES mutant and the lack of conservation in animals suggest that ERMES is a promising antifungal drug target. Our data further indicate that NLRP3 inhibition by MCC950 could modulate C. albicans-induced inflammation. IMPORTANCE The yeast Candida albicans causes human infections that have mortality rates approaching 50%. The key to developing improved therapeutics is to understand the host-pathogen interface. A critical interaction is that with macrophages: intracellular Candida triggers the NLRP3/caspase-1 inflammasome

  9. Hotspot-ridge interaction in the Indian Ocean: constraints from Geosat/ERM altimetry

    NASA Astrophysics Data System (ADS)

    Grevemeyer, Ingo

    1996-09-01

    Upper-mantle structure of Indian Ocean spreading ridges was investigated by track segments of Geosat/ERM altimeter measurements. To determine the upper-mantle structure of the Earth's gravity field, a low-degree and -order spherical harmonic representation of the geoid was removed. A test of several reference fields suggested that a degree 2-25 field with gradually rolled off coefficients (Sandwell & Renkin 1988) offers an adequate representation of the long-wavelength geoidal undulations. Filtered profiles of three individual ridge segments display a strong asymmetry in geoid versus age trends of conjugated rift flanks. The unusually low geoid slopes on one flank can perhaps best be explained as a dynamic or thermal phenomenon reflecting a flow connection between a neighbouring off-axis hotspot plume and the ridge axis, while the other flank simply cools as it spreads away from the axial zone. It seems reasonable to hypothesize that the Southwest Indian Ridge and the Southeast Indian Ridge act as sinks for plumes beneath Agulhas Plateau and Kerguelen Islands, respectively. The Carlsberg Ridge data suggest that the Réunion hotspot contaminated northwestern African lithosphere until 15 Ma. Moreover, symmetric flattening of geoid versus age trends of conjugated ridge flanks offers evidence that plume events affect geoid versus age trends

  10. Structure of the ERM protein moesin reveals the FERM domain fold masked by an extended actin binding tail domain.

    PubMed

    Pearson, M A; Reczek, D; Bretscher, A; Karplus, P A

    2000-04-28

    The ezrin-radixin-moesin (ERM) protein family link actin filaments of cell surface structures to the plasma membrane, using a C-terminal F-actin binding segment and an N-terminal FERM domain, a common membrane binding module. ERM proteins are regulated by an intramolecular association of the FERM and C-terminal tail domains that masks their binding sites. The crystal structure of a dormant moesin FERM/tail complex reveals that the FERM domain has three compact lobes including an integrated PTB/PH/ EVH1 fold, with the C-terminal segment bound as an extended peptide masking a large surface of the FERM domain. This extended binding mode suggests a novel mechanism for how different signals could produce varying levels of activation. Sequence conservation suggests a similar regulation of the tumor suppressor merlin. PMID:10847681

  11. Experiments and simulated calculations on the resistance to low-velocity impact of layered plates with a sandwiched ERM

    NASA Astrophysics Data System (ADS)

    Zhao, Yinyan; Xiao, Tianyuan; Xue, Sixin; Dong, Janhua

    2004-04-01

    Comparison experiments and simulated calculations are conducted on the resistance to low-velocity (or low-energy) impact of layered structures with and without an electro-rheological material (ERM) sandwiched, under different electric voltages applied to the ERM layer. From the experiments, it is found that the stiffness of the specimen under different electric intensities applied to the ERM layer is approximately a constant. From the calculations, within the range of 0.0 kV/mm⩽ E⩽3.5 kV/mm, the resistance to impact decreases somewhat with the increasing electric intensity for the layered composite specimens. The same conclusion is obtained for a layered aluminum plate within the range of 0.0 kV/mm⩽ E⩽0.75 kV/mm. Meanwhile, data from the experiments and calculations show that these results are repeatable under different impact velocities. Further analysis by computations shows that the change of viscous proportional damping, [ C]= α[ M]+ β[ C], is the main cause of the reduction of the specimen's resistance to impact, where the stiffness coefficient β is the key factor.

  12. Time-to-Detection of Inducible Macrolide Resistance in Mycobacterium abscessus Subspecies and Its Association with the Erm(41) Sequevar

    PubMed Central

    Christianson, Sara; Grierson, William; Kein, Daniel; Tyler, Andrea D.; Wolfe, Joyce; Sharma, Meenu K.

    2016-01-01

    Mutations in the erm(41) gene of M.abscessus group organisms are associated with differences in inducible macrolide resistance, with current recommendations being to hold rapidly growing isolates for up to 14 days in order to ensure that resistance which develops more slowly can be detected. This study aimed to determine the ideal incubation time for accurate identification of inducible macrolide resistance as well as to determine if there was an association between the time taken to detect inducible resistance in M.abscessus group organisms and their erm(41) sequevar. We amplified and sequenced the erm(41) genes of a total of 104 M.abscessus group isolates and determined their sequevars. The isolates were tested for phenotypic clarithromycin resistance at days 7, 10, 14 and 21, using Trek Diagnostics Sensititre RAPMYCO microbroth dilution plates. Associations between erm(41) gene sequevars and time to detection of resistance were evaluated using Fisher’s exact test in R. The samples included in this study fell into 14 sequevars, with the majority of samples falling into Sequevar02 (16), Sequevar06 (15), Sequevar08 (7) and Sequvar 15 (31), and several isolates that were in small clusters, or unique. The majority (82.7%) of samples exhibiting inducible macrolide resistance were interpreted as resistant by day 7. Two isolates in Sequevar02, which has a T28C mutation that is associated with sensitivity, showed intermediate resistance at day 14, though the majority (13) were sensitive at day 14. The majority of isolates with inducible macrolide resistance fell into Sequevars 06,08 and 15, none of which contain the T28C mutation. These sequevars were analyzed to determine if there was any correlation between sequevar and time to detection of resistance. None was found. Based on these findings, we recommend the addition of a day 7 read to the CLSI guidelines to improve turn-around-times for these isolates. It is also recommended that erm(41) gene sequencing be added to

  13. Advanced order management in ERM systems: the tic-tac-toe algorithm

    NASA Astrophysics Data System (ADS)

    Badell, Mariana; Fernandez, Elena; Puigjaner, Luis

    2000-10-01

    The concept behind improved enterprise resource planning systems (ERP) systems is the overall integration of the whole enterprise functionality into the management systems through financial links. Converting current software into real management decision tools requires crucial changes in the current approach to ERP systems. This evolution must be able to incorporate the technological achievements both properly and in time. The exploitation phase of plants needs an open web-based environment for collaborative business-engineering with on-line schedulers. Today's short lifecycles of products and processes require sharp and finely tuned management actions that must be guided by scheduling tools. Additionally, such actions must be able to keep track of money movements related to supply chain events. Thus, the necessary outputs require financial-production integration at the scheduling level as proposed in the new approach of enterprise management systems (ERM). Within this framework, the economical analysis of the due date policy and its optimization become essential to manage dynamically realistic and optimal delivery dates with price-time trade-off during the marketing activities. In this work we propose a scheduling tool with web-based interface conducted by autonomous agents when precise economic information relative to plant and business actions and their effects are provided. It aims to attain a better arrangement of the marketing and production events in order to face the bid/bargain process during e-commerce. Additionally, management systems require real time execution and an efficient transaction-oriented approach capable to dynamically adopt realistic and optimal actions to support marketing management. To this end the TicTacToe algorithm provides sequence optimization with acceptable tolerances in realistic time.

  14. Protein kinase C regulates the phosphorylation and oligomerization of ERM binding phosphoprotein 50

    SciTech Connect

    Fouassier, Laura; Nichols, Matthew T.; Gidey, Elizabeth; McWilliams, Ryan R.; Robin, Helene; Finnigan, Claire; Howell, Kathryn E.; Housset, Chantal; Doctor, R. Brian . E-mail: brian.doctor@uchsc.edu

    2005-05-15

    Ezrin-Radixin-Moesin (ERM) binding phosphoprotein 50 (EBP50, a.k.a. NHERF-1) is a scaffold protein essential for the localization and coordinated activity of apical transporters, enzymes and receptors in epithelial cells. EBP50 acts via multiple protein binding interactions, including oligomerization through interactions of its PSD95-Dlg-ZO1 (PDZ) domains. EBP50 can be phosphorylated on multiple sites and phosphorylation of specific sites modulates the extent of oligomerization. The aim of the present study was to test the capacity of protein kinase C (PKC) to phosphorylate EBP50 and to regulate its oligomerization. In vitro experiments showed that the catalytic subunit of PKC directly phosphorylates EBP50. In HEK-293 cells transfected with rat EBP50 cDNA, a treatment with 12 myristate 13-acetate (PMA) induced a translocation of PKC{alpha} and {beta} isoforms to the membrane and increased {sup 32}P incorporation into EBP50. In co-transfection/co-precipitation studies, PMA treatment stimulated EBP50 oligomerization. Mass spectrometry analysis of full-length EBP50 and phosphorylation analyses of specific domains, and of mutated or truncated forms of EBP50, indicated that PKC-induced phosphorylation of EBP50 occurred on the Ser{sup 337}/Ser{sup 338} residue within the carboxyl-tail domain of the protein. Truncation of Ser{sup 337}/Ser{sup 338} also diminished PKC-induced oligomerization of EBP50. These results suggest the PKC signaling pathway can impact EBP50-dependent cellular functions by regulating EBP50 oligomerization.

  15. Genome-Wide Localization Study of Yeast Pex11 Identifies Peroxisome–Mitochondria Interactions through the ERMES Complex

    PubMed Central

    Mattiazzi Ušaj, M.; Brložnik, M.; Kaferle, P.; Žitnik, M.; Wolinski, H.; Leitner, F.; Kohlwein, S.D.; Zupan, B.; Petrovič, U.

    2015-01-01

    Pex11 is a peroxin that regulates the number of peroxisomes in eukaryotic cells. Recently, it was found that a mutation in one of the three mammalian paralogs, PEX11β, results in a neurological disorder. The molecular function of Pex11, however, is not known. Saccharomyces cerevisiae Pex11 has been shown to recruit to peroxisomes the mitochondrial fission machinery, thus enabling proliferation of peroxisomes. This process is essential for efficient fatty acid β-oxidation. In this study, we used high-content microscopy on a genome-wide scale to determine the subcellular localization pattern of yeast Pex11 in all non-essential gene deletion mutants, as well as in temperature-sensitive essential gene mutants. Pex11 localization and morphology of peroxisomes was profoundly affected by mutations in 104 different genes that were functionally classified. A group of genes encompassing MDM10, MDM12 and MDM34 that encode the mitochondrial and cytosolic components of the ERMES complex was analyzed in greater detail. Deletion of these genes caused a specifically altered Pex11 localization pattern, whereas deletion of MMM1, the gene encoding the fourth, endoplasmic-reticulum-associated component of the complex, did not result in an altered Pex11 localization or peroxisome morphology phenotype. Moreover, we found that Pex11 and Mdm34 physically interact and that Pex11 plays a role in establishing the contact sites between peroxisomes and mitochondria through the ERMES complex. Based on these results, we propose that the mitochondrial/cytosolic components of the ERMES complex establish a direct interaction between mitochondria and peroxisomes through Pex11. PMID:25769804

  16. A combined cryo-EM and molecular dynamics approach reveals the mechanism of ErmBL-mediated translation arrest

    PubMed Central

    Arenz, Stefan; Bock, Lars V.; Graf, Michael; Innis, C. Axel; Beckmann, Roland; Grubmüller, Helmut; Vaiana, Andrea C.; Wilson, Daniel N.

    2016-01-01

    Nascent polypeptides can induce ribosome stalling, regulating downstream genes. Stalling of ErmBL peptide translation in the presence of the macrolide antibiotic erythromycin leads to resistance in Streptococcus sanguis. To reveal this stalling mechanism we obtained 3.6-Å-resolution cryo-EM structures of ErmBL-stalled ribosomes with erythromycin. The nascent peptide adopts an unusual conformation with the C-terminal Asp10 side chain in a previously unseen rotated position. Together with molecular dynamics simulations, the structures indicate that peptide-bond formation is inhibited by displacement of the peptidyl-tRNA A76 ribose from its canonical position, and by non-productive interactions of the A-tRNA Lys11 side chain with the A-site crevice. These two effects combine to perturb peptide-bond formation by increasing the distance between the attacking Lys11 amine and the Asp10 carbonyl carbon. The interplay between drug, peptide and ribosome uncovered here also provides insight into the fundamental mechanism of peptide-bond formation. PMID:27380950

  17. A combined cryo-EM and molecular dynamics approach reveals the mechanism of ErmBL-mediated translation arrest.

    PubMed

    Arenz, Stefan; Bock, Lars V; Graf, Michael; Innis, C Axel; Beckmann, Roland; Grubmüller, Helmut; Vaiana, Andrea C; Wilson, Daniel N

    2016-01-01

    Nascent polypeptides can induce ribosome stalling, regulating downstream genes. Stalling of ErmBL peptide translation in the presence of the macrolide antibiotic erythromycin leads to resistance in Streptococcus sanguis. To reveal this stalling mechanism we obtained 3.6-Å-resolution cryo-EM structures of ErmBL-stalled ribosomes with erythromycin. The nascent peptide adopts an unusual conformation with the C-terminal Asp10 side chain in a previously unseen rotated position. Together with molecular dynamics simulations, the structures indicate that peptide-bond formation is inhibited by displacement of the peptidyl-tRNA A76 ribose from its canonical position, and by non-productive interactions of the A-tRNA Lys11 side chain with the A-site crevice. These two effects combine to perturb peptide-bond formation by increasing the distance between the attacking Lys11 amine and the Asp10 carbonyl carbon. The interplay between drug, peptide and ribosome uncovered here also provides insight into the fundamental mechanism of peptide-bond formation. PMID:27380950

  18. A combined cryo-EM and molecular dynamics approach reveals the mechanism of ErmBL-mediated translation arrest

    NASA Astrophysics Data System (ADS)

    Arenz, Stefan; Bock, Lars V.; Graf, Michael; Innis, C. Axel; Beckmann, Roland; Grubmüller, Helmut; Vaiana, Andrea C.; Wilson, Daniel N.

    2016-07-01

    Nascent polypeptides can induce ribosome stalling, regulating downstream genes. Stalling of ErmBL peptide translation in the presence of the macrolide antibiotic erythromycin leads to resistance in Streptococcus sanguis. To reveal this stalling mechanism we obtained 3.6-Å-resolution cryo-EM structures of ErmBL-stalled ribosomes with erythromycin. The nascent peptide adopts an unusual conformation with the C-terminal Asp10 side chain in a previously unseen rotated position. Together with molecular dynamics simulations, the structures indicate that peptide-bond formation is inhibited by displacement of the peptidyl-tRNA A76 ribose from its canonical position, and by non-productive interactions of the A-tRNA Lys11 side chain with the A-site crevice. These two effects combine to perturb peptide-bond formation by increasing the distance between the attacking Lys11 amine and the Asp10 carbonyl carbon. The interplay between drug, peptide and ribosome uncovered here also provides insight into the fundamental mechanism of peptide-bond formation.

  19. Absence of a functional erm gene in isolates of Mycobacterium immunogenum and the Mycobacterium mucogenicum group, based on in vitro clarithromycin susceptibility.

    PubMed

    Brown-Elliott, Barbara A; Hanson, Kimberly; Vasireddy, Sruthi; Iakhiaeva, Elena; Nash, Kevin A; Vasireddy, Ravikiran; Parodi, Nicholas; Smith, Terry; Gee, Martha; Strong, Anita; Barker, Adam; Cohen, Samuel; Muir, Haleina; Slechta, E Susan; Wallace, Richard J

    2015-03-01

    Macrolide resistance has been linked to the presence of a functional erythromycin ribosomal methylase (erm) gene in most species of pathogenic rapidly growing mycobacteria (RGM). For these Mycobacterium isolates, extended incubation in clarithromycin is necessary to determine macrolide susceptibility. In contrast, the absence of a detectable erm gene in isolates of M. chelonae, M. senegalense, and M. peregrinum and a nonfunctional erm gene in M. abscessus subsp. massiliense and 15% to 20% of M. abscessus subsp. abscessus isolates renders these species intrinsically macrolide susceptible. Not all RGM species have been screened for the presence of an erm gene, including the Mycobacterium mucogenicum group (M. mucogenicum, M. phocaicum, and M. aubagnense) and Mycobacterium immunogenum. A total of 356 isolates of these two pathogenic RGM taxa from two reference laboratories (A.R.U.P. Reference Laboratories and the Mycobacteria/Nocardia Laboratory at the University of Texas Health Science Center at Tyler) underwent clarithromycin susceptibility testing with readings at 3 to 5 days and 14 days. Only 13 of the 356 isolates had resistant clarithromycin MICs at initial extended MIC readings, and repeat values on all available isolates were ≤2 μg/ml. These studies suggest that these two additional RGM groups do not harbor functional erm genes and, like M. chelonae, do not require extended clarithromycin susceptibility testing. We propose to the Clinical Laboratory and Standards Institute that isolates belonging to these above-mentioned six rapidly growing mycobacterial groups based on molecular identification with no known functional erm genes undergo only 3 to 5 days of susceptibility testing (to exclude mutational resistance). PMID:25568437

  20. Evidence for the translational attenuation model: ribosome-binding studies and structural analysis with an in vitro run-off transcript of ermC.

    PubMed Central

    Narayanan, C S; Dubnau, D

    1985-01-01

    Several features of the translational attenuation model of ermC regulation were tested. This model predicts two possible secondary structures for the leader of the ermC transcript and requires that the leader contains two Shine-Dalgarno (SD) sequences. The ribosome binding site for a leader peptide (SD1) is predicted to be accessible, whereas that for the rRNA methylase protein that confers erythromycin (Em) resistance (SD2) is sequestered by base pairing. The model suggests that in the presence of inducer (Em), a ribosome stalls while translating the peptide, altering the mRNA conformation, thereby exposing SD2. The results of our ribosome binding studies demonstrate that SD1 is exposed and binds to ribosomes, whereas SD2 is unavailable. Also, the secondary structure of the 5' region of the ermC transcript was analyzed using methidium propyl-EDTA.Fe (II), T1 nuclease, and nucleases from cobra venom and mung bean sprouts as structure probes. Our results support the previously proposed model for folding of ermC mRNA, and demonstrate that SD1 is single-stranded, while SD2 and its neighboring sequences are largely base paired, consistent with the ribosome-binding results. Images PMID:3903662

  1. Mutational analysis defines the roles of conserved amino acid residues in the predicted catalytic pocket of the rRNA:m6A methyltransferase ErmC'.

    PubMed

    Maravić, Gordana; Feder, Marcin; Pongor, Sándor; Flögel, Mirna; Bujnicki, Janusz M

    2003-09-01

    Methyltransferases (MTases) from the Erm family catalyze S-adenosyl-L-methionine-dependent modification of a specific adenine residue in bacterial 23S rRNA, thereby conferring resistance to clinically important macrolide, lincosamide and streptogramin B antibiotics. Despite the available structural data and functional analyses on the level of the RNA substrate, still very little is known about the mechanism of rRNA:adenine-N(6) methylation. Only predictions regarding various aspects of this reaction have been made based on the analysis of the crystal structures of methyltransferase ErmC' (without the RNA) and their comparison with the crystallographic and biochemical data for better studied DNA:m(6)A MTases. To validate the structure-based predictions of presumably essential residues in the catalytic pocket of ErmC', we carried out the site-directed mutagenesis and studied the function of the mutants in vitro and in vivo. Our results indicate that the active site of rRNA:m(6)A MTases is much more tolerant to amino acid substitutions than the active site of DNA:m(6)A MTases. Only the Y104 residue implicated in stabilization of the target base was found to be indispensable. Remarkably, the N101 residue from the "catalytic" motif IV and two conserved residues that form the floor (F163) and one of the walls (N11) of the base-binding site are not essential for catalysis in ErmC'. This somewhat surprising result is discussed in the light of the available structural data and in the phylogenetic context of the Erm family. PMID:12946350

  2. Characterization of Enterococcus faecium with macrolide resistance and reduced susceptibility to quinupristin/dalfopristin in a Japanese hospital: detection of extensive diversity in erm(B)-regulator regions.

    PubMed

    Isogai, Nayuta; Urushibara, Noriko; Kawaguchiya, Mitsuyo; Ghosh, Souvik; Suzaki, Keisuke; Watanabe, Naoki; Quiñones, Dianelys; Kobayashi, Nobumichi

    2013-08-01

    Cross-resistance to macrolide, lincosamide, and streptogramin B (MLSB) antibiotics is mainly mediated by the erm (erythromycin ribosome methylation) genes that encode 23S rRNA methylases in enterococi, and various mechanisms are involved in the streptogramin B resistance. Prevalence of MLSB resistance and its genetic mechanisms were analyzed for a total of 159 strains of Enterococcus faecium isolated from clinical specimens in a university hospital in Japan from 1997 to 2006. Resistance to erythromycin (EM) and clindamycin was detected in 88.1% and 89.9% of all the strains examined, respectively, and expression of resistance was totally constitutive. Although none of the strain was resistant to quinupristin/dalfopristin (Q/D), 28 strains (17.6%) showed intermediate resistance to Q/D (MIC: 2 μg/ml). The erm(B) gene was detected in 139 strains (87.4%), and msrC was found in all the strains examined, whereas no other known MLSB resistance genes were identified. The erm(B) regulator region (RR) containing a coding region of the leader peptide was classified into 13 genetic variations (L1-L3, M, S1-S7, D, and R genotypes) in 56 strains. However, no relatedness was identified between the erm(B) RR genotype and EM resistance, or reduced susceptibility to Q/D, although most of Q/D-intermediate strains were assigned to the L1, L2, and S1 genotypes. Q/D-intermediate strains were classified into five multiple-locus variable-number tandem-repeat analysis (MLVA) types, including four types of clonal complex (CC)-C1, five sequence types (STs), including four STs of CC-17, and several resistance gene/virulence factor profiles. The present study revealed the occurrence of Q/D-intermediate E. faecium, which are composed of heterogeneous strains in Japan, and more genetic diversity in the erm(B) RRs than those reported previously. PMID:23442208

  3. Mutational analysis of basic residues in the N-terminus of the rRNA:m6A methyltransferase ErmC'.

    PubMed

    Maravić, G; Bujnicki, J M; Flögel, M

    2004-01-01

    Erm methyltransferases mediate the resistance to the macrolide-lincosamide-streptogramin B antibiotics via dimethylation of a specific adenine residue in 23S rRNA. The role of positively charged N-terminal residues of the ErmC' methyltransferase in RNA binding and/or catalysis was determined. Mutational analysis of amino acids K4 and K7 was performed and the mutants were characterized in in vivo and in vitro experiments. The K4 and K7 residues were suggested not to be essential for the enzyme activity but to provide a considerable support for the catalytic step of the reaction, probably by maintaining the optimum conformation of the transition state through interactions with the phosphate backbone of RNA. PMID:15114858

  4. Novel erm(T)-Carrying Multiresistance Plasmids from Porcine and Human Isolates of Methicillin-Resistant Staphylococcus aureus ST398 That Also Harbor Cadmium and Copper Resistance Determinants

    PubMed Central

    Gómez-Sanz, Elena; Kadlec, Kristina; Feßler, Andrea T.; Zarazaga, Myriam; Schwarz, Stefan

    2013-01-01

    This study describes three novel erm(T)-carrying multiresistance plasmids that also harbor cadmium and copper resistance determinants. The plasmids, designated pUR1902, pUR2940, and pUR2941, were obtained from porcine and human methicillin-resistant Staphylococcus aureus (MRSA) of the clonal lineage ST398. In addition to the macrolide-lincosamide-streptogramin B (MLSB) resistance gene erm(T), all three plasmids also carry the tetracycline resistance gene tet(L). Furthermore, plasmid pUR2940 harbors the trimethoprim resistance gene dfrK and the MLSB resistance gene erm(C), while plasmids pUR1902 and pUR2941 possess the kanamycin/neomycin resistance gene aadD. Sequence analysis of approximately 18.1 kb of the erm(T)-flanking region from pUR1902, 20.0 kb from pUR2940, and 20.8 kb from pUR2941 revealed the presence of several copies of the recently described insertion sequence ISSau10, which is probably involved in the evolution of the respective plasmids. All plasmids carried a functional cadmium resistance operon with the genes cadD and cadX, in addition to the multicopper oxidase gene mco and the ATPase copper transport gene copA, which are involved in copper resistance. The comparative analysis of S. aureus RN4220 and the three S. aureus RN4220 transformants carrying plasmid pUR1902, pUR2940, or pUR2941 revealed an 8-fold increase in CdSO4 and a 2-fold increase in CuSO4 MICs. The emergence of multidrug resistance plasmids that also carry heavy metal resistance genes is alarming and requires further surveillance. The colocalization of antimicrobial resistance genes and genes that confer resistance to heavy metals may facilitate their persistence, coselection, and dissemination. PMID:23629701

  5. Anchoring of Protein Kinase A by ERM (Ezrin-Radixin-Moesin) Proteins Is Required for Proper Netrin Signaling through DCC (Deleted in Colorectal Cancer)*

    PubMed Central

    Deming, Paula B.; Campbell, Shirley L.; Stone, Jamie B.; Rivard, Robert L.; Mercier, Alison L.; Howe, Alan K.

    2015-01-01

    Netrin-1, acting through its principal receptor DCC (deleted in colorectal cancer), serves as an axon guidance cue during neural development and also contributes to vascular morphogenesis, epithelial migration, and the pathogenesis of some tumors. Several lines of evidence suggest that netrin-DCC signaling can regulate and be regulated by the cAMP-dependent protein kinase, PKA, although the molecular details of this relationship are poorly understood. Specificity in PKA signaling is often achieved through differential subcellular localization of the enzyme by interaction with protein kinase A anchoring proteins (AKAPs). Here, we show that AKAP function is required for DCC-mediated activation of PKA and phosphorylation of cytoskeletal regulatory proteins of the Mena/VASP (vasodilator-stimulated phosphoprotein) family. Moreover, we show that DCC and PKA physically interact and that this association is mediated by the ezrin-radixin-moesin (ERM) family of plasma membrane-actin cytoskeleton cross-linking proteins. Silencing of ERM protein expression inhibits DCC-PKA interaction, DCC-mediated PKA activation, and phosphorylation of Mena/VASP proteins as well as growth cone morphology and neurite outgrowth. Finally, although expression of wild-type radixin partially rescued growth cone morphology and tropism toward netrin in ERM-knockdown cells, expression of an AKAP-deficient mutant of radixin did not fully rescue growth cone morphology and switched netrin tropism from attraction to repulsion. These data support a model in which ERM-mediated anchoring of PKA activity to DCC is required for proper netrin/DCC-mediated signaling. PMID:25575591

  6. Analysis of mutations in Neurospora crassa ERMES components reveals specific functions related to β-barrel protein assembly and maintenance of mitochondrial morphology.

    PubMed

    Wideman, Jeremy G; Lackey, Sebastian W K; Srayko, Martin A; Norton, Kacie A; Nargang, Frank E

    2013-01-01

    The endoplasmic reticulum mitochondria encounter structure (ERMES) tethers the er to mitochondria and contains four structural components: Mmm1, Mdm12, Mdm10, and Mmm2 (Mdm34). The Gem1 protein may play a role in regulating ERMES function. Saccharomyces cerevisiae and Neurospora crassa strains lacking any of Mmm1, Mdm12, or Mdm10 are known to show a variety of phenotypic defects including altered mitochondrial morphology and defects in the assembly of β-barrel proteins into the mitochondrial outer membrane. Here we examine ERMES complex components in N. crassa and show that Mmm1 is an ER membrane protein containing a Cys residue near its N-terminus that is conserved in the class Sordariomycetes. The residue occurs in the ER-lumen domain of the protein and is involved in the formation of disulphide bonds that give rise to Mmm1 dimers. Dimer formation is required for efficient assembly of Tom40 into the TOM complex. However, no effects are seen on porin assembly or mitochondrial morphology. This demonstrates a specificity of function and suggests a direct role for Mmm1 in Tom40 assembly. Mutation of a highly conserved region in the cytosolic domain of Mmm1 results in moderate defects in Tom40 and porin assembly, as well as a slight morphological phenotype. Previous reports have not examined the role of Mmm2 with respect to mitochondrial protein import and assembly. Here we show that absence of Mmm2 affects assembly of β-barrel proteins and that lack of any ERMES structural component results in defects in Tom22 assembly. Loss of N. crassa Gem1 has no effect on the assembly of these proteins but does affect mitochondrial morphology. PMID:23940790

  7. Molecular cloning of a novel NF2/ERM/4.1 superfamily gene, ehm2, that is expressed in high-metastatic K1735 murine melanoma cells.

    PubMed

    Shimizu, K; Nagamachi, Y; Tani, M; Kimura, K; Shiroishi, T; Wakana, S; Yokota, J

    2000-04-15

    We have cloned a novel gene, Ehm2, that is expressed in high-metastatic but not in low-metastatic K-1735 murine melanoma cells. The Ehm2 gene encodes a protein of 527 amino acid residues, showing up to 41% amino acid identity with the FERM domain of NF2/ERM/4.1 superfamily proteins, which have the function of connecting cell surface transmembrane proteins to cytoskeletal molecules. The Ehm2 gene was mapped to chromosome 4 and was expressed in the liver, lung, kidney, and testis and in 7- to 17-day embryos. The highest level of homology was observed with NBL4, which is a new subfamily protein of the NF2/ERM/4.1 superfamily. A human homologue of the mouse Ehm2 gene, showing significant homology (83% identity), was identified in the genomic DNA and EST databases. Furthermore, seven rat EST clones and one pig EST clone in the GenBank EST database were identified as having 83-92% sequence homology with the cDNA sequence of the mouse Ehm2 gene. Thus, Ehm2 is a highly conserved gene that encodes a novel member of the NF2/ERM/4.1 superfamily proteins. PMID:10783258

  8. Genome sequence and virulence factors of a group G Streptococcus dysgalactiae subsp. equisimilis strain with a new element carrying erm(B)

    PubMed Central

    Wang, Xiaohui; Zhang, Xiaoxia; Zong, Zhiyong

    2016-01-01

    A Streptococcus dysgalactiae subsp. equisimilis (SDSE) strain WCHSDSE-1, which caused an outbreak of tonsillopharyngitis among healthcare workers in China, was subjected to genome sequencing and analysis. WCHSDSE-1 belongs to the Lancefield group G, emm type stG211.1 and sequence type 44. WCHSDSE-1 has virulence factors for adherence, impairing the recruitment of neutrophils to infection sites and toxins including streptolysins O and S and exotoxin G. WCHSDSE-1 has a 45.4-kb element resembling a conjugative transposon. This element is absent from other known SDSE genomes and contains the macrolide-resistant gene erm(B). Conjugative transfer of erm(B) was not successful in mating experiments, suggesting that the element might have lost its ability of conjugation. An almost identical element, which contains the tetracycline-resistant gene tet(M) instead of erm(B), is present on the genome of Filifactor alocis ATCC 35896. The boundaries and insertion sites of the two elements were identified and both were flanked by a 3-bp direct repeat, which is characteristic of transposition. In conclusion, the spectrum of virulence factors of WCHSDSE-1 is similar to other SDSE strains causing invasive diseases. WCHSDSE-1 possesses a new transposable element encoding macrolide resistance, which could pick up different resistance genes and could be transferred across species in oral microflora. PMID:26843282

  9. Alanine-scanning mutagenesis of the predicted rRNA-binding domain of ErmC' redefines the substrate-binding site and suggests a model for protein-RNA interactions.

    PubMed

    Maravić, Gordana; Bujnicki, Janusz M; Feder, Marcin; Pongor, Sándor; Flögel, Mirna

    2003-08-15

    The Erm family of adenine-N(6) methyltransferases (MTases) is responsible for the development of resistance to macrolide-lincosamide-streptogramin B antibiotics through the methylation of 23S ribosomal RNA. Hence, these proteins are important potential drug targets. Despite the availability of the NMR and crystal structures of two members of the family (ErmAM and ErmC', respectively) and extensive studies on the RNA substrate, the substrate-binding site and the amino acids involved in RNA recognition by the Erm MTases remain unknown. It has been proposed that the small C-terminal domain functions as a target-binding module, but this prediction has not been tested experimentally. We have undertaken structure-based mutational analysis of 13 charged or polar residues located on the predicted rRNA-binding surface of ErmC' with the aim to identify the area of protein-RNA interactions. The results of in vivo and in vitro analyses of mutant protein suggest that the key RNA-binding residues are located not in the small domain, but in the large catalytic domain, facing the cleft between the two domains. Based on the mutagenesis data, a preliminary three-dimensional model of ErmC' complexed with the minimal substrate was constructed. The identification of the RNA-binding site of ErmC' may be useful for structure-based design of novel drugs that do not necessarily bind to the cofactor-binding site common to many S-adenosyl-L- methionine-dependent MTases, but specifically block the substrate-binding site of MTases from the Erm family. PMID:12907737

  10. ERM Ideas and Innovations

    ERIC Educational Resources Information Center

    Schmidt, Kari

    2012-01-01

    In this column, the author discusses how the management of e-books has introduced, at many libraries and in varying degrees, the challenges of maintaining effective technical services workflows. Four different e-book workflows are identified and explored, and the author takes a closer look at how particular variables for each are affected, such as…

  11. The NHERF2 sequence adjacent and upstream of the ERM-binding domain affects NHERF2-ezrin binding and dexamethasone stimulated NHE3 activity.

    PubMed

    Yang, Jianbo; Sarker, Rafiquel; Singh, Varsha; Sarker, Prateeti; Yin, Jianyi; Chen, Tian-E; Chaerkady, Raghothama; Li, Xuhang; Tse, C Ming; Donowitz, Mark

    2015-08-15

    In the brush border of intestinal and kidney epithelial cells, scaffolding proteins ezrin, Na(+)-H(+) exchanger regulatory factor (NHERF)1 and NHERF2 play important roles in linking transmembrane proteins to the cytoskeleton and assembling signalling regulatory complexes. The last 30 carboxyl residues of NHERF1 and NHERF2 form the EBDs [ezrin, radixin and moesin (ERM)-binding domain]. The current study found that NHERF1/2 contain an ERM-binding regulatory sequence (EBRS), which facilitates the interaction between the EBD and ezrin. The EBRSs are located within 24 and 19 residues immediately upstream of EBDs for NHERF1 and NHERF2 respectively. In OK (opossum kidney) epithelial cells, EBRSs are necessary along with the EBD to distribute NHERF1 and NHERF2 exclusively to the apical domain. Furthermore, phosphorylation of Ser(303) located in the EBRS of NHERF2, decreases the binding affinity for ezrin, dislocates apical NHERF2 into the cytosol and increases the NHERF2 microvillar mobility rate. Moreover, increased phosphorylation of Ser(303) was functionally significant preventing acute stimulation of NHE3 (Na(+)-H(+) exchanger 3) activity by dexamethasone. PMID:26251448

  12. Phylogeography of Yersinia ruckeri reveals effects of past evolutionary events on the current strain distribution and explains variations in the global transmission of enteric redmouth (ERM) disease

    PubMed Central

    Bastardo, Asmine; Ravelo, Carmen; Romalde, Jesús L.

    2015-01-01

    Phylogeographic patterns and population genetic structure of Yersinia ruckeri, the pathological agent of enteric redmouth disease (ERM) in salmonids, were investigated on the basis of concatenated multiloci sequences from isolates of different phenotypes obtained between 1965 and 2009 from diverse areas and hosts. Sequence analyses revealed genetic differentiation among subpopulations with the largest genetic distance occurring between subpopulations of Europe and Canada and/or South America. Bayesian analysis indicated the presence of three ancestral population clusters. Mismatch distribution displayed signatures characteristic of changes in size due to demographic and spatial expansions in the overall Y. ruckeri population, and also in the geographically separate subpopulations. Furthermore, a weak signal of isolation by distance was determined. A significant positive correlation between genetic and geographical distances was observed. These results revealed that the population of Y. ruckeri has undergone both ancient and recent population changes that were probably induced by biogeography forces in the past and, much more recently, by adaptive processes forced by aquaculture expansion. These findings have important implications for future studies on Y. ruckeri population dynamics, on the potential role of genetic structure to explain variations in ERM transmission, and on the effect of past evolutionary events on current estimations of gene flow. PMID:26579104

  13. The NHERF2 sequence adjacent and upstream of the ERM-binding domain affects NHERF2–ezrin binding and dexamethasone stimulated NHE3 activity

    PubMed Central

    Yang, Jianbo; Sarker, Rafiquel; Singh, Varsha; Sarker, Prateeti; Yin, Jianyi; Chen, Tian-E; Chaerkady, Raghothama; Li, Xuhang; Tse, C. Ming; Donowitz, Mark

    2015-01-01

    In the brush border of intestinal and kidney epithelial cells, scaffolding proteins ezrin, Na+-H+ exchanger regulatory factor (NHERF)1 and NHERF2 play important roles in linking transmembrane proteins to the cytoskeleton and assembling signalling regulatory complexes. The last 30 carboxyl residues of NHERF1 and NHERF2 form the EBDs [ezrin, radixin and moesin (ERM)-binding domain]. The current study found that NHERF1/2 contain an ERM-binding regulatory sequence (EBRS), which facilitates the interaction between the EBD and ezrin. The EBRSs are located within 24 and 19 residues immediately upstream of EBDs for NHERF1 and NHERF2 respectively. In OK (opossum kidney) epithelial cells, EBRSs are necessary along with the EBD to distribute NHERF1 and NHERF2 exclusively to the apical domain. Furthermore, phosphorylation of Ser303 located in the EBRS of NHERF2, decreases the binding affinity for ezrin, dislocates apical NHERF2 into the cytosol and increases the NHERF2 microvillar mobility rate. Moreover, increased phosphorylation of Ser303 was functionally significant preventing acute stimulation of NHE3 (Na+-H+ exchanger 3) activity by dexamethasone. PMID:26251448

  14. ICAM-2 regulates vascular permeability and N-cadherin localization through ezrin-radixin-moesin (ERM) proteins and Rac-1 signalling

    PubMed Central

    2014-01-01

    Background Endothelial junctions control functions such as permeability, angiogenesis and contact inhibition. VE-Cadherin (VECad) is essential for the maintenance of intercellular contacts. In confluent endothelial monolayers, N-Cadherin (NCad) is mostly expressed on the apical and basal membrane, but in the absence of VECad it localizes at junctions. Both cadherins are required for vascular development. The intercellular adhesion molecule (ICAM)-2, also localized at endothelial junctions, is involved in leukocyte recruitment and angiogenesis. Results In human umbilical vein endothelial cells (HUVEC), both VECad and NCad were found at nascent cell contacts of sub-confluent monolayers, but only VECad localized at the mature junctions of confluent monolayers. Inhibition of ICAM-2 expression by siRNA caused the appearance of small gaps at the junctions and a decrease in NCad junctional staining in sub-confluent monolayers. Endothelioma lines derived from WT or ICAM-2-deficient mice (IC2neg) lacked VECad and failed to form junctions, with loss of contact inhibition. Re-expression of full-length ICAM-2 (IC2 FL) in IC2neg cells restored contact inhibition through recruitment of NCad at the junctions. Mutant ICAM-2 lacking the binding site for ERM proteins (IC2 ΔERM) or the cytoplasmic tail (IC2 ΔTAIL) failed to restore junctions. ICAM-2-dependent Rac-1 activation was also decreased in these mutant cell lines. Barrier function, measured in vitro via transendothelial electrical resistance, was decreased in IC2neg cells, both in resting conditions and after thrombin stimulation. This was dependent on ICAM-2 signalling to the small GTPase Rac-1, since transendothelial electrical resistance of IC2neg cells was restored by constitutively active Rac-1. In vivo, thrombin-induced extravasation of FITC-labeled albumin measured by intravital fluorescence microscopy in the mouse cremaster muscle showed that permeability was increased in ICAM-2-deficient mice compared to controls

  15. Human Immunodeficiency virus (HIV-1) Vpr induced downregulation of NHE1 induces alteration in intracellular pH and loss of ERM complex in target cells

    PubMed Central

    Janket, Michelle L.; DeRicco, Jeremy S.; Borowski, LuAnn; Ayyavoo, Velpandi

    2007-01-01

    Human Immunodeficiency Virus type 1 (HIV-1) Vpr is known to dysregulate host cellular functions through its interaction with cellular proteins. Using a protein array we assessed Vpr-mediated differential regulation of host cellular proteins expression. Results demonstrated that Vpr differentially regulated host factors that are involved in functions such as cell proliferation, differentiation and apoptosis. One of the most highly downregulated proteins attained was the sodium hydrogen exchanger, isoform 1 (NHE1), which showed a significant (60%) decrease in HIV-1 vpr(+) virus infected cells as compared to HIV-1 vpr(−) virus infected control. NHE1 downregulation further led to acidification of cells and was directly correlated with loss of ezrin, radixin and moesin (ERM) protein complex and decreased AKT phosphorylation. Vpr-mediated NHE1 dyregulation is in part through GR pathway as GR antagonist, mifepristone reversed Vpr-induced NHE1 downregulation. PMID:17349711

  16. Resistin, a fat-derived secretory factor, promotes metastasis of MDA-MB-231 human breast cancer cells through ERM activation

    PubMed Central

    Lee, Jung Ok; Kim, Nami; Lee, Hye Jeong; Lee, Yong Woo; Kim, Su Jin; Park, Sun Hwa; Kim, Hyeon Soo

    2016-01-01

    Resistin, an adipocyte-secreted factor, is known to be elevated in breast cancer patients. However, the molecular mechanism by which resistin acts is not fully understood. The aim of this study was to investigate whether resistin could stimulate invasion and migration of breast cancer cells. Here, we report that resistin stimulated invasion and migration of breast cancer cells as well as phosphorylation of c-Src. Inhibition of c-Src blocked resistin-induced breast cancer cell invasion. Resistin increased intracellular calcium concentration, and chelation of intracellular calcium blocked resistin-mediated activation of Src. Resistin also induced phosphorylation of protein phosphatase 2A (PP2A). Inhibition of c-Src blocked resistin-mediated PP2A phosphorylation. In addition, resistin increased phosphorylation of PKCα. Inhibition of PP2A enhanced resistin-induced PKCα phosphorylation, demonstrating that PP2A activity is critical for PKCα phosphorylation. Resistin also increased phosphorylation of ezrin, radixin, and moesin (ERM). Additionally, ezrin interacted with PKCα, and resistin promoted co-localization of ezrin and PKCα. Either inhibition of c-Src and PKCα or knock-down of ezrin blocked resistin-induced breast cancer cells invasion. Moreover, resistin increased expression of vimentin, a key molecule for cancer cell invasion. Knock-down of ezrin abrogated resistin-induced vimentin expression. These results suggest that resistin play as a critical regulator of breast cancer metastasis. PMID:26729407

  17. ERM 593 Applied Project_Guidance for Reviewing and Approving a Waste Stream Profile in the Waste Compliance and Tracking System_Final_05-05-15

    SciTech Connect

    Elicio, Andy U.

    2015-05-05

    My ERM 593 applied project will provide guidance for the Los Alamos National Laboratory Waste Stream Profile reviewer (i.e. RCRA reviewer) in regards to Reviewing and Approving a Waste Stream Profile in the Waste Compliance and Tracking System. The Waste Compliance and Tracking system is called WCATS. WCATS is a web-based application that “supports the generation, characterization, processing and shipment of LANL radioactive, hazardous, and industrial waste.” The LANL generator must characterize their waste via electronically by filling out a waste stream profile (WSP) in WCATS. Once this process is completed, the designated waste management coordinator (WMC) will perform a review of the waste stream profile to ensure the generator has completed their waste stream characterization in accordance with applicable state, federal and LANL directives particularly P930-1, “LANL Waste Acceptance Criteria,” and the “Waste Compliance and Tracking System User's Manual, MAN-5004, R2,” as applicable. My guidance/applied project will describe the purpose, scope, acronyms, definitions, responsibilities, assumptions and guidance for the WSP reviewer as it pertains to each panel and subpanel of a waste stream profile.

  18. C. difficile 630Δerm Spo0A Regulates Sporulation, but Does Not Contribute to Toxin Production, by Direct High-Affinity Binding to Target DNA

    PubMed Central

    Rosenbusch, Katharina E.; Bakker, Dennis; Kuijper, Ed J.; Smits, Wiep Klaas

    2012-01-01

    Clostridium difficile is a Gram positive, anaerobic bacterium that can form highly resistant endospores. The bacterium is the causative agent of C. difficile infection (CDI), for which the symptoms can range from a mild diarrhea to potentially fatal pseudomembranous colitis and toxic megacolon. Endospore formation in Firmicutes, including C. difficile, is governed by the key regulator for sporulation, Spo0A. In Bacillus subtilis, this transcription factor is also directly or indirectly involved in various other cellular processes. Here, we report that C. difficile Spo0A shows a high degree of similarity to the well characterized B. subtilis protein and recognizes a similar binding sequence. We find that the laboratory strain C. difficile 630Δerm contains an 18bp-duplication near the DNA-binding domain compared to its ancestral strain 630. In vitro binding assays using purified C-terminal DNA binding domain of the C. difficile Spo0A protein demonstrate direct binding to DNA upstream of spo0A and sigH, early sporulation genes and several other putative targets. In vitro binding assays suggest that the gene encoding the major clostridial toxin TcdB may be a direct target of Spo0A, but supernatant derived from a spo0A negative strain was no less toxic towards Vero cells than that obtained from a wild type strain, in contrast to previous reports. These results identify for the first time direct (putative) targets of the Spo0A protein in C. difficile and make a positive effect of Spo0A on production of the large clostridial toxins unlikely. PMID:23119071

  19. Certification of new Pb iCRM (Candidate ERM-38xx series) via Gravimetric Isotope Mixtures and MC-ICP-MS Measurements

    NASA Astrophysics Data System (ADS)

    Ponzevera, E.; Quetel, C. R.

    2008-12-01

    Lead is known to be a particularly toxic chemical element. Mining and smelting of Pb and its domestic use over the centuries have contaminated the surface of Earth and jeopardized the health of humans, domestic animals and wildlife. Omnipresence of Pb has however an advantage. It can be used as an isotopic tracer of pollution sources and pathways in the environment. Pb isotopic Certified Reference Materials (iCRM) with undisputed characteristics are then required to validate Pb isotope ratio measurements. The materials currently available worldwide were produced in the 1960's by the National Bureau of Standards (now NIST, USA) and are now the object of polemics regarding the accuracy of some of the accompanying certified values. Moreover, new materials with lower relative uncertainty statements are demanded by users. This presentation is centred on the production and the certification at the EC-JRC-Institute for Reference Materials and Measurements of a new series of Pb iCRM (candidate ERM-38xx series). The production included six Pb gravimetric isotope mixtures, a common Pb material dedicated to routine calibration work and a series of four 207Pb slightly enriched natural-like Pb materials for the validation of the δ-scale method below 0.01%. The gravimetric isotope mixtures method, originally supported by mass spectrometry measurements performed on GS-MS or TI-MS instruments, was applied successfully to MC-ICP-MS. The measurements and certification methods we developed were validated in several ways, including a systematic investigation on possible significant sources of uncertainty and comparisons of results obtained by different laboratories on identical samples. Relative uncertainties on isotope ratios obtained for the newly produced Pb iCRM are as low as 0.017%, which is between 2 and 4 times smaller than the uncertainties carried by the NIST-981 material. Going below 0.01% was not possible mainly because of uncompressible uncertainties coming from the

  20. A Locally Created ERM: How and Why We Did It

    ERIC Educational Resources Information Center

    Doering, William; Chilton, Galadriel

    2008-01-01

    The University of Wisconsin-La Crosse's Murphy Library provides access to approximately 200 subscription databases. As with many academic libraries, methods and tools for managing these resources were sorely needed. However, the budget has been cut repeatedly over the past few years--thus the possibility of purchasing a commercial electronic…

  1. Shorter-Duration Therapy Using Vincristine, Dactinomycin, and Lower-Dose Cyclophosphamide With or Without Radiotherapy for Patients With Newly Diagnosed Low-Risk Rhabdomyosarcoma: A Report From the Soft Tissue Sarcoma Committee of the Children's Oncology Group

    PubMed Central

    Walterhouse, David O.; Pappo, Alberto S.; Meza, Jane L.; Breneman, John C.; Hayes-Jordan, Andrea A.; Parham, David M.; Cripe, Timothy P.; Anderson, James R.; Meyer, William H.; Hawkins, Douglas S.

    2014-01-01

    Purpose Intergroup Rhabdomyosarcoma Study Group (IRSG) studies III and IV showed improved failure-free survival (FFS) rates with vincristine, dactinomycin, and cyclophosphamide (VAC; total cumulative cyclophosphamide dose, 26.4 g/m2) compared with vincristine and dactinomycin (VA) for patients with subset-one low-risk embryonal rhabdomyosarcoma (ERMS; stage 1/2 group I/II ERMS or stage 1 group III orbit ERMS). The objective of Children's Oncology Group ARST0331 was to reduce the length of therapy without compromising FFS for this subset of low-risk patients by using VA in combination with lower-dose cyclophosphamide (total cumulative dose, 4.8 g/m2) plus radiotherapy (RT). Patients and Methods This noninferiority prospective clinical trial enrolled newly diagnosed patients with subset-one clinical features. Therapy included four cycles of VAC followed by four cycles of VA over 22 weeks. Patients with microscopic or gross residual disease at study entry received RT. Results With a median follow-up of 4.3 years, we observed 35 failures among 271 eligible patients versus 48.4 expected failures, calculated using a fixed outcome based on the FFS expected for similar patients treated on the IRSG D9602 protocol. The estimated 3-year FFS rate was 89% (95% CI, 85% to 92%), and the overall survival rate was 98% (95% CI, 95% to 99%). Patients with paratesticular tumors had the most favorable outcome. Three-year cumulative incidence rates for any local, regional, or distant failures were 7.6%, 1.5%, and 3.4%, respectively. Conclusion Shorter-duration therapy that included lower-dose cyclophosphamide and RT did not compromise FFS for patients with subset-one low-risk ERMS. PMID:25267746

  2. Tylosin-resistant Enterococci, erm genes, and tylosin in drained fields receiving swine manure

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The use of tylosin at subtherapeutic levels by the swine industry provides selective pressure for the development of antibiotic resistance in gastrointestinal bacteria. The land application of swine manure to drained agricultural fields might introduce elevated levels of total and tylosin-resistant ...

  3. Note: Focus error detection device for thermal expansion-recovery microscopy (ThERM).

    PubMed

    Domené, E A; Martínez, O E

    2013-01-01

    An innovative focus error detection method is presented that is only sensitive to surface curvature variations, canceling both thermoreflectance and photodefelection effects. The detection scheme consists of an astigmatic probe laser and a four-quadrant detector. Nonlinear curve fitting of the defocusing signal allows the retrieval of a cutoff frequency, which only depends on the thermal diffusivity of the sample and the pump beam size. Therefore, a straightforward retrieval of the thermal diffusivity of the sample is possible with microscopic lateral resolution and high axial resolution (~100 pm). PMID:23387710

  4. The actin-binding ERM protein Moesin directly regulates spindle assembly and function during mitosis.

    PubMed

    Vilmos, Péter; Kristó, Ildikó; Szikora, Szilárd; Jankovics, Ferenc; Lukácsovich, Tamás; Kari, Beáta; Erdélyi, Miklós

    2016-06-01

    Ezrin-Radixin-Moesin proteins are highly conserved, actin-binding cytoskeletal proteins that play an essential role in microvilli formation, T-cell activation, and tumor metastasis by linking actin filaments to the plasma membrane. Recent studies demonstrated that the only Ezrin-Radixin-Moesin protein of Drosophila melanogaster, Moesin, is involved in mitotic spindle function through stabilizing cell shape and microtubules at the cell cortex. We previously observed that Moesin localizes to the mitotic spindle; hence, we tested for the biological significance of this surprising localization and investigated whether it plays a direct role in spindle function. To separate the cortical and spindle functions of Moesin during mitosis we combined cell biological and genetic methods. We used early Drosophila embryos, in which mitosis occurs in the absence of a cell cortex, and found in vivo evidence for the direct requirement of Moesin in mitotic spindle assembly and function. We also found that the accumulation of Moesin precedes the construction of the microtubule spindle, and the fusiform structure formed by Moesin persists even after the microtubules have disassembled. PMID:27006187

  5. Persistence of erythromycin resistance gene erm(B) in cattle feedlot pens over time

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The prevalence of antibiotic resistance was evaluated in cattle feedlot surface soil samples collected prior to and after a 13-month feedlot trial of two common diets, a 0% wet distillers grains plus solubles (WDGS) corn-based ration versus a 35% WDGS ration. Soil samples were collected from the mo...

  6. Extreme Precipitation Events in the Middle East: Dynamics of the Active Red Sea Trough

    NASA Astrophysics Data System (ADS)

    de Vries, Andries Jan; Lelieveld, Jos; Tyrlis, Evangelos; Krichak, Shimon; Edry, Dudi; Steil, Benedikt

    2013-04-01

    Extreme precipitation events in the Middle East can cause flash floods with serious societal impacts. For example, in Egypt in November 1994, and in Jeddah (Saudi Arabia) in November 2009, flash floods caused almost 600 and more than 120 casualties, respectively. A major meteorological phenomenon involved is the Active Red Sea Trough (ARST), a midlatitude-tropical interaction. Previous ARST studies addressed the dynamics, however, do not provide a complete understanding of the mechanism, and moreover, focus only on the Levant. This study presents a generalized concept of the ARST dynamics and aims for an improved understanding of its geographical extent and seasonality. We used ERA-Interim data to analyze several ARST events affecting the Levant, the Jeddah flooding, and the seasonality of the ARST associated dynamics. The synoptics around 25 November 2009 show a similar dynamical evolution as the ARST events over the Levant, revealing that the Jeddah flooding was in fact caused by an ARST. An ARST concept is defined, involving six important dynamical features: (1) a semi-permanent low-level trough (i.e. the Red Sea Trough), (2) a semi-permanent anticyclone at mid-levels over the Arabian Peninsula, (3) a mid-latitude upper level trough, (4) an intensified subtropical jet stream, (5) moisture transport and convergence, and (6) ascending motions resulting from the large scale forcing and tropospheric instability, leading to mesoscale convective systems and (local) extreme precipitation. Our analysis emphasizes the significance of the semi-permanent anticyclone over the Arabian Peninsula, predominantly causing moisture transport from the Arabian and Red Seas. The seasonal cycle of these dynamical factors explains why the ARSTs over the Levant region occur primarily in autumn and to a lesser extent in spring, and suggests a temporal shift further to the south (e.g. around Jeddah), starting later in autumn, ending earlier in spring, and favorable conditions for ARSTs

  7. Efficacy testing of a 35 year old commercially produced Yersinia ruckeri bacterin for the control of Enteric Redmouth ERM) Disease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In 1976 the United States Department of Agriculture, Veterinary Services, granted the first veterinary product license for a biologic to be administered to fish. That license was issued for Enteric Redmouth Bacterin (Yerisina ruckeri) to the first commercial fish vaccine company, Wildlife Vaccines. ...

  8. Assessment of altered binding specificity of bacteriophage for ciprofloxacin-induced antibiotic-resistant Salmonella Typhimurium.

    PubMed

    Kim, Jeongjin; Jo, Ara; Ding, Tian; Lee, Hyeon-Yong; Ahn, Juhee

    2016-08-01

    This study describes a new effort toward understanding the interaction mechanisms between antibiotic-resistant Salmonella Typhimurium and phages. The antibiotic susceptibility, β-lactamase activity, bacterial motility, gene expression, and lytic activity were evaluated in ciprofloxacin-induced antibiotic-sensitive Salmonella Typhimurium (ASST(CIP)) and ciprofloxacin-induced antibiotic-resistant S. Typhimurium (ARST(CIP)), which were compared to the wild-type strains (ASST(WT) and ARST(WT)). The MIC values of ampicillin, norfloxacin, chloramphenicol, and tetracycline were significantly increased to > 512, 16, 16, and 256 μg/ml, respectively, in the ARST(CIP). The lowest and highest extracellular lactamase activities were observed in ASST(WT) (6.85 μmol/min/ml) and ARST(CIP) (48.83 μmol/min/ml), respectively. The acrA, lpfE, and hilA genes were significantly upregulated by more than tenfold in both ASST(CIP) and ARST(CIP). The induction of multiple antibiotic resistance resulted from the increased efflux pump activity (AcrAB-TolC). The highest phage adsorption rates were more than 95 % for ASST(WT), ASST(CIP), and ARST(WT), while the lowest adsorption rate was 52 % for ARST(CIP) at 15 min of infection. The least lytic activity of phage was 20 % against the ARST(CIP), followed by ASST(CIP) (30 %). The adsorption rate of phage against ARST(CIP) was 52 % at 15 min of infection, which resulted in the decrease in lytic activity (12 %). Understanding the interaction of phage and bacteria is essential for the practical application of phage to control and detect antibiotic-resistant bacteria. The results provide useful information for understanding the binding specificity of phages for multiple antibiotic-resistant pathogens. PMID:27000396

  9. Extreme precipitation events in the Middle East: Dynamics of the Active Red Sea Trough

    NASA Astrophysics Data System (ADS)

    Vries, A. J.; Tyrlis, E.; Edry, D.; Krichak, S. O.; Steil, B.; Lelieveld, J.

    2013-07-01

    The Active Red Sea Trough (ARST) is an infrequent weather phenomenon that is associated with extreme precipitation, flash floods, and severe societal impacts in the Middle East (ME). Using reanalysis (ERA-Interim) and observational precipitation (Aphrodite and stations) data, we investigate its underlying dynamics, geographical extent, and seasonality. Twelve ARST events affecting the Levant have the same dynamical characteristics as those associated with a major flood in Jeddah (Saudi Arabia) on 25 November 2009. Hence, the Jeddah flooding was caused by an ARST, which implies that ARSTs can affect a much larger part of the ME than previously assumed. We present an ARST concept involving six dynamical factors: (1) a low-level trough; the Red Sea Trough (RST), (2) an anticyclone over the Arabian Peninsula; the Arabian Anticyclone (AA), (3) a transient midlatitude upper trough, (4) an intensified subtropical jet stream, (5) moisture transport pathways, and (6) strong ascent resulting from tropospheric instability and the synoptic-scale dynamical forcing. We explain the ARST as the interaction of a persistent stationary wave in the tropical easterlies (i.e., the RST) with a superimposed amplifying Rossby wave, resulting in northward propagating moist air masses over the Red Sea. Our findings emphasize the relevance of the AA, causing moisture transport from the Arabian and Red Seas. The particular topography in the Red Sea region and associated low-level circulation makes the ARST unique among tropical-extratropical interactions. The ARST seasonality is explained by the large-scale circulation and in particular the seasonal cycle of the semipermanent quasi-stationary RST and AA.

  10. Differential regulation of phagosome maturation in macrophages and dendritic cells mediated by Rho GTPases and ezrin–radixin–moesin (ERM) proteins

    PubMed Central

    Erwig, Lars-Peter; McPhilips, Kathleen A.; Wynes, Murray W.; Ivetic, Alexander; Ridley, Anne J.; Henson, Peter M.

    2006-01-01

    Deletion of apoptotic cells from tissues involves their phagocytosis by macrophages, dendritic cells, and tissue cells. Although much attention has been focused on the participating ligands, receptors, and mechanisms of uptake, little is known of the disposition of the ingested cell within the phagosome. Here we show that uptake of apoptotic cells by macrophages or fibroblasts results in rapid phagosome maturation, whereas macrophage phagosomes containing Ig-opsonized target cells mature at a slower rate. The early maturation was shown to depend on activation of Rho acting through Rho kinase on ezrin–radixin–moesin proteins. Blockade of Rho signaling or inhibition of moesin both delayed maturation rates to those seen with opsonized targets. By contrast, phagosome maturation in dendritic cells was slower, similar between apoptotic and opsonized target cells, and unaffected by Rho inhibition. These observations have direct implications for the clearance of dying cells and the roles played by different phagocytes in antigen digestion and presentation. PMID:16908865

  11. Trimming the threshold dispersion below 10 e-rms in a large area readout IC working in a single photon counting mode

    NASA Astrophysics Data System (ADS)

    Kmon, P.; Maj, P.; Gryboś, P.; Szczygieł, R.

    2016-01-01

    We present a new method of an in-pixel threshold dispersion correction implemented in a prototype readout integrated circuit (IC) operating in a single photon counting mode. The new threshold correction method was implemented in a readout IC of area 9.6× 14.9 mm2 containing 23552 square pixels with the pitch of 75 μm designed and fabricated in CMOS 130 nm technology. Each pixel of the IC consists of a charge sensitive amplifier, a shaper, two discriminators, two 14-bit counters and a low-area trim DACs for threshold correction. The user can either control the range of the trim DAC globally for all the pixels in the integrated circuit or modify the trim DACs characteristics locally in each pixel independently. Using a simulation tool based on the Monte-Carlo methods, we estimated how much we could improve the offset trimming by increasing the number of bits in the trim DACs or implementing additional bits in a pixel to modify the characteristics of the trim DACs. The measurements of our IC prototype show that it is possible to reduce the effective threshold dispersion in large-area single-photon counting chips below 10 electrons rms.

  12. Automatic Radiated Susceptibility Test System for Payload Equipment

    NASA Technical Reports Server (NTRS)

    Ngo, Hoai T.; Sturman, John C.; Sargent, Noel B.

    1995-01-01

    An automatic radiated susceptibility test system (ARSTS) was developed for NASA Lewis Research Center's Electro-magnetic Interference laboratory. According to MSFC-SPEC 521B, any electrical or electronic equipment that will be transported by the spacelab and space shuttle must be tested for susceptibility to electromagnetic interference. This state-of-the-art automatic test system performs necessary calculations; analyzes, processes, and records a great quantity of measured data; and monitors the equipment being tested in real-time and with minimal user intervention. ARSTS reduces costly test time, increases test accuracy, and provides reliable test results.

  13. Automatic radiated susceptibility test system for payload equipment

    NASA Astrophysics Data System (ADS)

    Ngo, Hoai T.; Sturman, John C.; Sargent, Noel B.

    1995-09-01

    An automatic radiated susceptibility test system (ARSTS) was developed for NASA Lewis Research Center's Electro-magnetic Interference laboratory. According to MSFC-SPEC 521B, any electrical or electronic equipment that will be transported by the spacelab and space shuttle must be tested for susceptibility to electromagnetic interference. This state-of-the-art automatic test system performs necessary calculations; analyzes, processes, and records a great quantity of measured data; and monitors the equipment being tested in real-time and with minimal user intervention. ARSTS reduces costly test time, increases test accuracy, and provides reliable test results.

  14. KARST HYDROLOGY AND CHEMICAL CONTAMINATION

    EPA Science Inventory

    Ground-water flow in karst aquifers is very different from flow in granular or fractured aquifers. arst ground-water flow is often turbulent within discrete conduits that are convergent in their upper reaches and may be divergent in their very lower reaches, simulating the flow p...

  15. VULNERABILITY OF KARST AQUIFERS TO CHEMICAL CONTAMINATION

    EPA Science Inventory

    Ground water flow in karst aquifers is very different from flow in granular or fractured aquifers. arst ground water flow is often turbulent within discrete conduits that are convergent in the upper reaches and divergent in the lower, simulating discharge to one or more springs. ...

  16. Comparative Genomic Analysis of the ICESa2603 Family ICEs and Spread of erm(B)- and tet(O)-Carrying Transferable 89K-Subtype ICEs in Swine and Bovine Isolates in China.

    PubMed

    Huang, Jinhu; Liang, Yuan; Guo, Dawei; Shang, Kexin; Ge, Lin; Kashif, Jam; Wang, Liping

    2016-01-01

    Integrative and conjugative elements (ICEs) of the ICESa2603 family have been isolated from several species of Streptococcus spp.; however, the comparative genomic and evolutionary analyses of these particular ICEs are currently only at their initial stages. By investigating 13 ICEs of the ICESa2603 family and two ICESa2603 family-like ICEs derived from diverse hosts and locations, we have determined that ICEs comprised a backbone of 30 identical syntenic core genes and accessory genes that were restricted to the intergenic sites or the 3'-end of the non-conserved domain of core genes to maintain its function. ICESa2603 family integrase IntICE Sa 2603 specifically recognized a 15-bp att sequence (TTATTTAAGAGTAAC) at the 3'-end of rplL, which was highly conserved in genus Streptococcus. Phylogenetic analyses suggest that extensive recombination/insertion and the occurrence of a hybrid/mosaic in the ICESa2603 family were responsible for the significant increase in ICE diversity, thereby broadening its host range. Approximately 42.5 and 38.1% of the tested Streptococcus suis and Streptococcus agalactiae clinical isolates respectively contained ICESa2603 family Type IV secretion system (T4SS) genes, and 80.5 and 62.5% of which also respectively carried int ICE Sa 2603, indicating that ICESa2603 family is widely distributed across these bacteria. Sequencing and conjugation transfer of a novel sequence type ST303 clinical S. suis isolate HB1011 demonstrated that the 89K-subtype ICESsuHB1011 retained its transferrable function, thereby conferring tetracycline and macrolide resistance. PMID:26870017

  17. Comparative Genomic Analysis of the ICESa2603 Family ICEs and Spread of erm(B)- and tet(O)-Carrying Transferable 89K-Subtype ICEs in Swine and Bovine Isolates in China

    PubMed Central

    Huang, Jinhu; Liang, Yuan; Guo, Dawei; Shang, Kexin; Ge, Lin; Kashif, Jam; Wang, Liping

    2016-01-01

    Integrative and conjugative elements (ICEs) of the ICESa2603 family have been isolated from several species of Streptococcus spp.; however, the comparative genomic and evolutionary analyses of these particular ICEs are currently only at their initial stages. By investigating 13 ICEs of the ICESa2603 family and two ICESa2603 family-like ICEs derived from diverse hosts and locations, we have determined that ICEs comprised a backbone of 30 identical syntenic core genes and accessory genes that were restricted to the intergenic sites or the 3′-end of the non-conserved domain of core genes to maintain its function. ICESa2603 family integrase IntICESa2603 specifically recognized a 15-bp att sequence (TTATTTAAGAGTAAC) at the 3′-end of rplL, which was highly conserved in genus Streptococcus. Phylogenetic analyses suggest that extensive recombination/insertion and the occurrence of a hybrid/mosaic in the ICESa2603 family were responsible for the significant increase in ICE diversity, thereby broadening its host range. Approximately 42.5 and 38.1% of the tested Streptococcus suis and Streptococcus agalactiae clinical isolates respectively contained ICESa2603 family Type IV secretion system (T4SS) genes, and 80.5 and 62.5% of which also respectively carried intICESa2603, indicating that ICESa2603 family is widely distributed across these bacteria. Sequencing and conjugation transfer of a novel sequence type ST303 clinical S. suis isolate HB1011 demonstrated that the 89K-subtype ICESsuHB1011 retained its transferrable function, thereby conferring tetracycline and macrolide resistance. PMID:26870017

  18. Fate and transport of tylosin-resistant bacteria and macrolide resistance genes in artificially drained agricultural fields receiving swine manure

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Application of manure from swine treated with antibiotics introduces antibiotics and antibiotic resistance genes to soil with the potential for further movement in drainage water. Manure concentrations for ermB, ermC and ermF were all >109 copy g-1. Manure contained 1.76 x 105 CFUg-1 enterococci w...

  19. Inhibition of cell adhesion by phosphorylated Ezrin/Radixin/Moesin

    PubMed Central

    Tachibana, Kouichi; Haghparast, Seyed Mohammad Ali; Miyake, Jun

    2015-01-01

    Altered phosphorylation status of the C-terminal Thr residues of Ezrin/Radixin/Moesin (ERM) is often linked to cell shape change. To determine the role of phophorylated ERM, we modified phosphorylation status of ERM and investigated changes in cell adhesion and morphology. Treatment with Calyculin-A (Cal-A), a protein phosphatase inhibitor, dramatically augmented phosphorylated ERM (phospho-ERM). Cal-A-treatment or expression of phospho-mimetic Moesin mutant (Moesin-TD) induced cell rounding in adherent cells. Moreover, reattachment of detached cells to substrate was inhibited by either treatment. Phospho-ERM, Moesin-TD and actin cytoskeleton were observed at the plasma membrane of such round cells. Augmented cell surface rigidity was also observed in both cases. Meanwhile, non-adherent KG-1 cells were rather rich in phospho-ERM. Treatment with Staurosporine, a protein kinase inhibitor that dephosphorylates phospho-ERM, up-regulated the integrin-dependent adhesion of KG-1 cells to substrate. These findings strongly suggest the followings: (1) Phospho-ERM inhibit cell adhesion, and therefore, dephosphorylation of ERM proteins is essential for cell adhesion. (2) Phospho-ERM induce formation and/or maintenance of spherical cell shape. (3) ERM are constitutively both phosphorylated and dephosphorylated in cultured adherent and non-adherent cells. PMID:26555866

  20. Charting a Course through CORAL: Texas A&M University Libraries' Experience Implementing an Open-Source Electronic Resources Management System

    ERIC Educational Resources Information Center

    Hartnett, Eric; Beh, Eugenia; Resnick, Taryn; Ugaz, Ana; Tabacaru, Simona

    2013-01-01

    In 2010, after two previous unsuccessful attempts at electronic resources management system (ERMS) implementation, Texas A&M University (TAMU) Libraries set out once again to find an ERMS that would fit its needs. After surveying the field, TAMU Libraries selected the University of Notre Dame Hesburgh Libraries-developed, open-source ERMS, CORAL…

  1. Pathway and sink activity for photosynthate translocation in Pisolithus extraradical mycelium of ectomycorrhizal Pinus thunbergii seedlings.

    PubMed

    Teramoto, Munemasa; Wu, Bingyun; Hogetsu, Taizo

    2016-07-01

    The purpose of this study was to identify the pathway and sink activity of photosynthate translocation in the extraradical mycelium (ERM) of a Pisolithus isolate. We labelled ectomycorrhizal (ECM) Pinus thunbergii seedlings with (14)CO2 and followed (14)C distribution within the ERM by autoradiography. (14)C photosynthate translocation in the ERM resulted in (14)C distribution in rhizomorphs throughout the ERM, with (14)C accumulation at the front. When most radial mycelial connections between ECM root tips and the ERM front were cut, the whole allocation of (14)C photosynthates to the ERM was reduced. However, the overall pattern of (14)C distribution in the ERM was maintained even in regions immediately above and below the cut, with no local (14)C depletion or accumulation. We inferred from this result that every portion in the ERM has a significant sink activity and a definite sink capacity for photosynthates and that photosynthates detour the cut and reach throughout the ERM by translocation in every direction. Next, we prepared paired ECM seedlings, ERMs of which had been connected with each other by hyphal fusion, alongside, labelled the left seedling with (14)CO2, and shaded none, one or both of them. (14)C photosynthates were acropetally and basipetally translocated from the left ERM to ECM root tips of the right seedling through rhizomorphs in the left and right ERMs, respectively. With the left seedling illuminated, (14)C translocation from the left to the right ERM increased by shading the right seedling. This result suggests that reduced photosynthate transfer from the host to its ERM increased sink activity of the ERM. PMID:26861479

  2. Glycogen synthase kinase 3 inhibitors induce the canonical WNT/β-catenin pathway to suppress growth and self-renewal in embryonal rhabdomyosarcoma

    PubMed Central

    Chen, Eleanor Y.; DeRan, Michael T.; Ignatius, Myron S.; Grandinetti, Kathryn Brooke; Clagg, Ryan; McCarthy, Karin M.; Lobbardi, Riadh M.; Brockmann, Jillian; Keller, Charles; Wu, Xu; Langenau, David M.

    2014-01-01

    Embryonal rhabdomyosarcoma (ERMS) is a common pediatric malignancy of muscle, with relapse being the major clinical challenge. Self-renewing tumor-propagating cells (TPCs) drive cancer relapse and are confined to a molecularly definable subset of ERMS cells. To identify drugs that suppress ERMS self-renewal and induce differentiation of TPCs, a large-scale chemical screen was completed. Glycogen synthase kinase 3 (GSK3) inhibitors were identified as potent suppressors of ERMS growth through inhibiting proliferation and inducing terminal differentiation of TPCs into myosin-expressing cells. In support of GSK3 inhibitors functioning through activation of the canonical WNT/β-catenin pathway, recombinant WNT3A and stabilized β-catenin also enhanced terminal differentiation of human ERMS cells. Treatment of ERMS-bearing zebrafish with GSK3 inhibitors activated the WNT/β-catenin pathway, resulting in suppressed ERMS growth, depleted TPCs, and diminished self-renewal capacity in vivo. Activation of the canonical WNT/β-catenin pathway also significantly reduced self-renewal of human ERMS, indicating a conserved function for this pathway in modulating ERMS self-renewal. In total, we have identified an unconventional tumor suppressive role for the canonical WNT/β-catenin pathway in regulating self-renewal of ERMS and revealed therapeutic strategies to target differentiation of TPCs in ERMS. PMID:24706870

  3. Epiretinal membrane: optical coherence tomography-based diagnosis and classification.

    PubMed

    Stevenson, William; Prospero Ponce, Claudia M; Agarwal, Daniel R; Gelman, Rachel; Christoforidis, John B

    2016-01-01

    Epiretinal membrane (ERM) is a disorder of the vitreomacular interface characterized by symptoms of decreased visual acuity and metamorphopsia. The diagnosis and classification of ERM has traditionally been based on clinical examination findings. However, modern optical coherence tomography (OCT) has proven to be more sensitive than clinical examination for the diagnosis of ERM. Furthermore, OCT-derived findings, such as central foveal thickness and inner segment ellipsoid band integrity, have shown clinical relevance in the setting of ERM. To date, no OCT-based ERM classification scheme has been widely accepted for use in clinical practice and investigation. Herein, we review the pathogenesis, diagnosis, and classification of ERMs and propose an OCT-based ERM classification system. PMID:27099458

  4. Epiretinal membrane: optical coherence tomography-based diagnosis and classification

    PubMed Central

    Stevenson, William; Prospero Ponce, Claudia M; Agarwal, Daniel R; Gelman, Rachel; Christoforidis, John B

    2016-01-01

    Epiretinal membrane (ERM) is a disorder of the vitreomacular interface characterized by symptoms of decreased visual acuity and metamorphopsia. The diagnosis and classification of ERM has traditionally been based on clinical examination findings. However, modern optical coherence tomography (OCT) has proven to be more sensitive than clinical examination for the diagnosis of ERM. Furthermore, OCT-derived findings, such as central foveal thickness and inner segment ellipsoid band integrity, have shown clinical relevance in the setting of ERM. To date, no OCT-based ERM classification scheme has been widely accepted for use in clinical practice and investigation. Herein, we review the pathogenesis, diagnosis, and classification of ERMs and propose an OCT-based ERM classification system. PMID:27099458

  5. An ethanol extract of Ramulus mori improves blood circulation by inhibiting platelet aggregation.

    PubMed

    Lee, Jiyun; Kwon, Gayeung; Park, Jieun; Kim, Jeong-Keun; Choe, Soo Young; Seo, Yoonhee; Lim, Young-Hee

    2016-07-01

    Inappropriate platelet aggregation can cause blood coagulation and thrombosis. In this study, the effect of an ethanol extract of Ramulus mori (ERM) on blood circulation was investigated. The antithrombotic activity of ERM on rat carotid arterial thrombosis was evaluated in vivo, and the effect of ERM on platelet aggregation and blood coagulation time was evaluated ex vivo. To evaluate the safety of ERM, its cytotoxicity to platelets and its effect on tail bleeding time were assessed; ERM was not toxic to rat platelets and did not prolong bleeding time. Moreover, administering ERM to rats had a significant preventive effect on carotid arterial thrombosis in vivo, and significantly inhibited adenosine diphosphate- and collagen-induced platelet aggregation ex vivo, whereas it did not prolong coagulation periods, such as prothrombin time and activated partial thromboplastin time. The results suggest that ERM is effective in improving blood circulation via antiplatelet activity rather than anticoagulation activity. PMID:26967156

  6. Fate and transport of tylosin-resistant bacteria and macrolide resistance genes in artificially drained agricultural fields receiving swine manure.

    PubMed

    Luby, Elizabeth M; Moorman, Thomas B; Soupir, Michelle L

    2016-04-15

    Application of manure from swine treated with antibiotics introduces antibiotics and antibiotic resistance genes to soil with the potential for further movement in drainage water, which may contribute to the increase in antibiotic resistance in non-agricultural settings. We compared losses of antibiotic-resistant Enterococcus and macrolide-resistance (erm and msrA) genes in water draining from plots with or without swine manure application under chisel plow and no till conditions. Concentrations of ermB, ermC and ermF were all >10(9)copies g(-1) in manure from tylosin-treated swine, and application of this manure resulted in short-term increases in the abundance of these genes in soil. Abundances of ermB, ermC and ermF in manured soil returned to levels identified in non-manured control plots by the spring following manure application. Tillage practices yielded no significant differences (p>0.10) in enterococci or erm gene concentrations in drainage water and were therefore combined for further analysis. While enterococci and tylosin-resistant enterococci concentrations in drainage water showed no effects of manure application, ermB and ermF concentrations in drainage water from manured plots were significantly higher (p<0.01) than concentrations coming from non-manured plots. ErmB and ermF were detected in 78% and 44%, respectively, of water samples draining from plots receiving manure. Although ermC had the highest concentrations of the three genes in drainage water, there was no effect of manure application on ermC abundance. MsrA was not detected in manure, soil or water. This study is the first to report significant increases in abundance of resistance genes in waters draining from agricultural land due to manure application. PMID:26874610

  7. [Effects of Thermophilic Composting on Antibiotic Resistance Genes (ARGs) of Swine Manure Source].

    PubMed

    Zheng, Ning-guo; Huang, Nan; Wang, Wei-wei; Yu, Man; Chen, Xiao-yang; Yao, Yan-lai; Wang, Wei-ping; Hong, Chun-lai

    2016-05-15

    To investigate the effects of thermophilic composting process on antibiotic resistance genes (ARGs) of swine manure source at a field scale, the abundance of four erythromycin resistance genes (ermA, ermB, ermC and ermF), three β-lactam resistance genes (blaTEM, blaCTX and blaSHV) and two quinolone resistance genes (qnrA and qnrS) were quantified by quantitative PCR ( qPCR) during the composting process. The results suggested that the erm genes' copy numbers were significantly higher than those of the bla and qnr genes in the early stage of composting (P < 0.01). The maximum abundance of erm genes was ermB (9.88 x 10⁸ copies · g⁻¹), following by ermF (9.4 x 10⁸ copies · g⁻¹). At the end of the composting process, bla and qnr genes were at low levels, while erm genes were still at high levels. Even through ermF was proliferated comparing with the initial copies. These results indicated that thermophilic composting process could not effectively remove all ARGs. For some ARGs, compost may be a good bioreactor resulting in their proliferation. Application of composting products on farmland may cause transference of ARGs. PMID:27506057

  8. ER-mitochondrial junctions can be bypassed by dominant mutations in the endosomal protein Vps13.

    PubMed

    Lang, Alexander B; John Peter, Arun T; Walter, Peter; Kornmann, Benoît

    2015-09-14

    The endoplasmic reticulum-mitochondria encounter structure (ERMES) complex tethers the endoplasmic reticulum and the mitochondria. It is thought to facilitate interorganelle lipid exchange and influence mitochondrial dynamics and mitochondrial DNA maintenance. Despite this important role, ERMES is not found in metazoans. Here, we identified single amino acid substitutions in Vps13 (vacuolar protein sorting 13), a large universally conserved eukaryotic protein, which suppress all measured phenotypic consequences of ERMES deficiency. Combined loss of VPS13 and ERMES is lethal, indicating that Vps13 and ERMES function in redundant pathways. Vps13 dynamically localizes to vacuole-mitochondria and to vacuole-nucleus contact sites depending on growth conditions, suggesting that ERMES function can be bypassed by the activity of other contact sites, and that contact sites establish a growth condition-regulated organelle network. PMID:26370498

  9. Local Control and Outcome in Children with Localized Vaginal Rhabdomyosarcoma: A Report from the Soft Tissue Sarcoma Committee of the Children’s Oncology Group

    PubMed Central

    Walterhouse, David O.; Meza, Jane L.; Breneman, John C.; Donaldson, Sarah S.; Hayes-Jordan, Andrea; Pappo, Alberto S.; Arndt, Carola; Raney, R. Beverly; Meyer, William H.; Hawkins, Douglas S.

    2012-01-01

    Background The local control approach for girls with non-resected vaginal rhabdomyosarcoma (RMS) enrolled onto Intergroup RMS Study Group (IRSG)/Children’s Oncology Group (COG) studies has differed from that used at other primary sites by delaying or eliminating radiotherapy (RT) based on response achieved with chemotherapy and delayed primary resection. Procedures We reviewed locoregional treatment and outcome for patients with localized RMS of the vagina on the two most recent COG low-risk RMS studies. Results Forty-one patients with localized vaginal RMS were enrolled: 25 onto D9602 and 16 onto Subset 2 of ARST0331. Only four of the 39 with non-resected tumors received RT. The 5-year cumulative incidence of local recurrence was 26% on D9602, and the 2-year cumulative incidence of local recurrence was 43% on ARST0331. Increased local failure rates appeared to correlate with chemotherapy regimens that incorporated lower cumulative doses of cyclophosphamide. Estimated 5-year and 2-year failure free survival rates were 70% (95% CI: 46%, 84%) on D9602 and 42% (95% CI: 11%, 70%) on ARST0331, respectively. Conclusions To prevent local recurrence, we recommend a local control approach for patients with non-resected RMS of the vagina that is similar to that used for other primary sites and includes RT. We recognize that potential long-term effects of RT are sometimes unacceptable, especially for children less than 24 months of age. However, when making the decision to eliminate RT, the risk of local recurrence must be considered especially when using a chemotherapy regimen with a total cumulative cyclophosphamide dose of ≤ 4.8 g/m2. PMID:21298768

  10. Rapid molecular detection of inducible macrolide resistance in Mycobacterium chelonae and M. abscessus strains: a replacement for 14-day susceptibility testing?

    PubMed

    Hanson, Kimberly E; Slechta, E Susan; Muir, Haleina; Barker, Adam P

    2014-05-01

    The erm(41) gene causes inducible macrolide resistance in Mycobacterium abscessus but not Mycobacterium chelonae. erm(41) sequencing of 285 M. abscessus and 45 M. chelonae isolates was compared to 14-day susceptibility; agreement percentages were 98.9% and 100%, respectively. Extended incubation may not be necessary for M. chelonae, and the erm(41) genotype is a useful adjunct for M. abscessus. PMID:24554745

  11. Establishment of Hertwig's epithelial root sheath/epithelial rests of Malassez cell line from human periodontium.

    PubMed

    Nam, Hyun; Kim, Ji-Hye; Kim, Jae-Won; Seo, Byoung-Moo; Park, Joo-Cheol; Kim, Jung-Wook; Lee, Gene

    2014-07-01

    Human Hertwig's epithelial root sheath/epithelial rests of Malassez (HERS/ERM) cells are epithelial remnants of teeth residing in the periodontium. Although the functional roles of HERS/ERM cells have yet to be elucidated, they are a unique epithelial cell population in adult teeth and are reported to have stem cell characteristics. Therefore, HERS/ERM cells might play a role as an epithelial component for the repair or regeneration of dental hard tissues; however, they are very rare population in periodontium and the primary isolation of them is considered to be difficult. To overcome these problems, we immortalized primary HERS/ERM cells isolated from human periodontium using SV40 large T antigen (SV40 LT) and performed a characterization of the immortalized cell line. Primary HERS/ERM cells could not be maintained for more than 6 passages; however, immortalized HERS/ERM cells were maintained for more than 20 passages. There were no differences in the morphological and immunophenotypic characteristics of HERS/ERM cells and immortalized HERS/ERM cells. The expression of epithelial stem cell and embryonic stem cell markers was maintained in immortalized HERS/ERM cells. Moreover, immortalized HERS/ERM cells could acquire mesenchymal phenotypes through the epithelial-mesenchymal transition via TGF-β1. In conclusion, we established an immortalized human HERS/ERM cell line with SV40 LT and expect this cell line to contribute to the understanding of the functional roles of HERS/ERM cells and the tissue engineering of teeth. PMID:25081036

  12. Effects of advanced glycation end products on ezrin-dependent functions in LLC-PK1 proximal tubule cells.

    PubMed

    Bach, Leon A; Gallicchio, Marisa A; McRobert, E Anne; Tikoo, Anjali; Cooper, Mark E

    2005-06-01

    We have recently shown that advanced glycation products (AGEs) bind to the ERM (ezrin, radixin, moesin) family of proteins. ERM proteins act as cross-linkers between cell membrane proteins and the actin cytoskeleton. They are also involved in signal transduction pathways. They therefore have a critical role in normal cell processes, including modulation of cell shape, adhesion, and motility. We postulate that AGEs may contribute to diabetic complications by disrupting ERM function. In support of this hypothesis, AGEs inhibit ezrin-dependent tubulogenesis of proximal tubule cells. Phosphorylation is an important activating mechanism for ERM proteins, and AGEs inhibit ezrin phosphorylation mediated by the epidermal growth factor receptor. PMID:16037284

  13. Histone Deacetylase Inhibitors Antagonize Distinct Pathways to Suppress Tumorigenesis of Embryonal Rhabdomyosarcoma

    PubMed Central

    Vleeshouwer-Neumann, Terra; Phelps, Michael; Bammler, Theo K.; MacDonald, James W.; Jenkins, Isaac; Chen, Eleanor Y.

    2015-01-01

    Embryonal rhabdomyosarcoma (ERMS) is the most common soft tissue cancer in children. The prognosis of patients with relapsed or metastatic disease remains poor. ERMS genomes show few recurrent mutations, suggesting that other molecular mechanisms such as epigenetic regulation might play a major role in driving ERMS tumor biology. In this study, we have demonstrated the diverse roles of histone deacetylases (HDACs) in the pathogenesis of ERMS by characterizing effects of HDAC inhibitors, trichostatin A (TSA) and suberoylanilide hydroxamic acid (SAHA; also known as vorinostat) in vitro and in vivo. TSA and SAHA suppress ERMS tumor growth and progression by inducing myogenic differentiation as well as reducing the self-renewal and migratory capacity of ERMS cells. Differential expression profiling and pathway analysis revealed downregulation of key oncogenic pathways upon HDAC inhibitor treatment. By gain-of-function, loss-of-function, and chromatin immunoprecipitation (ChIP) studies, we show that Notch1- and EphrinB1-mediated pathways are regulated by HDACs to inhibit differentiation and enhance migratory capacity of ERMS cells, respectively. Our study demonstrates that aberrant HDAC activity plays a major role in ERMS pathogenesis. Druggable targets in the molecular pathways affected by HDAC inhibitors represent novel therapeutic options for ERMS patients. PMID:26636678

  14. Ezrin/radixin/moesin proteins differentially regulate endothelial hyperpermeability after thrombin.

    PubMed

    Adyshev, Djanybek M; Dudek, Steven M; Moldobaeva, Nurgul; Kim, Kyung-mi; Ma, Shwu-Fan; Kasa, Anita; Garcia, Joe G N; Verin, Alexander D

    2013-08-01

    Endothelial cell (EC) barrier disruption induced by inflammatory agonists such as thrombin leads to potentially lethal physiological dysfunction such as alveolar flooding, hypoxemia, and pulmonary edema. Thrombin stimulates paracellular gap and F-actin stress fiber formation, triggers actomyosin contraction, and alters EC permeability through multiple mechanisms that include protein kinase C (PKC) activation. We previously have shown that the ezrin, radixin, and moesin (ERM) actin-binding proteins differentially participate in sphingosine-1 phosphate-induced EC barrier enhancement. Phosphorylation of a conserved threonine residue in the COOH-terminus of ERM proteins causes conformational changes in ERM to unmask binding sites and is considered a hallmark of ERM activation. In the present study we test the hypothesis that ERM proteins are phosphorylated on this critical threonine residue by thrombin-induced signaling events and explore the role of the ERM family in modulating thrombin-induced cytoskeletal rearrangement and EC barrier function. Thrombin promotes ERM phosphorylation at this threonine residue (ezrin Thr567, radixin Thr564, moesin Thr558) in a PKC-dependent fashion and induces translocation of phosphorylated ERM to the EC periphery. Thrombin-induced ERM threonine phosphorylation is likely synergistically mediated by protease-activated receptors PAR1 and PAR2. Using the siRNA approach, depletion of either moesin alone or of all three ERM proteins significantly attenuates thrombin-induced increase in EC barrier permeability (transendothelial electrical resistance), cytoskeletal rearrangements, paracellular gap formation, and accumulation of phospho-myosin light chain. In contrast, radixin depletion exerts opposing effects on these indexes. These data suggest that ERM proteins play important differential roles in the thrombin-induced modulation of EC permeability, with moesin promoting barrier dysfunction and radixin opposing it. PMID:23729486

  15. Cognitive architectures and autonomy: Commentary and Response

    NASA Astrophysics Data System (ADS)

    2012-11-01

    Editors: Włodzisław Duch / Ah-Hwee Tan / Stan Franklin Autonomy for AGI Cristiano Castelfranchi 31 Are Disembodied Agents Really Autonomous? Antonio Chella 33 The Perception-…-Action Cycle Cognitive Architecture and Autonomy: the View from the Brain Vassilis Cutsuridis 36 Autonomy Requires Creativity and Meta-Learning Włodzisław Duch 39 Meta Learning, Change of Internal Workings, and LIDA Ryan McCall / Stan Franklin 42 An Appeal for Declaring Research Goals Brandon Rohrer 45 The Development of Cognition as the Basis for Autonomy Frank van der Velde 47 Autonomy and Intelligence Pei Wang 49 Autonomy, Isolation, and Collective Intelligence Nikolaos Mavridis 51 Response to Comments Kristinn R. Thórisson / Helgi Páll Helgasson 56

  16. Molecular epidemiology and genetic linkage of macrolide and aminoglycoside resistance in Staphylococcus intermedius of canine origin.

    PubMed

    Boerlin, P; Burnens, A P; Frey, J; Kuhnert, P; Nicolet, J

    2001-03-20

    A collection of 77 Staphylococcus intermedius isolates from dogs and cats in Switzerland was examined for resistance to erythromycin. Resistance profiles for 14 additional antibiotics were compared between erythromycin-resistant and susceptible isolates. A resistance prevalence of 27% for erythromycin was observed in the population under study. Complete correlation between resistance to erythromycin, and to spiramycin, streptomycin, and neomycin was observed. The erythromycin-resistant isolates all had a reduced susceptibility to clindamycin when compared to the erythromycin-susceptible isolates. Both constitutive and inducible resistance phenotypes were observed for clindamycin. Ribotyping showed that macrolide-aminoglycoside resistance was randomly distributed among unrelated strains. This suggests that this particular resistance profile is not related to a single bacterial clone but to the horizontal transfer of resistance gene clusters in S. intermedius populations. The erythromycin-resistant isolates were all carrying erm(B), but not erm(A), erm(C), or msr(A). The erm(B) gene was physically linked to Tn5405-like elements known as resistance determinants for streptomycin, streptothricin, neomycin and kanamycin. Analysis of the region flanking erm(B) showed the presence of two different groups of erm(B)-Tn5405-like elements in the S. intermedius population examined and of elements found in Gram-positive species other than staphylococci. This strongly suggests that erm(B) or the whole erm(B)-Tn5405-like elements in S. intermedius originate from other bacterial species, possibly from enterococci. PMID:11230937

  17. Think Locally: A Prudent Approach to Electronic Resource Management Systems

    ERIC Educational Resources Information Center

    Gustafson-Sundell, Nat

    2011-01-01

    A few articles have drawn some amount of attention specifically to the local causes of the success or failure of electronic resource management system (ERMS) implementations. In fact, it seems clear that local conditions will largely determine whether any given ERMS implementation will succeed or fail. This statement might seem obvious, but the…

  18. Multicenter Study of the Mechanisms of Resistance and Clonal Relationships of Streptococcus agalactiae Isolates Resistant to Macrolides, Lincosamides, and Ketolides in Spain

    PubMed Central

    Gonzalez, J. J.; Andreu, A.

    2005-01-01

    Macrolide, lincosamide, and ketolide mechanisms of resistance and clonal relationships were characterized in a collection of 79 resistant group B streptococcus isolates obtained from neonates or pregnant women. The erm(B), erm(TR), and mef(A) genes were present in 62%, 30.4%, and 3.8% of the isolates, respectively. There was considerable clonal diversity among them. PMID:15917563

  19. Ericoid mycorrhizal root fungi and their multicopper oxidases from a temperate forest shrub

    PubMed Central

    Wurzburger, Nina; Higgins, Brian P; Hendrick, Ronald L

    2012-01-01

    Ericoid mycorrhizal fungi (ERM) may specialize in capturing nutrients from their host's litter as a strategy for regulating nutrient cycles in terrestrial ecosystems. In spite of their potential significance, we know little about the structure of ERM fungal communities and the genetic basis of their saprotrophic traits (e.g., genes encoding extracellular enzymes). Rhododendron maximum is a model ERM understory shrub that influences the nutrient cycles of montane hardwood forests in the southern Appalachians (North Carolina, USA). We sampled ERM roots of R. maximum from organic and mineral soil horizons and identified root fungi by amplifying and sequencing internal transcribed spacer (ITS) ribosomal DNA (rDNA) collected from cultures and clones. We observed 71 fungal taxa on ERM roots, including known symbionts Rhizoscyphus ericae and Oidiodendron maius, putative symbionts from the Helotiales, Chaetothyriales, and Sebacinales, ectomycorrhizal symbionts, and saprotrophs. Supporting the idea that ERM fungi are adept saprotrophs, richness of root-fungi was greater in organic than in mineral soil horizons. To study the genetic diversity of oxidative enzymes that contribute to decomposition, we amplified and sequenced a portion of genes encoding multicopper oxidases (MCOs) from ERM ascomycetes. Most fungi possessed multiple copies of MCO sequences with strong similarities to known ferroxidases and laccases. Our findings indicate that R. maximum associates with a taxonomically and ecologically diverse fungal community. The study of MCO gene diversity and expression may be useful for understanding how ERM root fungi regulate the cycling of nutrients between the host plant and the soil environment. PMID:22408727

  20. Thinking beyond "the Way We've Always Done It"

    ERIC Educational Resources Information Center

    Ohler, Lila

    2013-01-01

    In this article, the author discusses the future potential of electronic resources management (ERM) organization, within the development of the next-generation integrated library systems (ILS), as a very promising strategy. This also presents enormous challenges to ERM librarians though, especially on how to determine standards and workflows. The…

  1. Vitiating Vulnerability

    ERIC Educational Resources Information Center

    Abraham, Janice M.

    2007-01-01

    Enterprise Risk Management (ERM) is an institutionwide approach to identifying and responding to risks that threaten an institution's ability to achieve its objectives. Board members may use ERM to clarify the role of governance in risk management and decide whether the institution should take on new risks or reduce its current risks. Establishing…

  2. Leveraging Enterprise Risk Management: Opportunity for Greater Relevance

    ERIC Educational Resources Information Center

    Abraham, Janice M.; Baird, Robert; Neugebauer, Frank

    2013-01-01

    Enterprise Risk Management (ERM) gained a foothold in higher education during the first decade of the 21st century. College and university governing board members came to their board service with the perspective and experience of having served on corporate boards and in corporate leadership positions where ERM was in active use. Trustees asked…

  3. In human T cells mifepristone antagonizes glucocorticoid non-genomic rapid responses in terms of Na(+)/H(+)-exchange 1 activity, but not ezrin/radixin/moesin phosphorylation.

    PubMed

    Chien, Eileen Jea; Hsu, Ching-Hui; Chang, Vincent Han-Jhih; Lin, Enoch Pin-Yi; Kuo, Trista Pin-Tsun; Chien, Chau-Heng; Lin, Hsiao-Yi

    2016-07-01

    Glucocorticoids (GCs) and progesterone have been employed as immunosuppressive agents during pregnancy for many years. Intracellular acidification by GCs is due to a rapid non-genomic inhibition of membrane Na(+)/H(+)-exchange 1 (NHE1) activity and is followed by immunosuppression of PHA-stimulated proliferation. NHE1 is tethered to the cortical actin cytoskeleton through ezrin/radixin/moesin (ERM) proteins within lipid rafts; these regulate cell shape, migration and resistance to apoptosis. We explored whether mifepristone (RU486), an antagonist of GCs in T cells, is able to completely block rapid non-genomic responses, namely NHE1 activity and the phosphorylation C-terminal residues of ERM proteins at threonine (cp-ERM). GCs stimulate a rapid non-genomic cp-ERM response in cells within 5min. RU486 antagonized the GC-induced rapid decrease in NHE1 activity, and arrested PHA-stimulated T cells at G0/G1 phase but had no effect on the rapid increase in cp-ERM, which persisted for 24h. However, the cp-ERM response was blocked by staurosporine in both resting and GC stimulated cells. The results of RU486 antagonized the GC induced rapid decrease in NHE1 ion transport activity, but not the increase cp-ERM. This suggests that RU486 in T cells exerts its antagonistic effects at NHE1 containing plasma membrane sites and not where cp-ERM links lipid rafts to cortical cytoskeletons. PMID:26773750

  4. Emergent Motivation to Read in Prekindergarten Children

    ERIC Educational Resources Information Center

    Zheng, Guoguo; Schwanenflugel, Paula J.; Rogers, Samantha M.

    2016-01-01

    This study aimed to develop and validate a measure of emergent reading motivation designed for prekindergarten children, called the Emergent Reading Motivation Scale (ERMS). The development of the ERMS was to overcome the limitation that current existing reading motivation measures are not developmentally appropriate for young children. Fifty-six…

  5. Tools, Techniques, and Training: Results of an E-Resources Troubleshooting Survey

    ERIC Educational Resources Information Center

    Rathmel, Angela; Mobley, Liisa; Pennington, Buddy; Chandler, Adam

    2015-01-01

    A primary role of any e-resources librarian or staff is troubleshooting electronic resources (e-resources). While much progress has been made in many areas of e-resources management (ERM) to understand the ERM lifecycle and to manage workflows, troubleshooting access remains a challenge. This collaborative study is the result of the well-received…

  6. The conserved GTPase Gem1 regulates endoplasmic reticulum–mitochondria connections

    PubMed Central

    Kornmann, Benoît; Osman, Christof; Walter, Peter

    2011-01-01

    Mitochondria are connected to the endoplasmic reticulum (ER) through specialized protein complexes. We recently identified the ER–mitochondria encounter structure (ERMES) tethering complex, which plays a role in phospholipid exchange between the two organelles. ERMES also has been implicated in the coordination of mitochondrial protein import, mitochondrial DNA replication, and mitochondrial dynamics, suggesting that these interorganelle contact sites play central regulatory roles in coordinating various aspects of the physiology of the two organelles. Here we purified ERMES complexes and identified the Ca2+-binding Miro GTPase Gem1 as an integral component of ERMES. Gem1 regulates the number and size of the ERMES complexes. In vivo, association of Gem1 to ERMES required the first of Gem1’s two GTPase domains and the first of its two functional Ca2+-binding domains. In contrast, Gem1’s second GTPase domain was required for proper ERMES function in phospholipid exchange. Our results suggest that ERMES is not a passive conduit for interorganellar lipid exchange, but that it can be regulated in response to physiological needs. Furthermore, we provide evidence that the metazoan Gem1 ortholog Miro-1 localizes to sites of ER–mitochondrial contact, suggesting that some of the features ascribed to Gem1 may be evolutionarily conserved. PMID:21825164

  7. Prevalence of inducible clindamycin resistance among community-associated staphylococcal isolates in central Serbia.

    PubMed

    Aleksandra, A D; Misic, M S; Mira, Z V; Violeta, N M; Dragana, I T; Zoran, B M; Dejan, V S; Milanko, S D; Dejan, B D

    2014-01-01

    The emergence of resistance to most antimicrobial agents in staphylococci indicates the need for new effective agents in the treatment of staphylococcal infections. Clindamycin is considered to be one safe, effective and less costly agent. We analysed 482 staphylococcal isolates. Detection of inducible clindamycin resistance was performed by the D-test, while the presence of methylases genes: erm (A), erm (B) and erm (C), as well as, macrolide efflux gene mef was determined by polymerase chain reaction. Inducible clindamycin resistance phenotype was significantly higher in Staphylococcus aureus (S. aureus) strains then in coagulase-negative staphylococci (CNS). Among analysed S. aureus isolates, the predominance of the erm (C) gene, followed by the erm (A) gene were detected. These results indicate that the D-test should be routinely performed on each staphylococcal isolates. PMID:24399388

  8. Artesunate induces ROS- and p38 MAPK-mediated apoptosis and counteracts tumor growth in vivo in embryonal rhabdomyosarcoma cells.

    PubMed

    Beccafico, Sara; Morozzi, Giulio; Marchetti, Maria Cristina; Riccardi, Carlo; Sidoni, Angelo; Donato, Rosario; Sorci, Guglielmo

    2015-09-01

    Rhabdomyosarcoma represents about 50% of soft-tissue sarcomas and 10% of malignant solid tumors in childhood. Embryonal rhabdomyosarcoma (ERMS) is the most frequent subtype, suggested to have an origin in muscle precursor cells that fail to exit the cell cycle and terminally differentiate mainly because of overexpression of the transcription factor, PAX7, which sustains proliferation, migration and invasiveness in ERMS cells. Artesunate (ARS) is a semi-synthetic derivative of artemisinin (ART), a natural compound well known as an antimalarial drug. However, ART and its derivatives have been found efficacious even as anticancer drugs that induce cell cycle arrest and/or apoptosis in several kinds of cancer. Here, we show that ARS dose-dependently induces DNA damage and apoptosis in ERMS cell lines. Production of reactive oxygen species (ROS) and activation of p38 MAPK have a central role in triggering ARS-mediated apoptosis in ERMS cells; indeed either the antioxidant, N-acetylcysteine or the p38 MAPK inhibitor, SB203580, protects ERMS cells from ARS-induced apoptosis. Moreover, ARS treatment in ERMS cells ROS-dependently induces the expression of the myo-miRs, miR-133a and miR-206, which are down-regulated in RMS, and reduces PAX7 protein levels. Finally, ARS upregulates the expression of the adhesion molecules, NCAM and integrin β1, and reduces migration and invasiveness of ERMS cells in vitro, and ARS treatment reduces of about 50% the growth of ERMS xenografts in vivo. Our results are the first evidence of efficacy of ART derivatives in restraining ERMS growth in vivo, and suggest ARS as a potential candidate for therapeutic treatment of ERMS. PMID:26153023

  9. Disruption of mycorrhizal extraradical mycelium and changes in leaf water status and soil aggregate stability in rootbox-grown trifoliate orange

    PubMed Central

    Zou, Ying-Ning; Srivastava, A. K.; Ni, Qiu-Dan; Wu, Qiang-Sheng

    2015-01-01

    Arbuscular mycorrhizas possess well developed extraradical mycelium (ERM) network that enlarge the surrounding soil for better acquisition of water and nutrients, besides soil aggregation. Distinction in ERM functioning was studied under a rootbox system, which consisted of root+hyphae and root-free hyphae compartments separated by 37-μm nylon mesh with an air gap. Trifoliate orange (Poncirus trifoliata) seedlings were inoculated with Funneliformis mosseae in root+hyphae compartment, and the ERM network was established between the two compartments. The ERM network of air gap was disrupted before 8 h of the harvest (one time disruption) or multiple disruptions during seedlings acclimation. Our results showed that mycorrhizal inoculation induced a significant increase in growth (plant height, stem diameter, and leaf, stem, and root biomass) and physiological characters (leaf relative water content, leaf water potential, and transpiration rate), irrespective of ERM status. Easily-extractable glomalin-related soil protein (EE-GRSP) and total GRSP (T-GRSP) concentration and mean weight diameter (MWD, an indicator of soil aggregate stability) were significantly higher in mycorrhizosphere of root+hyphae and root-free hyphae compartments than non-mycorrhizosphere. One time disruption of ERM network did not influence plant growth and soil properties but only notably decreased leaf water. Periodical disruption of ERM network at weekly interval markedly inhibited the mycorrhizal roles on plant growth, leaf water, GRSP production, and MWD in root+hyphae and hyphae chambers. EE-GRSP was the most responsive GRSP fraction to changes in leaf water and MWD under root+hyphae and hyphae conditions. It suggests that effect of peridical disruption of ERM network was more impactful than one-time disruption of ERM network with regard to leaf water, plant growth, and aggregate stability responses, thereby, implying ERM network aided in developing the host plant metabolically more active

  10. Disruption of mycorrhizal extraradical mycelium and changes in leaf water status and soil aggregate stability in rootbox-grown trifoliate orange.

    PubMed

    Zou, Ying-Ning; Srivastava, A K; Ni, Qiu-Dan; Wu, Qiang-Sheng

    2015-01-01

    Arbuscular mycorrhizas possess well developed extraradical mycelium (ERM) network that enlarge the surrounding soil for better acquisition of water and nutrients, besides soil aggregation. Distinction in ERM functioning was studied under a rootbox system, which consisted of root+hyphae and root-free hyphae compartments separated by 37-μm nylon mesh with an air gap. Trifoliate orange (Poncirus trifoliata) seedlings were inoculated with Funneliformis mosseae in root+hyphae compartment, and the ERM network was established between the two compartments. The ERM network of air gap was disrupted before 8 h of the harvest (one time disruption) or multiple disruptions during seedlings acclimation. Our results showed that mycorrhizal inoculation induced a significant increase in growth (plant height, stem diameter, and leaf, stem, and root biomass) and physiological characters (leaf relative water content, leaf water potential, and transpiration rate), irrespective of ERM status. Easily-extractable glomalin-related soil protein (EE-GRSP) and total GRSP (T-GRSP) concentration and mean weight diameter (MWD, an indicator of soil aggregate stability) were significantly higher in mycorrhizosphere of root+hyphae and root-free hyphae compartments than non-mycorrhizosphere. One time disruption of ERM network did not influence plant growth and soil properties but only notably decreased leaf water. Periodical disruption of ERM network at weekly interval markedly inhibited the mycorrhizal roles on plant growth, leaf water, GRSP production, and MWD in root+hyphae and hyphae chambers. EE-GRSP was the most responsive GRSP fraction to changes in leaf water and MWD under root+hyphae and hyphae conditions. It suggests that effect of peridical disruption of ERM network was more impactful than one-time disruption of ERM network with regard to leaf water, plant growth, and aggregate stability responses, thereby, implying ERM network aided in developing the host plant metabolically more active

  11. Different Distribution Patterns between Putative Ercoid Mycorrhizal and Other Fungal Assemblages in Roots of Rhododendron decorum in the Southwest of China

    PubMed Central

    Sun, Lifu; Pei, Kequan; Wang, Fang; Ding, Qiong; Bing, Yanhong; Gao, Bo; Zheng, Yu; Liang, Yu; Ma, Keping

    2012-01-01

    Fungal diversity within plant roots is affected by several factors such as dispersal limitation, habitat filtering, and plant host preference. Given the differences in life style between symbiotic and non-symbiotic fungi, the main factors affecting these two groups of fungi may be different. We assessed the diversity of root associated fungi of Rhododendron decorum using internal transcribed spacer (ITS) sequencing and terminal restriction fragment length polymorphism (T-RFLP) analysis, and our aim was to evaluate the role of different factors in structuring ericoid mycorrhizal (ERM) and non-ericoid mycorrhizal (NEM) fungal communities. Thirty-five fungal operational taxonomic units (OTUs) were found in roots of R. decorum, of which 25 were putative ERM fungal species. Of the two main groups of known ERM, helotialean fungi were more abundant and common than sebacinalean species. Geographic and host patterning of the fungal assemblages were different for ERM and NEM. The distribution of putative ERM fungal terminal restriction fragments (TRFs) showed that there were more common species within ERM than in the NEM fungal assemblages. Results of Mantel tests indicated that the composition of NEM fungal assemblages correlated with geographic parameters while ERM fungal assemblages lacked a significant geographic pattern and instead were correlated with host genotype. Redundancy analysis (RDA) showed that the NEM fungal assemblages were significantly correlated with latitude, longitude, elevation, mean annual precipitation (MAP), and axis 2 of a host-genetic principle component analysis (PCA), while ERM fungal assemblages correlated only with latitude and axis 1 of the host-genetic PCA. We conclude that ERM and NEM assemblages are affected by different factors, with the host genetic composition more important for ERM and geographic factors more important for NEM assemblages. Our results contribute to understanding the roles of dispersal limitation, abiotic factors and

  12. Emergence of Multidrug-Resistant Campylobacter Species Isolates with a Horizontally Acquired rRNA Methylase

    PubMed Central

    Wang, Yang; Zhang, Maojun; Deng, Fengru; Shen, Zhangqi; Wu, Congming; Zhang, Jianzhong

    2014-01-01

    Antibiotic-resistant Campylobacter constitutes a serious threat to public health, and resistance to macrolides is of particular concern, as this class of antibiotics is the drug of choice for clinical therapy of campylobacteriosis. Very recently, a horizontally transferrable macrolide resistance mediated by the rRNA methylase gene erm(B) was reported in a Campylobacter coli isolate, but little is known about the dissemination of erm(B) among Campylobacter isolates and the association of erm(B)-carrying isolates with clinical disease. To address this question and facilitate the control of antibiotic-resistant Campylobacter, we determined the distribution of erm(B) in 1,554 C. coli and Campylobacter jejuni isolates derived from food-producing animals and clinically confirmed human diarrheal cases. The results revealed that 58 of the examined isolates harbored erm(B) and exhibited high-level resistance to macrolides, and most were recent isolates, derived in 2011-2012. In addition, the erm(B)-positive isolates were all resistant to fluoroquinolones, another clinically important antibiotic used for treating campylobacteriosis. The erm(B) gene is found to be associated with chromosomal multidrug resistance genomic islands (MDRGIs) of Gram-positive origin or with plasmids of various sizes. All MDRGIs were transferrable to macrolide-susceptible C. jejuni by natural transformation under laboratory conditions. Molecular typing of the erm(B)-carrying isolates by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) identified diverse genotypes and outbreak-associated diarrheal isolates. Molecular typing also suggested zoonotic transmission of erm(B)-positive Campylobacter. These findings reveal an emerging and alarming trend of dissemination of erm(B) and MDRGIs in Campylobacter and underscore the need for heightened efforts to control their further spread. PMID:24982085

  13. Distribution of Genes Encoding Resistance to Macrolides Among Staphylococci Isolated From the Nasal Cavity of Hospital Employees in Khorramabad, Iran

    PubMed Central

    Goudarzi, Gholamreza; Tahmasbi, Farzad; Anbari, Khatereh; Ghafarzadeh, Masoumeh

    2016-01-01

    Background Epidemiological data on antibiotic susceptibility of Staphylococcus strains isolated from nasal carriers in each region can be helpful to select appropriate drugs to eradicate carriage states, control nosocomial infections and also treat patients. Objectives The current study aimed to investigate the antibiotic resistance profile and the molecular prevalence of the ermA, ermB, ermC and msrA genes among Staphylococcus strains isolated from the anterior nares of hospital employees. Patients and Methods In this cross-sectional study, a total of 100 Staphylococcus isolates, 51 Staphylococcus aureus, 49 coagulase-negative staphylococci (CoNS) were isolated from the anterior nares of hospital employees in Khorramabad, Iran. Susceptibility pattern to macrolide antibiotics were determined using the disk diffusion method. The polymerase chain reaction (PCR) assay was applied to determine the major erythromycin-resistant genes (ermA, ermB, ermC and msrA). Results Fifty-three (53%) isolates were simultaneously resistant to erythromycin, azithromycin and clarithromycin (cross-resistance); while 8 (8%) isolates had variable macrolide susceptibility pattern. Among the S. aureus isolates, the difference in prevalence of resistance to erythromycin between males and females was significant (P = 0.011). The frequency of ermA, ermB, ermC, and msrA genes were 3%, 5%, 33% and 20%, respectively. It was also found that out of 53 isolates resistant to erythromycin, 44 (83%) isolates (eight S. aureus and thirty-six CoNS strains) carried at least one of the four tested genes. Eight (8%) isolates had intermediate phenotype to erythromycin, in which 4 (50%) isolates carried ermB or ermC genes. In addition, out of 39 erythromycin-susceptible isolates, 3 (7.7%) isolates were positive for ermB or ermC genes. Conclusions No entire association was found between genotype and phenotype methods to detect macrolides-resistant isolates. In addition, distribution of genetically erythromycin

  14. Genetic engineering techniques for lactic acid bacteria: construction of a stable shuttle vector and expression vector for β-glucuronidase.

    PubMed

    Chang, Shiao-Ming; Yan, Tsong-Rong

    2014-02-01

    The shuttle vector, pUL6erm, was constructed by using a replicon from pL2, a multiple cloning site, colE1 ori, the ori of Gram-negative bacteria from vector pUC19, and the erythromycin resistance gene from pVA838 as a selection marker. pUL6erm could be transformed easily and maintained stably in Lactococcus lactis, Streptococcus thermophilus, Lactobacillus plantarum and Lactobacillus casei. Transformation assays of pUL6erm indicated that it had a narrow host range. β-Glucuronidase was induced in the presence of 0.3 M NaCl and 50 mM glutamate and expressed at 2.4 U mg(-1) with the expression vector (pUL6erm-gadR-GUS) constructed based on pUL6erm carrying β-glucuronidase gene wuth a chloride-inducible (gadR) expression cassette using Pgad as promoter. Therefore, pUL6erm and pUL6erm-gadR-GUS might be a safe and useful genetic tool for the improvement of lactic acid bacteria. PMID:24101246

  15. Evidence for a putative biomarker for substance dependence.

    PubMed

    Taylor, Jeanette; James, Lisa M

    2009-09-01

    Electrodermal response modulation (ERM) reflects the reduction in skin conductance response to an aversive stimulus that is temporally predictable relative to when it is unpredictable. Poor ERM is associated with substance dependence (SD). It was hypothesized that ERM is a putative biomarker for SD rather than for externalizing disorders generally. Participants included 83 controls (no SD, antisocial personality disorder [PD] or borderline PD), 52 participants with SD only (SD and no PD), 12 with PD only (antisocial and/or borderline PD and no SD), and 35 comorbid (having SD and PD). Diagnoses at definite and probable certainty levels were used and were determined by semistructured clinical interviews. ERM was calculated from skin conductance responses to predictable and unpredictable 2-s 110-dB white noise blasts. As expected, the SD-only and comorbid groups had significantly lower ERM scores than the control group, which did not differ significantly from the PD-only group. Results provide preliminary evidence that ERM is a putative biomarker for SD. Future research should examine cognitive correlates of ERM in an effort to understand why it relates to SD. PMID:19769433

  16. G Protein-coupled Receptor Kinase 2–mediated Phosphorylation of Ezrin Is Required for G Protein-coupled Receptor–dependent Reorganization of the Actin Cytoskeleton

    PubMed Central

    Cant, Sarah H.; Pitcher, Julie A.

    2005-01-01

    G protein-coupled receptor kinase 2 (GRK2) phosphorylates and desensitizes activated G protein-coupled receptors (GPCRs). Here, we identify ezrin as a novel non-GPCR substrate of GRK2. GRK2 phosphorylates glutathione S-transferase (GST)-ezrin, but not an ezrin fusion protein lacking threonine 567 (T567), in vitro. These results suggest that T567, the regulatory phosphorylation site responsible for maintaining ezrin in its active conformation, represents the principle site of GRK2-mediated phosphorylation. Two lines of evidence indicate that GRK2-mediated ezrin-radixinmoesin (ERM) phosphorylation serves to link GPCR activation to cytoskeletal reorganization. First, in Hep2 cells muscarinic M1 receptor (M1MR) activation causes membrane ruffling. This ruffling response is ERM dependent and is accompanied by ERM phosphorylation. Inhibition of GRK2, but not rho kinase or protein kinase C, prevents ERM phosphorylation and membrane ruffling. Second, agonist-induced internalization of the β2-adrenergic receptor (β2AR) and M1MR is accompanied by ERM phosphorylation and localization of phosphorylated ERM to receptor-containing endocytic vesicles. The colocalization of internalized β2AR and phosphorylated ERM is not dependent on Na+/H+ exchanger regulatory factor binding to the β2AR. Inhibition of ezrin function impedes β2AR internalization, further linking GPCR activation, GRK activity, and ezrin function. Overall, our results suggest that GRK2 serves not only to attenuate but also to transduce GPCR-mediated signals. PMID:15843435

  17. High Incidence of Macrolide and Tetracycline Resistance among Streptococcus Agalactiae Strains Isolated from Clinical Samples in Tehran, Iran

    PubMed Central

    EMANEINI, Mohammad; MIRSALEHIAN, Akbar; BEIGVIERDI, Reza; FOOLADI, Abbas Ali Imani; ASADI, Fatemeh; JABALAMELI, Fereshteh; TAHERIKALANI, Morovat

    2014-01-01

    Background: Streptococcus agalactiae or Group B Streptococci (GBS) is an important bacterial pathogen that causes a wide range of infections including neonatal sepsis, meningitis, pneumonia and soft tissue or urinary tract infections. Material and methods: One hundred and fifteen isolates of Streptococcus agalactiae collected from urine specimens of patients attending a hospital in Tehran. All isolates were screened for their capsular types and genes encoding resistance to the macrolide and tetracycline antibiotics by PCR and multiplex PCR–based methods. Results: Most of isolates belonged to capsular types III (49%), V (19%), II (16%), and Ib (6%). Twelve isolates (10%) were nontypable. All isolates were susceptible to penicillin and Quinupristin-dalfopristin, but were resistant to clindamycin (35%), chloramphenicol (45%), erythromycin (35%), linezolid (1%) and tetracycline (96%). The most prevalent antimicrobial resistance gene was tetM found in 93% of the isolates followed by ermTR, ermB, and tetK, found in 23%, 16%, and 16% of isolates, respectively. The genes, tetL, tetO, ermA, ermC and mefA were not detected in any of the S. agalactiae isolates. Of the 110 tetracycline resistant S. agalactiae, 89 isolates harbored the tetM gene alone and eighteen isolates carried the tetM gene with the tetK gene. All erythromycin-resistant isolates exhibited cMLSB resistance phenotype, 22 isolates harbored the ermTR gene alone and five isolates carried the ermTR gene with the ermB gene. The rate of coexistence of genes encoding the erythromycin and tetracycline resistance determinants was 34%. Conclusion: The present study demonstrated that S. agalactiae isolates obtained from urine samples showed a high rate of resistance to tetracycline, chloramphenicol and macrolide antibiotics and were commonly associated with the resistance genes temM, ermTR or ermB. PMID:25705271

  18. Prevalence and mechanisms of erythromycin resistance in Streptococcus agalactiae from healthy pregnant women.

    PubMed

    Pinheiro, Sandra; Radhouani, Hajer; Coelho, Céline; Gonçalves, Alexandre; Carvalho, Eulália; Carvalho, José António; Ruiz-Larrea, Fernanda; Torres, Carmen; Igrejas, Gilberto; Poeta, Patrícia

    2009-06-01

    We sought to determine the resistance phenotypes for erythromycin and clindamycin and the mechanisms implicated in 93 Streptococcus agalactiae isolates recovered from healthy pregnant women. Susceptibility testing for erythromycin, clindamycin, penicillin, cefotaxime, vancomycin, quinupristin-dalfopristin, choramphenicol, ofloxacin, and meropenen was carried out by disc-diffusion test, and the E-test was also applied for erythromycin and clindamycin. The constitutive MLS(B) resistance (cMLS(B)) and inducible MLS(B) resistance (iMLS(B)) phenotypes, respectively, as well as the M resistance phenotype were determined by the erythromycin-clindamycin double-disc test. The presence of ermA, ermB, ermC, msrA, and mef(A/E) macrolide resistance genes was studied by PCR. Resistance to erythromycin and clindamycin was found in 15% and 9.6% of the isolates, respectively. The resistance phenotypes detected among the 14 erythromycin-resistant isolates were as follows (number of isolates): cMLS(B) (9), iMLS(B) (3), and M (2). The MICs for erythromycin and clindamycin were as follows: cMLS(B) isolates (128-256 and >or=32 mg/L, respectively), iMLS(B) isolates (16-256 and 1 mg/L), and M isolates (2-8 and 1 mg/L). The following combination of genes were detected among isolates with cMLS(B) or iMLS(B) phenotypes: erm(B) (6 isolates), ermA + ermTR (3), ermA + ermB + ermTR (1), and none of these genes (2). The two isolates with M phenotype harbored the mef(A/E), and msrA gene was also found in one of them. PMID:19432524

  19. [Fate of ARB and ARGs During Wastewater Treatment Process of Spiramycin Production].

    PubMed

    Qin, Cai-xia; Tong, Juan; Shen, Pei-hong; Wei, Yuan-song

    2015-09-01

    Antibiotic resistant bacteria (ARB) and antibiotic resistance gene(ARG) pose great risk to both environment and human health. This study aimed to investigate the fate of macrolide resistant bacteria, six macrolide resistance genes ermB, ermF, ermX, mefA, ereA, mphB and three transfer elements ISCR1, intIl and Tn916/1545 during wastewater biological treatment processes of spiramycin production. Samples were collected from an antibiotic wastewater treatment station in different seasons. Results showed that the total heterotrophs and Enterococci were mostly removed during wastewater biological treatment, with the reduction of 1. 6-2. 1 logs for total heterotrophs and of 3. 7 logs for Enterococci, respectively. For 94 antibiotic resistant Enterococci individually isolated from four different treatment units including adjusting tank, anaerobic tank, anoxic tank, and aerobic tank, all of these strains showed resistance to spiramycin, azithromycin, erythromycin, and clarithromycin; moreover, the antibiotic resistance rates was not reduced in the effluent. Results of PCR and quantitative PCR showed that 80% of antibiotic resistant Enterococci were positive for PCR amplification of erAB, but negative for PCR amplification of other genes. Concentrations of ermB and ermF were peaked in the spring and autumn samples. Resistance genes of ermB, ermF, mefA, ereA, mphB and transfer element of Tn916/1545 were reduced to some degree during antibiotic production wastewater treatment, but concentrations of ermX, intIl, ISCRl in the effluent were higher than those in the influent. The abundance of mefA, ereA and Tn916/1545 were reduced during wastewater treatment process, and the better removal performance for mefA, ereA, Tn916/1545 occurred in spring than in autumn; however, the abundance of ermX, intI1 and ISCR1 were increased. PMID:26717693

  20. Molecular basis of resistance to macrolides, lincosamides and streptogramins in Staphylococcus hominis strains isolated from clinical specimens.

    PubMed

    Szczuka, Ewa; Makowska, Nicoletta; Bosacka, Karolina; Słotwińska, Anna; Kaznowski, Adam

    2016-03-01

    Coagulase-negative staphylococci (CoNS) are the most frequently isolated bacteria from the blood and the predominant cause of nosocomial infections. Macrolides, lincosamides and streptogramin B (MLSB) antibiotics, especially erythromycin and clindamycin, are important therapeutic agents in the treatment of methicillin-resistant staphylococci infections. Among CoNS, Staphylococcus hominis represents the third most common organism. In spite of its clinical significance, very little is known about its mechanisms of resistance to antibiotics, especially MLSB. Fifty-five S. hominis isolates from the blood and the surgical wounds of hospitalized patients were studied. The erm(C) gene was predominant in erythromycin-resistant S. hominis isolates. The methylase genes, erm(A) and erm(B), were present in 15 and 25 % of clinical isolates, respectively. A combination of various erythromycin resistance methylase (erm) genes was detected in 15 % S. hominis isolates. The efflux gene msr(A) was detected in 18 % of isolates, alone in four isolates, and in different combinations in a further six. The lnu(A) gene, responsible for enzymatic inactivation of lincosamides was carried by 31 % of the isolates. No erythromycin resistance that could not be attributed to the genes erm(A), erm(B), erm(C) and msr(A) was detected. In S. hominis, 75 and 84 %, respectively, were erythromycin resistant and clindamycin susceptible. Among erythromycin-resistant S. hominis isolates, 68 % of these strains showed the inducible MLSB phenotype. Four isolates harbouring the msr(A) genes alone displayed the MSB phenotype. These studies indicated that resistance to MLSB in S. hominis is mostly based on the ribosomal target modification mechanism mediated by erm genes, mainly the erm(C), and enzymatic drug inactivation mediated by lnu(A). PMID:26253583

  1. Detection of macrolide and disinfectant resistance genes in clinical Staphylococcus aureus and coagulase-negative staphylococci

    PubMed Central

    2011-01-01

    Background Staphylococcus aureus and Coagulase-negative staphylococci (CoNS) are a major source of infections associated with indwelling medical devices. Many antiseptic agents are used in hygienic handwash to prevent nosocomial infections by Staphylococci. Our aim was to determine the antibiotic susceptibility and resistance to quaternary ammonium compound of 46 S. aureus strains and 71 CoNS. Methods S. aureus (n = 46) isolated from auricular infection and CoNS (n = 71), 22 of the strains isolated from dialysis fluids and 49 of the strains isolated from needles cultures were investigated. Erythromycin resistance genes (ermA, ermB, ermC, msrA and mef) were analysed by multiplex PCR and disinfectant-resistant genes (qacA, qacB, and qacC) were studied by PCR-RFLP. Results The frequency of erythromycin resistance genes in S. aureus was: ermA+ 7.7%, ermB+ 13.7%, ermC+ 6% and msrA+ 10.2%. In addition, the number of positive isolates in CoNS was respectively ermA+ (9.4%), ermB+ (11.1%), ermC+ (27.4%), and msrA+ (41%). The MIC analyses revealed that 88 isolates (74%) were resistant to quaternary ammonium compound-based disinfectant benzalkonium chloride (BC). 56% of the BC-resistant staphylococcus isolates have at least one of the three resistant disinfectants genes (qacA, qacB and qacC). Nine strains (7.7%) among the CoNS species and two S. aureus strains (2%) harboured the three-qac genes. In addition, the qacC were detected in 41 strains. Conclusions Multi-resistant strains towards macrolide and disinfectant were recorded. The investigation of antibiotics and antiseptic-resistant CoNS may provide crucial information on the control of nosocomial infections. PMID:22032892

  2. Localisation of vascular endothelial growth factor and its receptors to cells of vascular and avascular epiretinal membranes

    PubMed Central

    Chen, Y.; Hackett, S.; Schoenfeld, C.; Vinores, M.; Vinores, S.; Campochiaro, P.

    1997-01-01

    AIMS/BACKGROUND—Epiretinal membranes (ERMs) arise from a variety of causes or, in some cases, for unknown reasons. Once established, ERMs tend to progress, becoming more extensive and exerting increasing traction along the inner surface of the retina. One possible cause for their progression is the production of growth factors by cells within ERMs that may provide autocrine or paracrine stimulation. Platelet derived growth factor (PDGF) and its receptors have been localised to cells of ERMs and may play such a role. In this study, comparative data were sought for several other growth factors that have been implicated in ERM formation.
METHODS—Immunohistochemical staining of ERMs was done for PDGF-A, PDGF-B, basic fibroblast growth factor (bFGF), three isoforms of transforming growth factor β (TGF-β), and vascular endothelial growth factor (VEGF) and its receptors, flt-1 and flk-1/KDR. Expression of flt-1 and flk-1/KDR was examined in cultured retinal pigmented epithelial (RPE) cells and retinal glia from postmortem eyes by immunohistochemistry and by reverse transcription coupled to polymerase chain reaction (RT-PCR).
RESULTS—Staining was most intense and most frequently observed for VEGF and PDGF-A, both in vascular and avascular ERMs. The majority of cells stained for VEGF in nine of 11 (81.8%) diabetic ERMs and in 14 of 24 (58.3%) proliferative vitreoretinopathy ERMs. The receptors for VEGF, flt-1, and flk-1/KDR were also identified on cells in ERMs and on cultured RPE cells. By RT-PCR, mRNA for flt-1 was identified in RPE cells and retinal glia, and mRNA for flk-1/KDR was identified in RPE cells.
CONCLUSIONS—These data show that VEGF and its receptors are localised to both vascular and avascular ERMs and suggest that VEGF, like PDGF-A, may be an autocrine and paracrine stimulator that may contribute to progression of vascular and avascular ERMs.

 PMID:9486038

  3. Identifying risks in the realm of enterprise risk management.

    PubMed

    Carroll, Roberta

    2016-01-01

    An enterprise risk management (ERM) discipline is comprehensive and organization-wide. The effectiveness of ERM is governed in part by the strength and breadth of its practices and processes. An essential element in decision making is a thorough process by which organizational risks and value opportunities can be identified. This article will offer identification techniques that go beyond those used in traditional risk management programs and demonstrate how these techniques can be used to identify risks and opportunity in the ERM environment. PMID:26789745

  4. Mechanism of resistance to macrolide-lincosamide-streptogramin antibiotics in Streptococcus thermophilus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Resistance to macrolide-lincosamide-streptogramin (MLS) group antibiotics in the dairy bacterium Streptococcus thermophilus (ST) is documented but the mechanism of resistance has not been elucidated. MIC values for erythromycin (Erm), azithromycin (Azm), tylosin (Tyl), spiramycin (Spm), pristinamyci...

  5. Adaptor Protein Cerebral Cavernous Malformation 3 (CCM3) Mediates Phosphorylation of the Cytoskeletal Proteins Ezrin/Radixin/Moesin by Mammalian Ste20-4 to Protect Cells from Oxidative Stress*

    PubMed Central

    Fidalgo, Miguel; Guerrero, Ana; Fraile, María; Iglesias, Cristina; Pombo, Celia M.; Zalvide, Juan

    2012-01-01

    While studying the functions of CCM3/PDCD10, a gene encoding an adaptor protein whose mutation results in vascular malformations, we have found that it is involved in a novel response to oxidative stress that results in phosphorylation and activation of the ezrin/radixin/moesin (ERM) family of proteins. This phosphorylation protects cells from accidental cell death induced by oxidative stress. We also present evidence that ERM phosphorylation is performed by the GCKIII kinase Mst4, which is activated and relocated to the cell periphery after oxidative stress. The cellular levels of Mst4 and its activation after oxidative stress depend on the presence of CCM3, as absence of the latter impairs the phosphorylation of ERM proteins and enhances death of cells exposed to reactive oxygen species. These findings shed new light on the response of cells to oxidative stress and identify an important pathophysiological situation in which ERM proteins and their phosphorylation play a significant role. PMID:22291017

  6. Environmental Enterprise Risk Management Benefits for a Government Contractor

    SciTech Connect

    Linda Guinn

    2012-05-01

    An often overlooked advantage that an Environmental Enterprise Risk Management System (ERMS) has to organizations is the added protection from the Civil False Claims Act (FCA) for activities under a government contract.

  7. Novel Root-Fungus Symbiosis in Ericaceae: Sheathed Ericoid Mycorrhiza Formed by a Hitherto Undescribed Basidiomycete with Affinities to Trechisporales

    PubMed Central

    Vohník, Martin; Sadowsky, Jesse J.; Kohout, Petr; Lhotáková, Zuzana; Nestby, Rolf; Kolařík, Miroslav

    2012-01-01

    Ericaceae (the heath family) are widely distributed calcifuges inhabiting soils with inherently poor nutrient status. Ericaceae overcome nutrient limitation through symbiosis with ericoid mycorrhizal (ErM) fungi that mobilize nutrients complexed in recalcitrant organic matter. At present, recognized ErM fungi include a narrow taxonomic range within the Ascomycota, and the Sebacinales, basal Hymenomycetes with unclamped hyphae and imperforate parenthesomes. Here we describe a novel type of basidiomycetous ErM symbiosis, termed ‘sheathed ericoid mycorrhiza’, discovered in two habitats in mid-Norway as a co-dominant mycorrhizal symbiosis in Vaccinium spp. The basidiomycete forming sheathed ErM possesses clamped hyphae with perforate parenthesomes, produces 1- to 3-layer sheaths around terminal parts of hair roots and colonizes their rhizodermis intracellularly forming hyphal coils typical for ErM symbiosis. Two basidiomycetous isolates were obtained from sheathed ErM and molecular and phylogenetic tools were used to determine their identity; they were also examined for the ability to form sheathed ErM and lignocellulolytic potential. Surprisingly, ITS rDNA of both conspecific isolates failed to amplify with the most commonly used primer pairs, including ITS1 and ITS1F + ITS4. Phylogenetic analysis of nuclear LSU, SSU and 5.8S rDNA indicates that the basidiomycete occupies a long branch residing in the proximity of Trechisporales and Hymenochaetales, but lacks a clear sequence relationship (>90% similarity) to fungi currently placed in these orders. The basidiomycete formed the characteristic sheathed ErM symbiosis and enhanced growth of Vaccinium spp. in vitro, and degraded a recalcitrant aromatic substrate that was left unaltered by common ErM ascomycetes. Our findings provide coherent evidence that this hitherto undescribed basidiomycete forms a morphologically distinct ErM symbiosis that may occur at significant levels under natural conditions, yet remain

  8. Antimicrobial Susceptibility of Bacillus Strains Isolated from Primary Starters for African Traditional Bread Production and Characterization of the Bacitracin Operon and Bacitracin Biosynthesis

    PubMed Central

    Sørensen, Kim I.; Thorsen, Line; Stuer-Lauridsen, Birgitte; Abdelgadir, Warda S.; Nielsen, Dennis S.; Derkx, Patrick M. F.; Jespersen, Lene

    2012-01-01

    Bacillus spp. are widely used as feed additives and probiotics. However, there is limited information on their resistance to various antibiotics, and there is a growing concern over the transfer of antibiotic resistance genes. The MIC for 8 antibiotics was determined for 85 Bacillus species strains, Bacillus subtilis subsp. subtilis (n = 29), Bacillus licheniformis (n = 38), and Bacillus sonorensis (n = 18), all of which were isolated from starters for Sudanese bread production. All the strains were sensitive to tetracycline (8.0 mg/liter), vancomycin (4.0 mg/liter), and gentamicin (4.0 mg/liter) but resistant to streptomycin. Sensitivity to clindamycin, chloramphenicol, and kanamycin was species specific. The erythromycin resistance genes ermD and ermK were detected by PCR in all of the erythromycin-resistant (MIC, ≥16.0 mg/liter) B. licheniformis strains and one erythromycin-sensitive (MIC, 4.0 mg/liter) B. licheniformis strain. Several amino acid changes were present in the translated ermD and ermK nucleotide sequences of the erythromycin-sensitive strain, which could indicate ErmD and ErmK protein functionalities different from those of the resistance strains. The ermD and ermK genes were localized on an 11.4-kbp plasmid. All of the B. sonorensis strains harbored the bacitracin synthetase gene, bacA, and the transporter gene bcrA, which correlated with their observed resistance to bacitracin. Bacitracin was produced by all the investigated species strains (28%), as determined by ultra-high-definition quadrupole time-of-flight liquid chromatography-mass spectrometry (UHD-QTOF LC/MS). The present study has revealed species-specific variations in the antimicrobial susceptibilities of Bacillus spp. and provides new information on MIC values, as well as the occurrence of resistance genes in Bacillus spp., including the newly described species B. sonorensis. PMID:22941078

  9. MicroRNA-200b Impacts Breast Cancer Cell Migration and Invasion by Regulating Ezrin-Radixin-Moesin

    PubMed Central

    Hong, Hong; Yu, Haizhong; Yuan, Jianfen; Guo, Chunyan; Cao, Hongyan; Li, Weibing; Xiao, Chunhong

    2016-01-01

    Background Ezrin-radixin-moesin (ERM) plays an important role in multiple links of tumors. It also involved in breast cancer invasion and metastasis, and might be a potential biomarker of breast cancer. Another study suggested that ERM expression was regulated directly by miR-200c, and had a critical role in miR-200c suppressing cell migration. This study aimed to investigate the effect of miR-200b on ERM expression in a breast cancer cell line and its influence on invasion and metastasis ability in vitro. Material/Methods Breast cancer cell lines MCF-7 and MDA-MB-231 with different metastatic potentials were selected as a model. MiR-200b overexpression or inhibition was achieved by Lipofectamine™ 2000-mediated miRNA transfection. RT-PCR was used to test miR-200b level, while Western blot was selected to detect ERM protein expression. Wound healing assay and Transwell assay were performed to determine cell migration and invasion ability. Results RT-PCR revealed that miR-200b level in MDA-MB-231 was obviously lower than that in MCF-7, while Western blot analysis showed that ERM expression was significantly higher. MiR-200b inhibition by transfection in MCF-7 markedly decreased miR-200b level, elevated ERM expression, and enhanced cell migration and invasion. MiR-200b overexpression in MDA-MB-231 obviously increased miR-200b level, reduced ERM expression, and weakened cell migration and invasion. Conclusions MiR-200b participates in breast cancer cell migration and invasion through regulating ERM in MCF-7 and MDA-MB-231. PMID:27276064

  10. DNA in Antibiotic Preparations: Absence of Intact Resistance Genes

    PubMed Central

    Woegerbauer, Markus; Lagler, Heimo; Graninger, Wolfgang; Burgmann, Heinz

    2005-01-01

    Fragments of erm(E2), otrA, and aph(6) shorter than 400 bp and producer strain-specific rRNA genes were amplified from various antibiotics. The amount of genetic material and the sizes of amplicons recovered from murine feces after oral administration of a β-lactamase-encoding plasmid indicated substantial DNA degradation in the mammalian gastrointestinal tract. These observations imply that antibiotics are no major source for horizontal resistance gene transfer in clinical settings. PMID:15917552

  11. In vitro antimicrobial susceptibility of Helcococcus kunzii and molecular analysis of macrolide and tetracycline resistance.

    PubMed

    Vergne, A; Guérin, F; Lienhard, R; Le Coustumier, A; Daurel, C; Isnard, C; Marty, N; Poyart, C; Cattoir, V

    2015-10-01

    Thanks to the recent advent of matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) technology, Helcococcus kunzii is now easily identifiable and considered as an opportunistic pathogen. However, data about antimicrobial susceptibilities remain very limited. The aim of the study was, then, to assess its in vitro susceptibility to 18 antimicrobial agents and to investigate the genetic basis of macrolide and tetracycline resistance. Thirty-nine human clinical isolates of H. kunzii collected from 2008 to 2013 were studied, as well as the type strain ATCC 51366(T). Minimum inhibitory concentrations (MICs) of penicillin G, amoxicillin, cefotaxime, imipenem, gentamicin, erythromycin, clindamycin, quinupristin-dalfopristin, ciprofloxacin, levofloxacin, tetracycline, tigecycline, vancomycin, teicoplanin, linezolid, daptomycin, cotrimoxazole and rifampin were determined by the microdilution method. Screening for macrolide [erm(A) including erm(TR), erm(B), erm(C), erm(F), erm(T), erm(X), msr(A) and mef(A)] and tetracycline [tet(L), tet(M) and tet(O)] resistance genes was performed, as well as the detection of mutations in 23S rRNA. Except for one strain resistant to cefotaxime, all strains were categorised as susceptible to β-lactams, glycopeptides, linezolid, daptomycin and tigecycline. Whereas ciprofloxacin and gentamicin exhibited limited activity, 95% of strains were categorised as susceptible to levofloxacin. Concerning erythromycin, a bimodal distribution was observed, with 29 'wild-type' strains (MICs from 0.25 to 2 mg/L) and 11 'resistant' strains (MICs ≥ 256 mg/L), including ten harbouring erm(TR). Two isolates exhibited acquired tetracycline resistance (MICs of 16 mg/L) by the production of tet(M). This large study on the in vitro antimicrobial susceptibility of H. kunzii suggests that β-lactams (especially penicillins) should be preferred for the treatment. PMID:26194694

  12. Predicting stress in benthic communities of southeastern U.S. estuaries in relation to chemical contamination of sediments

    SciTech Connect

    Hyland, J.L.; Dolah, R.F. van; Snoots, T.R.

    1999-11-01

    Matching data on sediment contaminants (metals, PAHs, PCBs) and macroinfaunal community structure from 231 subtidal stations in southeastern US estuaries were used to develop a framework for evaluating risks of benthic impacts from multiple-contaminant exposure. Sediment contamination was expressed as the mean ratio of individual contaminant concentrations relative to corresponding sediment quality guidelines, that is, to effects range-median (ERM) values, probable effects level (PEL) values, or an aggregate of the two. The probability of a degraded benthos was relatively low in samples with mean ERM quotients {le}0.020, PEL quotients {le}0.035, or combined ERM/PEL quotients {le}0.024. Only 5% of stations within these ranges had degraded benthic assemblages, while 95% had healthy assemblages. A higher probability of benthic impacts was observed in samples with mean ERM quotients >0.058, PEL quotients >0.096, or ERM/PEL quotients >0.077. Seventy-three to 78% of stations with values in these upper ranges had degraded benthic assemblages, while 22 to 27% had healthy assemblages. Only four stations (three with degraded, one with healthy assemblages) had mean ERM or PEL quotients >1.0, which is the beginning of the range associated with a high probability of mortality in short-term laboratory toxicity tests using amphipods.

  13. Ovarian embryonal rhabdomyosarcoma is a rare manifestation of the DICER1 syndrome.

    PubMed

    de Kock, Leanne; Druker, Harriet; Weber, Evan; Hamel, Nancy; Traubici, Jeffrey; Malkin, David; Arseneau, Jocelyne; Stewart, Colin J R; Bouron-Dal Soglio, Dorothée; Priest, John R; Foulkes, William D

    2015-06-01

    Embryonal rhabdomyosarcoma (ERMS), a soft tissue sarcoma, is one of the most common pediatric cancers. Certain ERMSs are associated with the DICER1 syndrome, a tumor predisposition syndrome caused by germ-line DICER1 mutations. Characteristic somatic mutations have also been identified in DICER1-associated tumor types. These "hotspot" mutations affect the catalytic activity of the DICER1 ribonuclease IIIb domain. Primary ovarian ERMS (oERMS) is extremely rare. We present a case of a 6-year-old girl with an oERMS harboring 2 DICER1 mutations. The girl also exhibited other DICER1 phenotypes: cystic nephroma (CN) and multinodular goiter. Somatic investigations of the CN identified a hotspot DICER1 mutation different from that in the oERMS. Significantly, the CN presented at 12 years of age, which is much older than the previously reported age range of susceptibility. This report documents the occurrence of DICER1 mutations in a case of oERMS, expanding the spectrum of DICER1-associated tumors. PMID:25836323

  14. Significant groundwater input to a coastal plain estuary: assessment from excess radon

    NASA Astrophysics Data System (ADS)

    Schwartz, M. C.

    2003-01-01

    A submarine groundwater discharge (SGD) zone was identified through the investigation of persistent excess radon in the Delaware River and Bay Estuary. This zone, the excess radon maximum (ERM), was located approximately 82 km upstream from the mouth of the Delaware Bay. Water column radon activity at the ERM was elevated relative to the radon activity observed outside of this zone and was as high as 2.73 dpm l -1. A radon budget model developed to assess the contribution of multiple radon sources to the ERM indicated that simple molecular diffusion from sediments could not support the observed excess radon activity; an additional source of radon is required to sustain the excess radon activity in the ERM. A groundwater flux of 14-29 m 3 s -1 from the two aquifers that subcrop on the Delmarva Peninsula immediately west of the ERM can support the excess radon observed in the water column. The stratigraphy and hydrology of these aquifers make them likely sources of the groundwater discharge that results in the ERM. The calculated SGD flux is equivalent to the surface water discharge of the second and third largest tributary rivers of the Delaware Estuary.

  15. Notch maintains Drosophila type II neuroblasts by suppressing expression of the Fez transcription factor Earmuff.

    PubMed

    Li, Xiaosu; Xie, Yonggang; Zhu, Sijun

    2016-07-15

    Notch signaling is crucial for maintaining neural stem cell (NSC) self-renewal and heterogeneity; however, the underlying mechanism is not well understood. In Drosophila, loss of Notch prematurely terminates the self-renewal of larval type II neuroblasts (NBs, the Drosophila NSCs) and transforms type II NBs into type I NBs. Here, we demonstrate that Notch maintains type II NBs by suppressing the activation of earmuff (erm) by Pointed P1 (PntP1). We show that loss of Notch or components of its canonical pathway leads to PntP1-dependent ectopic Erm expression in type II NBs. Knockdown of Erm significantly rescues the loss-of-Notch phenotypes, and misexpression of Erm phenocopies the loss of Notch. Ectopically expressed Erm promotes the transformation of type II NBs into type I NBs by inhibiting PntP1 function and expression in type II NBs. Our work not only elucidates a key mechanism of Notch-mediated maintenance of type II NB self-renewal and identity, but also reveals a novel function of Erm. PMID:27151950

  16. Mycoparasitism of arbuscular mycorrhizal fungi: a pathway for the entry of saprotrophic fungi into roots.

    PubMed

    De Jaeger, Nathalie; Declerck, Stéphane; de la Providencia, Ivan E

    2010-08-01

    Within the rhizosphere, arbuscular mycorrhizal (AM) fungi interact with a cohort of microorganisms, among which is the biological control agent, Trichoderma spp. This fungus parasitizes a wide range of phytopathogenic fungi, a phenomenon also reported in the extraradical mycelium (ERM) of AM fungi. Here, we question whether the mycoparasitism of the ERM could be extended to the intraradical mycelium (IRM), thus representing a pathway for the entry of Trichoderma harzianum within the root. Microcosm experiments allowing interactions between Glomus sp. MUCL 41833 placed in a clade that contains the recently described species Glomus irregulare and T. harzianum were set up under in vitro autotrophic culture conditions using potato as a host. A microscope camera-imaging system, coupled with succinate dehydrogenase staining, was used to assess the mycoparasitism in the ERM and IRM. Trichoderma harzianum colonized the ERM of the AM fungus and spread into the IRM, before exiting into the root cells. Intrahyphal growth of T. harzianum caused protoplasm degradation, decreasing the ERM and IRM viability. ERM of the AM fungus represented a pathway for the entry of T. harzianum into the roots of potato. It further sets off the debate on the susceptibility of the AM fungi of being infected by microorganisms from the rhizosphere. PMID:20533946

  17. The Mediator complex subunit MED25 is targeted by the N-terminal transactivation domain of the PEA3 group members

    PubMed Central

    Verger, Alexis; Baert, Jean-Luc; Verreman, Kathye; Dewitte, Frédérique; Ferreira, Elisabeth; Lens, Zoé; de Launoit, Yvan; Villeret, Vincent; Monté, Didier

    2013-01-01

    PEA3, ERM and ER81 belong to the PEA3 subfamily of Ets transcription factors and play important roles in a number of tissue-specific processes. Transcriptional activation by PEA3 subfamily factors requires their characteristic amino-terminal acidic transactivation domain (TAD). However, the cellular targets of this domain remain largely unknown. Using ERM as a prototype, we show that the minimal N-terminal TAD activates transcription by contacting the activator interacting domain (ACID)/Prostate tumor overexpressed protein 1 (PTOV) domain of the Mediator complex subunit MED25. We further show that depletion of MED25 disrupts the association of ERM with the Mediator in vitro. Small interfering RNA-mediated knockdown of MED25 as well as the overexpression of MED25-ACID and MED25-VWA domains efficiently inhibit the transcriptional activity of ERM. Moreover, mutations of amino acid residues that prevent binding of MED25 to ERM strongly reduce transactivation by ERM. Finally we show that siRNA depletion of MED25 diminishes PEA3-driven expression of MMP-1 and Mediator recruitment. In conclusion, this study identifies the PEA3 group members as the first human transcriptional factors that interact with the MED25 ACID/PTOV domain and establishes MED25 as a crucial transducer of their transactivation potential. PMID:23531547

  18. Change in refraction after lens-sparing vitrectomy for rhegmatogenous retinal detachment and epiretinal membrane

    PubMed Central

    Iwase, Takeshi; Yamamoto, Kentaro; Yanagida, Kosei; Kobayashi, Misato; Ra, Eimei; Murotani, Kenta; Terasaki, Hiroko

    2016-01-01

    Abstract The aim of this study was to compare changes in refraction following lens-sparing vitrectomy between patients with rhegmatogenous retinal detachment (RRD) and epiretinal membrane (ERM) and to investigate factors associated with the change in refraction. We reviewed medical records of 49 eyes of 49 patients with RRD (53.6 ± 7.8 years, mean ± standard deviation) and 24 eyes of 24 patients with ERM (50.9 ± 15.7 years) who underwent lens-sparing vitrectomy. Spherical equivalent refractive power was evaluated before and up to 18 months after surgery. The relationship between the change in refraction and several parameters was evaluated. A significant progressive myopic shift in refractive power was observed after vitrectomy in operated RRD and ERM eyes (P < 0.001, P = 0.016, respectively), with no significant difference in fellow eyes. The refraction values observed at ≥3 and ≥12 months following vitrectomy were significantly different as compared with those observed at baseline in the RRD group (P < 0.001) and the ERM group (P < 0.05), respectively. The change in refraction between the RRD and ERM groups was significant (P = 0.030). The multiple linear regression analysis showed that only age was significantly correlated with the change in refraction in RRD (P = 0.018) and ERM (P < 0.001) groups. The change in refraction was significantly and positively correlated with age in RRD (r = −0.461, P = 0.001) and ERM (r = −0.687, P < 0.001) groups. Following lens-sparing vitrectomy, cataract surgery was performed on 30 eyes after 0.89 ± 0.26 years in the RRD group and on 10 eyes after 1.11 ± 0.14 years in the ERM group; there was a significant difference in time to cataract surgery between the groups (P = 0.007). Kaplan–Meier survival analysis demonstrated that there was a significant difference in the rate of cataract surgeries between the RRD and ERM groups (P = 0

  19. Change in refraction after lens-sparing vitrectomy for rhegmatogenous retinal detachment and epiretinal membrane.

    PubMed

    Iwase, Takeshi; Yamamoto, Kentaro; Yanagida, Kosei; Kobayashi, Misato; Ra, Eimei; Murotani, Kenta; Terasaki, Hiroko

    2016-08-01

    The aim of this study was to compare changes in refraction following lens-sparing vitrectomy between patients with rhegmatogenous retinal detachment (RRD) and epiretinal membrane (ERM) and to investigate factors associated with the change in refraction.We reviewed medical records of 49 eyes of 49 patients with RRD (53.6 ± 7.8 years, mean ± standard deviation) and 24 eyes of 24 patients with ERM (50.9 ± 15.7 years) who underwent lens-sparing vitrectomy. Spherical equivalent refractive power was evaluated before and up to 18 months after surgery. The relationship between the change in refraction and several parameters was evaluated.A significant progressive myopic shift in refractive power was observed after vitrectomy in operated RRD and ERM eyes (P < 0.001, P = 0.016, respectively), with no significant difference in fellow eyes. The refraction values observed at ≥3 and ≥12 months following vitrectomy were significantly different as compared with those observed at baseline in the RRD group (P < 0.001) and the ERM group (P < 0.05), respectively. The change in refraction between the RRD and ERM groups was significant (P = 0.030). The multiple linear regression analysis showed that only age was significantly correlated with the change in refraction in RRD (P = 0.018) and ERM (P < 0.001) groups. The change in refraction was significantly and positively correlated with age in RRD (r = -0.461, P = 0.001) and ERM (r = -0.687, P < 0.001) groups. Following lens-sparing vitrectomy, cataract surgery was performed on 30 eyes after 0.89 ± 0.26 years in the RRD group and on 10 eyes after 1.11 ± 0.14 years in the ERM group; there was a significant difference in time to cataract surgery between the groups (P = 0.007). Kaplan-Meier survival analysis demonstrated that there was a significant difference in the rate of cataract surgeries between the RRD and ERM groups (P = 0.022).Following lens

  20. Reflectivity and thickness analysis of epiretinal membranes using spectral-domain optical coherence tomography

    PubMed Central

    Kuriyan, Ajay E.; DeBuc, Delia Cabrera; Smiddy, William E.

    2016-01-01

    AIM To compare thickness and reflectivity spectral domain optical coherence tomography (SD-OCT) findings in patients with idiopathic epiretinal membranes (ERMs), before and after ERM peeling surgery, with normal controls. METHODS A retrospective study analyzed SD-OCTs of eyes with ERMs undergoing ERM peeling surgery by one surgeon from 2008 to 2010 and normal control eyes. SD-OCTs were analyzed using a customized algorithm to measure reflectivity and thickness. The relationship between the SD-OCT findings and best corrected visual acuity (BCVA) outcomes was also studied. RESULTS Thirty-four ERM eyes and 12 normal eyes were identified. Preoperative eyes had high reflectivity and thickness of the group of layers from the internal limiting membrane (ILM) to the retinal pigment epithelium (RPE) and the group of layers from the ILM to the external limiting membrane (ELM). The values of reflectivity of these two groups of layers decreased postoperatively, but were still higher than normal eyes. In contrast, preoperative eyes had lower reflectivity of two 10×15 pixel regions of interest (ROIs) incorporating: 1) ELM + outer nuclear layer (ONL) and 2) photoreceptor layer (PRL) + RPE, compared to controls. The values of reflectivity of these ROIs increased postoperatively, but were still lower than normal controls. A larger improvement in BCVA postoperatively was correlated with a greater degree of abnormal preoperative reflectivity and thickness findings. CONCLUSION Quantitative differences in reflectivity and thickness between preoperative, postoperative, and normal SD-OCTs allow assessment of changes in the retina secondary to ERM. Our study identified hyperreflective inner retina changes and hyporeflective outer retina changes in patients with ERMs. SD-OCT quantitative measures of reflectivity and/or thickness of specific groups of retinal layers and/or ROIs correlate with improvement in BCVA. PMID:26949617

  1. New Real-Time PCR Assays for Detection of Inducible and Acquired Clarithromycin Resistance in the Mycobacterium abscessus Group

    PubMed Central

    Shallom, Shamira J.; Moura, Natalia S.; Olivier, Kenneth N.; Sampaio, Elizabeth P.; Holland, Steven M.

    2015-01-01

    Members of the Mycobacterium abscessus group (MAG) cause lung, soft tissue, and disseminated infections. The oral macrolides clarithromycin and azithromycin are commonly used for treatment. MAG can display clarithromycin resistance through the inducible erm(41) gene or via acquired mutations in the rrl (23S rRNA) gene. Strains harboring a truncation or a T28C substitution in erm(41) lose the inducible resistance trait. Phenotypic detection of clarithromycin resistance requires extended incubation (14 days), highlighting the need for faster methods to detect resistance. Two real-time PCR-based assays were developed to assess inducible and acquired clarithromycin resistance and tested on a total of 90 clinical and reference strains. A SYBR green assay was designed to distinguish between a full-length and truncated erm(41) gene by temperature shift in melting curve analysis. Single nucleotide polymorphism (SNP) allele discrimination assays were developed to distinguish T or C at position 28 of erm(41) and 23S rRNA rrl gene mutations at position 2058 and/or 2059. Truncated and full-size erm(41) genes were detected in 21/90 and 69/90 strains, respectively, with 64/69 displaying T at nucleotide position 28 and 5/69 containing C at that position. Fifteen isolates showed rrl mutations conferring clarithromycin resistance, including A2058G (11 isolates), A2058C (3 isolates), and A2059G (1 isolate). Targeted sequencing and phenotypic assessment of resistance concurred with molecular assay results. Interestingly, we also noted cooccurring strains harboring an active erm(41), inactive erm(41), and/or acquired mutational resistance, as well as slowly growing MAG strains and also strains displaying an inducible resistance phenotype within 5 days, long before the recommended 14-day extended incubation. PMID:26269619

  2. Genetic basis of resistance waves among methicillin resistant Staphylococcus aureus isolates recovered from milk and meat products in Egypt.

    PubMed

    Ammar, A M; Attia, A M; Abd El-Hamid, M I; El-Shorbagy, I M; Abd El-Kader, S A

    2016-01-01

    Antimicrobial resistance of methicillin-resistant Staphylococcus aureus (MRSA) poses a serious problem for clinicians worldwide. The present study attempted to evaluate the susceptibility patterns of MRSA to various antimicrobials and the prevalence of inducible clindamycin resistance as well as the relevant antibiotic and antiseptic resistance genes among these isolates. Totally, 40 MRSA isolates were recovered from examined milk and meat product samples (18.60%). Multi-drug resistance (MDR) was remarkably observed among 85% of these isolates. There was a good correlation between phenotypic determination of methicillin, amoxicillin/clavulinic acid and tetracycline resistances and PCR detections of mecA, blaZ and tet(K) genes, respectively, but norA gene was not detected in the four ciprofloxacin resistant isolates. Although, 55% of MRSA expressed resistance to benzalkonium chloride (BC), neither qacA/B nor smr gene was detected. Of 20 isolates exhibiting erythromycin- clindamycin discordant resistance pattern, 8 displayed positive double disk diffusion (D-zone) test denoting inducible macrolide-lincosamide-streptogramin B (MLSB) resistance phenotype with the inducibly expressed erm(A) and erm(C) genes in 87.5% of these isolates. Besides, the remaining 12 isolates showed MS phenotype (resistant to macrolides and type B streptogramins only) with a variety of erm(A), mph(C), msr(A) or a combination of these genes including erm(C). Finally, the constitutive MLSB phenotype with the constitutive expression of erm(A), erm(B) and erm(C) genes was comprised in 2 isolates with higher minimum inhibitory concentration (MIC) values for erythromycin (512 and 1024 µg/ml) and clindamycin (16 and 32 µg/ml). These findings suggested the importance of monitoring the evolution of MRSA resistance. PMID:27609468

  3. Isolation and molecular characterization of antibiotic-resistant lactic acid bacteria from poultry and swine meat products.

    PubMed

    Aquilanti, Lucia; Garofalo, Cristiana; Osimani, Andrea; Silvestri, Gloria; Vignaroli, Carla; Clementi, Francesca

    2007-03-01

    The transfer via the food chain from animals to humans of microbes that are resistant to antimicrobial agents is of increasing concern. To determine the contributions of nonpathogenic microflora to the occurrence and spread of antibiotic resistance (AR) genes in the food chain, 123 lactic acid bacteria were isolated from 29 samples of raw and processed pork and chicken meat products that had previously tested positive for one or more AR genes that encode clinically relevant ARs: tet(M), tet(O), tet(K), erm(A), erm(B), erm(C), aac (6')-Ie aph (2")-Ia, mecA, and blaZ. All of the isolates were initially tested for their AR gene profiles by PCR. The 59 isolates carrying a tet, erm, or blaZ gene were taken through molecular identification, analyzed by determination of the MIC, and subjected to genetic fingerprinting. Lactococcus garvieae was the predominant species (28 isolates), followed by Lactobacillus plantarum (11 isolates) and L. salivarius (6 isolates), whereas Lactococcus lactis subsp. lactis, Lactobacillus johnsonii, L. reuteri, L. crispatus, and L. brevis were identified at lower frequencies. The tet(M) and erm(B) genes were the most frequently detected. Assessment of multiple resistances in 18 tet positive (tet+) isolates revealed that tet(M) plus erm(B) and tet(K) plus erm(B) were the most frequent AR gene patterns. Partial sequencing of the tet(M) open reading frame of three selected strains showed high sequence similarities (> 99%) with tet(M) genes previously found in human pathogens (Listeria monocytogenes and Neisseria meningitidis). Southern hybridization with plasmid profiles revealed these strains contained tet(M)-carrying plasmids. PMID:17388042

  4. Effects of RAS on the genesis of embryonal rhabdomyosarcoma

    PubMed Central

    Langenau, David M.; Keefe, Matthew D.; Storer, Narie Y.; Guyon, Jeffrey R.; Kutok, Jeffery L.; Le, Xiuning; Goessling, Wolfram; Neuberg, Donna S.; Kunkel, Louis M.; Zon, Leonard I.

    2007-01-01

    Embryonal rhabdomyosarcoma (ERMS) is a devastating cancer with specific features of muscle differentiation that can result from mutational activation of RAS family members. However, to date, RAS pathway activation has not been reported in a majority of ERMS patients. Here, we have created a zebrafish model of RAS-induced ERMS, in which animals develop externally visible tumors by 10 d of life. Microarray analysis and cross-species comparisons identified two conserved gene signatures found in both zebrafish and human ERMS, one associated with tumor-specific and tissue-restricted gene expression in rhabdomyosarcoma and a second comprising a novel RAS-induced gene signature. Remarkably, our analysis uncovered that RAS pathway activation is exceedingly common in human RMS. We also created a new transgenic coinjection methodology to fluorescently label distinct subpopulations of tumor cells based on muscle differentiation status. In conjunction with fluorescent activated cell sorting, cell transplantation, and limiting dilution analysis, we were able to identify the cancer stem cell in zebrafish ERMS. When coupled with gene expression studies of this cell population, we propose that the zebrafish RMS cancer stem cell shares similar self-renewal programs as those found in activated satellite cells. PMID:17510286

  5. INTRAOPERATIVE SPECTRAL DOMAIN OPTICAL COHERENCE TOMOGRAPHY IMAGING AFTER INTERNAL LIMITING MEMBRANE PEELING IN IDIOPATHIC EPIRETINAL MEMBRANE WITH CONNECTING STRANDS

    PubMed Central

    NAM, DONG HEUN; DESOUZA, PHILIP J.; HAHN, PAUL; TAI, VINCENT; SEVILLA, MONICA B.; TRAN-VIET, DU; CUNEFARE, DAVID; FARSIU, SINA; IZATT, JOSEPH A.; TOTH, CYNTHIA A.

    2015-01-01

    Purpose To report the intraoperative optical coherence tomography findings in idiopathic epiretinal membrane (ERM) with connecting strands and to describe the postoperative outcomes. Methods A retrospective, case series study within a prospective observational intraoperative optical coherence tomography imaging study was performed. Epiretinal membranes with connecting strands were characterized on preoperative spectral domain optical coherence tomography images and assessed against corresponding intraoperative (after internal limiting membrane [ILM] peeling) and postoperative spectral domain optical coherence tomography images. Results Eleven locations of the connecting strands in 7 eyes were studied. The connecting strands had visible connections from the inner retinal surface to the ERM in all locations, and the reflectivity was moderate in 8 locations and high in 3 locations. After ERM and ILM peeling, disconnected strands were identified in all of the intraoperative optical coherence tomography images. The reflectivity of the remaining intraoperative strands was higher than that of the preoperative lesions and appeared as “finger-like” and branching projections. The remaining disconnected lesions were contiguous with the inner retinal layers. Postoperatively, the intraoperative lesions disappeared completely in all locations, and recurrent formation of ERM was not identified in any eyes. Conclusion In ERM eyes with connecting strands, intraoperative spectral domain optical coherence tomography imaging showed moderately to highly reflective sub-ILM finger-like lesions that persist immediately after membrane and ILM peeling. Postoperatively, the hyperreflective lesions disappeared spontaneously without localized nerve fiber layer loss. The sub-ILM connecting strands may represent glial retinal attachments. PMID:25829349

  6. Ezrin and Moesin Are Required for Efficient T Cell Adhesion and Homing to Lymphoid Organs

    PubMed Central

    Chen, Emily J. H.; Shaffer, Meredith H.; Williamson, Edward K.; Huang, Yanping; Burkhardt, Janis K.

    2013-01-01

    T cell trafficking between the blood and lymphoid organs is a complex, multistep process that requires several highly dynamic and coordinated changes in cyto-architecture. Members of the ezrin, radixin and moesin (ERM) family of actin-binding proteins have been implicated in several aspects of this process, but studies have yielded conflicting results. Using mice with a conditional deletion of ezrin in CD4+ cells and moesin-specific siRNA, we generated T cells lacking ERM proteins, and investigated the effect on specific events required for T cell trafficking. ERM-deficient T cells migrated normally in multiple in vitro and in vivo assays, and could undergo efficient diapedesis in vitro. However, these cells were impaired in their ability to adhere to the β1 integrin ligand fibronectin, and to polarize appropriately in response to fibronectin and VCAM-1 binding. This defect was specific for β1 integrins, as adhesion and polarization in response to ICAM-1 were normal. In vivo, ERM-deficient T cells showed defects in homing to lymphoid organs. Taken together, these results show that ERM proteins are largely dispensable for T cell chemotaxis, but are important for β1 integrin function and homing to lymphoid organs. PMID:23468835

  7. Diversity of antimicrobial resistance and virulence genes in methicillin-resistant non-Staphylococcus aureus staphylococci from veal calves.

    PubMed

    Argudín, M Angeles; Vanderhaeghen, Wannes; Butaye, Patrick

    2015-04-01

    In this study we determined whether methicillin-resistant non-Staphylococcus aureus (MRNAS) from veal calves may be a potential reservoir of antimicrobial-resistance and virulence genes. Fifty-eight MRNAS were studied by means of DNA-microarray and PCR for detection of antimicrobial resistance and virulence genes. The isolates carried a variety of antimicrobial-resistance genes [aacA-aphD, aadD, aph3, aadE, sat, spc, ampA, erm(A), erm(B), erm(C), erm(F), erm(T), lnu(A), msr(A)-msr(B), vga(A), mph(C), tet(K), tet(M), tet(L), cat, fexA, dfrA, dfrD, dfrG, dfrK, cfr, fusB, fosB, qacA, qacC, merA-merB]. Some isolates carried resistance genes without showing the corresponding resistance phenotype. Most MRNAS carried typical S. aureus virulence factors like proteases (sspP) and enterotoxins (seg) genes. Most Staphylococcus epidermidis isolates carried the arginine catabolic element, and nearly 40% of the Staphylococcus sciuri isolates carried leukocidins, and/or fibronectin-binding protein genes. MRNAS were highly multi-resistant and represent an important reservoir of antimicrobial resistance and virulence genes. PMID:25637268

  8. Identification and characterization of proliferative retinopathy-related long noncoding RNAs

    SciTech Connect

    Zhou, Rong-Mei; Wang, Xiao-Qun; Yao, Jin; Shen, Yi; Chen, Sai-Nan; Yang, Hong; Jiang, Qin; Yan, Biao

    2015-09-25

    Proliferative vitreoretinopathy (PVR) is a serious complication of retinal detachment and vitreoretinal surgery, which can lead to severe vision reduction. Long non-coding RNAs (lncRNAs) play critical roles in many biological processes and disease development. We attempted to determine the role of lncRNAs in the setting of PVR. Microarray analysis revealed that 78 lncRNAs were abnormally expressed in the epiretinal membranes (ERMs) of PVR patients, including 48 up-regulated and 30 down-regulated lncRNA transcripts. We subsequently focus on one lncRNA, MALAT1, and investigated its expression pattern in the biofluid of PVR patients. MALAT1 was significantly up-regulated in the cellular and plasma fraction of peripheral blood in PVR patients. MALAT1 expression was obviously reduced after PVR operation. In vitro experiments revealed the role of MALAT1 in regulating RPE proliferation and migration, which is critical for ERMs formation. This study suggests that lncRNAs are the potential regulators of PVR pathology. MALAT1 is a potential prognostic indicator and a target for the diagnosis and gene therapy for PVR diseases. - Highlights: • 78 lncRNAs are differentially expressed between PVR-ERMs and secondary ERMs. • MALAT1 level is elevated in the ERMs of PVR patients. • Circulating MALAT1 level is up-regulated in PVR patients. • MALAT1 knockdown regulates RPE proliferation and migration.

  9. Role of epithelial-mesenchymal transition in proliferative vitreoretinopathy.

    PubMed

    Tamiya, Shigeo; Kaplan, Henry J

    2016-01-01

    Proliferative vitreoretinopathy (PVR) is a potentially blinding fibrotic complication. It is caused by the formation and contraction of epiretinal membranes (ERMs) that ultimately lead to retinal folds and traction retinal detachments. While multiple cell types have been identified in ERMs, retinal pigment epithelial (RPE) cells have long been implicated as a key player in the pathophysiology of PVR. Clinical and experimental evidence has shown that RPE cells undergo epithelial-mesenchymal transition (EMT) to adopt a fibroblastic phenotype. Cell-cell adhesions maintained by adherens and tight junctions are important for the maintenance of RPE phenotype, and disruption of these junctional complexes results in EMT via activation of signaling pathways such as β-catenin/Wnt and Hippo signaling, as well as transcription factors involving Zeb1, Snail, and ZONAB. Upon EMT, RPE cells can further differentiate into myofibroblasts in the presence of TGF-β with cytoskeletal tension mediated by RhoGTPase. These fibroblasts and myofibroblasts derived from RPE cells can contribute to ERM formation by cell migration, proliferation and matrix modification, and play a key role in ERM contraction. It is not solely the proliferation of these cells that results in PVR but rather the contraction of these cells in the ERM. PMID:26675400

  10. Variability in the Gulf of Alaska from Geosat altimetry data

    NASA Technical Reports Server (NTRS)

    Bhaskaran, Shyam; Lagerloef, Gary S. E.; Born, George H.; Emery, W. J.; Leben, Robert R.

    1993-01-01

    Satellite altimetry was used to examine annual and interannual variability in the Gulf of Alaska region. Crossover data from the Geosat Geodetic Mission (GM) and collinear data from the Exact Repeat Mission (ERM) were processed separately to form sea surface height anomalies at grid points. The time series from the GM and ERM were then combined to produce a 3.75-year data set. The time series from the ERM data set agreed fairly well with hydrographic dynamic heights at several locations, with an average correlation of 0.70 between the two data sets. The combined and ERM altimetric data sets were analyzed using empirical orthogonal functions (EOFs). These revealed variability that occurs primarily on annual and interannual time scales. A comparison with EOF analysis of the atmospheric pressure field during the same time periods showed that the annual variation in pressure seemed to be reflected in both the combined and ERM altimetric data sets. The amplitude time series of the first mode in the combined data set was very similar to the North Pacific pressure index during the 1985-1989 time frame. The maximum correlation was at a lag of 250 days. Finally, an interannual mode was present in all three data sets which was closely linked to the baroclinic variations measured by the hydrographic data.

  11. Effects of tylosin use on erythromycin resistance in enterococci isolated from swine.

    PubMed

    Jackson, Charlene R; Fedorka-Cray, Paula J; Barrett, John B; Ladely, Scott R

    2004-07-01

    The effect of tylosin on erythromycin-resistant enterococci was examined on three farms; farm A used tylosin for growth promotion, farm B used tylosin for treatment of disease, and farm C did not use tylosin for either growth promotion or disease treatment. A total of 1,187 enterococci were isolated from gestation, farrowing, suckling, nursery, and finishing swine from the farms. From a subset of those isolates (n = 662), 59% (124 out of 208), 28% (80 out of 281), and 2% (4 out of 170) were resistant to erythromycin (MIC >/= 8 microg/ml) from farms A, B, and C, respectively. PCR analysis and Southern blotting revealed that 95% (65 out of 68) of isolates chosen from all three farms for further study were positive for ermB, but all were negative for ermA and ermC. By using Southern blotting, ermB was localized to the chromosome in 56 of the isolates while 9 isolates from farms A and B contained ermB on two similar-sized plasmid bands (12 to 16 kb). Pulsed-field gel electrophoresis revealed that the isolates were genetically diverse and represented a heterogeneous population of enterococci. This study suggests that although there was resistance to a greater number of enterococcal isolates on a farm where tylosin was used as a growth promotant, resistant enterococci also existed on a farm where no antimicrobial agents were used. PMID:15240302

  12. A Patient-Derived Xenograft Model of Parameningeal Embryonal Rhabdomyosarcoma for Preclinical Studies

    PubMed Central

    Hooper, Jody E.; Cantor, Emma L.; Ehlen, Macgregor S.; Banerjee, Avirup; Malempati, Suman; Stenzel, Peter; Woltjer, Randy L.; Gandour-Edwards, Regina; Goodwin, Neal C.; Yang, Yan; Kaur, Pali; Bult, Carol J.; Airhart, Susan D.; Keller, Charles

    2015-01-01

    Embryonal rhabdomyosarcoma (eRMS) is one of the most common soft tissue sarcomas in children and adolescents. Parameningeal eRMS is a variant that is often more difficult to treat than eRMS occurring at other sites. A 14-year-old female with persistent headaches and rapid weight loss was diagnosed with parameningeal eRMS. She progressed and died despite chemotherapy with vincristine, actinomycin-D, and cyclophosphamide plus 50.4 Gy radiation therapy to the primary tumor site. Tumor specimens were acquired by rapid autopsy and tumor tissue was transplanted into immunodeficient mice to create a patient-derived xenograft (PDX) animal model. As autopsy specimens had an ALK R1181C mutation, PDX tumor bearing animals were treated with the pan-kinase inhibitor lestaurtinib but demonstrated no decrease in tumor growth, suggesting that single agent kinase inhibitor therapy may be insufficient in similar cases. This unique parameningeal eRMS PDX model is publicly available for preclinical study. PMID:26696773

  13. Embryonal Rhabdomyosarcoma of the Adult’s Vocal Cord: A Case Report

    PubMed Central

    Li, Yining; Fu, Zhongying; Chen, Weilun; Yang, Lixin; Zhu, Wei

    2015-01-01

    Introduction: Although embryonal rhabdomyosarcomas (ERMS) represent the most common (50% - 60%) clinical form of rhabdomyosarcoma, these are extremely rare in adult’s vocal cords. To date, only five cases of laryngeal ERMS have been reported in adults and only one of them involved the vocal cords. Case Presentation: Herein, we reported a case of a 22-year-old woman diagnosed with ERMS, T1N0M0 stage, localized at the right vocal cord. Tumor was successfully treated with a radical excision and selective neck dissection and followed-up for 5 years with no evidence of local recurrence. Conclusions: Immunohistochemistry combined with computerized tomography (CT) and color Doppler solography of the neck and thorax should be performed. This study suggested that surgical resection was suitable for the treatment of this disease and reported the evaluations of an extremely rare case for five years. PMID:26430531

  14. A mouse model of rhabdomyosarcoma originating from the adipocyte lineage

    PubMed Central

    Hatley, Mark E.; Tang, Wei; Garcia, Matthew R.; Finkelstein, David; Millay, Douglas P.; Liu, Ning; Graff, Jonathan; Galindo, Rene L.; Olson, Eric N.

    2012-01-01

    SUMMARY Rhabdomyosarcoma (RMS) is an aggressive skeletal muscle-lineage tumor composed of malignant myoblasts that fail to exit the cell cycle and are blocked from fusing into syncytial muscle. Rhabdomyosarcoma includes two histolopathologic subtypes: alveolar rhabdomyosarcoma, driven by the fusion protein PAX3-FOXO1 or PAX7-FOXO1, and embryonal rhabdomyosarcoma (ERMS), which is genetically heterogeneous. Here, we show that adipocyte-restricted activation of Sonic Hedgehog signaling through expression of a constitutively active Smoothened allele in mice gives rise to aggressive skeletal muscle tumors that display the histologic and molecular characteristics of human ERMS with high penetrance. Our findings suggest that adipocyte progenitors can be a cell of origin for Sonic Hedgehog-driven ERMS, showing that RMS can originate from non-skeletal muscle precursors. PMID:23079662

  15. Ezrin-Radixin-Moesin family proteins in the regulation of B cell immune response

    PubMed Central

    Pore, Debasis; Gupta, Neetu

    2015-01-01

    Dynamic reorganization of the cortical cytoskeleton is essential for numerous cellular processes including B and T cell activation and migration. The Ezrin, Radixin and Moesin (ERM) family proteins play structural and regulatory roles in the rearrangement of plasma membrane flexibility and protrusions through their reversible interaction with cortical actin filaments and plasma membrane. Recent studies demonstrate that ERM proteins are not only involved in cytoskeletal organization but also offer a platform for the transmission of signals in response to a variety of extracellular stimuli through their ability to crosslink transmembrane receptors with downstream signaling components. In this review, we summarize the present knowledge and recent progress made towards elucidating a novel role of ERM proteins in the regulation of B function in health and disease. PMID:25746045

  16. Pursuing enterprise risk management: a local road map for Canadian healthcare leaders.

    PubMed

    Haney, James R; Church, John; Cockerill, Rhonda

    2013-01-01

    An in-depth analysis of organizational risk management in healthcare, and in particular the concepts of Enterprise Risk Management (ERM), has identified a 5-part model that can be used by Canadian healthcare leaders as an evidence-supported approach to successful organizational risk management. The Model for Organizational Risk Management, termed "the Model," has been developed as a basis for linking the components of an ERM Framework into a Canadian health organization to overcome the barriers that commonly disrupt strategic risk management. The Model addresses how an ERM Framework can fit within an existing health organization by building off and enhancing existing processes and resources to ensure familiarity, acceptance, and sustainability of the risk management program. By approaching the Model in a stepwise fashion (based on individual organizational context), healthcare leaders are provided with a road map from which to advance their own organizational risk management program. PMID:24409582

  17. Investigation of postoperative intraocular pressure in cases of silicone oil removal using 25-gauge transconjunctival sutureless vitrectomy with oblique incisions

    PubMed Central

    Takashina, Hirotsugu; Watanabe, Akira; Tsuneoka, Hiroshi

    2015-01-01

    Background The purpose of this study was to investigate postoperative intraocular pressure (IOP) in cases of silicone oil (SO) removal when using 25-gauge transconjunctival sutureless vitrectomy (TSV) with oblique incisions. Methods We enrolled ten consecutive eyes with SO removal (SO group) and eleven consecutive eyes with idiopathic epiretinal membrane (ERM) as the initial vitrectomy (ERM group) in cases using 25-gauge TSV with oblique incisions. Postoperative IOPs were compared between the two groups at each of the four examination periods. Results No significant differences were identified in any of the periods examined. Conclusion The use of 25-gauge TSV with oblique incisions resulted in almost equivalent postoperative IOPs between cases with SO removal and idiopathic ERM as the initial operation. Self-sealing sclerotomy in 25-gauge TSV with oblique incisions may primarily involve the valve architecture, and be complemented by vitreous incarceration. PMID:26508831

  18. Evidence for an Unanticipated Relationship Between Undifferentiated Pleomorphic Sarcoma and Embryonal Rhabdomyosarcoma

    PubMed Central

    Rubin, Brian P.; Nishijo, Koichi; Chen, Hung-I Harry; Yi, Xiaolan; Schuetze, David P.; Pal, Ranadip; Prajapati, Suresh I.; Abraham, Jinu; Arenkiel, Benjamin R.; Chen, Qing-Rong; Davis, Sean; McCleish, Amanda T.; Capecchi, Mario R.; Michalek, Joel E.; Zarzabal, Lee Ann; Khan, Javed; Yu, Zhongxin; Parham, David M.; Barr, Frederic G.; Meltzer, Paul S.; Chen, Yidong; Keller, Charles

    2011-01-01

    SUMMARY Embryonal rhabdomyosarcoma (eRMS) shows the most myodifferentiation amongst sarcomas, yet the precise cell of origin remains undefined. Using Ptch1, p53 and/or Rb1 conditional mouse models and controlling prenatal or postnatal myogenic cell of origin, we demonstrate that eRMS and undifferentiated pleomorphic sarcoma (UPS) lie in a continuum, with satellite cells predisposed to giving rise to UPS. Conversely, p53 loss in maturing myoblasts gives rise to eRMS, which have the highest myodifferentiation potential. Irrespective of origin, Rb1 loss modifies tumor phenotype to mimic UPS. In human sarcomas that lack pathognomic chromosomal translocations, p53 loss of function is prevalent whereas Shh or Rb1 alterations likely act primarily as modifiers. Thus, sarcoma phenotype is strongly influenced by cell of origin and mutational profile. PMID:21316601

  19. Another Estimate for the Percentage of Partly Embayed Craters in Equilibrium Resurfacing

    NASA Astrophysics Data System (ADS)

    Muinonen, P. J.

    1996-03-01

    When choosing between different scenarios of Venus' resurfacing history, one of the objections against an Equilibrium Resurfacing Model (ERM) is an apparent lack of craters that have suffered a partial embayment by lava flows of outside origin. Any test of ERM so far, be it numeric or analytic in nature, has failed to predict anything even near the observed ~5% of partly embayed craters (PECs) without violating the statistics of crater distribution. This study is mainly based on straightening out as many as possible of the simplifications that have been employed while estimating the statistical implications of the ERM. As a result, the expectation value for PECs falls to less than 10% from previous ~30%.

  20. Estimation of quality factors by energy ratio method

    NASA Astrophysics Data System (ADS)

    Wang, Zong-Jun; Cao, Si-Yuan; Zhang, Hao-Ran; Qu, Ying-Ming; Yuan, Dian; Yang, Jin-Hao; Shao, Guan-Ming

    2015-03-01

    The quality factor Q, which reflects the energy attenuation of seismic waves in subsurface media, is a diagnostic tool for hydrocarbon detection and reservoir characterization. In this paper, we propose a new Q extraction method based on the energy ratio before and after the wavelet attenuation, named the energy-ratio method (ERM). The proposed method uses multipoint signal data in the time domain to estimate the wavelet energy without invoking the source wavelet spectrum, which is necessary in conventional Q extraction methods, and is applicable to any source wavelet spectrum; however, it requires high-precision seismic data. Forward zero-offset VSP modeling suggests that the ERM can be used for reliable Q inversion after nonintrinsic attenuation (geometric dispersion, reflection, and transmission loss) compensation. The application to real zero-offset VSP data shows that the Q values extracted by the ERM and spectral ratio methods are identical, which proves the reliability of the new method.

  1. Precise orbit computation for the Geosat Exact Repeat Mission

    NASA Technical Reports Server (NTRS)

    Haines, Bruce J.; Born, George H.; Rosborough, George W.; Marsh, James G.; Williamson, Ronald G.

    1990-01-01

    Results are reported from an extensive investigation of orbit-determination strategies for the Geosat Exact Repeat Mission (ERM). The goal is to establish optimum geodetic parameters and procedures for the computation of the most accurate Geosat orbits possible and to apply these procedures for routine computation during the ERM for the following purposes: (1) to enhance the value of the Geosat oceanographic investigations by providing the user community with improved ephemerides, (2) to develop orbit determination techniques for the upcoming altimetric mission Topex/Poseidon, and (3) to assess the radial orbit accuracy obtainable with recently developed gravity models. To this end, ephemerides for the entire first year of the ERM have been computed using the GEODYN II orbit program on the Cyber 205 supercomputer system at the NASA Goddard.

  2. Hepatocyte Growth Factor-mediated satellite cells niche perturbation promotes development of distinct sarcoma subtypes.

    PubMed

    Morena, Deborah; Maestro, Nicola; Bersani, Francesca; Forni, Paolo Emanuele; Lingua, Marcello Francesco; Foglizzo, Valentina; Šćepanović, Petar; Miretti, Silvia; Morotti, Alessandro; Shern, Jack F; Khan, Javed; Ala, Ugo; Provero, Paolo; Sala, Valentina; Crepaldi, Tiziana; Gasparini, Patrizia; Casanova, Michela; Ferrari, Andrea; Sozzi, Gabriella; Chiarle, Roberto; Ponzetto, Carola; Taulli, Riccardo

    2016-01-01

    Embryonal Rhabdomyosarcoma (ERMS) and Undifferentiated Pleomorphic Sarcoma (UPS) are distinct sarcoma subtypes. Here we investigate the relevance of the satellite cell (SC) niche in sarcoma development by using Hepatocyte Growth Factor (HGF) to perturb the niche microenvironment. In a Pax7 wild type background, HGF stimulation mainly causes ERMS that originate from satellite cells following a process of multistep progression. Conversely, in a Pax7 null genotype ERMS incidence drops, while UPS becomes the most frequent subtype. Murine EfRMS display genetic heterogeneity similar to their human counterpart. Altogether, our data demonstrate that selective perturbation of the SC niche results in distinct sarcoma subtypes in a Pax7 lineage-dependent manner, and define a critical role for the Met axis in sarcoma initiation. Finally, our results provide a rationale for the use of combination therapy, tailored on specific amplifications and activated signaling pathways, to minimize resistance emerging from sarcomas heterogeneity. PMID:26987019

  3. Direct detection of antibiotic resistance genes in specimens of chicken and pork meat.

    PubMed

    Garofalo, Cristiana; Vignaroli, Carla; Zandri, Giada; Aquilanti, Lucia; Bordoni, Donatella; Osimani, Andrea; Clementi, Francesca; Biavasco, Francesca

    2007-01-01

    Antibiotic resistance (AR) in bacteria, a major threat to human health, has emerged in the last few decades as a consequence of the selective pressure exerted by the widespread use of antibiotics in medicine, agriculture and veterinary practice and as growth promoters in animal husbandry. The frequency of 11 genes [tet(M), tet(O), tet(K), erm(A), erm(B), erm(C), vanA, vanB, aac (6')-Ie aph (2'')-Ia, mecA, blaZ] encoding resistance to some antibiotics widely used in clinical practice was analysed in raw pork and chicken meat and in fermented sausages as well as in faecal samples from the relevant farm animals using a molecular approach based on PCR amplification of bacterial DNA directly extracted from specimens. Some of the 11 AR genes were highly prevalent, the largest number being detected in chicken meat and pig faeces. The genes found most frequently in meat were tet(K) and erm(B); vanB and mecA were the least represented. All 11 determinants were detected in faecal samples except mecA, which was found only in chicken faeces. erm(B) and erm(C) were detected in all faecal samples. The frequency of AR genes was not appreciably different in meat compared to faecal specimens of the relevant animal except for vanB, which was more prevalent in faeces. Our findings suggest that AR genes are highly prevalent in food-associated bacteria and that AR contamination is likely related to breeding rather than processing techniques. Finally, the cultivation-independent molecular method used in this work to determine the prevalence of AR genes in foods proved to be a rapid and reliable alternative to traditional tools. PMID:17005283

  4. Inducible and Acquired Clarithromycin Resistance in the Mycobacterium abscessus Complex

    PubMed Central

    Rubio, Marc; March, Francesca; Garrigó, Montserrat; Moreno, Carmen; Español, Montserrat; Coll, Pere

    2015-01-01

    Purpose Clarithromycin was considered the cornerstone for the treatment of Mycobacterium abscessus complex infections. Genetic resistance mechanisms have been described and many experts propose amikacin as an alternative. Nevertheless, clarithromycin has several advantages; therefore, it is necessary to identify the non-functional erm(41) allele to determine the most suitable treatment. The aims of this study were to characterize the molecular mechanisms of clarithromycin resistance in a collection of Mycobacterium abscessus complex isolates and to verify the relationship between these mechanisms and the antibiogram. Materials and Methods Clinical isolates of M. abscessus complex (n = 22) from 16 patients were identified using four housekeeping genes (rpoB, secA1, sodA and hsp65), and their genetic resistance was characterized by studying erm(41) and rrl genes. Nine strains were recovered from the clinical isolates and subjected to E-test and microdilution clarithromycin susceptibility tests, with readings at 3, 7 and 14 days. Results We classified 11/16 (68.8%) M. abscessus subsp. abscessus, 4/16 (25.0%) M. abscessus subsp. bolletii, and 1/16 (6.3%) M. abscessus subsp. massiliense. T28 erm(41) allele was observed in 8 Mycobacterium abscessus subps. abscessus and 3 Mycobacterium abscessus subsp. bolletii. One strain of M. abscessus subsp. bolletii had an erm(41) gene truncated and was susceptible to clarithromycin. No mutations were observed in rrl gene first isolates. In three patients, follow-up of initial rrl wild-type strains showed acquired resistance. Conclusions Most clinical isolates of M. abscessus complex had inducible resistance to clarithromycin and total absence of constitutive resistance. Our findings showed that the acquisition of resistance mutations in rrl gene was associated with functional and non-functional erm(41) gene. Caution is needed when using erm(41) sequencing alone to identify M. abscessus subspecies. This study reports an acquired

  5. Evaluation of the in vitro activity of tigecycline against multiresistant Gram-positive cocci containing tetracycline resistance determinants.

    PubMed

    Borbone, Sonia; Lupo, Agnese; Mezzatesta, Maria Lina; Campanile, Floriana; Santagati, Maria; Stefani, Stefania

    2008-03-01

    This study was undertaken to test the in vitro activity of tigecycline against 117 clinically relevant Gram-positive pathogens and to correlate this activity with their resistance gene content. Overall, tigecycline showed a minimal inhibitory concentration (MIC) range of 0.015-0.5mg/L, able to inhibit potently all multiresistant streptococci, enterococci and MR staphylococci. Tigecycline was active against methicillin-resistant Staphylococcus aureus (MRSA) and enterococci, with MICs for 90% of the organisms (MIC(90)) of 0.25 mg/L and 0.12 mg/L, respectively, being more active than linezolid (MIC(90)=2 mg/L) and quinupristin/dalfopristin (MIC(90)=0.5 and 2-4 mg/L, respectively). Molecular characterisation of resistance determinants demonstrated the concomitant presence of different classes of genes: in particular, tet(M), erm(B) and erm(C) in Streptococcus agalactiae; tet(O), variably associated with different erm genes, in Streptococcus pyogenes; vanA, tet(M) and erm(B) in Enterococcus faecalis, and vanA and erm(B) in Enterococcus faecium. All the MRSA strains harboured SCCmec and erm genes and 50% possessed tet(K) with tet(M) genes. Staphylococcus epidermidis strains were only resistant to erythromycin. These results clearly demonstrate that tigecycline has a MIC(90) range of 0.015-0.5 mg/L both against tetracycline-susceptible and -resistant isolates carrying other resistance determinants, suggesting that this drug could play an important role in the treatment of infections caused by these multiresistant Gram-positive pathogens. PMID:17646087

  6. The Prevalence and Incidence of Epiretinal Membranes in Eyes With Inactive Extramacular CMV Retinitis

    PubMed Central

    Kozak, Igor; Vaidya, Vijay; Van Natta, Mark L.; Pak, Jeong W.; May, K. Patrick; Thorne, Jennifer E.

    2014-01-01

    Purpose. To determine the prevalence and incidence of epiretinal membranes (ERM) in eyes with inactive extramacular cytomegalovirus (CMV) retinitis in patients with acquired immune deficiency syndrome (AIDS). Methods. A case–control report from a longitudinal multicenter observational study by the Studies of the Ocular Complications of AIDS (SOCA) Research Group. A total of 357 eyes of 270 patients with inactive CMV retinitis and 1084 eyes of 552 patients with no ocular opportunistic infection (OOI) were studied. Stereoscopic views of the posterior pole from fundus photographs were assessed at baseline and year 5 visits for the presence of macular ERM. Generalized estimating equations (GEE) logistic regression was used to compare the prevalence and 5-year incidence of ERM in eyes with and without CMV retinitis at enrollment. Crude and adjusted logistic regression was performed adjusting for possible confounders. Main outcome measures included the prevalence, incidence, estimated prevalence, and incidence odds ratios. Results. The prevalence of ERM at enrollment was 14.8% (53/357) in eyes with CMV retinitis versus 1.8% (19/1084) in eyes with no OOI. The incidence of ERM at 5 years was 18.6% (16/86) in eyes with CMV retinitis versus 2.4% (6/253) in eyes with no OOI. The crude odds ratio (OR) (95% confidence interval, CI) for prevalence was 9.8 (5.5–17.5) (P < 0.01). The crude OR (95% CI) for incidence was 9.4 (3.2–27.9) (P < 0.01). Conclusions. A history of extramacular CMV retinitis is associated with increased prevalence and incidence of ERM formation compared to what is seen in eyes without ocular opportunistic infections in AIDS patients. PMID:24925880

  7. Statin medication in patients with epiretinal membrane is associated with low intravitreal EPO, TGF-beta-1, and VEGF levels

    PubMed Central

    Tuuminen, Raimo; Loukovaara, Sirpa

    2016-01-01

    Background In eyes with idiopathic epiretinal membrane (iERM), the intravitreal growth factor and cytokine levels may associate with postvitrectomy outcomes. Here, we have analyzed the perioperative intravitreal protein levels of potent vasoactive, proinflammatory, and extracellular matrix-remodeling factors in iERM eyes and evaluated the postvitrectomy outcomes. Methods This was an institutional, observational study. Eyes operated on for iERM (n=26) were analyzed according to the use of statin medication. Vitreous samples were subjected to protein measurements of angiopoietin-1 and -2, erythropoietin, transforming growth factor-β1, and vascular endothelial growth factor by enzyme-linked immunosorbent assay, and of matrix metalloproteinase-2 and -9 by gelatin zymography. One-month visual outcomes and 1-year revitrectomy rates were recorded. Results In iERM eyes of patients taking statins, intravitreal levels of erythropoietin (mean ± standard deviation, 10.8±4.9 vs 82.9±119.5 mIU/mg, P=0.003), transforming growth factor-β1 (2.3±4.7 vs 15.8±16.3 pg/mg, P=0.035), and vascular endothelial growth factor (5.5±9.9 vs 236.6±491.6 pg/mg, P=0.006) were lower than in nonstatin-treated patients. At 1-month, visual gain did not significantly differ between iERM eyes of patients with statins and those without (improvement 0.27±0.20 vs 0.16±0.38 logarithm of the minimum angle of resolution units, P=0.118). Conclusion Systemic statin therapy might have a favorable effect on intravitreal factors involved in vascular permeability, inflammation, and fibroproliferation in aging human iERM eyes. PMID:27284236

  8. Detection of a Common and Persistent tet(L)-Carrying Plasmid in Chicken-Waste-Impacted Farm Soil

    PubMed Central

    Hilpert, Markus; Ward, Mandy J.

    2012-01-01

    The connection between farm-generated animal waste and the dissemination of antibiotic resistance in soil microbial communities, via mobile genetic elements, remains obscure. In this study, electromagnetic induction (EMI) surveying of a broiler chicken farm assisted soil sampling from a chicken-waste-impacted site and a marginally affected site. Consistent with the EMI survey, a disparity existed between the two sites with regard to soil pH, tetracycline resistance (Tcr) levels among culturable soil bacteria, and the incidence and prevalence of several tet and erm genes in the soils. No significant difference was observed in these aspects between the marginally affected site and several sites in a relatively pristine regional forest. When the farm was in operation, tet(L), tet(M), tet(O), erm(A), erm(B), and erm(C) genes were detected in the waste-affected soil. Two years after all waste was removed from the farm, tet(L), tet(M), tet(O), and erm(C) genes were still detected. The abundances of tet(L), tet(O), and erm(B) were measured using quantitative PCR, and the copy numbers of each were normalized to eubacterial 16S rRNA gene copy numbers. tet(L) was the most prevalent gene, whereas tet(O) was the most persistent, although all declined over the 2-year period. A mobilizable plasmid carrying tet(L) was identified in seven of 14 Tcr soil isolates. The plasmid's hosts were identified as species of Bhargavaea, Sporosarcina, and Bacillus. The plasmid's mobilization (mob) gene was quantified to estimate its prevalence in the soil, and the ratio of tet(L) to mob was shown to have changed from 34:1 to 1:1 over the 2-year sampling period. PMID:22389375

  9. In vivo spread of macrolide-lincosamide-streptogramin B (MLSB) resistance--a model study in chickens.

    PubMed

    Marosevic, D; Cervinkova, D; Vlkova, H; Videnska, P; Babak, V; Jaglic, Z

    2014-07-16

    The influence of specific and non-specific antibiotic pressure on in vivo spread of macrolide-lincosamide-streptogramin B (MLSB) resistance was evaluated in this study. Chickens repeatedly inoculated with Enterococcus faecalis harbouring the plasmid pAMβ1 carrying the erm(B) gene were perorally treated for one week with tylosin, lincomycin (both specific antibiotic pressure) and chlortetracycline (non-specific antibiotic pressure). Antibiotic non-treated but E. faecalis inoculated chickens served as a control. To quantify the erm(B) gene and characterise intestinal microflora, faecal DNA was analysed by qPCR and 454-pyrosequencing. Under the pressure of antibiotics, a significant increase in erm(B) was observed by qPCR. However, at the final stage of the experiment, an increase in erm(B) was also observed in two out of five non-treated chickens. In chickens treated with tylosin and chlortetracycline, the increase in erm(B) was accompanied by an increase in enterococci. However, E. faecalis was at the limit of detection in all animals. This suggests that the erm(B) gene spread among the gut microbiota other than E. faecalis. Pyrosequencing results indicated that, depending on the particular antibiotic pressure, different bacteria could be responsible for the spread of MLSB resistance. Different species of MLSB-resistant enterococci and streptococci were isolated from cloacal swabs during and after the treatment. PFGE analysis of MLSB-resistant enterococci revealed four clones, all differing from the challenge strain. All of the MLSB-resistant isolates harboured a plasmid of the same size as pAMβ1. This study has shown that MLSB resistance may spread within the gut microbiota under specific and non-specific pressure and even in the absence of any antimicrobial pressure. Finally, depending on the particular antibiotic pressure, different bacterial species seems to be involved in the spread of MLSB resistance. PMID:24467930

  10. Protection against methoxyacetic-acid-induced spermatocyte apoptosis with calcium channel blockers in cultured rat seminiferous tubules: possible mechanisms.

    PubMed

    Li, L H; Wine, R N; Miller, D S; Reece, J M; Smith, M; Chapin, R E

    1997-05-01

    A calcium-mediated mechanism underlying spermatocyte apoptosis induced by 2-methoxyethanol (2-ME) has been previously proposed. This hypothesis was tested in vitro in the present study using cultured juvenile (25 days old) and adult rat seminiferous tubules (JRST and ARST, respectively) with methoxyacetic acid (MAA, the active metabolite of 2-ME). In JRST, spermatocyte degeneration was morphologically obvious 19 hr after a 5-hr exposure to 5 mM MAA. The lesion was unaffected by the presence or absence of extratubular Ca2+. However, MAA-induced cell death was significantly prevented by cotreatment with the dihydropyridines (DHP) nifedipine (50 microM) and nicardipine (20 microM), as well as verapamil (50 microM) and TMB-8 (50 microM), all of which are able to inhibit calcium movement through plasma membranes. However, neither ryanodine, dantrolene, nor cyclosporin A and ruthenium red, which inhibit Ca2+ mobilization from intracellular stores (endoplasmic reticulum and mitochondria), affected the MAA-induced cell death. Inhibition of calcium mobilization through IP3-sensitive pathways by blocking the product of IP3 with manoalide, neomycin, and U73122 did not block the MAA-induced lesion. The protective effects of 50 microM nifedipine and 50 microM TMB-8 were also observed in ARSTs treated with 10 mM MAA for 5 hr. However, when rat testicular sections were immunohistochemically stained with monoclonal antibodies specific for the alpha 1 (the DHP receptor) or the alpha 2 subunits of DHP-sensitive calcium channels, no positive staining was found. Finally, in an attempt to see whether the intracellular free calcium concentrations ([Ca2+]i) in germ cells were increased after the MAA treatment, intact seminiferous tubules were loaded with indo-1 and were measured using laser-scanning confocal microscopy. No detectable increase in the signal in MA A-sensitive spermatocytes was observed, while a 34-54% increase in the signal could be detected in the same cell types when

  11. [Juvenile idiopathic epiretinal membrane].

    PubMed

    Kontopoulou, K; Krause, S; Fili, S; Hayvazov, S; Schilling, H; Kohlhaas, M

    2016-07-01

    Idiopathic epiretinal membrane (iERM) is very rare in adolescent patients. The pathogenesis remains unclear although the role of hyalocytes is of major importance. The clinical features in young patients are different from those in older patients. We describe a case of iERM in a 15-year-old girl who presented with metamorphopsia of the right eye. This case report presents the basis for the decision for surgical treatment as well as the clinical features at follow-up examination 9 months after surgery. PMID:26458892

  12. Sicheres Navigieren in dynamischen Umgebungen mit 3D-Kollisionsvermeidung

    NASA Astrophysics Data System (ADS)

    Stiene, Stefan; Hertzberg, Joachim

    Diese Papier stellt eine neuartige Methode vor, die 3D-Hindernisvermeidung ermöglicht. Die Sensorkonfiguration beruht auf sechs Laserscannern, die die gesamte Roboteroberfläche abdecken. Die Daten der sechs Laserscanner werden zu einem virtuellen, zweidimensionalen, horizontal ausgerichteten 360°-Laserscanner kombiniert. Da der virtuelle Laserscanner die 3D-Umgebung in einem zweidimensionalen Datensatz repräsentiert, ermöglicht er in Kombination mit klassischen Hindernisvermeidungsalgorithmen wie der Vector Field Histogram Methode eine 3D-Hindernisvermeidung.

  13. A Novel Erythromycin Resistance Plasmid from Bacillus Sp. Strain HS24, Isolated from the Marine Sponge Haliclona Simulans

    PubMed Central

    Leong, Dara; Morrissey, John P.; Adams, Claire; Dobson, Alan D. W.; O’Gara, Fergal

    2014-01-01

    A better understanding of the origin and natural reservoirs of resistance determinants is fundamental to efficiently tackle antibiotic resistance. This paper reports the identification of a novel 5.8 kb erythromycin resistance plasmid, from Bacillus sp. HS24 isolated from the marine sponge Haliclona simulans. pBHS24B has a mosaic structure and carries the erythromycin resistance gene erm(T). This is the first report of an erythromycin resistance plasmid from a sponge associated bacteria and of the Erm(T) determinant in the genus Bacillus. PMID:25548909

  14. Serotype and serovar distribution of Neisseria gonorrhoeae isolated from high-risk populations in Bangladesh.

    PubMed

    Alam, M A; Chowdhury, M Z; Ahmed, F; Alam, A; Hossain, M A

    2012-12-01

    Neisseria gonorrhoeae, the causative agent of gonococcal infection, is known to frequently change their characteristics to evade host immune mechanism. Characterization of the clinical isolates of the organism can lead to identification of the circulating strains and often a sexual network in a community to help in designing the control strategy. Keeping in mind the above consideration, a total of 239 N. gonorrhoeae, isolated from high-risk populations, were characterized for serotypes and serovars by monoclonal antibodies against protein 1 of the organism. Majority of the serotypes were serotype B (142, 59.4%). Majority of the isolates showing resistance to at least one of the antibiotics tested were also serotype B (139, 59.2%), whereas, majority of the isolates showing resistance to any three of the antibiotics (multidrug resistant, MDR) (63%) was serotype A. A total of 41 different serovars were also identified and five of which (Arst, Bropt, Bopt, Arost, and Brop) included the highest percent (49.3%) of the isolates. Many serovars (23/41, 56.1%) were new emergent and included 58 (24.3%) of the isolates investigated. All of the new serovars were resistant to at least one of the antibiotics tested and the highest rate (40/102, 39.2%) was MDR. Serotyping and serovar determination was found contributory to understand the microepidemics of the N. gonorrhoeae isolates. Further studies including antibiogram and contact tracing can efficiently help in control of the disease. PMID:23540188

  15. The correlation between visual acuity outcomes and optical coherence tomography parameters following surgery for diabetic epiretinal membrane and taut posterior hyaloid

    PubMed Central

    Güngel, Hülya; Öztürker, Zeynep Kayaarası; Zırtıloğlu, Sibel; Eltutar, Kadir

    2015-01-01

    Purpose To evaluate the relationship between visual outcomes and the determinants detected by spectral domain optical coherence tomography (OCT) in eyes with epiretinal membrane (ERM) and/or taut posterior hyaloid (TPH) that underwent pars plana vitrectomy (PPV). Materials and methods A total of 30 participants with diabetic ERM and TPH were included in the study. All study participants underwent PPV. Preoperative and postoperative best corrected visual acuity (BCVA), peripapillary retinal nerve fiber layer (RNFL), macular RNFL, ganglion cell layer, inner plexiform layer, and ganglion cell complex thicknesses were measured in each participant. Linear regression analyses were performed to determine the association between the OCT parameters and the visual acuity measured at the time of the OCT measurement. Results The postoperative BCVA logarithm of the minimum angle of resolution (logMAR) values were statistically higher than the preoperative values in the ERM group and TPH group (P=0.001 and P<0.001, respectively). The postoperative BCVA logMAR value was negatively correlated with average RNFL, inferior RNFL thicknesses, and image quality (P=0.002, P=0.004, and P=0.006, respectively). The preoperative and postoperative BCVA logMAR value difference was not correlated with age and all of the OCT parameters measured (P>0.05). Conclusion This study shows that achievement of better peripapillary RNFL thickness results in better visual outcome after PPV and ERM/TPH removal. PMID:26316690

  16. Targeting hedgehog signaling reduces self-renewal in embryonal rhabdomyosarcoma.

    PubMed

    Satheesha, S; Manzella, G; Bovay, A; Casanova, E A; Bode, P K; Belle, R; Feuchtgruber, S; Jaaks, P; Dogan, N; Koscielniak, E; Schäfer, B W

    2016-04-21

    Current treatment regimens for rhabdomyosarcoma (RMS), the most common pediatric soft tissue cancer, rely on conventional chemotherapy, and although they show clinical benefit, there is a significant risk of adverse side effects and secondary tumors later in life. Therefore, identifying and targeting sub-populations with higher tumorigenic potential and self-renewing capacity would offer improved patient management strategies. Hedgehog signaling has been linked to the development of embryonal RMS (ERMS) through mouse genetics and rare human syndromes. However, activating mutations in this pathway in sporadic RMS are rare and therefore the contribution of hedgehog signaling to oncogenesis remains unclear. Here, we show by genetic loss- and gain-of-function experiments and the use of clinically relevant small molecule modulators that hedgehog signaling is important for controlling self-renewal of a subpopulation of RMS cells in vitro and tumor initiation in vivo. In addition, hedgehog activity altered chemoresistance, motility and differentiation status. The core stem cell gene NANOG was determined to be important for ERMS self-renewal, possibly acting downstream of hedgehog signaling. Crucially, evaluating the presence of a subpopulation of tumor-propagating cells in patient biopsies identified by GLI1 and NANOG expression had prognostic significance. Hence, this work identifies novel functional aspects of hedgehog signaling in ERMS, redefines the rationale for its targeting as means to control ERMS self-renewal and underscores the importance of studying functional tumor heterogeneity in pediatric cancers. PMID:26189795

  17. Usability Testing for e-Resource Discovery: How Students Find and Choose e-Resources Using Library Web Sites

    ERIC Educational Resources Information Center

    Fry, Amy; Rich, Linda

    2011-01-01

    In early 2010, library staff at Bowling Green State University (BGSU) in Ohio designed and conducted a usability study of key parts of the library web site, focusing on the web pages generated by the library's electronic resources management system (ERM) that list and describe the library's databases. The goal was to discover how users find and…

  18. A consolidated environmental monitoring plan for Aberdeen Proving Ground, Maryland

    SciTech Connect

    Ebinger, M.H.; Hansen, W.R.

    1997-04-01

    The US Army operates facilities in Edgewood and Aberdeen under several licenses from the Nuclear Regulatory Commission (NRC). Compliance with each license is time consuming and could potentially result in duplicated efforts to demonstrate compliance with existing environmental regulations. The goal of the ERM plan is to provide the sampling necessary to ensure that operations at Edgewood and Aberdeen are within applicable regulatory guidelines and to provide a means of ensuring that adverse effects to the environment are minimized. Existing sampling plans and environmental data generated from those plans are briefly reviewed as part of the development of the present ERM plan. The new ERM plan was designed to provide data that can be used for assessing risks to the environment and to humans using Aberdeen and Edgewood areas. Existing sampling is modified and new sampling is proposed based on the results of the long-term DU fate study. In that study, different environmental pathways were identified that would show transport of DU at Aberdeen. Those pathways would also be impacted by other radioactive constituents from Aberdeen and Edgewood areas. The ERM plan presented in this document includes sampling from Edgewood and Aberdeen facilities. The main radioactive constituents of concern at Edgewood are C, P, N, S, H, I, Co, Cs, Ca, Sr and U that are used in radiolabeling different compounds and tracers for different reactions and syntheses. Air and water sampling are the thrust of efforts at the Edgewood area.

  19. Transport of tylosin and tylosin-resistance genes in subsurface drainage water from manured fields

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Animal agriculture appears to contribute to the spread of antibiotic resistance genes, but few studies have quantified gene transport in agricultural fields. The transport of tylosin, tylosin-resistance genes (erm B, F, A) and tylosin-resistant Enterococcus were measured in tile drainage water from ...

  20. Getting insight into the prevalence of antibiotic resistance genes in specimens of marketed edible insects.

    PubMed

    Milanović, Vesna; Osimani, Andrea; Pasquini, Marina; Aquilanti, Lucia; Garofalo, Cristiana; Taccari, Manuela; Cardinali, Federica; Riolo, Paola; Clementi, Francesca

    2016-06-16

    This study was aimed at investigating the occurrence of 11 transferable antibiotic resistance (AR) genes [erm(A), erm(B), erm(C), vanA, vanB, tet(M), tet(O), tet(S), tet(K), mecA, blaZ] in 11 species of marketed edible insects (small crickets powder, small crickets, locusts, mealworm larvae, giant waterbugs, black ants, winged termite alates, rhino beetles, mole crickets, silkworm pupae, and black scorpions) in order to provide a first baseline for risk assessment. Among the AR genes under study, tet(K) occurred with the highest frequency, followed by erm(B), tet(S) and blaZ. A high variability was seen among the samples, in terms of occurrence of different AR determinants. Cluster Analysis and Principal Coordinates Analysis allowed the 11 samples to be grouped in two main clusters, one including all but one samples produced in Thailand and the other including those produced in the Netherlands. PMID:27046628

  1. Synthesis and antibacterial activity of novel lincomycin derivatives. I. Enhancement of antibacterial activities by introduction of substituted azetidines.

    PubMed

    Kumura, Ko; Wakiyama, Yoshinari; Ueda, Kazutaka; Umemura, Eijiro; Watanabe, Takashi; Shitara, Eiki; Fushimi, Hideki; Yoshida, Takuji; Ajito, Keiichi

    2016-06-01

    The synthesis and antibacterial activity of (7S)-7-sulfur-azetidin-3-yl lincomycin derivatives are described. Modification was achieved by a simple reaction of (7R)-7-O-methanesulfonyllincomycin and the corresponding substituted azetidine-2-thiol. Several compounds first showed moderate antibacterial activity against Streptococcus pneumoniae and Streptococcus pyogenes with erm gene as lincomycin derivatives. PMID:26758495

  2. Species Identification and Clarithromycin Susceptibility Testing of 278 Clinical Nontuberculosis Mycobacteria Isolates.

    PubMed

    Nie, Wenjuan; Duan, Hongfei; Huang, Hairong; Lu, Yu; Chu, Naihui

    2015-01-01

    Purpose of this paper is to analyze different species' proportion of nontuberculosis mycobacteria (NTM) and susceptibility to clarithromycin of different species. 278 clinical NTM isolates were identified into species by using 16S rRNA, rpoB and hsp65. Then clarithromycin susceptibility testing against different species was done separately, using microplate Alamar Blue assay. Finally, resistance isolates' erm(41) of M. abscessus were sequenced in order to analyze mechanisms for clarithromycin resistant. In this test, 131 isolates (47%) belonged to M. avium complex (MAC), and 70 isolates (25%) belonged to M. abscessus. Nearly all the M. abscessus subsp. abscessus resistant to clarithromycin had T28 in erm(41). However, all the M. abscessus subsp. abscessus susceptible to clarithromycin had C28 in erm(41). In this study, we find that MAC was the most common pathogens of NTM, and the second one was M. abscessus. However, M. chelonei, M. fuerth, and M. gordon were rare. Clarithromycin had a good inhibition activity against all the NTM species except M. abscessus subsp. abscessus. The erm(41) genotype is of high relevance to clarithromycin resistance. PMID:26146620

  3. Species Identification and Clarithromycin Susceptibility Testing of 278 Clinical Nontuberculosis Mycobacteria Isolates

    PubMed Central

    Nie, Wenjuan; Duan, Hongfei; Huang, Hairong; Lu, Yu; Chu, Naihui

    2015-01-01

    Purpose of this paper is to analyze different species' proportion of nontuberculosis mycobacteria (NTM) and susceptibility to clarithromycin of different species. 278 clinical NTM isolates were identified into species by using 16S rRNA, rpoB and hsp65. Then clarithromycin susceptibility testing against different species was done separately, using microplate Alamar Blue assay. Finally, resistance isolates' erm(41) of M. abscessus were sequenced in order to analyze mechanisms for clarithromycin resistant. In this test, 131 isolates (47%) belonged to M. avium complex (MAC), and 70 isolates (25%) belonged to M. abscessus. Nearly all the M. abscessus subsp. abscessus resistant to clarithromycin had T28 in erm(41). However, all the M. abscessus subsp. abscessus susceptible to clarithromycin had C28 in erm(41). In this study, we find that MAC was the most common pathogens of NTM, and the second one was M. abscessus. However, M. chelonei, M. fuerth, and M. gordon were rare. Clarithromycin had a good inhibition activity against all the NTM species except M. abscessus subsp. abscessus. The erm(41) genotype is of high relevance to clarithromycin resistance. PMID:26146620

  4. Electronic Resource Management 2.0: Using Web 2.0 Technologies as Cost-Effective Alternatives to an Electronic Resource Management System

    ERIC Educational Resources Information Center

    Murray, Adam

    2008-01-01

    Designed to assist with the management of e-resources, electronic resource management (ERM) systems are time- and fund-consuming to purchase and maintain. Questions of system compatibility, data population, and workflow design/redesign can be difficult to answer; sometimes those answers are not what we'd prefer to hear. The two primary functions…

  5. Correlation of agar dilution and VITEK2 system for detection of resistance to macrolides, lincosamides and pristinamycin among Staphylococcus aureus and Staphylococcus epidermidis: association with genotypes.

    PubMed

    Bémer, P; Juvin, M-E; Corvec, S; Ros, A; Drugeon, H

    2005-08-01

    The performance of the VITEK2 system was evaluated against the agar dilution reference procedure for testing susceptibility of Staphylococcus aureus and Staphylococcus epidermidis to macrolides, lincosamides and streptogramins (MLS). Eighty clinical isolates were selected according to their resistance phenotype and genotype. Results for erythromycin and clindamycin showed 100% agreement; results for lincomycin showed agreement of 78%, with one very major error and 17 minor errors; and results for pristinamycin showed agreement of 46%, with one major error and 43 minor errors. Most isolates resistant to lincomycin and streptogramin A (L SgAr phenotype) were falsely susceptible to lincomycin, and intermediately-resistant or resistant to pristinamycin, with the VITEK2 system. No resistance gene was detected. Most (80%) isolates resistant constitutively to MLS (MLS(r)BC phenotype) were falsely intermediately-resistant to pristinamycin with the VITEK2 system. The erm(A) gene was more common than erm(C) in MLS(r)BC strains. Resistance to pristinamycin alone (SgA SgB PTr phenotype), or associated with either lincomycin resistance (L SgA SgB PTr phenotype) or constitutive MLS(B) resistance (MLS(BC) SgA PTr phenotype), was well-characterised without discordant results. Resistance to pristinamycin was always associated with resistance to streptogramin A, encoded by the vga(A), vga(B), vgb(A) and vat(A) genes in association with the erm(A) or erm(C) genes. PMID:16008619

  6. Yersinia ruckeri lipopolysaccharide is necessary and sufficient for eliciting a protective immune response in rainbow trout (Oncorhynchus mykiss, Walbaum)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Enteric redmouth disease (ERM), caused by Yersinia ruckeri, has been controlled successfully using immersion-applied bacterin vaccines for several decades. While the host response to vaccination and the mechanism of protection of this vaccine have been elucidated, the bacterial components eliciting ...

  7. Restoration of retinal layers after epiretinal membrane peeling

    PubMed Central

    Hartmann, K. I.; Schuster, A. K.; Bartsch, D.-U.; Kim, J. S.; Chhablani, J.; Freeman, W. R.

    2014-01-01

    Purpose To evaluate the morphological restoration of retinal anatomy after surgery for epiretinal membrane (ERM) peeling using spectral domain optical coherence tomography (SD-OCT). Correlation of retinal structure with visual outcome. Design Retrospective consecutive case series Methods 34 consecutive eyes with epiretinal membrane underwent surgery with one year follow up examination. Spectral domain optical coherence tomography (SD-OCT) scans were analyzed pre-operatively and 1 3, 6, 9 and 12 months post-operative. Best-corrected visual acuity (BCVA) using ETDRS charts was measured at each visit. Results All eyes showed a significant improvement of BCVA after ERM-peeling (p=0.002). The timepoint of BCVA and retinal restoration seen on SD-OCT occurred simultaneously and varied between individuals (occurrence of BCVA: mean 4.82 months; retinal restoration: mean 4.24 months). At 3-months, the retinal anatomic restoration rate was 70% and 88% at 6-months. Conclusion Restoration of the retinal anatomic structure predominantly occurs within the first three months post ERM peeling. An improvement of BCVA and anatomic retinal restoration after ERM-removal varies in individuals. If retinal layers fully restore in their anatomic structure, BCVA improves at the same time-point. PMID:24077089

  8. Immunolocalization of calbindin D28k and vitamin D receptor during root formation of murine molar teeth.

    PubMed

    Onishi, Tomoyuki; Okawa, Rena; Murakami, Hiroaki; Ogawa, Tomohiro; Ooshima, Takashi; Wakisaka, Satoshi

    2003-08-01

    Cells in the epithelial rest of Malassez (ERM cells) express calbindin D28k (CB); however, the hormonal regulation of CB in ERM cells remains to be elucidated. We investigated the immunohistochemical localization of CB and 1,25-dihydroxyvitamin D3 receptor (VDR) during root formation of mouse molar teeth in order to clarify whether the expression of CB in ERM cells is dependent on vitamin D. At the early stage of root formation (postnatal (PN) days 10-14), both CB- and VDR-immunoreactive cells were observed intermittently along the root surface. In the apical portion, almost all CB-immunoreactive cells showed VDR immunoreactivity; however, VDR-immunoreactive cells in the most apical portion were immunonegative for CB. In the middle and cervical portions, the distributions of the two proteins were completely different. At the late stage of root formation (PN28d) and in adult animals, CB immunoreactivity was distributed in cells found along the acellular cementum at the bifurcation region, as well as between the dentin and cellular cementum in the apical portion (although these lacked immunoreactivity for VDR). The present results indicate that CB expression in newly disrupted cells from Hertwig's epithelial root sheath occurs in a vitamin-D dependent manner, whereas the expression of CB in mature ERM cells may be independent of vitamin D. PMID:12845706

  9. Electronic Resource Management Systems in Practice

    ERIC Educational Resources Information Center

    Grogg, Jill E.

    2008-01-01

    Electronic resource management (ERM) systems have inundated the library marketplace. Both integrated library systems (ILS) vendors and subscription agents are now offering products and service enhancements that claim to help libraries efficiently manage their electronic resources. Additionally, some homegrown and open-source solutions have emerged…

  10. Combined Exact-Repeat and Geodetic Mission Altimetry for High-Resolution Empirical Tide Mapping

    NASA Astrophysics Data System (ADS)

    Zaron, E. D.

    2014-12-01

    The configuration of present and historical exact-repeat mission (ERM) altimeter ground tracks determines the maximum resolution of empirical tidal maps obtained with ERM data. Although the mode-1 baroclinic tide is resolvable at mid-latitudes in the open ocean, the ability to detect baroclinic and barotropic tides near islands and complex coastlines is limited, in part, by ERM track density. In order to obtain higher resolution maps, the possibility of combining ERM and geodetic mission (GM) altimetry is considered, using a combination of spatial thin-plate splines and temporal harmonic analysis. Given the present spatial and temporal distribution of GM missions, it is found that GM data can contribute to resolving tidal features smaller than 75 km, provided the signal amplitude is greater than about 1 cm. Uncertainties in the mean sea surface and environmental corrections are significant components of the GM error budget, and methods to optimize data selection and along-track filtering are still being optimized. Application to two regions, Monterey Bay and Luzon Strait, finds evidence for complex tidal fields in agreement with independent observations and modeling studies.

  11. Europäisches Organ der Festkörperforschung und DDR-Devisenbringer Die Zeitschrift Physica Status Solidi im Kalten Krieg

    NASA Astrophysics Data System (ADS)

    Hoffmann, Dieter

    Mit dem Ziele, ein einheitliches internationales Organ der Festkörperphysik für den europäischen Raum zu schaffen, das eine rasche Publikation der für das Festkörpergebiet repräsentative Arbeiten ermöglicht, wird durch ein internationales Herausgebergremium eine neue wissenschaftliche Zeitschrift gegründet.

  12. Isolation and Characterization of Human Adult Epithelial Stem Cells from the Periodontal Ligament.

    PubMed

    Athanassiou-Papaefthymiou, M; Papagerakis, P; Papagerakis, S

    2015-11-01

    We report a novel method for the isolation of adult human epithelial stem cells (hEpiSCs) from the epithelial component of the periodontal ligament-the human epithelial cell rests of Malassez (hERM). hEpiSC-rich integrin-α6(+ve) hERM cells derived by fluorometry can be clonally expanded, can grow organoids, and express the markers of pluripotency (OCT4, NANOG, SOX2), polycomb protein RING1B, and the hEpiSC supermarker LGR5. They maintain the growth profile of their originating hERM in vitro. Subcutaneous cotransplantation with mesenchymal stem cells from the dental pulp on poly-l-lactic acid scaffolds in nude mice gave rise to perfect heterotopic ossicles in vivo with ultrastructure of dentin, enamel, cementum, and bone. These remarkable fully mineralized ossicles underscore the importance of epithelial-mesenchymal crosstalk in tissue regeneration using human progenitor stem cells, which may have already committed to lineage despite maintaining hallmarks of pluripotency. In addition, we report the clonal expansion and isolation of human LGR5(+ve) cells from the hERM in xeno-free culture conditions. The genetic profile of LGR5(+ve) cells includes both markers of pluripotency and genes important for secretory epithelial and dental epithelial cell differentiation, giving us a first insight into periodontal ligament-derived hEpiSCs. PMID:26392003

  13. Multiple independent emergence of biotype 2 Yersinia ruckeri in the United States and Europe

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Biotype 2 (BT2) variants of the bacterium Yersinia ruckeri are an emerging disease problem in US and European salmonid aquaculture. The emergence of this biotype has been associated with an increased frequency of enteric redmouth disease (ERM) outbreaks in previously vaccinated salmonid fish. We use...

  14. Meeting the Challenges of Enterprise Risk Management in Higher Education

    ERIC Educational Resources Information Center

    Mattie, John

    2007-01-01

    In the summer of 2007, the Association of Governing Boards of Universities and Colleges (AGB) and the National Association of College and University Business Officers (NACUBO) held a summit on enterprise risk management (ERM) in Washington DC with senior officers and trustees from several leading colleges and universities. Approximately 40…

  15. In vitro activity of beta-lactams, macrolides, telithromycin, and fluoroquinolones against clinical isolates of Streptococcus pneumoniae: correlation between drug resistance and genetic characteristics.

    PubMed

    Yamaguchi, Toshiyuki; Hashikita, Giichi; Takahashi, Shun; Itabashi, Akira; Yamazaki, Tsutomu; Maesaki, Shigefumi

    2005-10-01

    The in vitro activity of antimicrobial agents against Streptococcus pneumoniae was determined using 16 strains of penicillin-susceptible S. pneumoniae (PSSP) and 26 strains of penicillin intermediately resistant S. pneumoniae (PISP) + penicillin-resistant S. pneumoniae (PRSP) in Japan. The minimum inhibitory concentrations (MICs) of potent antibiotics, including eight beta-lactams (benzylpenicillin, ampicillin, cefotiam, cefepime, cefditoren, faropenem, panipenem, and biapenem), three macrolides (erythromycin, clarithromycin, and azithromycin), telithromycin, and three fluoroquinolones (ciprofloxacin, levofloxacin, and gatifloxacin), were determined. Twenty-three strains exhibited genetic variations at pbp1a + pbp2x + pbp2b, which are genetic-PRSP (g-PRSP). g-PISP strains accounted for 62.5% (10/16) of the PSSP strains. The existence of an abnormal pbp gene conferred not only penicillin resistance but resistance to cephems; however, panipenem and biapenem had potent in vitro efficacy against alterations. Regarding the macrolide resistance mechanisms (mefA or ermB): 16 isolates had only mefA, 18 isolates had ermB, and 2 isolates had both mefA and ermB. There was no correlation between the existence of an abnormal pbp gene and the existence of the mefA gene or the ermB gene. PMID:16258826

  16. The prevalence of genotypes that determine resistance to macrolides, lincosamides, and streptogramins B compared with spiramycin susceptibility among erythromycin-resistant Staphylococcus epidermidis

    PubMed Central

    Juda, Marek; Chudzik-Rzad, Beata; Malm, Anna

    2016-01-01

    Coagulase-negative staphylococci, particularly Staphylococcus epidermidis, can be regarded as potential reservoirs of resistance genes for pathogenic strains, e.g., Staphylococcus aureus. The aim of this study was to assess the prevalence of different resistance phenotypes to macrolide, lincosamide, and streptogramins B (MLSB) antibiotics among erythromycin-resistant S. epidermidis, together with the evaluation of genes promoting the following different types of MLSB resistance:ermA, ermB, ermC,msrA, mphC, and linA/A’. Susceptibility to spiramycin was also examined. Among 75 erythromycin-resistantS. epidermidis isolates, the most frequent phenotypes were macrolides and streptogramins B (MSB) and constitutive MLSB (cMLSB). Moreover, all strains with the cMLSB phenotype and the majority of inducible MLSB (iMLSB) isolates were resistant to spiramycin, whereas strains with the MSB phenotype were sensitive to this antibiotic. The D-shape zone of inhibition around the clindamycin disc near the spiramycin disc was found for some spiramycin-resistant strains with the iMLSB phenotype, suggesting an induction of resistance to clindamycin by this 16-membered macrolide. The most frequently isolated gene was ermC, irrespective of the MLSB resistance phenotype, whereas the most often noted gene combination wasermC, mphC, linA/A’. The results obtained showed that the genes responsible for different mechanisms of MLSB resistance in S. epidermidis generally coexist, often without the phenotypic expression of each of them. PMID:27008373

  17. Expression, Localization, and Binding Activity of the Ezrin/Radixin/Moesin Proteins in the Mouse Testis

    PubMed Central

    Wakayama, Tomohiko; Nakata, Hiroki; Kurobo, Miho; Sai, Yoshimichi; Iseki, Shoichi

    2009-01-01

    The ezrin, radixin, and moesin (ERM) proteins represent a family of adaptor proteins linking transmembrane proteins to the cytoskeleton. The seminiferous epithelium undergoes extensive changes in cellular composition, location, and shape, implicating roles of the membrane–cytoskeleton interaction. It remains unknown, however, whether the ERM proteins are expressed and play significant roles in the testis. In the present study, we examined the spatiotemporal expression of ERM proteins in the mouse testis by Western blotting and immunohistochemistry. Ezrin immunoreactivity was demonstrated in the cytoplasm of steps 15 and 16 spermatids from 5 weeks postpartum through adulthood, whereas radixin immunoreactivity was in the apical cytoplasm of Sertoli cells from 1 week through 2 weeks postpartum. No immunoreactivity for moesin was detected at any age. Immunoprecipitation demonstrated that ezrin was bound to the cytoskeletal component actin, whereas radixin was bound to both actin and tubulin. Of the transmembrane proteins known to interact with ERM proteins, only cystic fibrosis transmembrane conductance regulator, a chloride transporter, was bound to ezrin in elongated spermatids. These results suggest that ezrin is involved in spermiogenesis whereas radixin is involved in the maturation of Sertoli cells, through interaction with different sets of membrane proteins and cytoskeletal components. (J Histochem Cytochem 57:351–362, 2009) PMID:19064715

  18. LKB1 Is Required for the Development and Maintenance of Stereocilia in Inner Ear Hair Cells in Mice

    PubMed Central

    Men, Yuqin; Zhang, Aizhen; Li, Haixiang; Zhang, Tingting; Jin, Yecheng; Li, Huashun

    2015-01-01

    The LKB1 gene, which encodes a serine/threonine kinase, was discovered to play crucial roles in cell differentiation, proliferation, and the establishment of cell polarity. In our study, LKB1 conditional knockout mice (Atoh1-LKB1-/- mice) were generated to investigate LKB1 function in the inner ear. Tests of auditory brainstem response and distortion product otoacoustic emissions revealed significant decreases in the hearing sensitivities of the Atoh1-LKB1-/- mice. In Atoh1-LKB1-/- mice, malformations of hair cell stereocilliary bundles were present as early as postnatal day 1 (P1), a time long before the maturation of the hair cell bundles. In addition, we also observed outer hair cell (OHC) loss starting at P14. The impaired stereocilliary bundles occurred long before the presence of hair cell loss. Stereociliary cytoskeletal structure depends on the core actin-based cytoskeleton and several actin-binding proteins. By Western blot, we examined actin-binding proteins, specifically ERM (ezrin/radixin/moesin) proteins involved in the regulation of the actin cytoskeleton of hair cell stereocilia. Our results revealed that the phosphorylation of ERM proteins (pERM) was significantly decreased in mutant mice. Thus, we propose that the decreased pERM may be a key factor for the impaired stereocillia function, and the damaged stereocillia may induce hair cell loss and hearing impairments. Taken together, our data indicates that LKB1 is required for the development and maintenance of stereocilia in the inner ear. PMID:26274331

  19. Flagella biosynthesis and regulation by the Rcs pathway within the fish pathogen Yersinia ruckeri during infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The gram-negative Enterobacterium Yersinia ruckeri is the etiologic agent of enteric redmouth disease (ERM) within farmed rainbow trout (Oncorhynchus mykiss, Walbaum). Over the past decade, there has been an increase in the prevalence of non-motile variants of Y. ruckeri and the appearance of these ...

  20. Fate of antimicrobials and antimicrobial resistance genes in simulated swine manure storage

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The behavior of three antibiotics (bacitracin, chlortetracycline, and tylosin) and two classes of antibiotic resistance genes (ARGs), tet and erm, were monitored in swine manure slurry under anaerobic conditions. First-order decay rates were determined for each antibiotic with half-lives ranging fr...

  1. Soil Identification using Field Electrical Resistivity Method

    NASA Astrophysics Data System (ADS)

    Hazreek, Z. A. M.; Rosli, S.; Chitral, W. D.; Fauziah, A.; Azhar, A. T. S.; Aziman, M.; Ismail, B.

    2015-06-01

    Geotechnical site investigation with particular reference to soil identification was important in civil engineering works since it reports the soil condition in order to relate the design and construction of the proposed works. In the past, electrical resistivity method (ERM) has widely being used in soil characterization but experienced several black boxes which related to its results and interpretations. Hence, this study performed a field electrical resistivity method (ERM) using ABEM SAS (4000) at two different types of soils (Gravelly SAND and Silty SAND) in order to discover the behavior of electrical resistivity values (ERV) with type of soils studied. Soil basic physical properties was determine thru density (p), moisture content (w) and particle size distribution (d) in order to verify the ERV obtained from each type of soil investigated. It was found that the ERV of Gravelly SAND (278 Ωm & 285 Ωm) was slightly higher than SiltySAND (223 Ωm & 199 Ωm) due to the uncertainties nature of soils. This finding has showed that the results obtained from ERM need to be interpreted based on strong supported findings such as using direct test from soil laboratory data. Furthermore, this study was able to prove that the ERM can be established as an alternative tool in soil identification provided it was being verified thru other relevance information such as using geotechnical properties.

  2. "Not" a One-Size-Fits-All Solution: Lessons Learned from Implementing an Electronic Resources Management System in Three Days

    ERIC Educational Resources Information Center

    Pan, Denise

    2009-01-01

    Auraria Library purchased Innovative Interfaces, Inc.'s Millennium Electronic Resources Management (ERM) to manage data about acquisitions, licensing, troubleshooting, and usage statistics of electronic resources. After 3 days of implementation, the software vendor enabled resources records to display. As a result, the electronic resources team…

  3. Checklist Manifesto for Electronic Resources: Getting Ready for the Fiscal Year and Beyond

    ERIC Educational Resources Information Center

    England, Lenore; Fu, Li; Miller, Stephen

    2011-01-01

    Organization of electronic resources workflow is critical in the increasingly complicated and complex world of library management. A simple organizational tool that can be readily applied to electronic resources management (ERM) is the use of checklists. Based on the principles discussed in The Checklist Manifesto: How to Get Things Right, the…

  4. Teaching Electronic Records Management in the Archival Curriculum

    ERIC Educational Resources Information Center

    Zhang, Jane

    2016-01-01

    Electronic records management has been incorporated into the archival curriculum in North America since the 1990s. This study reported in this paper provides a systematic analysis of the content of electronic records management (ERM) courses currently taught in archival education programs. Through the analysis of course combinations and their…

  5. Enteric Redmouth Disease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Yersinia ruckeri, the causative agent of Enteric Redmouth Disease (ERM), is a disease of salmonid fish species that is endemic in areas of the world where salmonids are intensively cultured. The disease causes a chronic to acute hemorrhagic septicemia which can lead to high rates of mortality partic...

  6. Temporal and pathogen-load dependent changes in rainbow trout (Oncorhynchus mykiss) immune response traits following challenge with biotype 2 Yersinia ruckeri

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rainbow trout infected with Yersinia ruckeri, the causative agent of enteric redmouth disease (ERM), produce a pro-inflammatory and acute-phase response attributed in part to the innate recognition of bacterial-produced flagellin. Recently, variants of Yersinia ruckeri have been identified that lac...

  7. Linear Logistic Test Modeling with R

    ERIC Educational Resources Information Center

    Baghaei, Purya; Kubinger, Klaus D.

    2015-01-01

    The present paper gives a general introduction to the linear logistic test model (Fischer, 1973), an extension of the Rasch model with linear constraints on item parameters, along with eRm (an R package to estimate different types of Rasch models; Mair, Hatzinger, & Mair, 2014) functions to estimate the model and interpret its parameters. The…

  8. Streptococcus pneumoniae Isolates Resistant to Telithromycin

    PubMed Central

    Rantala, M.; Haanperä-Heikkinen, M.; Lindgren, M.; Seppälä, H.; Huovinen, P.; Jalava, J.

    2006-01-01

    The telithromycin susceptibility of 210 erythromycin-resistant pneumococci was tested with the agar diffusion method. Twenty-six erm(B)-positive isolates showed heterogeneous resistance to telithromycin, which was manifested by the presence of colonies inside the inhibition zone. When these cells were cultured and tested, they showed stable, homogeneous, and high-level resistance to telithromycin. PMID:16641460

  9. The prevalence of genotypes that determine resistance to macrolides, lincosamides, and streptogramins B compared with spiramycin susceptibility among erythromycin-resistant Staphylococcus epidermidis.

    PubMed

    Juda, Marek; Chudzik-Rzad, Beata; Malm, Anna

    2016-03-01

    Coagulase-negative staphylococci, particularly Staphylococcus epidermidis, can be regarded as potential reservoirs of resistance genes for pathogenic strains, e.g., Staphylococcus aureus. The aim of this study was to assess the prevalence of different resistance phenotypes to macrolide, lincosamide, and streptogramins B (MLSB) antibiotics among erythromycin-resistant S. epidermidis, together with the evaluation of genes promoting the following different types of MLSB resistance:ermA, ermB, ermC,msrA, mphC, and linA/A'. Susceptibility to spiramycin was also examined. Among 75 erythromycin-resistantS. epidermidis isolates, the most frequent phenotypes were macrolides and streptogramins B (MSB) and constitutive MLSB (cMLSB). Moreover, all strains with the cMLSB phenotype and the majority of inducible MLSB (iMLSB) isolates were resistant to spiramycin, whereas strains with the MSB phenotype were sensitive to this antibiotic. The D-shape zone of inhibition around the clindamycin disc near the spiramycin disc was found for some spiramycin-resistant strains with the iMLSB phenotype, suggesting an induction of resistance to clindamycin by this 16-membered macrolide. The most frequently isolated gene was ermC, irrespective of the MLSB resistance phenotype, whereas the most often noted gene combination wasermC, mphC, linA/A'. The results obtained showed that the genes responsible for different mechanisms of MLSB resistance in S. epidermidis generally coexist, often without the phenotypic expression of each of them. PMID:27008373

  10. Somewhere over the Verde Rainbow

    ERIC Educational Resources Information Center

    Ekart, Donna F.

    2008-01-01

    When the electronic resource management system (ERM) at Kansas State University Libraries suffered a horrible data loss, the "contract db" presented a challenge for the librarians responsible for electronic resources. It was a decent data repository, but it had no ability to manage the tangled process of licensing, acquiring, activating, using,…

  11. 46 CFR 11.325 - Requirements to qualify for an STCW endorsement as chief engineer officer on vessels powered by...

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...) Engineroom resource management (ERM) if not completed at the operational level. (ii) Leadership and... at the operational level. (2) Leadership and managerial skills. (3) Management of electrical and... more (management level). 11.325 Section 11.325 Shipping COAST GUARD, DEPARTMENT OF HOMELAND...

  12. Characterisation of geographically and temporally diverse Yersinia ruckeri isolates: evidence that UK and mainland European biotype 2 isolates represent different clonal groups

    Technology Transfer Automated Retrieval System (TEKTRAN)

    There have been increased reports of outbreaks of Enteric Redmouth Disease (ERM) caused by Yersinia ruckeri in previously-vaccinated salmonids in Europe, with some of these outbreaks attributed to emergent non-motile, Tween 80 negative, biotype 2 isolates. To gain information about their likely orig...

  13. Draft Genome Sequences of the Two Unrelated Macrolide-Resistant Corynebacterium argentoratense Strains CNM 463/05 and CNM 601/08, Isolated from Patients in the University Hospital of León, Spain

    PubMed Central

    Soriano, Francisco; Acedo, Alberto; Hernandez, Marta; Tauch, Andreas

    2015-01-01

    Corynebacterium argentoratense has been associated mainly with infections in the human respiratory tract. Genome sequencing of two unrelated clinical macrolide-resistant strains, CNM 463/05 and CNM 601/08, revealed the presence of the antibiotic resistance gene erm(X) allocated to a specific genomic region with 100% similarity to the widely distributed transposable element Tn5432. PMID:26159536

  14. Fate of antimicrobials and antimicrobial resistance genes in simulated swine manure storage.

    PubMed

    Joy, Stacey R; Li, Xu; Snow, Daniel D; Gilley, John E; Woodbury, Bryan; Bartelt-Hunt, Shannon L

    2014-05-15

    The behavior of three antibiotics (bacitracin, chlortetracycline, and tylosin) and two classes of antibiotic resistance genes (ARGs), tet and erm, were monitored in swine manure slurry under anaerobic conditions. First-order decay rates were determined for each antibiotic with half-lives ranging from 1 day (chlortetracycline) to 10 days (tylosin). ARGs were monitored in the swine manure slurry, and losses of approximately 1 to 3 orders of magnitude in relative abundance were observed during the 40 day storage period. First-order degradation profiles were observed for chlortetracycline and its corresponding resistance genes, tet(X) and tet(Q). Tylosin was degraded to approximately 10% of the starting concentration by day 40; however, the relative abundance of erm(B) remained at 50-60% of the initial relative abundance while the relative abundance of erm(F) decreased by 80-90%, consistent with tylosin. These results indicate that tet resistance genes respond primarily to chlortetracycline antimicrobials, and may be lost when the parent tetracycline compound is degraded. In contrast, erm(B) resistance gene may respond to a range of antimicrobials in animal manure, and may persist despite losses of tylosin. PMID:24583946

  15. Community Environmental Response Facilitation Act (CERFA) report. New Orleans Military Ocean Terminal (NOMOT), New Orleans, LA. Final report

    SciTech Connect

    Crossman, M.; Ward, L.

    1994-04-11

    This report presents the results of the Community Environmental Response Facilitation Act (CERFA) investigation conducted by Environmental Resources Management (ERM) at New Orleans Military Ocean Terminal (NOMOT), a U.S. Government property selected for closure by the Base Realignment and Closure (BRAC) Commission under Public Laws 100-526 and 101-510. Under CERFA (Public Law 102-426), Federal agencies are required to identify expeditiously real property that can be immediately reused and redeveloped. Satisfying this objective requires the identification of real property where no hazardous substances or petroleum products, regulated by the Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA), were stored for one year or more, known to have been released, or disposed NOMOT is a 17.6-acre site located in New Orleans, Louisiana. NOMOT has been used for warehousing and shipping of equipment since 1919. Environmentally significant operations include routine maintenance and hazardous material handling. ERM reviewed existing investigation documents; U.S. Environmental Protection Agency (EPA) , State, and county regulatory records; environmental data bases; and title documents pertaining to NOMOT during this investigation. In addition, ERM conducted interviews and visual inspections of NOMOT as well as visual inspections of and data base searches for the surrounding properties. Information in this CERFA report was current as of the site visit by ERM in October 1993. This information was used to divide the installation into two categories of parcels: CERFA Disqualified Parcels and CERFA Parcels, as defined by the Army. New Orleans military ocean terminal, CERF.

  16. Quantitation of Bacillus clausii in biological samples by real-time polymerase chain reaction.

    PubMed

    Perotti, Mario; Mancini, Nicasio; Cavallero, Annalisa; Carletti, Silvia; Canducci, Filippo; Burioni, Roberto; Clementi, Massimo

    2006-06-01

    A real-time PCR assay targeting the highly specific erm34 sequence of Bacillus clausii DNA was developed and optimized. The quantitative assay showed a sensitivity level of 10(2) CFU/microl of sample. The method may represent a useful tool for monitoring the role of B. clausii as probiotic in vivo. PMID:16318892

  17. Serotypes, Clones, and Mechanisms of Resistance of Erythromycin-Resistant Streptococcus pneumoniae Isolates Collected in Spain▿

    PubMed Central

    Calatayud, Laura; Ardanuy, C.; Cercenado, E.; Fenoll, A.; Bouza, E.; Pallares, R.; Martín, R.; Liñares, J.

    2007-01-01

    The aim of this study was to analyze the distributions of antibiotic susceptibility patterns, serotypes, phenotypes, genotypes, and macrolide resistance genes among 125 nonduplicated erythromycin-resistant Streptococcus pneumoniae clinical isolates collected in a Spanish point prevalence study. The prevalence of resistance to macrolides in this study was 34.7%. Multiresistance (to three or more antimicrobials) was observed in 81.6% of these strains. Among 15 antimicrobials studied, cefotaxime, moxifloxacin, telithromycin, and quinupristin-dalfopristin were the most active drugs. The most frequent serotypes of erythromycin-resistant isolates were 19F (25%), 19A (17%), 6B (12%), 14 (10%), and 23F (10%). Of the 125 strains, 109 (87.2%) showed the MLSB phenotype [103 had the erm(B) gene and 6 had both erm(B) and mef(E) genes]. Sixteen (12.8%) strains showed the M phenotype [14 with mef(E) and 2 with mef(A)]. All isolates were tested by PCR for the presence of the int, xis, tnpR, and tnpA genes associated with conjugative transposons (Tn916 family and Tn917). Positive detection of erm(B), tet(M), int, and xis genes related to the Tn916 family was found in 77.1% of MLSB phenotype strains. In 16 strains, only the tndX, erm(B), and tet(M) genes were detected, suggesting the presence of Tn1116, a transposon recently described for Streptococcus pyogenes. Five clones, namely, Sweden15A-25, clone19F ST87, Spain23F-1, Spain6B-2, and clone19A ST276, accounted for half of the MLSB strains. In conclusion, the majority of erythromycin-resistant pneumococci isolated in Spain had the MLSB phenotype, belonged to multiresistant international clones, and carried the erm(B), tet(M), xis, and int genes, suggesting the spread of transposons of the Tn916 family. PMID:17606677

  18. Genetic heterogeneity in rhabdomyosarcoma revealed by SNP array analysis.

    PubMed

    Walther, Charles; Mayrhofer, Markus; Nilsson, Jenny; Hofvander, Jakob; Jonson, Tord; Mandahl, Nils; Øra, Ingrid; Gisselsson, David; Mertens, Fredrik

    2016-01-01

    Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in children and adolescents. Alveolar (ARMS) and embryonal (ERMS) histologies predominate, but rare cases are classified as spindle cell/sclerosing (SRMS). For treatment stratification, RMS is further subclassified as fusion-positive (FP-RMS) or fusion-negative (FN-RMS), depending on whether a gene fusion involving PAX3 or PAX7 is present or not. We investigated 19 cases of pediatric RMS using high resolution single-nucleotide polymorphism (SNP) array. FP-ARMS displayed, on average, more structural rearrangements than ERMS; the single FN-ARMS had a genomic profile similar to ERMS. Apart from previously known amplification (e.g., MYCN, CDK4, and MIR17HG) and deletion (e.g., NF1, CDKN2A, and CDKN2B) targets, amplification of ERBB2 and homozygous loss of ASCC3 or ODZ3 were seen. Combining SNP array with cytogenetic data revealed that most cases were polyploid, with at least one case having started as a near-haploid tumor. Further bioinformatic analysis of the SNP array data disclosed genetic heterogeneity, in the form of subclonal chromosomal imbalances, in five tumors. The outcome was worse for patients with FP-ARMS than ERMS or FN-ARMS (6/8 vs. 1/9 dead of disease), and the only children with ERMS showing intratumor diversity or with MYOD1 mutation-positive SRMS also died of disease. High resolution SNP array can be useful in evaluating genomic imbalances in pediatric RMS. PMID:26482321

  19. Intraocular Lens Power Estimation in Combined Phacoemulsification and Pars Plana Vitrectomy in Eyes with Epiretinal Membranes: A Case-Control Study

    PubMed Central

    Kim, Min; Kim, Hyoung Eun; Lee, Dong Hyun; Koh, Hyoung Jun; Lee, Sung Chul

    2015-01-01

    Purpose To evaluate the accuracy of postoperative refractive outcomes of combined phacovitrectomy for epiretinal membrane (ERM) in comparison to cataract surgery alone. Materials and Methods Thirty-nine eyes that underwent combined phacovitrectomy with intraocular lens (IOL) implantation for cataract and ERM (combined surgery group) and 39 eyes that received phacoemulsification for cataract (control group) were analyzed, retrospectively. The predicted preoperative refractive aim was compared with the results of postoperative refraction. Results In the combined surgery group, refractive prediction error by A-scan and IOLMaster were -0.305±0.717 diopters (D) and -0.356±0.639 D, respectively, compared to 0.215±0.541 and 0.077±0.529 in the control group, showing significantly more myopic change compared to the control group (p=0.001 and p=0.002, respectively). Within each group, there was no statistically significant difference in refractive prediction error between A-scan and IOLMaster (all p>0.05). IOL power calculation using adjusted A-scan measurement of axial length based on the macular thickness of the normal contralateral eye still resulted in significant postoperative refractive error (all p<0.05). Postoperative refraction calculated with adjusted axial length based on actual postoperative central foveal thickness change showed the closest value to the actual postoperative achieved refraction (p=0.599). Conclusion Combined phacovitrectomy for ERM resulted in significantly more myopic shift of postoperative refraction, compared to the cataract surgery alone, for both A-scan and IOLMaster. To improve the accuracy of IOL power estimation in eyes with cataract and ERM, sequential surgery for ERM and cataract may need to be considered. PMID:25837189

  20. Estrogen response element and the promoter context of the human and mouse lactoferrin genes influence estrogen receptor alpha-mediated transactivation activity in mammary gland cells.

    PubMed

    Stokes, Kenya; Alston-Mills, Brenda; Teng, Christina

    2004-10-01

    A critical step in estrogen action is the recognition of estrogen responsive elements (EREs) by liganded estrogen receptor. Our current studies were designed to determine whether an extended estrogen response element half-site (ERRE) contributes to the differential estrogen responses of the human and mouse lactoferrin overlapping chicken ovalbumin upstream promoter/ERE sequences (estrogen response modules, ERMs) in the context of their natural promoters. Transient transfections of MCF-7 cells show that liganded estrogen receptor alpha (ERalpha) activates transcription of the human lactoferrin ERM fourfold higher than the mouse lactoferrin ERM in the context of their natural promoters. Since the ERRE of the human lactoferrin gene naturally occurs 18 bp upstream from the ERM and is absent in the mouse lactoferrin gene promoter, we created a chimeric mouse lactoferrin CAT reporter, which now encodes the ERRE in the identical location as in the human lactoferrin gene. The addition of the ERRE in the mouse lactoferrin gene rendered this reporter extremely responsive to estrogen stimulation. Using limited protease digestions and electrophoretic mobility shift assays, we showed that the binding and protease sensitivity of ERalpha bound to the mouse ERM with or without the ERRE, differed. Importantly, occupancy of additional nuclear receptors at the ERRE may contribute to ERalpha binding and activation. Furthermore, the presence of ERRE influences the selectivity of coactivators in liganded ERalpha-mediated transcriptional activity. When the receptor is bound to human and mouse plus genes, which contain the ERRE, steroid receptor coactivator (SRC)-2 was preferred, while SRC-1 and SRC-3 coactivators selectively enhanced the mouse lactoferrin gene activity. Moreover, peroxisome proliferator activated receptor-gamma coactivator-1 (PGC-1alpha) and PGC-1-related estrogen receptor coactivator (PERC) robustly increase the transcriptional function of ERalpha in the presence of the

  1. Different levels of hyphal self-incompatibility modulate interconnectedness of mycorrhizal networks in three arbuscular mycorrhizal fungi within the Glomeraceae.

    PubMed

    Pepe, Alessandra; Giovannetti, Manuela; Sbrana, Cristiana

    2016-05-01

    Arbuscular mycorrhizal fungi (AMF) live in symbiosis with most plant species and produce underground extraradical hyphal networks functional in the uptake and translocation of mineral nutrients from the soil to host plants. This work investigated whether fungal genotype can affect patterns of interconnections and structural traits of extraradical mycelium (ERM), by comparing three Glomeraceae species growing in symbiosis with five plant hosts. An isolate of Funneliformis coronatus consistently showed low ability to form interconnected ERM and self-incompatibility that represented up to 21 % of hyphal contacts. The frequency of post-fusion self-incompatible interactions, never detected before in AMF extraradical networks, was 8.9 %. In F. coronatus ERM, the percentage of hyphal contacts leading to perfect hyphal fusions was 1.2-7.7, while it ranged from 25.8-48 to 35.6-53.6 in Rhizophagus intraradices and Funneliformis mosseae, respectively. Low interconnectedness of F. coronatus ERM resulted also from a very high number of non-interacting contacts (83.2 %). Such findings show that AMF genotypes in Glomeraceae can differ significantly in anastomosis behaviour and that ERM interconnectedness is modulated by the fungal symbiont, as F. coronatus consistently formed poorly interconnected networks when growing in symbiosis with five different host plants and in the asymbiotic stage. Structural traits, such as extent, density and hyphal self-compatibility/incompatibility, may represent key factors for the differential performance of AMF, by affecting fungal absorbing surface and foraging ability and thus nutrient flow from soil to host roots. PMID:26630971

  2. Accuracy of the GEM-T2 geopotential from Geosat and ERS 1 crossover altimetry

    NASA Astrophysics Data System (ADS)

    Wagner, C. A.; Klokočník, J.

    1994-05-01

    Extensive analyses of altimetrically determined sea height differences at crossovers have been used to assess the accuracy of the GEM-T2 geopotential. The orbits used were determined with GEM-T2 for Geosat in its 17-day Exact Repeat Mission (ERM) in 1986-1989 and ERS 1 in both its 3-day ERM in 1991-1992 and its 35-day ERM in 1992. The data examined are completely independent of the data used in GEM-T2's development though GEM-T2 had considerable use of Doppler tracking information on Geosat. The test of the radial accuracy of the ERS 1 orbit (98.5° inclination) is especially significant because it is not ``close'' to any other orbit well represented in GEM-T2. The assessment consists of a comparison of observed mean height differences at thousands of distinct geographic locations with error projections from the GEM-T2 covariance matrix which was estimated from other data sources. This first comprehensive, independent test of the purely radial accuracy of an orbit-geopotential model clearly shows that the covariant predictions for GEM-T2 are broadly reliable for this purpose. Thus, the agreement of crossover predictions and observations suggests that the total radial errors for these ERMs, due to only to GEM-T2 (but excluding the effects of initial state error) are about 23 cm for Geosat and 115 cm (rms) for ERS 1. However, there is little detailed agreement of measurements and predictions for ERS 1 and only partial agreement in detail for Geosat. Our 30,000 mean crossover discrepancies for Geosat (derived from ERM cycles 1-44) are also shown to reduce substantially the crossover height differences in cycles 45-61, almost exactly as predicted if these are the true GEM-T2 errors for this orbit.

  3. Simultaneous observation of VHF radio wave transmission anomaly propagated beyond line of site prior to earthquakes in multiple sites

    NASA Astrophysics Data System (ADS)

    Yamashita, H.; Mogi, T.; Moriya, T.; Takada, M.; Morisada, M.

    2010-12-01

    The VHF radio wave transmission anomalies propagated beyond line of site prior to earthquakes (M>4), (hereafter termed EQ-echo) have been observed more than 20 times from 2004 at the Erimo observatory (ERM) in Hokkaido, Northern Japan. A statistical relationship between magnitude of preceding earthquake and total duration time of the EQ-echo has been proposed (Moriya et al.2009). To confirm a region where the EQ-echo simultaneously observed for each earthquake, we installed another 3 observatory with approximately 5 km spacing in the surroundings of ERM. The EQ-echoes have been observed simultaneously at two observatories prior to four earthquakes since 2008. The initial time and duration of each EQ echo were same time in several cases but different at some minutes each other in other cases. The wave forms of the EQ-echoes were similar in both records. In the Fuyushima observatory (FYS, 10km away from ERM) , three-way antennas were installed at every 120 degree to detect an arrival direction of EQ-echoes. Simultaneous observations of EQ-echoes at ERM and FYS for the preceding EQ (M=4.7) that occurred in the Hidaka mountains revealed that this EQ-echo came from direction of the epicenter based on the FYS observation and this direction was consistent with that of EQ-echo observed simultaneously in ERM. Although some of simultaneous observed EQ-echoes were observed in same time completely at both observatories, but some of them were with time rag of duration of each EQ-echo between multiple observed sites. We discussed what these time rags mean by considering possibilities of moving of scattering objects, generation of a radio duct, and so on, as in response to this fact.

  4. Abundance and distribution of Macrolide-Lincosamide-Streptogramin resistance genes in an anaerobic-aerobic system treating spiramycin production wastewater.

    PubMed

    Liu, Miaomiao; Ding, Ran; Zhang, Yu; Gao, Yingxin; Tian, Zhe; Zhang, Tong; Yang, Min

    2014-10-15

    The behaviors of the Macrolide-Lincosamide-Streptogramin (MLS) resistance genes were investigated in an anaerobic-aerobic pilot-scale system treating spiramycin (SPM) production wastewater. After screening fifteen typical MLS resistance genes with different mechanisms using conventional PCR, eight detected genes were determined by quantitative PCR, together with three mobile elements. Aerobic sludge in the pilot system exhibited a total relative abundance of MLS resistance genes (per 16S rRNA gene) 2.5 logs higher than those in control samples collected from sewage and inosine wastewater treatment systems (P < 0.05), implying the presence of SPM could induce the production of MLS resistance genes. However, the total relative gene abundance in anaerobic sludge (4.3 × 10(-1)) was lower than that in aerobic sludge (3.7 × 10(0)) despite of the higher SPM level in anaerobic reactor, showing the advantage of anaerobic treatment in reducing the production of MLS resistance genes. The rRNA methylase genes (erm(B), erm(F), erm(X)) were the most abundant in the aerobic sludge (5.3 × 10(-1)-1.7 × 10(0)), followed by esterase gene ere(A) (1.3 × 10(-1)) and phosphorylase gene mph(B) (5.7 × 10(-2)). In anaerobic sludge, erm(B), erm(F), ere(A), and msr(D) were the major ones (1.2 × 10(-2)-3.2 × 10(-1)). These MLS resistance genes (except for msr(D)) were positively correlated with Class 1 integron (r(2) = 0.74-0.93, P < 0.05), implying the significance of horizontal transfer in their proliferation. PMID:24973730

  5. Intracellular sphingosine kinase 2-derived sphingosine-1-phosphate mediates epidermal growth factor-induced ezrin-radixin-moesin phosphorylation and cancer cell invasion.

    PubMed

    Adada, Mohamad M; Canals, Daniel; Jeong, Nara; Kelkar, Ashwin D; Hernandez-Corbacho, Maria; Pulkoski-Gross, Michael J; Donaldson, Jane C; Hannun, Yusuf A; Obeid, Lina M

    2015-11-01

    The bioactive sphingolipid sphingosine-1-phosphate (S1P) mediates cellular proliferation, mitogenesis, inflammation, and angiogenesis. These biologies are mediated through S1P binding to specific GPCRs [sphingosine-1-phosphate receptor (S1PR)1-5] and some other less well-characterized intracellular targets. Ezrin-radixin-moesin (ERM) proteins, a family of adaptor molecules linking the cortical actin cytoskeleton to the plasma membrane, are emerging as critical regulators of cancer invasion via regulation of cell morphology and motility. Recently, we identified S1P as an acute ERM activator (via phosphorylation) through its action on S1PR2. In this work, we dissect the mechanism of S1P generation downstream of epidermal growth factor (EGF) leading to ERM phosphorylation and cancer invasion. Using pharmacologic inhibitors, small interfering RNA technologies, and genetic approaches, we demonstrate that sphingosine kinase (SK)2, and not SK1, is essential and sufficient in EGF-mediated ERM phosphorylation in HeLa cells. In fact, knocking down SK2 decreased ERM activation 2.5-fold. Furthermore, we provide evidence that SK2 is necessary to mediate EGF-induced invasion. In addition, overexpressing SK2 causes a 2-fold increase in HeLa cell invasion. Surprisingly, and for the first time, we find that this event, although dependent on S1PR2 activation, does not generate and does not require extracellular S1P secretion, therefore introducing a potential novel model of autocrine/intracrine action of S1P that still involves its GPCRs. These results define new mechanistic insights for EGF-mediated invasion and novel actions of SK2, therefore setting the stage for novel targets in the treatment of growth factor-driven malignancies. PMID:26209696

  6. Antimicrobial Resistance and Molecular Characteristics of Nasal Staphylococcus aureus Isolates From Newly Admitted Inpatients.

    PubMed

    Chen, Xu; Sun, Kangde; Dong, Danfeng; Luo, Qingqiong; Peng, Yibing; Chen, Fuxiang

    2016-05-01

    Staphylococcus aureus, or methicillin-resistant S. aureus (MRSA), is a significant pathogen in both nosocomial and community infections. Community-associated MRSA (CA-MRSA) strains tend to be multi-drug resistant and to invade hospital settings. This study aimed to assess the antimicrobial resistance and molecular characteristicsof nasal S. aureus among newlyadmitted inpatients.In the present study, 66 S. aureus isolates, including 10 healthcare-associated MRSA (HA-MRSA), 8 CA-MRSA, and 48 methicillin-sensitive S. aureus (MSSA) strains, were found in the nasal cavities of 62 patients by screening 292 newlyadmitted patients. Antimicrobial resistance and molecular characteristics of these isolates, including spa-type, sequence type (ST) and SCCmec type, were investigated. All isolates were sensitive to linezolid, teicoplanin, and quinupristin/dalfopristin, but high levels of resistance to penicillin and erythromycin were detected. According to D-test and erm gene detection results, the cMLS(B) and iMLS(B) phenotypes were detected in 24 and 16 isolates, respectively. All 10 HA-MRSA strains displayed the cMLS(B) phenotypemediated by ermA or ermA/ermC, while the cMLS(B) CA-MRSA and MSSA strains carried the ermB gene. Molecular characterization revealedall 10 HA-MRSA strains were derived from the ST239-SCCmec III clone, and four out of eight CA-MRSA strains were t437-ST59-SCCmec V. The results suggest that patients play an indispensable role in transmitting epidemic CA-MRSA and HA-MRSA strains. PMID:26915614

  7. Antimicrobial Resistance and Molecular Characteristics of Nasal Staphylococcus aureus Isolates From Newly Admitted Inpatients

    PubMed Central

    Chen, Xu; Sun, Kangde; Luo, Qingqiong; Peng, Yibing

    2016-01-01

    Staphylococcus aureus, or methicillin-resistant S. aureus (MRSA), is a significant pathogen in both nosocomial and community infections. Community-associated MRSA (CA-MRSA) strains tend to be multi-drug resistant and to invade hospital settings. This study aimed to assess the antimicrobial resistance and molecular characteristicsof nasal S. aureus among newlyadmitted inpatients.In the present study, 66 S. aureus isolates, including 10 healthcare-associated MRSA (HA-MRSA), 8 CA-MRSA, and 48 methicillin-sensitive S. aureus (MSSA) strains, were found in the nasal cavities of 62 patients by screening 292 newlyadmitted patients. Antimicrobial resistance and molecular characteristics of these isolates, including spa-type, sequence type (ST) and SCCmec type, were investigated. All isolates were sensitive to linezolid, teicoplanin, and quinupristin/dalfopristin, but high levels of resistance to penicillin and erythromycin were detected. According to D-test and erm gene detection results, the cMLSB and iMLSB phenotypes were detected in 24 and 16 isolates, respectively. All 10 HA-MRSA strains displayed the cMLSB phenotypemediated by ermA or ermA/ermC, while the cMLSB CA-MRSA and MSSA strains carried the ermB gene. Molecular characterization revealedall 10 HA-MRSA strains were derived from the ST239-SCCmec III clone, and four out of eight CA-MRSA strains were t437-ST59-SCCmec V. The results suggest that patients play an indispensable role in transmitting epidemic CA-MRSA and HA-MRSA strains. PMID:26915614

  8. The potential of Dark Septate Endophytes to form root symbioses with ectomycorrhizal and ericoid mycorrhizal middle European forest plants.

    PubMed

    Lukešová, Tereza; Kohout, Petr; Větrovský, Tomáš; Vohník, Martin

    2015-01-01

    The unresolved ecophysiological significance of Dark Septate Endophytes (DSE) may be in part due to existence of morphologically indistinguishable cryptic species in the most common Phialocephala fortinii s. l.--Acephala applanata species complex (PAC). We inoculated three middle European forest plants (European blueberry, Norway spruce and silver birch) with 16 strains of eight PAC cryptic species and other DSE and ectomycorrhizal/ericoid mycorrhizal fungi and focused on intraradical structures possibly representing interfaces for plant-fungus nutrient transfer and on host growth response. The PAC species Acephala applanata simultaneously formed structures resembling ericoid mycorrhiza (ErM) and DSE microsclerotia in blueberry. A. macrosclerotiorum, a close relative to PAC, formed ectomycorrhizae with spruce but not with birch, and structures resembling ErM in blueberry. Phialocephala glacialis, another close relative to PAC, formed structures resembling ErM in blueberry. In blueberry, six PAC strains significantly decreased dry shoot biomass compared to ErM control. In birch, one A. macrosclerotiorum strain increased root biomass and the other shoot biomass in comparison with non-inoculated control. The dual mycorrhizal ability of A. macrosclerotiorum suggested that it may form mycorrhizal links between Ericaceae and Pinaceae. However, we were unable to detect this species in Ericaceae roots growing in a forest with presence of A. macrosclerotiorum ectomycorrhizae. Nevertheless, the diversity of Ericaceae mycobionts was high (380 OTUs) with individual sites often dominated by hitherto unreported helotialean and chaetothyrialean/verrucarialean species; in contrast, typical ErM fungi were either absent or low in abundance. Some DSE apparently have a potential to form mycorrhizae with typical middle European forest plants. However, except A. applanata, the tested representatives of all hitherto described PAC cryptic species formed typical DSE colonization without

  9. Global 3D Imaging of Yersinia ruckeri Bacterin Uptake in Rainbow Trout Fry

    PubMed Central

    Ohtani, Maki; Villumsen, Kasper Rømer; Koppang, Erling Olaf; Raida, Martin Kristian

    2015-01-01

    Yersinia ruckeri is the causative agent of enteric redmouth disease (ERM) in rainbow trout, and the first commercially available fish vaccine was an immersion vaccine against ERM consisting of Y. ruckeri bacterin. The ERM immersion vaccine has been successfully used in aquaculture farming of salmonids for more than 35 years. The gills and the gastrointestinal (GI) tract are believed to be the portals of antigen uptake during waterborne vaccination against ERM; however, the actual sites of bacterin uptake are only partly understood. In order to obtain insight into bacterin uptake during waterborne vaccination, optical projection tomography (OPT) together with immunohistochemistry (IHC) was applied to visualize bacterin uptake and processing in whole rainbow trout fry. Visualization by OPT revealed that the bacterin was initially taken up via gill lamellae from within 30 seconds post vaccination. Later, bacterin uptake was detected on other mucosal surfaces such as skin and olfactory bulb from 5 to 30 minutes post vaccination. The GI tract was found to be filled with a complex of bacterin and mucus at 3 hours post vaccination and the bacterin remained in the GI tract for at least 24 hours. Large amounts of bacterin were present in the blood, and an accumulation of bacterin was found in filtering lymphoid organs such as spleen and trunk kidney where the bacterin accumulates 24 hours post vaccination as demonstrated by OPT and IHC. These results suggest that bacterin is taken up via the gill epithelium in the earliest phases of the bath exposure and from the GI tract in the later phase. The bacterin then enters the blood circulatory system, after which it is filtered by spleen and trunk kidney, before finally accumulating in lymphoid organs where adaptive immunity against ERM is likely to develop. PMID:25658600

  10. Macrolide-lincosamide-resistant phenotypes and genotypes of Staphylococcus aureus isolated from bovine clinical mastitis.

    PubMed

    Wang, Yang; Wu, Cong-Ming; Lu, Li-Ming; Ren, Gao-Wa Na; Cao, Xing-Yuan; Shen, Jian-Zhong

    2008-07-27

    The present study aimed to determine the prevalence and mechanisms of macrolide-lincosamide (ML) resistance in 72 Staphylococcus aureus isolates from cows with clinical mastitis. Minimum inhibitory concentrations (MIC) of ML antibiotics were determined by the broth microdilution technique, inducible ML resistance phenotype by the D test, and ML resistance genes by PCR assay. The isolates showed a high level of resistance to erythromycin (93.1%), azithromycin (93.1%), spiramycin (41.7%), tylosin (40.3%), tilmicosin (27.8%), and clindamycin (36.1%). Macrolide-lincosamide MIC(90) values were > or = 128 mg/L. Inducible ML resistance (iML) phenotype was detected in 52.8% (38/72) of isolates. In erythromycin-resistant (ER-R) strains, methylase genes ermB and ermC, efflux gene msrA/msrB, and inactivating enzyme genes lnuA and mphC were present alone or in various combinations, with ermB and ermC genes predominating. This is the first report of ML resistance genes ermB, mrsA/mrsB and mphC in S. aureus isolated from bovine mastitis. The occurrence of high levels of resistance to ML antibiotics among the S. aureus isolates, and the high rate of iML phenotype, indicate that appropriate alternative antibiotics should be prescribed for treating bovine mastitis caused by S. aureus. Furthermore, significant differences in the conformations of lactone rings of 16- and 14-membered macrolides could explain why some isolates with a constitutive ML resistance (cML) phenotype were sensitive to 16-membered macrolides alone. The different interaction of the 16-membered macrolides with the 50S ribosomal subunit is also presumably the reason why the susceptibility results of tilmcosin differed from those of tylosin and spiramycin. PMID:18272297

  11. Susceptibility to tulathromycin in Mannheimia haemolytica isolated from feedlot cattle over a 3-year period

    PubMed Central

    Alexander, Trevor W.; Cook, Shaun; Klima, Cassidy L.; Topp, Ed; McAllister, Tim A.

    2013-01-01

    Mannheimia haemolytica isolated from feedlot cattle were tested for tulathromycin resistance. Cattle were sampled over a 3-year period, starting 12 months after approval of tulathromycin for prevention and treatment of bovine respiratory disease. Nasopharyngeal samples from approximately 5,814 cattle were collected when cattle entered feedlots (N = 4) and again from the same cattle after ≥60 days on feed. The antimicrobial use history for each animal was recorded. Mannheimia haemolytica was isolated from 796 (13.7%) entry samples and 1,038 (20.6%) ≥ 60 days samples. Of the cattle positive for M. haemolytica, 18.5, 2.9, and 2.4% were administered therapeutic concentrations of tulathromycin, tilmicosin, or tylosin tartrate, respectively. In addition, 13.2% were administered subtherapeutic concentrations of tylosin phosphate in feed. In years one and two, no tulathromycin-resistant M. haemolytica were detected, whereas five isolates (0.4%) were resistant in year three. These resistant isolates were collected from three cattle originating from a single pen, were all serotype 1, and were genetically related (≥89% similarity) according to pulsed-field gel electrophoreses patterns. The five tulathromycin-resistant isolates were multi-drug resistant also exhibiting resistance to oxytetracycline, tilmicosin, ampicillin, or penicillin. The macrolide resistance genes erm(42), erm(A), erm(B), erm(F), erm(X) and msr(E)-mph(E), were not detected in the tulathromycin-resistant M. haemolytica. This study showed that tulathromycin resistance in M. haemolytica from a general population of feedlot cattle in western Canada was low and did not change over a 3-year period after tulathromycin was approved for use in cattle. PMID:24130555

  12. The Hippo effector TAZ (WWTR1) transforms myoblasts and TAZ abundance is associated with reduced survival in embryonal rhabdomyosarcoma.

    PubMed

    Mohamed, Abdalla; Sun, Congshan; De Mello, Vanessa; Selfe, Joanna; Missiaglia, Edoardo; Shipley, Janet; Murray, Graeme I; Zammit, Pete S; Wackerhage, Henning

    2016-09-01

    The Hippo effector YAP has recently been identified as a potent driver of embryonal rhabdomyosarcoma (ERMS). Most reports suggest that the YAP paralogue TAZ (gene symbol WWTR1) functions as YAP but, in skeletal muscle, TAZ has been reported to promote myogenic differentiation, whereas YAP inhibits it. Here, we investigated whether TAZ is also a rhabdomyosarcoma oncogene or whether TAZ acts as a YAP antagonist. Immunostaining of rhabdomyosarcoma tissue microarrays revealed that TAZ is significantly associated with poor survival in ERMS. In 12% of fusion gene-negative rhabdomyosarcomas, the TAZ locus is gained, which is correlated with increased expression. Constitutively active TAZ S89A significantly increased proliferation of C2C12 myoblasts and, importantly, colony formation on soft agar, suggesting transformation. However, TAZ then switches to enhance myogenic differentiation in C2C12 myoblasts, unlike YAP. Conversely, lentiviral shRNA-mediated TAZ knockdown in human ERMS cells reduced proliferation and anchorage-independent growth. While TAZ S89A or YAP1 S127A similarly activated the 8XGTIIC-Luc Hippo reporter, only YAP1 S127A activated the Brachyury (T-box) reporter. Consistent with its oncogene function, TAZ S89A induced expression of the ERMS cancer stem cell gene Myf5 and the serine biosynthesis pathway (Phgdh, Psat1, Psph) in C2C12 myoblasts. Thus, TAZ is associated with poor survival in ERMS and could act as an oncogene in rhabdomyosarcoma. © 2016 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland. PMID:27184927

  13. FOXM1 expression in rhabdomyosarcoma: a novel prognostic factor and therapeutic target.

    PubMed

    Kuda, Masaaki; Kohashi, Kenichi; Yamada, Yuichi; Maekawa, Akira; Kinoshita, Yoshiaki; Nakatsura, Tetsuya; Iwamoto, Yukihide; Taguchi, Tomoaki; Oda, Yoshinao

    2016-04-01

    The transcription factor Forkhead box M1 (FOXM1) is known to play critical roles in the development and progression of various types of cancer, but the clinical significance of FOXM1 expression in rhabdomyosarcoma (RMS) is unknown. This study aimed to determine the role of FOXM1 in RMS. We investigated the expression levels of FOXM1 and vascular endothelial growth factor (VEGF) and angiogenesis in a large series of RMS clinical cases using immunohistochemistry (n = 92), and we performed clinicopathologic and prognostic analyses. In vitro studies were conducted to examine the effect of FOXM1 knock-down on VEGF expression, cell proliferation, migration, and invasion in embryonal RMS (ERMS) and alveolar RMS (ARMS) cell lines, using small interference RNA (siRNA). High FOXM1 expression was significantly increased in the cases of ARMS, which has an adverse prognosis compared to ERMS (p = 0.0310). The ERMS patients with high FOXM1 expression (n = 25) had a significantly shorter survival than those with low FOXM1 expression (n = 24; p = 0.0310). FOXM1 expression was statistically correlated with VEGF expression in ERMS at the protein level as shown by immunohistochemistry and at the mRNA level by RT-PCR. The in vitro study demonstrated that VEGF mRNA levels were decreased in the FOXM1 siRNA-transfected ERMS and ARMS cells. FOXM1 knock-down resulted in a significant decrease of cell proliferation and migration in all four RMS cell lines and invasion in three of the four cell lines. Our results indicate that FOXM1 overexpression may be a prognostic factor of RMS and that FOXM1 may be a promising therapeutic target for the inhibition of RMS progression. PMID:26553361

  14. The over-expression of cell migratory genes in alveolar rhabdomyosarcoma could contribute to metastatic spread.

    PubMed

    Rapa, Elizabeth; Hill, Sophie K; Morten, Karl J; Potter, Michelle; Mitchell, Chris

    2012-06-01

    Alveolar (ARMS) and Embryonal (ERMS) rhabdomyosarcoma differ in their response to current treatments. The ARMS subtype has a less favourable prognosis and often presents with widespread metastases, while the less metastatic ERMS has a 5 year survival rate of more than 80 %. In this study we investigate gene expression differences that could contribute to the high frequency of metastasis in ARMS. Microarray analysis identified significant differences in DNA repair, cell cycle and cell migration between the two RMS subtypes. Two genes up regulated in ARMS and involved in cell migration; the engulfment and cell motility gene 1 (ELMO1) and NEL-like 1 gene (NELL1) were selected for further investigation. Over-expression of ELMO1 significantly increased cell invasion from 24.70 ± 7% to 93 ± 5.4% in primary myoblasts and from 29.43 ± 2.1% to 87.33 ± 4.1% in the ERMS cell line RD. siRNA knockout of ELMO1 in the ARMS cell line RH30 significantly reduced cell invasion from 88.2 ± 3.8% to 35.2 ± 2.5%. Over-expression of NELL1 significantly increased myoblast invasion from 23.6 ± 6.9% to 100 ± 0.1%, but had no effect on invasion of the ERMS cell line RD. These findings suggest that ELMO1 may play a key role in ARMS metastasis. NELL1 increased invasion in primary myoblasts, but other factors required for it to enhance motility were not present in the RD ERMS cell line. Impairing ELMO1 function by pharmacological or siRNA knockdown could be a highly effective approach to reduce the metastatic spread of RMS. PMID:22415709

  15. The Potential of Dark Septate Endophytes to Form Root Symbioses with Ectomycorrhizal and Ericoid Mycorrhizal Middle European Forest Plants

    PubMed Central

    Lukešová, Tereza; Kohout, Petr; Větrovský, Tomáš; Vohník, Martin

    2015-01-01

    The unresolved ecophysiological significance of Dark Septate Endophytes (DSE) may be in part due to existence of morphologically indistinguishable cryptic species in the most common Phialocephala fortinii s. l.—Acephala applanata species complex (PAC). We inoculated three middle European forest plants (European blueberry, Norway spruce and silver birch) with 16 strains of eight PAC cryptic species and other DSE and ectomycorrhizal/ericoid mycorrhizal fungi and focused on intraradical structures possibly representing interfaces for plant-fungus nutrient transfer and on host growth response. The PAC species Acephala applanata simultaneously formed structures resembling ericoid mycorrhiza (ErM) and DSE microsclerotia in blueberry. A. macrosclerotiorum, a close relative to PAC, formed ectomycorrhizae with spruce but not with birch, and structures resembling ErM in blueberry. Phialocephala glacialis, another close relative to PAC, formed structures resembling ErM in blueberry. In blueberry, six PAC strains significantly decreased dry shoot biomass compared to ErM control. In birch, one A. macrosclerotiorum strain increased root biomass and the other shoot biomass in comparison with non-inoculated control. The dual mycorrhizal ability of A. macrosclerotiorum suggested that it may form mycorrhizal links between Ericaceae and Pinaceae. However, we were unable to detect this species in Ericaceae roots growing in a forest with presence of A. macrosclerotiorum ectomycorrhizae. Nevertheless, the diversity of Ericaceae mycobionts was high (380 OTUs) with individual sites often dominated by hitherto unreported helotialean and chaetothyrialean/verrucarialean species; in contrast, typical ErM fungi were either absent or low in abundance. Some DSE apparently have a potential to form mycorrhizae with typical middle European forest plants. However, except A. applanata, the tested representatives of all hitherto described PAC cryptic species formed typical DSE colonization without

  16. Tylosin susceptibility of Staphylococci from bovine mastitis.

    PubMed

    Entorf, Monika; Feßler, Andrea T; Kadlec, Kristina; Kaspar, Heike; Mankertz, Joachim; Peters, Thomas; Schwarz, Stefan

    2014-07-16

    Although the 16-membered macrolide tylosin is commonly used for the treatment of bovine mastitis, little information is currently available about the susceptibility of mastitis pathogens to tylosin. In the present study, 112 Staphylococcus aureus and 110 coagulase-negative Staphylococcus (CoNS) spp. isolates from cases of bovine mastitis were tested by broth microdilution and agar disk diffusion with 30 μg tylosin disks. Susceptibility to erythromycin was tested by broth microdilution and disk diffusion using 15 μg disks. Both test populations showed bimodal distributions of minimal inhibitory concentrations (MICs) and zone diameters with eleven S. aureus and eight CoNS isolates showing tylosin MICs of ≥ 256 μg/ml and no zones of growth inhibition around the tylosin 30 μg disks. All 19 isolates with tylosin MICs of ≥ 256 μg/ml were also resistant to erythromycin. For six additional erythromycin-resistant isolates, tylosin MICs of 1-8 μg/ml were observed. One S. aureus and two CoNS isolates showed inducible macrolide resistance. PCR analysis of the 25 erythromycin-resistant staphylococcal isolates identified the resistance genes erm(A), erm(B), erm(C), erm(T), mph(C) and msr(A) alone or in different combinations. An excellent correlation between the results of the different tylosin susceptibility tests (broth microdilution versus disk diffusion) was seen for S. aureus and CoNS isolates. Since tylosin does not induce the expression of the aforementioned erm genes, isolates with an inducible resistance phenotype may - if only tylosin is tested - be falsely classified as tylosin-susceptible. Thus, erythromycin should be tested in parallel and tylosin should only be used for the treatment of infections caused by erythromycin-susceptible staphylococci. PMID:24461550

  17. Erythromycin resistance and virulence genes in Enterococcus faecalis from swine in China.

    PubMed

    Zou, Li-Kou; Wang, Hong-Ning; Zeng, Bo; Li, Jin-Niang; Li, Xu-Ting; Zhang, An-Yun; Zhou, Ying-Shun; Yang, Xin; Xu, Chang-Wen; Xia, Qing-Qing

    2011-01-01

    This study aims to describe the erythromycin resistance phenotypes and genotypes, and the prevalence of virulence genes of Enterococcus faecalis isolated from swine in China. A total of 117 nonreplicate E. faecalis isolates, obtained from 502 clinical samples taken from different pig farms between 2007 and 2009 were included in the study. Minimum inhibitory concentrations were determined using the broth microdilution method. All of the isolates were screened for the presence of seven virulence genes (ace, asa1, cylA, efaA, esp, gelE, and hyl). In addition, the DNA from rythromycin-resistant isolates were amplified with primers specific for erythromycin resistance erm(A), erm(B), erm(C), mef(A/E), and msr(C) genes. Results show that erythromycin, tylosin, and ciprofloxacin resistance rates in E. faecalis were 66.67% (n=78), 66.67% (n=78), and 64.10% (n=75), respectively. About 69.23% of isolates (n=81) were positive for gelE, 48.72% (n=57) for ace, 15.38% (n=18) for efa, 7.69% (n=9) for asa1, and 6.84% (n=8) for esp. Among the erythromycin-resistant isolates, erm(B) (n=54) was the most prevalent resistance gene, followed by erm(A) (n=37). A significant correlation was found between the presence of the gelE virulence gene and erythromycin resistance (P<0.05). These findings suggest that enterococci from swine should be regarded with caution because they can be reservoirs for antimicrobial resistance and virulence genes. PMID:21344149

  18. Modeling gross primary production and ecosystem respiration for terrestrial ecosystems in North China and Tibet Plateau using MODIS imagery

    NASA Astrophysics Data System (ADS)

    Gao, Y.; Yu, G.; Yan, H.; Zhu, X.; Li, S.; Wang, Q.; Zhang, J.; Wang, Y.; Li, Y.; Zhao, L.; Shi, P.

    2013-12-01

    Gross primary production (GPP) and ecosystem respiration (Re) are two large components in the studying of regional and global carbon cycles. Accurate quantification of spatio-temporal variations of GPP and Re for terrestrial ecosystems is of great importance to research carbon budget on regional and global scales. In this study, we proposed two satellite-based models, i.e. Photosynthetic Capacity Model (PCM) and Ecosystem Respiration Model (ERM), to simulate GPP and Re of terrestrial ecosystems, respectively. Multi-year eddy CO2 flux data from five vegetation types in North China (temperate mixed forest, temperate steppe) and Tibet Plateau (alpine shrubland, alpine marsh and alpine meadow-steppe) were used for assessing the model performances. The PCM model was driven by the Enhanced Vegetation Index (EVI) and the Land Surface Water Index (LSWI) from MODIS imagery. In most cases, the PCM-simulated GPP and the observed GPP displayed very consistent seasonal and inter-seasonal variability regardless of vegetation types. The PCM predicted versus observed GPP performed better than the MODIS GPP products, and was compatible with the Vegetation Photosynthesis Model (VPM). Moreover, the model parameter of the PCM could be gained from the linear function of mean annual remote sensing data. Based on this linear function, the PCM model simulated 93% variations of the observed GPP across all five vegetation types. The ERM model was developed based on both GPP and temperature, and was driven by EVI, LSWI and the Land Surface Temperature (LST) from MODIS imagery. In most cases, the seasonal and interannual variations of the simulated Re matched well with the observed Re. Compared with the model driven by temperature, and the model further added GPP in the reference respiration, the ERM model was optimal in each vegetation type. The model parameters of the ERM could also be presented by the liner functions of mean annual remote sensing data. Based on these linear functions, 90

  19. Emergence of quinupristin/dalfopristin resistance among livestock-associated Staphylococcus aureus ST9 clinical isolates.

    PubMed

    Yu, Fangyou; Lu, Chaohui; Liu, Yunling; Sun, He; Shang, Yongpeng; Ding, Yu; Li, Dan; Qin, Zhiqiang; Parsons, Chris; Huang, Xiaoying; Li, Yuping; Hu, Longhua; Wang, Liangxing

    2014-11-01

    Quinupristin/dalfopristin (Q/D) is a valuable alternative to vancomycin for the treatment of meticillin-resistant Staphylococcus aureus (MRSA) infections. However, not long after Q/D was approved, bacteria with resistance to this newer antimicrobial agent were reported. To investigate the prevalence of Q/D resistance, a total of 1476 non-duplicate S. aureus isolates, including 775 MRSA, from a Chinese tertiary hospital were selected randomly from 2003 to 2013. Of the 775 MRSA, 3 (0.4%) were resistant to Q/D. All meticillin-susceptible S. aureus were susceptible to Q/D. The prevalence of Q/D resistance among S. aureus was 0.2% (3/1476). The three isolates with Q/D resistance had the same antimicrobial resistance profile, except for cefaclor and chloramphenicol. All three Q/D-resistant MRSA were positive for five streptogramin B resistance genes (ermA, ermB, ermC, msrA and msrB) and two streptogramin A resistance genes (vatC and vgaA) as determined by PCR and DNA sequencing. MRSA WZ1031 belonged to ST9-MRSA-SCCmecV-t899, whilst MRSA WZ414 and WZ480 belonged to ST9-MRSA-SCCmecNT(non-typeable)-t899. ST9 has been reported predominantly in livestock-associated (LA) MRSA in some Asian countries. The three patients with these MRSA isolates were not livestock handlers and did not keep close contact with livestock. The origin of these important LA-MRSA isolates causing human infections is not known. Taken together, Q/D resistance, which was caused by a combination of ermA-ermB-ermC-msrA-msrB-vatC-vgaA, was first found among S. aureus clinical isolates in China. The present study is the first report of the emergence of human infections caused by ST9 LA-MRSA isolates with Q/D resistance. PMID:25218154

  20. Molecular Cytogenetics Detect an Unbalanced t(2;13)(q36;q14) and PAX3-FOXO1 Fusion in Rhabdomyosarcoma With Mixed Embryonal/Alveolar Features.

    PubMed

    La Starza, Roberta; Nofrini, Valeria; Pierini, Tiziana; Pierini, Valentina; Zin, Angelica; Bisogno, Gianni; Cerri, Carla; Caniglia, Maurizio; Sidoni, Angelo; Ludwig, Kathrin; Mecucci, Cristina

    2015-12-01

    Distinguishing between alveolar rhabdomyosarcoma (ARMS) and embryonal rhabdomyosarcoma (ERMS) is crucial because treatment and prognosis are different. We describe a case of paratesticular rhabdomyosarcoma (RMS), which was classified as mixed ERMS/ARMS. Fluorescence in situ hybridization (FISH) detected losses of 3'PAX3 and 5'FOXO1, suggesting they had undergone an unbalanced rearrangement that probably produced the PAX3-FOXO1 fusion. Double-color FISH and reverse transcription-polymerase chain reaction (RT-PCR) revealed PAX3-FOXO1, which is characteristic of high-risk RMS. This finding highlights the importance of supplementing histology with genetics so that atypical RMS is appropriately classified and patients are correctly stratified and treated. PMID:26179572

  1. Occurrence and distribution of antibiotic resistance genes in the coastal area of the Bohai Bay, China.

    PubMed

    Niu, Zhi-Guang; Zhang, Kai; Zhang, Ying

    2016-06-15

    Considering the abuse of antibiotics worldwide, we investigated the abundance of three classes of antibiotic resistance genes (ARGs) and the concentrations of corresponding antibiotics in water and sediments of Bohai Bay. The results showed that sulI and sulII were detected in all samples, and their abundance range was 10(-5)-10(-2)/16S gene copies. The abundance of tetM and ermB were relatively higher than the other genes of tet-ARGs and erm-ARGs. Sulfonamides were the most prevalent antibiotics, and the concentrations of antibiotic in sediments were higher than those in water. The correlation analysis revealed that antibiotics had pertinence with corresponding ARGs, indicating that antibiotics play an important role in the creation and transfer of ARGs. The results of regression analysis indicated that the propagation and maintenance of sulI and sulII were facilitated by class I integrons. PMID:27107623

  2. An investigation of TNAV equipped aircraft in a simulated en route metering environment

    NASA Technical Reports Server (NTRS)

    Groce, J. L.; Izumi, K. H.; Markham, C. H.; Schwab, R. W.; Taylor, J. A.

    1986-01-01

    This document presents the results of an effort to estimate how often a TNAV (Time Navigation) equipped aircraft could be given a TNAV clearance in the En Route Metering (ERM) system as a function of the percentage of arriving traffic which is TNAV equipped. A fast-time simulation of Denver Stapleton international arrival traffic in the Denver Air Route Traffic Control Center route structure, including en route metering operations, was used to develop data on estimated conflicts, clearance communications and fuel usage for traffic mixes of 25, 50, 75 and 100% TNAV equipped. This study supports an overall effort by NASA to assess the benefits and required technology for using TNAV-equipped aircraft in the ERM environment.

  3. Macrolide resistance determinants in erythromycin-resistant Streptococcus pneumoniae in Turkey.

    PubMed

    Gulay, Zeynep; Ozbek, Ozgen Alpay; Bicmen, Meral; Gur, Deniz

    2008-11-01

    To determine the major molecular mechanisms of macrolide resistance among Streptococcus pneumoniae isolates in Turkey, we examined a total of 151 isolates collected from different regions of Turkey. Overall, 40 (26.4%) isolates were resistant to erythromycin. The most common mechanism (38/40) was target site modification due to erm (B) genes. Only two isolates harbored the mef (A)/(E) efflux gene. A clonal spread of resistant strains could not be demonstrated by BOX-polymerase chain reaction. The results from this study have shown that the erm (B) gene is predominant in Turkish S. pneumoniae isolates, as in isolates from the rest of the world, and a clonal dissemination is not responsible for this resistance profile. PMID:19050364

  4. The drug susceptibility profile and inducible resistance to macrolides of Mycobacterium abscessus and Mycobacterium massiliense in Korea.

    PubMed

    Kim, Song Yee; Kim, Chang-Ki; Bae, Il Kwon; Jeong, Seok Hoon; Yim, Jae-Joon; Jung, Ji Ye; Park, Moo Suk; Kim, Young Sam; Kim, Se Kyu; Chang, Joon; Kang, Young Ae

    2015-02-01

    We conducted drug susceptibility testing (DST) against various antimicrobial agents, including new candidate drugs, and investigated the relationship between inducible resistance (IR) to macrolides and erm(41) gene in Mycobacterium abscessus complex. Sixty-two isolates of M. abscessus complex from 2 tertiary care hospitals in South Korea were tested against 10 antimicrobial agents. Thirty-five isolates were M. abscessus, and 27 were Mycobacterium massiliense. Amikacin, moxifloxacin, linezolid, clofazimine, and tigecycline were active against most isolates and cefoxitin and ciprofloxacin against moderate number of isolates. M. massiliense remained susceptible to macrolides; in contrast, M. abscessus became highly resistant on day 14 after incubation. DST pattern did not differ between clarithromycin and azithromycin. IR to clarithromycin was correlated with erm(41) genotype in M. abscessus. Variations in susceptibility to antimicrobial agents within species and the difference in DST patterns between M. abscessus and M. massiliense suggest that DST and identification of M. abscessus complex are significant before treatment. PMID:25467784

  5. Cloacal Lactobacillus isolates from broilers show high prevalence of resistance towards macrolide and lincosamide antibiotics.

    PubMed

    Cauwerts, K; Pasmans, F; Devriese, L A; Martel, A; Haesebrouck, F; Decostere, A

    2006-04-01

    Eighty-seven Lactobacillus strains isolated from cloacal swabs of broiler chickens derived from 20 different farms in Belgium were identified to species level and tested for susceptibility to macrolide and lincosamide antibiotics. Five different Lactobacillus species were identified as being predominantly present in the cloacae of broilers: Lactobacillus crispatus, Lactobacillus salivarius subsp. salivarius, Lactobacillus amylovorus, Lactobacillus gallinarum and Lactobacillu sreuteri. Acquired resistance prevalence to macrolides and lincosamides was very high in the investigated lactobacilli: 89% of the strains were resistant to either or both lincosamide and macrolide class antibiotics. The vast majority of these resistant strains (96%) displayed constitutive resistance. More than one-half of the macrolide and/or lincosamide resistant strains carried an erm(B), erm(C), mef(A), lnu(A) gene or a combination of these genes. PMID:16595310

  6. Thickness dependent fatigue life at microcrack nucleation for metal thin films on flexible substrates

    NASA Astrophysics Data System (ADS)

    Sun, X. J.; Wang, C. C.; Zhang, J.; Liu, G.; Zhang, G. J.; Ding, X. D.; Zhang, G. P.; Sun, J.

    2008-10-01

    For polymer-supported metal thin films used in flexible electronics, the definition of the fatigue lifetime at microcrack nucleation (FLMN) should be more physically meaningful than all the previous definitions at structural instability. In this paper, the FLMN of Cu films (with thickness from 100 nm to 3.75 µm) as well as Al thin films (from 80 to 800 nm) was experimentally characterized at different strain ranges and different thicknesses by using a simple electrical resistance measurement (ERM). A significant thickness dependence was revealed for the FLMN and a similar Coffin-Manson fatigue relationship observed commonly in bulk materials was found to be still operative in both the films. Microstructural analyses were carried out to verify the feasibility of ERM correspondingly.

  7. Pollution from organic contaminants in Greek marine areas, receiving anthropogenic pressures from intense activities in the coastal zone

    NASA Astrophysics Data System (ADS)

    Hatzianestis, Ioannis

    2014-05-01

    Polycyclic aromatic hydrocarbons (PAHs) are widespread pollutants in marine sediments, receiving the pressures from various anthropogenic activities in the coastal zone. Due to their mutagenic and carcinogenic behaviour, PAHs are classified as priority contaminants to be monitored in environmental quality control schemes. The purpose of this study was to determine the levels of PAHs in coastal areas of Greece directly influenced from the operation of major industrial units in the coastal zone, investigate their sources and evaluate their potential toxicity by comparison against effect - based sediment quality guidelines. Thirty two surface sediment samples were collected from three areas of the Hellenic coastline: a) Antikyra bay in Korinthiakos gulf, influenced from the operation of an alumina and aluminium production plant b) Larymna bay in Noth Evoikos gulf, influenced from the operation of a nickel production plant and c) Aliveri bay in South Evoikos Gulf, influenced from a cement production plant. In all the areas studied, aquaculture and fishing activities have been also developed in the coastal zone. PAH concentrations were determined by GC-MS, after soxhlet extraction and fractionation by silica column chromatography. PAH sources and origin were investigated by applying several isomeric ratio diagnostic criteria. The mean quotient Effect- Range Median (m-ERM) was used to evaluate the potential of adverse effects posed to benthic organisms. Three m-ERM-q values were used to differentiate the probability of observing toxicity and classify sites into four categories: sediments with m-ERM<0.1 have the lowest probability (9%) of being toxic, those with m-ERM from 0.11 to 0.5 have a 21% probability of toxicity, those with m-ERM from 0.51 to 1.5 a 49% probability of toxicity, while sediments with m-ERM >1.5 have the highest probability (76%) of toxicity. Extremely high PAH concentrations more than 100,000 ng/g were found in the close vicinity of the alumina

  8. Tandem Level II and Level III field analyses for in-field decision making

    SciTech Connect

    Gallis, D.E.; Christenson, J.

    1995-12-31

    In order to provide cost-effective and timely analytical support for a site investigation in central Pennsylvania, Environmental Resources Management, Inc. (ERM) used its mobile analytical service, the ERM-FAST{reg_sign} service to perform on-site soil analyses targeting both Level II and Level III data quality objectives. Of specific concern were volatile organic compounds. The soil concentration levels of these contaminants of interest were required on a rapid-turnaround basis in order to make decisions regarding subsequent boring locations and other analytical logistics. The strategy and analytical systems used for this investigation provided cost-effective, quick-turnaround results that would not have been possible through routine fixed-facility analyses.

  9. Retinal Changes Induced by Epiretinal Tangential Forces

    PubMed Central

    Romano, Mario R.; Comune, Chiara; Ferrara, Mariantonia; Cennamo, Gilda; De Cillà, Stefano; Toto, Lisa; Cennamo, Giovanni

    2015-01-01

    Two kinds of forces are active in vitreoretinal traction diseases: tangential and anterior-posterior forces. However, tangential forces are less characterized and classified in literature compared to the anterior-posterior ones. Tangential epiretinal forces are mainly due to anomalous posterior vitreous detachment (PVD), vitreoschisis, vitreopapillary adhesion (VPA), and epiretinal membranes (ERMs). Anomalous PVD plays a key role in the formation of the tangential vectorial forces on the retinal surface as consequence of gel liquefaction (synchysis) without sufficient and fast vitreous dehiscence at the vitreoretinal interface. The anomalous and persistent adherence of the posterior hyaloid to the retina can lead to vitreomacular/vitreopapillary adhesion or to a formation of avascular fibrocellular tissue (ERM) resulting from the proliferation and transdifferentiation of hyalocytes resident in the cortical vitreous remnants after vitreoschisis. The right interpretation of the forces involved in the epiretinal tangential tractions helps in a better definition of diagnosis, progression, prognosis, and surgical outcomes of vitreomacular interfaces. PMID:26421183

  10. Molecular Basis of Resistance to Selected Antimicrobial Agents in the Emerging Zoonotic Pathogen Streptococcus suis.

    PubMed

    Gurung, Mamata; Tamang, Migma Dorji; Moon, Dong Chan; Kim, Su-Ran; Jeong, Jin-Ha; Jang, Geum-Chan; Jung, Suk-Chan; Park, Yong-Ho; Lim, Suk-Kyung

    2015-07-01

    Characterization of 227 Streptococcus suis strains isolated from pigs during 2010 to 2013 showed high levels of resistance to clindamycin (95.6%), tilmicosin (94.7%), tylosin (93.8%), oxytetracycline (89.4%), chlortetracycline (86.8%), tiamulin (72.7%), neomycin (70.0%), enrofloxacin (56.4%), penicillin (56.4%), ceftiofur (55.9%), and gentamicin (55.1%). Resistance to tetracyclines, macrolides, aminoglycosides, and fluoroquinolone was attributed to the tet gene, erm(B), erm(C), mph(C), and mef(A) and/or mef(E) genes, aph(3')-IIIa and aac(6')-Ie-aph(2″)-Ia genes, and single point mutations in the quinolone resistance-determining region of ParC and GyrA, respectively. PMID:25903569

  11. Thermal expansion recovery microscopy: Practical design considerations

    SciTech Connect

    Mingolo, N. Martínez, O. E.

    2014-01-15

    A detailed study of relevant parameters for the design and operation of a photothermal microscope technique recently introduced is presented. The technique, named thermal expansion recovery microscopy (ThERM) relies in the measurement of the defocusing introduced by a surface that expands and recovers upon the heating from a modulated source. A new two lens design is presented that can be easily adapted to commercial infinite conjugate microscopes and the sensitivity to misalignment is analyzed. The way to determine the beam size by means of a focus scan and the use of that same scan to verify if a thermoreflectance signal is overlapping with the desired ThERM mechanism are discussed. Finally, a method to cancel the thermoreflectance signal by an adequate choice of a nanometric coating is presented.

  12. Molecular Basis of Resistance to Selected Antimicrobial Agents in the Emerging Zoonotic Pathogen Streptococcus suis

    PubMed Central

    Gurung, Mamata; Tamang, Migma Dorji; Moon, Dong Chan; Kim, Su-Ran; Jeong, Jin-Ha; Jang, Geum-Chan; Jung, Suk-Chan; Park, Yong-Ho

    2015-01-01

    Characterization of 227 Streptococcus suis strains isolated from pigs during 2010 to 2013 showed high levels of resistance to clindamycin (95.6%), tilmicosin (94.7%), tylosin (93.8%), oxytetracycline (89.4%), chlortetracycline (86.8%), tiamulin (72.7%), neomycin (70.0%), enrofloxacin (56.4%), penicillin (56.4%), ceftiofur (55.9%), and gentamicin (55.1%). Resistance to tetracyclines, macrolides, aminoglycosides, and fluoroquinolone was attributed to the tet gene, erm(B), erm(C), mph(C), and mef(A) and/or mef(E) genes, aph(3′)-IIIa and aac(6′)-Ie-aph(2″)-Ia genes, and single point mutations in the quinolone resistance-determining region of ParC and GyrA, respectively. PMID:25903569

  13. Current Trends about Inner Limiting Membrane Peeling in Surgery for Epiretinal Membranes

    PubMed Central

    Semeraro, Francesco; Morescalchi, Francesco; Duse, Sarah; Gambicorti, Elena; Russo, Andrea; Costagliola, Ciro

    2015-01-01

    The inner limiting membrane (ILM) is the basement membrane of the Müller cells and can act as a scaffold for cellular proliferation in the pathophysiology of disorders affecting the vitreomacular interface. The atraumatic removal of the macular ILM has been proposed for treating various forms of tractional maculopathy in particular for macular pucker. In the last decade, the removal of ILM has become a routine practice in the surgery of the epiretinal membranes (ERMs), with good anatomical results. However many recent studies showed that ILM peeling is a procedure that can cause immediate traumatic effects and progressive modification on the underlying inner retinal layers. Moreover, it is unclear whether ILM peeling is helpful to improve vision after surgery for ERM. In this review, we describe the current understanding about ILM peeling and highlight the beneficial and adverse effects associated with this surgical procedure. PMID:26425352

  14. Fate of antibiotic resistance genes and its drivers during anaerobic co-digestion of food waste and sewage sludge based on microwave pretreatment.

    PubMed

    Zhang, Junya; Chen, Meixue; Sui, Qianwen; Wang, Rui; Tong, Juan; Wei, Yuansong

    2016-10-01

    In this study, anaerobic digestion of mono-SS, MW-SS:FW and SS:MW-FW was investigated to understand the fate of ARGs and its drivers. Anaerobic digestion was effective for the reduction of metal resistance genes (MRGs), and could reduce the abundance of blaOXA-1, sulI and tetG, while sulII in co-digestion and blaTEM and ereA only in MW-SS. ARGs reduction could be partly attributed to the reduction of co-selective pressure from heavy metals reflected by MRGs. However, the abundance of mefA/E, ermB, ermF, tetM and tetX increased significantly. Anaerobic co-digestion, especially for MW-SS, could reduce total ARGs abundance compared with mono-SS, and evolution of bacterial community was the main driver for the fate of ARGs. PMID:26988135

  15. PAX3-FKHR sensitizes human alveolar rhabdomyosarcoma cells to camptothecin-mediated growth inhibition and apoptosis.

    PubMed

    Zeng, Fu-Yue; Cui, Jimmy; Liu, Lingling; Chen, Taosheng

    2009-11-01

    Patients with alveolar rhabdomyosarcoma (ARMS) have poorer response to conventional chemotherapy and lower survival rates than those with embryonal RMS (ERMS). By high-throughput screening, we identified camptothecin as an ARMS-selective inhibitor. Camptothecin more efficiently inhibited proliferation and induced apoptosis in Rh30 (ARMS) than RD (ERMS) cells. Ectopic expression of the PAX3-FKHR (PF) fusion protein in RD cells significantly increased sensitivity, whereas siRNA knockdown of PF decreased sensitivity of Rh30 cells to camptothecin. The sensitization required a transcriptionally active PF, and camptothecin downregulated levels of PF protein. These findings suggest that it is feasible to develop agents that preferentially block the growth of ARMS. PMID:19442434

  16. Mutations in 23S rRNA and Ribosomal Protein L4 Account for Resistance in Pneumococcal Strains Selected In Vitro by Macrolide Passage

    PubMed Central

    Tait-Kamradt, A.; Davies, T.; Cronan, M.; Jacobs, M. R.; Appelbaum, P. C.; Sutcliffe, J.

    2000-01-01

    The mechanisms responsible for macrolide resistance in Streptococcus pneumoniae mutants, selected from susceptible strains by serial passage in azithromycin, were investigated. These mutants were resistant to 14- and 15-membered macrolides, but resistance could not be explained by any clinically relevant resistance determinant [mef(A), erm(A), erm(B), erm(C), erm(TR), msr(A), mph(A), mph(B), mph(C), ere(A), ere(B)]. An investigation into the sequences of 23S rRNAs in the mutant and parental strains revealed individual changes of C2611A, C2611G, A2058G, and A2059G (Escherichia coli numbering) in four mutants. Mutations at these residues in domain V of 23S rRNA have been noted to confer erythromycin resistance in other species. Not all four 23S rRNA alleles have to contain the mutation to confer resistance. Some of the mutations also confer coresistance to streptogramin B (C2611A, C2611G, and A2058G), 16-membered macrolides (all changes), and clindamycin (A2058G and A2059G). Interestingly, none of these mutations confer high-level resistance to telithromycin (HMR-3647). Further, two of the mutants which had no changes in their 23S rRNA sequences had changes in a highly conserved stretch of amino acids (63KPWRQKGTGRAR74) in ribosomal protein L4. One mutant contained a single amino acid change (G69C), while the other mutant had a 6-base insert, resulting in two amino acids (S and Q) being inserted between amino acids Q67 and K68. To our knowledge, this is the first description of mutations in 23S rRNA genes or ribosomal proteins in macrolide-resistant S. pneumoniae strains. PMID:10898684

  17. Spectral-based inferential measurement of grey-body's temperature

    NASA Astrophysics Data System (ADS)

    Zheng, Feng; Liu, Liying; Zhu, Lingxi; Huan, Kewei; Li, Ye; Shi, Xiaoguang

    2015-11-01

    Aiming at the problems of temperature measurement and the defects of radiance thermometry theory, one method of spectral-based inferential measurement is proposed, which adopts the Empirical Risk Minimization (ERM) functional model as the temperature measurement model. Then, the radiance thermometry theory and inferential measurement technology are discussed comparatively. Temperatures of some targets, such and tungsten lamp and solar surface, are measured by spectral-based inferential measurement.

  18. Multimodal genetic diagnosis of solid variant alveolar rhabdomyosarcoma.

    PubMed

    Cerveira, Nuno; Torres, Lurdes; Ribeiro, Franclim R; Henrique, Rui; Pinto, Armando; Bizarro, Susana; Ferreira, Ana M; Lopes, Carlos; Teixeira, Manuel R

    2005-12-01

    The most common types of rhabdomyosarcoma (RMS) are alveolar RMS (ARMS), which are characterized by the specific translocation t(2;13)(q35;q14) or its rarer variant, t(1;13)(p36;q14), producing the fusion genes PAX3-FKHR and PAX7-FKHR, respectively, and embryonal RMS (ERMS), which is characterized by multiple numeric chromosome changes. A solid variant of ARMS that is morphologically indistinguishable from ERMS has been described recently. We present two cases with an initial histopathologic diagnosis of ERMS in which the combined findings by cytogenetic, reverse-transcriptase polymerase chain reaction (RT-PCR), and comparative genomic hybridization (CGH) analyses demonstrate that both tumors were in fact the solid variant of ARMS. The cytogenetic analysis of patient 1 revealed a t(2;13)(q35;q14) and the RT-PCR study detected the corresponding PAX3-FKHR chimeric transcript. In patient 2, the cytogenetic finding of multiple trisomies was compatible with the initial histopathologic diagnosis of ERMS, but the finding of a PAX7-FKHR fusion transcript by RT-PCR pointed to the diagnosis of ARMS. Interestingly, the CGH findings of this case reconciled the molecular and cytogenetic data by detecting, in addition to the trisomies, amplification of chromosomal bands 1p36 and 13q14, where the PAX7 and FKHR genes are located, respectively. Our data indicate that this multimodal genetic analysis could be important for the differential diagnosis of these tumors. Furthermore, our findings and previous studies indicate that there are no apparent genetic differences between solid variant and typical ARMS. PMID:16337856

  19. Nasal vaccination of young rainbow trout (Oncorhynchus mykiss) against infectious hematopoietic necrosis and enteric red mouth disease.

    PubMed

    Salinas, I; LaPatra, S E; Erhardt, E B

    2015-11-01

    Determining the earliest age at which farmed fish can be successfully vaccinated is a very important question for fish farmers. Nasal vaccines are novel mucosal vaccines that prevent aquatic infectious diseases of finfish. The present study investigates the ontogeny of the olfactory organ of rainbow trout by histology and aims to establish the earliest age for vaccination against infectious hematopoietic necrosis (IHN) and enteric red mouth (ERM) disease using the nasal route. Rainbow trout (Oncorhynchus mykiss) were vaccinated intranasally (I.N) at three different ages: 1050° days (DD) (group A); 450 DD (group B); and 360 DD (group C), or 70, 30 and 24 days post-hatch (dph), respectively. The mean weights of groups A, B and C were 4.69 g, 2.9 g and 2.37 g, respectively. Fish received either a live attenuated IHN virus vaccine, ERM formalin killed bacterin or saline (mock vaccinated). Fish were challenged to the corresponding live pathogen 28 days post-vaccination. IHN vaccine delivery at 360 DD resulted in 40% mortality likely due to residual virulence of the vaccine. No mortality was observed in the ERM nasal delivery groups. Following challenge, very high protection rates against IHN virus were recorded in all three age groups with survivals of 95%, 100% and 97.5% in groups A, B and C, respectively. Survival against ERM was 82.5%, 87.5% and 77.5% in groups A, B and C, respectively. Survival rates did not differ among ages for either vaccine. Our results indicate the feasibility and effectiveness of nasal vaccination as early as 360 DD and vaccination-related mortalities when a live attenuated viral vaccine was used in the youngest fish. PMID:26111996

  20. Sequence and properties of pIM13, a macrolide-lincosamide-streptogramin B resistance plasmid from Bacillus subtilis.

    PubMed Central

    Monod, M; Denoya, C; Dubnau, D

    1986-01-01

    We initiated a study of pIM13, a multicopy, macrolide-lincosamide-streptogramin B (MLS) plasmid first isolated from a strain of Bacillus subtilis and described by Mahler and Halvorson (J. Gen. Microbiol. 120:259-263, 1980). The copy number of this plasmid was about 200 in B. subtilis and 30 in Staphylococcus aureus. The MLS resistance determinant of pIM13 was shown to be highly homologous to ermC, an inducible element on the S. aureus plasmid pE194. The product of the pIM13 determinant was similar in size to that of ermC and immunologically cross-reactive with it. The MLS resistance of pIM13 was expressed constitutively. The complete base sequence of pIM13 is presented. The plasmid consisted of 2,246 base pairs and contained two open reading frames that specified products identified in minicell extracts. One was a protein of 16,000 molecular weight, possibly required for replication. The second was the 29,000-molecular-weight MLS resistance methylase. The regulatory region responsible for ermC inducibility was missing from pIM13, explaining its constitutivity. The remainder of the pIM13 MLS determinant was nearly identical to ermC. The ends of the region of homology between pIM13 and pE194 were associated with hyphenated dyad symmetries. A segment partially homologous to one of these termini on pIM13 and also associated with a dyad was found in pUB110 near the end of a region of homology between that plasmid and pBC16. The entire sequence of pIM13 was highly homologous to that of pE5, an inducible MLS resistance plasmid from S. aureus that differs from pIM13 in copy control. Images PMID:3087948

  1. Development of Biocompatibility Procedures for Assessment of Plant Growth in Ground Test Hardware for the EMCS

    NASA Technical Reports Server (NTRS)

    Bowman, R. N.; Steele, M. K.; Sun, S. (Technical Monitor)

    2002-01-01

    The European Modular Cultivation System (EMCS) is an European Space Agency-developed facility designed to support plant research in microgravity on the IS NASA is responsible for providing US specific hardware to use within the EMCS. In preparation for flight, research will be developed and tested at Ames Research Center in the EMCS ground test hardware, the Experiment Reference Module (ERM) In order to determine the acceptability of the ERM for such purposes, biocompatibility tests will be performed to determine that the hardware functions as intended and successfully supports the' growth of plants. In this report, we describe the development of procedures and the collection of baseline data against which to compare ERM function, e.g. biocompatibility testing. A simple and robust system was developed to grow whole Arabidopsis thaliana plants within the confined volumes characteristic of spaceflight hardware. Our system for growing plants eliminated the necessity of a water/nutrient delivery system and allowed for quantifiable assessment of individual plants, as well as entire population dynamics. To insure uniform germination, seeds were started in small straw segments and transplanted into modified scintillation vials. Seedlings were selected prior to transplantation to decrease genetic variability. Plants were grown for a total of 24 days in standard laboratory plant growth chambers under controlled conditions. Sequential digital still images were taken on a daily basis. Analysis of these images allowed for the quantification of even minute environmental effect, on growth dynamics whole plants. The data collected provide reliable growth curves against which to compare plants grown in the ERM.

  2. Molecular basis of antimicrobial drug resistance in Staphylococcus aureus isolates recovered from young healthy carriers in Spain.

    PubMed

    Argudín, M Angeles; Mendoza, M Carmen; Martín, M Cruz; Rodicio, M Rosario

    2014-09-01

    The frequency and genetic bases of antimicrobial drug resistance was determined for 111 Staphylococcus aureus recovered from young healthy carriers in a Spanish region. Resistances to ampicillin (84.7%), kanamycin (27%), erythromycin (25.2%), clindamycin (22.5%), tetracycline (11.7%), amikacin and tobramycin (6.3% each), gentamicin (5.4%), chloramphenicol (2.7%), ciprofloxacin (0.9%; MIC 4 μg/ml), moxifloxacin (0.9%) and mupirocin (0.9%; MIC 60 μg/ml) were found, and all were susceptible to methicillin (MSSA). Nearly 50% of the isolates were resistant to one antibiotic, 30% to two, 15.3% to three and 1.8% to four, while only 6.3% remained fully susceptible. A total of 31 profiles were found. For each phenotypic resistance, at least one gene accounting for it was identified. The detected genes were blaZ; erm(A)-erm(B)-erm(C)-msr(A)-msr(B)-lnu(A), aphA-aadE-sat4-aacA + aphD-aadD, tet(K), cat, and qacA/B, for resistance to ampicillin, macrolides and/or lincosamides, aminoglycosides, tetracycline, chloramphenicol, and quaternary ammonium compounds, respectively. In all isolates carrying cat genes, in all except one of the isolates positive for tet(K), and in most isolates with blaZ, erm(C), msr(A), or msr(B), the gene(s) mapped on resistance plasmids, which were detected in 69.2% of the resistant isolates (65% of the total). The S. aureus from young healthy carriers analysed in the present study do not constitute a reservoir of MRSA, but they represent a repository of multiple determinants conferring resistance to "old" antimicrobials. Some of these have still clinical applications and, considering the increasing resistance to recently introduced antimicrobials, none of them can be disregarded. PMID:24972115

  3. Rapid prediction of inducible clarithromycin resistance in Mycobacterium abscessus.

    PubMed

    Zhu, Yan Chun; Mitchell, Kara K; Nazarian, Elizabeth J; Escuyer, Vincent E; Musser, Kimberlee A

    2015-12-01

    We have developed a single tube TaqMan(®) real-time PCR assay that differentiates the full-length and truncated erm(41) gene to predict inducible resistance to clarithromycin in Mycobacterium abscessus. A study of 87 clinical isolates found this assay to be 90.8% concordant to conventional drug susceptibility testing results for the prediction of inducible clarithromycin drug resistance. PMID:26334290

  4. Activities of a New Fluoroketolide, HMR 3787, and Its (Des)-Fluor Derivative RU 64399 Compared to Those of Telithromycin, Erythromycin A, Azithromycin, Clarithromycin, and Clindamycin against Macrolide-Susceptible or -Resistant Streptococcus pneumoniae and S. pyogenes

    PubMed Central

    Nagai, Kensuke; Davies, Todd A.; Ednie, Lois M.; Bryskier, Andre; Palavecino, Elizabeth; Jacobs, Michael R.; Appelbaum, Peter C.

    2001-01-01

    Activities of HMR 3787 and RU 64399 were compared to those of three macrolides, telithromycin, and clindamycin against 175 Streptococcus pneumoniae isolates and 121 Streptococcus pyogenes isolates. HMR3787 and telithromycin were the most active compounds tested against pneumococci. Telithromycin and RU 64399 were equally active against macrolide-susceptible (MICs, 0.008 to 0.06 μg/ml) and -resistant S. pyogenes isolates, but HMR 3787 had lower MICs for ermB strains. PMID:11600391

  5. Marine gravity image available

    NASA Astrophysics Data System (ADS)

    The image below shows the gravity field from 30-72°S computed from Geosat geodetic mission (GM) and exact repeat mission (ERM) data. A color shaded-relief image of these gravity anomalies is available from NOAA in poster form (report MGG-8, [Marks et al., 1993] and also as a digital gridded data set on CD-ROM. To order, contact the National Geophysical Data Center, E/GC3, 325 Broadway, Boulder, CO 80303.

  6. On the deduction of chemical reaction pathways from measurements of time series of concentrations

    NASA Astrophysics Data System (ADS)

    Samoilov, Michael; Arkin, Adam; Ross, John

    2001-03-01

    We discuss the deduction of reaction pathways in complex chemical systems from measurements of time series of chemical concentrations of reacting species. First we review a technique called correlation metric construction (CMC) and show the construction of a reaction pathway from measurements on a part of glycolysis. Then we present two new improved methods for the analysis of time series of concentrations, entropy metric construction (EMC), and entropy reduction method (ERM), and illustrate (EMC) with calculations on a model reaction system.

  7. The four candidate Earth Explorer core missions. Report for mission selection. 3. Earth Radiation Mission.

    NASA Astrophysics Data System (ADS)

    Ingmann, P.; Poiares Baptista, J. P.

    1999-07-01

    The primary aim of the ERM is to determine world-wide the vertical profiles of aerosol and cloud field characteristics to provide basic input data for numerical modelling and atmospheric studies. The mission supports the goals of the World Climate Research Programme (WCRP) and, in particular, of its sub-programme Global Energy and Water Experiment (GEWEX), which is intended to develop an improved understanding of energy and water fluxes within the climate system, to secure reliable forecasts of weather and climate.

  8. Signatures of reionization on Lyα emitters

    NASA Astrophysics Data System (ADS)

    Dayal, Pratika; Ferrara, Andrea; Gallerani, Simona

    2008-10-01

    We use a semi-analytic model of Lyα emitters (LAEs) to constrain the reionization history. By considering two physically motivated scenarios in which reionization ends either early [early reionization model (ERM), zi ~ 7] or late [late reionization model (LRM), zi ~ 6], we fix the global value of the intergalactic medium neutral fraction (e.g. χHI = 3 × 10-4, 0.15 at z = 6.56 for the ERM and LRM, respectively) leaving only the star formation efficiency and the effective escape fraction of Lyα photons as free parameters. The ERM fits the observed LAE luminosity function (LF) at z = 5.7 and 6.56 requiring no redshift evolution or mass dependence of the star formation efficiency, and LAE star formation rates (SFR) of , contributing ~8 per cent of the cosmic SFR density at z = 5.7. The LRM requires a physically uncomfortable drop of ~4.5 times in the SFR of the emitters from z = 6.5 to 5.7. Thus, the data seem to imply that the Universe was already highly ionized at z = 6.56. The mass-dependent Lyα transmissivity is 0.36 <~ Tα <~ 0.51 (ERM) and Tα <~ 0.26 (LRM) at z = 6.56. The LF data at z = 4.5 imply an extra Lyα line damping factor of ~ 0.25 possibly due to dust; the presence of a (clumpy) dust component with E(B - V) <~ 0.28 is also required to reproduce the observed large Lyα equivalent widths at the same redshift. Additional useful information can be extracted from the line profile (weighted) skewness, found to be SW = 10-17 Å for the two reionization models, which shows an interesting Lα - χHI anti-correlation, holding under the model assumptions. The shortcomings of the model and strategies to overcome them are discussed.

  9. Umsetzung der Unternehmensstrategie mit der Balanced Scorecard

    NASA Astrophysics Data System (ADS)

    Crespo, Isabel; Bergmann, Lars; Portmann, Stefan; Lacker, Thomas; Lacker, Michael; Fleischmann, Jürgen; Kozó, Hans

    Die Balanced Scorecard (BSC) ist ein Ansatz zum strategischen Management, der neben der Ausrichtung des Unternehmens auf finanzielle Zielwerte ebenso großes Gewicht auf so genannte weiche Faktoren legt, die den wirtschaftlichen Erfolg eines Unternehmens erst ermöglichen. Das entscheidende Merkmal der Balanced Scorecard ist dabei, dass sie ein ausgewogenes System strategischer Ziele herstellt, welches das Unternehmen hinsichtlich der vier Perspektiven Finanzen, Kunden, interne Prozesse und Mitarbeiter und Potenziale strategisch ausrichtet (Kaplan u. Norton 1997).

  10. Drug Resistance Characteristics and Macrolide-Resistant Mechanisms of Streptococcus pneumoniae in Wenzhou City, China.

    PubMed

    Hu, Dakang; Sun, Zheng; Luo, Xinhua; Liu, Shuangchun; Yu, Lianhua; Qu, Ying; Yang, Jinhong; Yu, Jian; Li, Xiangyang; Zhang, Jin

    2016-01-01

    BACKGROUND Streptococcus pneumoniae (SP) is a Gram-positive, alpha-hemolytic, facultative anaerobic member of the genus Streptococcus. The erythromycin-resistant methylase (erm) gene and macrolide efflux (mef) gene are the 2 main genes that can mediate SP. Transposon (Tn) also plays an important role in the collection and metastasis of the gene. In the present study we investigated the drug resistance characteristics and the macrolide-resistant mechanisms of SP in Wenzhou City, China. MATERIAL AND METHODS Sixty-eight strains of SP were isolated from sputum samples of hospitalized children in the Second Affiliated Hospital of Wenzhou Medical University. These strains were analyzed using antimicrobial susceptibility tests to determine their drug resistance to 10 kinds of antibacterials. Macrolide-resistant phenotypes were identified using K-B method. PCR method was used to analyze the erm B gene, mef A gene, and int Tn gene. RESULTS Drug resistance rates of 68 strains of SP were 98.5%, 100.0%, 63.2%, 52.9%, 94.1%, 89.7%, 0.0%, 0.0%, 16.2%, and 14.7% for clindamycin, erythromycin, penicillin G, cefotaxime, tetracycline, sulfamethoxazole/trimethoprim, levofloxacin, vancomycin, chloramphenicol, and amoxicillin, respectively. Total detection rates of the erm B gene, mef A gene, and int Tn gene were 98.5%, 91.2%, and 100.0%, respectively. CONCLUSIONS SP shows significant multi-drug resistance in Wenzhou City, whereas there is no clinical value of macrolides antibiotics for SP. cMLSB mediated by erm B gene is the most predominant phenotype among macrolide-resistant SP. The int Tn gene may play an important role in horizontal transfer and clonal dissemination of SP drug resistance genes in Wenzhou City. PMID:27483416

  11. Improving the reproducibility of the NAP1/B1/027 epidemic strain R20291 in the hamster model of infection.

    PubMed

    Kelly, Michelle L; Ng, Yen Kuan; Cartman, Stephen T; Collery, Mark M; Cockayne, Alan; Minton, Nigel P

    2016-06-01

    Comparative analysis of the Clostridium difficile BI/NAP1/027 strain R20291 and ClosTron-derived ermB mutants in the hamster infection model are compromised by the clindamycin susceptibility of the parent. Mutants can appear more virulent. We have rectified this anomaly by genome engineering. The variant created (CRG20291) represents an ideal control strain for virulence assays of ClosTron mutants. PMID:26946361

  12. Salinity-dependent Toxicity Assay of Silver Nanocolloids Using Medaka Eggs.

    PubMed

    Kataoka, Chisato; Kashiwada, Shosaku

    2016-01-01

    Salinity is an important characteristic of the aquatic environment. For aquatic organisms it defines the habitats of freshwater, brackish water, and seawater. Tests of the toxicity of chemicals and assessments of their ecological risks to aquatic organisms are frequently performed in freshwater, but the toxicity of chemicals to aquatic organisms depends on pH, temperature, and salinity. There is no method, however, for testing the salinity dependence of toxicity to aquatic organisms. Here, we used medaka (Oryzias latipes) because they can adapt to freshwater, brackish water, and seawater. Different concentrations of embryo-rearing medium (ERM) (1x, 5x, 10x, 15x, 20x, and 30x) were employed to test the toxicity of silver nanocolloidal particles (SNCs) to medaka eggs (1x ERM and 30x ERM have osmotic pressures equivalent to freshwater and seawater, respectively). In six-well plastic plates, 15 medaka eggs in triplicate were exposed to SNCs at 10 mg/L(-1) in different concentrations of ERM at pH 7 and 25 °C in the dark. We used a dissecting microscope and a micrometer to measure heart rate per 15 sec and eye diameter on day 6 and full body length of the larvae on hatching day (section 4). The embryos were observed until hatching or day 14; we then counted the hatching rate every day for 14 days (section 4). To see silver accumulation in embryos, we used inductively coupled plasma mass spectrometry to measure the silver concentration of test solutions (section 5) and dechorionated embryos (section 6).The toxicity of the SNCs to medaka embryos obviously increased with increasing salinity. This new method allows us to test the toxicity of chemicals in different salinities. PMID:27023020

  13. Peroxisome Proliferator–Activated Receptor α Protects Renal Tubular Cells from Gentamicin-Induced Apoptosis via Upregulating Na+/H+ Exchanger NHE1

    PubMed Central

    Chen, Cheng-Hsien; Chen, Tso-Hsiao; Wu, Mei-Yi; Chen, Jia-Rung; Hong, Li-Yu; Zheng, Cai-Mei; Chiu, I-Jen; Lin, Yuh-Feng; Hsu, Yung-Ho

    2015-01-01

    Peroxisome proliferator–activated receptor (PPAR)-α is a transcription factor that has been reported to inhibit gentamicin-induced apoptosis in renal tubular cells. However, the antiapoptotic mechanism of PPARα is still unknown. In this study, we found that PPARα overexpression induced Na+/H+ exchanger-1 (NHE1) expression in the rat renal tubular cells NRK-52E. Beraprost, a PPARα ligand, also increased NHE1 expression in the renal tubules in normal mice, but not in PPARα knockout mice. Chromatin immunoprecipitation assays revealed that two PPARα binding elements were located in the rat NHE1 promoter region. Na+/H+ exchanger activity also increased in the PPARα-overexpressed cells. Flow cytometry showed that the PPARα-overexpressed cells were resistant to apoptosis-induced shrinkage. Cariporide, a selective NHE1 inhibitor, inhibited the antiapoptotic effect of PPARα in the gentamicin-treated cells. The interaction between NHE1 and ezrin/radixin/moesin (ERM) and between ERM and phosphatidylinositol 4,5-bisphosphate in the PPARα-overexpressed cells was more than in the control cells. ERM short interfering RNA (siRNA) transfection inhibited the PPARα-induced antiapoptotic effect. PPARα overexpression also increased the phosphoinositide 3-kinase (PI3K) expression, which is dependent on NHE1 activity. Increased PI3K further increased the phosphorylation of the prosurvival kinase Akt in the PPARα-overexpressed cells. Wortmannin, a PI3K inhibitor, inhibited PPARα-induced Akt activity and the antiapoptotic effect. We conclude that PPARα induces NHE1 expression and then recruits ERM to promote PI3K/Akt-mediated cell survival in renal tubular cells. The application of PPARα activation reduces the nephrotoxicity of gentamicin and may expand the clinical use of gentamicin. PMID:26623927

  14. The Engineering Radiation Monitor for the Radiation Belt Storm Probes Mission

    NASA Astrophysics Data System (ADS)

    Goldsten, J. O.; Maurer, R. H.; Peplowski, P. N.; Holmes-Siedle, A. G.; Herrmann, C. C.; Mauk, B. H.

    2013-11-01

    An Engineering Radiation Monitor (ERM) has been developed as a supplementary spacecraft subsystem for NASA's Radiation Belt Storm Probes (RBSP) mission. The ERM will monitor total dose and deep dielectric charging at each RBSP spacecraft in real time. Configured to take the place of spacecraft balance mass, the ERM contains an array of eight dosimeters and two buried conductive plates. The dosimeters are mounted under covers of varying shielding thickness to obtain a dose-depth curve and characterize the electron and proton contributions to total dose. A 3-min readout cadence coupled with an initial sensitivity of ˜0.01 krad should enable dynamic measurements of dose rate throughout the 9-hr RBSP orbit. The dosimeters are Radiation-sensing Field Effect Transistors (RadFETs) and operate at zero bias to preserve their response even when powered off. The range of the RadFETs extends above 1000 krad to avoid saturation over the expected duration of the mission. Two large-area (˜10 cm2) charge monitor plates set behind different thickness covers will measure the dynamic currents of weakly-penetrating electrons that can be potentially hazardous to sensitive electronic components within the spacecraft. The charge monitors can handle large events without saturating (˜3000 fA/cm2) and provide sufficient sensitivity (˜0.1 fA/cm2) to gauge quiescent conditions. High time-resolution (5 s) monitoring allows detection of rapid changes in flux and enables correlation of spacecraft anomalies with local space weather conditions. Although primarily intended as an engineering subsystem to monitor spacecraft radiation levels, real-time data from the ERM may also prove useful or interesting to a larger community.

  15. Survival of two bacterial fish pathogens (Aeromonas salmonicida and the Enteric Redmouth Bacterium) in ozonated, chlorinated, and untreated waters

    USGS Publications Warehouse

    Wedemeyer, Gary A.; Nelson, Nancy C.

    1977-01-01

    Ozone and chlorine inactivation curves were determined in three water types at 20 °C for the destruction of the fish pathogens Aeromonas salmonicida, the etiologic agent of furunculosis, and the enteric redmouth bacterium (ERM). In phosphate-buffered distilled water, 0.01 mg/ℓ ozone inactivated 103 cells/ml of ERM and A. salmonicida in 1/2 and 10 min, respectively. Chlorine at this concentration had little effect on either pathogen and a residual of at least 0.05 mg/ℓ was needed to achieve a complete kill within a 10-min contact time. In soft lake water (30 mg/ℓ as CaCO3) a chlorine residual of 0.1 mg/ℓ rapidly  inactivated A. salmonicida and ERM but in hard water (120 mg/ℓ) A. salmonicida was more resistant and 0.2 mg/ℓ chlorine was required. Ozonation of the two lake waters at 90 mg O3∙h−1∙ℓ−1 (equivalent to a 0.01 mg/ℓ residual in ozone demand-free water) was required to destroy both pathogens within 10 min.In untreated soft lake water 103 cells/ml of A. salmonicida survived only 2 days, while the ERM bacterium (103 cells/ml) survived even after 20 day s in soft and hard untreated lake waters.

  16. Complete Genome Sequence of Corynebacterium imitans DSM 44264, Isolated from a Five-Month-Old Boy with Suspected Pharyngeal Diphtheria

    PubMed Central

    Möllmann, Sabrina; Albersmeier, Andreas; Rückert, Christian

    2014-01-01

    The complete genome sequence of the type strain Corynebacterium imitans DSM 44264 comprises 2,565,321 bp with a mean G+C content of 64.26%. The detection of the antibiotic resistance genes erm(X), aphA1-IAB, strA-strB, and cmx is fully consistent with the previously observed multidrug-resistant pattern of C. imitans isolates. PMID:25414508

  17. Methylation of 23S rRNA Nucleotide G748 by RlmAII Methyltransferase Renders Streptococcus pneumoniae Telithromycin Susceptible

    PubMed Central

    Sato, Yoshiharu; Shoji, Tatsuma; Yamamoto, Tomoko

    2013-01-01

    Several posttranscriptional modifications of bacterial rRNAs are important in determining antibiotic resistance or sensitivity. In all Gram-positive bacteria, dimethylation of nucleotide A2058, located in domain V of 23S rRNA, by the dimethyltransferase Erm(B) results in low susceptibility and resistance to telithromycin (TEL). However, this is insufficient to produce high-level resistance to TEL in Streptococcus pneumoniae. Inactivation of the methyltransferase RlmAII, which methylates the N-1 position of nucleotide G748, located in hairpin 35 of domain II of 23S rRNA, results in increased resistance to TEL in erm(B)-carrying S. pneumoniae. Sixteen TEL-resistant mutants (MICs, 16 to 32 μg/ml) were obtained from a clinically isolated S. pneumoniae strain showing low TEL susceptibility (MIC, 2 μg/ml), with mutation resulting in constitutive dimethylation of A2058 because of nucleotide differences in the regulatory region of erm(B) mRNA. Primer extension analysis showed that the degree of methylation at G748 in all TEL-resistant mutants was significantly reduced by a mutation in the gene encoding RlmAII to create a stop codon or change an amino acid residue. Furthermore, RNA footprinting with dimethyl sulfate and a molecular modeling study suggested that methylation of G748 may contribute to the stable interaction of TEL with domain II of 23S rRNA, even after dimethylation of A2058 by Erm(B). This novel finding shows that methylation of G748 by RlmAII renders S. pneumoniae TEL susceptible. PMID:23716046

  18. Determination of the prevalence of antimicrobial resistance genes in canine Clostridium perfringens isolates.

    PubMed

    Kather, Elizabeth J; Marks, Stanley L; Foley, Janet E

    2006-03-10

    Clostridium perfringens is a well documented cause of a mild self-limiting diarrhea and a potentially fatal acute hemorrhagic diarrheal syndrome in the dog. A recent study documented that 21% of canine C. perfringens isolates had MIC's indicative of resistance to tetracycline, an antimicrobial commonly recommended for treatment of C. perfringens-associated diarrhea. The objective of the present study was to further evaluate the antimicrobial susceptibility profiles of these isolates by determining the prevalence of specific resistance genes, their expression, and ability for transference between bacteria. One hundred and twenty-four canine C. perfringens isolates from 124 dogs were evaluated. Minimum inhibitory concentrations of tetracycline, erythromycin, tylosin, and metronidazole were determined using the CLSI Reference Agar Dilution Method. All isolates were screened for three tetracycline resistance genes: tetA(P), tetB(P) and tetM, and two macrolide resistance genes: ermB and ermQ, via PCR using primer sequences previously described. Ninety-six percent (119/124) of the isolates were positive for the tetA(P) gene, and 41% (51/124) were positive for both the tetA(P) and tetB(P) genes. No isolates were positive for the tetB(P) gene alone. Highly susceptible isolates (MIC< or = 4 microg/ml) were significantly more likely to lack the tetB(P) gene. One isolate (0.8%) was positive for the ermB gene, and one isolate was positive for the ermQ gene. The tetM gene was not found in any of the isolates tested. Two out of 15 tested isolates (13%) demonstrated transfer of tetracycline resistance via bacterial conjugation. Tetracycline should be avoided for the treatment of C. perfringens-associated diarrhea in dogs because of the relatively high prevalence of in vitro resistance, and the potential for conjugative transfer of antimicrobial resistance. PMID:16330169

  19. Sialidase and Sialoglycoproteases Can Modulate Virulence in Porphyromonas gingivalis ▿ †

    PubMed Central

    Aruni, Wilson; Vanterpool, Elaine; Osbourne, Devon; Roy, Francis; Muthiah, Arun; Dou, Yuetan; Fletcher, Hansel M.

    2011-01-01

    The Porphyromonas gingivalis recombinant VimA can interact with the gingipains and several other proteins, including a sialidase. Sialylation can be involved in protein maturation; however, its role in virulence regulation in P. gingivalis is unknown. The three sialidase-related proteins in P. gingivalis showed the characteristic sialidase Asp signature motif (SXDXGXTW) and other unique domains. To evaluate the roles of the associated genes, randomly chosen P. gingivalis isogenic mutants created by allelic exchange and designated FLL401 (PG0778::ermF), FLL402 (PG1724::ermF), and FLL403 (PG0352::ermF-ermAM) were characterized. Similar to the wild-type strain, FLL402 and FLL403 displayed a black-pigmented phenotype in contrast to FLL401, which was not black pigmented. Sialidase activity in P. gingivalis FLL401 was reduced by approximately 70% in comparison to those in FLL402 and FLL403, which were reduced by approximately 42% and 5%, respectively. Although there were no changes in the expression of the gingipain genes, their activities were reduced by 60 to 90% in all the isogenic mutants compared to that for the wild type. Immunoreactive bands representing the catalytic domains for RgpA, RgpB, and Kgp were present in FLL402 and FLL403 but were missing in FLL401. While adhesion was decreased, the capacity for invasion of epithelial cells by the isogenic mutants was increased by 11 to 16% over that of the wild-type strain. Isogenic mutants defective in PG0778 and PG0352 were more sensitive to hydrogen peroxide than the wild type. Taken together, these results suggest that the P. gingivalis sialidase activity may be involved in regulating gingipain activity and other virulence factors and may be important in the pathogenesis of this organism. PMID:21502589

  20. Trypanosomal TAC40 constitutes a novel subclass of mitochondrial β-barrel proteins specialized in mitochondrial genome inheritance

    PubMed Central

    Schnarwiler, Felix; Niemann, Moritz; Doiron, Nicholas; Harsman, Anke; Käser, Sandro; Mani, Jan; Chanfon, Astrid; Dewar, Caroline E.; Oeljeklaus, Silke; Jackson, Christopher B.; Pusnik, Mascha; Schmidt, Oliver; Meisinger, Chris; Hiller, Sebastian; Warscheid, Bettina; Schnaufer, Achim C.; Ochsenreiter, Torsten; Schneider, André

    2014-01-01

    Mitochondria cannot form de novo but require mechanisms allowing their inheritance to daughter cells. In contrast to most other eukaryotes Trypanosoma brucei has a single mitochondrion whose single-unit genome is physically connected to the flagellum. Here we identify a β-barrel mitochondrial outer membrane protein, termed tripartite attachment complex 40 (TAC40), that localizes to this connection. TAC40 is essential for mitochondrial DNA inheritance and belongs to the mitochondrial porin protein family. However, it is not specifically related to any of the three subclasses of mitochondrial porins represented by the metabolite transporter voltage-dependent anion channel (VDAC), the protein translocator of the outer membrane 40 (TOM40), or the fungi-specific MDM10, a component of the endoplasmic reticulum–mitochondria encounter structure (ERMES). MDM10 and TAC40 mediate cellular architecture and participate in transmembrane complexes that are essential for mitochondrial DNA inheritance. In yeast MDM10, in the context of the ERMES, is postulated to connect the mitochondrial genomes to actin filaments, whereas in trypanosomes TAC40 mediates the linkage of the mitochondrial DNA to the basal body of the flagellum. However, TAC40 does not colocalize with trypanosomal orthologs of ERMES components and, unlike MDM10, it regulates neither mitochondrial morphology nor the assembly of the protein translocase. TAC40 therefore defines a novel subclass of mitochondrial porins that is distinct from VDAC, TOM40, and MDM10. However, whereas the architecture of the TAC40-containing complex in trypanosomes and the MDM10-containing ERMES in yeast is very different, both are organized around a β-barrel protein of the mitochondrial porin family that mediates a DNA–cytoskeleton linkage that is essential for mitochondrial DNA inheritance. PMID:24821793

  1. Nasal Carriage of Staphylococcus aureus: Frequency and Antibiotic Resistance in Healthy Ruminants

    PubMed Central

    Rahimi, Heidar; Dastmalchi Saei, Habib; Ahmadi, Malahat

    2015-01-01

    Background: Staphylococcus aureus is a significant pathogen that can colonize the nares of different animals, causing a wide range of infections in various hosts. Objectives: We intended to determine the prevalence of S. aureus in the nasal cavity of healthy ruminants and also to investigate the presence of antibiotic resistance genes. Materials and Methods: In the present study, healthy cattle (n = 79), sheep (n = 78) and goats (n = 44) were screened for nasal carriage of S. aureus by the Polymerase Chain Reaction (PCR). Staphylococcus aureus isolates were further assessed for the presence of blaZ (encoding penicillin resistance), mecA (encoding methicillin resistance), tetK and tetM (encoding tetracycline resistance), and ermA and ermC (encoding macrolide-lincosamide-streptogramin B resistance) genes. Results: The proportion of S. aureus-positive nasal swabs from cattle, sheep and goats were four (5.06%), 11 (14.1%) and 11 isolates (25%), respectively. The blaZ gene was detected in 20 out of 26 S. aureus isolates (76.9%), including four cattle (100%), nine sheep (81.8%) and seven goats (63.6%). Two of the four cattle isolates possessing the blaZ gene also had the tetK gene. Of the nine sheep isolates harboring the blaZ gene, one possessed the mecA and tetK genes together. Of the seven goat isolates with blaZ gene, one harbored the tetM gene. None of the S. aureus isolates were positive for the ermA and ermC genes. Conclusions: In contrast to cattle, S. aureus is frequently present in the nose of sheep and goats, which may represent the primary reservoir of S. aureus in small ruminant flocks. This study also showed that nasal isolates of S. aureus from healthy ruminants might be a potential reservoir of antimicrobial-resistance. PMID:26568802

  2. Combinations of Macrolide Resistance Determinants in Field Isolates of Mannheimia haemolytica and Pasteurella multocida▿

    PubMed Central

    Desmolaize, Benoit; Rose, Simon; Wilhelm, Cornelia; Warrass, Ralf; Douthwaite, Stephen

    2011-01-01

    Respiratory tract infections in cattle are commonly associated with the bacterial pathogens Mannheimia haemolytica and Pasteurella multocida. These infections can generally be successfully treated in the field with one of several groups of antibiotics, including macrolides. A few recent isolates of these species exhibit resistance to veterinary macrolides with phenotypes that fall into three distinct classes. The first class has type I macrolide, lincosamide, and streptogramin B antibiotic resistance and, consistent with this, the 23S rRNA nucleotide A2058 is monomethylated by the enzyme product of the erm(42) gene. The second class shows no lincosamide resistance and lacks erm(42) and concomitant 23S rRNA methylation. Sequencing of the genome of a representative strain from this class, P. multocida 3361, revealed macrolide efflux and phosphotransferase genes [respectively termed msr(E) and mph(E)] that are arranged in tandem and presumably expressed from the same promoter. The third class exhibits the most marked drug phenotype, with high resistance to all of the macrolides tested, and possesses all three resistance determinants. The combinations of erm(42), msr(E), and mph(E) are chromosomally encoded and intermingled with other exogenous genes, many of which appear to have been transferred from other members of the Pasteurellaceae. The presence of some of the exogenous genes explains recent reports of resistance to additional drug classes. We have expressed recombinant versions of the erm(42), msr(E), and mph(E) genes within an isogenic Escherichia coli background to assess their individually contributions to resistance. Our findings indicate what types of compounds might have driven the selection for these resistance determinants. PMID:21709086

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

    PubMed

    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

  4. Drug Resistance Characteristics and Macrolide-Resistant Mechanisms of Streptococcus pneumoniae in Wenzhou City, China

    PubMed Central

    Hu, Dakang; Sun, Zheng; Luo, Xinhua; Liu, Shuangchun; Yu, Lianhua; Qu, Ying; Yang, Jinhong; Yu, Jian; Li, Xiangyang; Zhang, Jin

    2016-01-01

    Background Streptococcus pneumoniae (SP) is a Gram-positive, alpha-hemolytic, facultative anaerobic member of the genus Streptococcus. The erythromycin-resistant methylase (erm) gene and macrolide efflux (mef) gene are the 2 main genes that can mediate SP. Transposon (Tn) also plays an important role in the collection and metastasis of the gene. In the present study we investigated the drug resistance characteristics and the macrolide-resistant mechanisms of SP in Wenzhou City, China. Material/Methods Sixty-eight strains of SP were isolated from sputum samples of hospitalized children in the Second Affiliated Hospital of Wenzhou Medical University. These strains were analyzed using antimicrobial susceptibility tests to determine their drug resistance to 10 kinds of antibacterials. Macrolide-resistant phenotypes were identified using K-B method. PCR method was used to analyze the erm B gene, mef A gene, and int Tn gene. Results Drug resistance rates of 68 strains of SP were 98.5%, 100.0%, 63.2%, 52.9%, 94.1%, 89.7%, 0.0%, 0.0%, 16.2%, and 14.7% for clindamycin, erythromycin, penicillin G, cefotaxime, tetracycline, sulfamethoxazole/trimethoprim, levofloxacin, vancomycin, chloramphenicol, and amoxicillin, respectively. Total detection rates of the erm B gene, mef A gene, and int Tn gene were 98.5%, 91.2%, and 100.0%, respectively. Conclusions SP shows significant multi-drug resistance in Wenzhou City, whereas there is no clinical value of macrolides antibiotics for SP. cMLSB mediated by erm B gene is the most predominant phenotype among macrolide-resistant SP. The int Tn gene may play an important role in horizontal transfer and clonal dissemination of SP drug resistance genes in Wenzhou City. PMID:27483416

  5. Quantification of lincomycin resistance genes associated with lincomycin residues in waters and soils adjacent to representative swine farms in China

    PubMed Central

    Li, Liang; Sun, Jian; Liu, Baotao; Zhao, Donghao; Ma, Jun; Deng, Hui; Li, Xue; Hu, Fengyang; Liao, Xiaoping; Liu, Yahong

    2013-01-01

    Lincomycin is commonly used on swine farms for growth promotion as well as disease treatment and control. Consequently, lincomycin may accumulate in the environment adjacent to the swine farms in many ways, thereby influencing antibiotic resistance in the environment. Levels of lincomycin-resistance genes and lincomycin residues in water and soil samples collected from multiple sites near wastewater discharge areas were investigated in this study. Sixteen lincomycin-resistance and 16S rRNA genes were detected using real-time PCR. Three genes, lnu(F), erm(A), and erm(B), were detected in all water and soil samples except control samples. Lincomycin residues were determined by rapid resolution liquid chromatography-tandem mass spectrometry, with concentrations detected as high as 9.29 ng/mL in water and 0.97 ng/g in soil. A gradual reduction in the levels of lincomycin-resistance genes and lincomycin residues in the waters and soils were detected from multiple sites along the path of wastewater discharging to the surrounding environment from the swine farms. Significant correlations were found between levels of lincomycin-resistance genes in paired water and soil samples (r = 0.885, p = 0.019), and between lincomycin-resistance genes and lincomycin residues (r = 0.975, p < 0.01). This study emphasized the potential risk of dissemination of lincomycin-resistance genes such as lnu(F), erm(A), and erm(B), associated with lincomycin residues in surrounding environments adjacent to swine farms. PMID:24348472

  6. Zebrafish Rhabdomyosarcoma.

    PubMed

    Phelps, Michael; Chen, Eleanor

    2016-01-01

    In vivo models of Rhabdomyosarcoma (RMS) have proven instrumental in understanding the development and progression of this devastating pediatric sarcoma. Both vertebrate and invertebrate model systems have been developed to study the tumor biology of both embryonal (ERMS) and alveolar (ARMS) RMS subtypes. Zebrafish RMS models have been particularly amenable for high-throughput studies to identify drug targetable pathways because of their short tumor latency, ease of ex vivo manipulation and conserved tumor biology. The transgenic KRASG12D-induced ERMS model allows for molecular and cellular characterization of distinct tumor cell subpopulations including the tumor propagating cells. Comparative genomic approaches have also been utilized in zebrafish ERMS to identify conserved candidate driver genes. Recent advances in zebrafish genome engineering have further enabled the ability to probe the functional significance of potential driver genes. Using the unique strengths of the zebrafish model organisms with the wealth of cellular and molecular tools currently available, zebrafish RMS models provide a powerful in vivo system for which to study RMS tumorigenesis. PMID:27165362

  7. Geostatistical evaluation of satellite radar altimetry for high-resolution mapping of Lambert Glacier, Antarctica

    NASA Technical Reports Server (NTRS)

    Herzfeld, Ute C.; Lingle, Craig S.; Lee, Li-Her

    1993-01-01

    The potential of satellite radar altimetry for high-resolution mapping of Antarctic ice streams is evaluated, using retracked and slope-corrected data from the Lambert Glacier and Amery Ice Shelf area, East Antarctica, acquired by Geosat during the Exact Repeat Mission (ERM), 1986-89. The map area includes lower Lambert Glacier north of 72.18 deg S, the southern Amery Ice Shelf, and the grounded inland ice sheet on both sides. The Geosat ERM altimetry is found to provide substantially more complete coverage than the 1978 Seasat altimetry, due to improved tracking. Variogram methods are used to estimate the noise levels in the data as a function of position throughout the map area. The spatial structure in the data is quantified by constructing experimental variograms using altimetry from the area of the grounding zone of Lambert Glacier, which is the area chiefly of interest in this topographically complex region. Kriging is employed to invert the along-track height measurements onto a fine-scale 3 km grid. The unsmoothed along-track Geosat ERM altimetry yields spatially continuous maps showing the main topographic features of lower Lambert Glacier, upper Amery Ice Shelf and the adjacent inland ice sheet. The probable position of the grounding line of Lambert Glacier is identified from a break in slope at the grounded ice/floating ice transition. The approximate standard error of the kriged map is inferred from the data noise levels.

  8. Constitutive activation of MET signaling impairs myogenic differentiation of rhabdomyosarcoma and promotes its development and progression

    PubMed Central

    Szewczyk, Barbara; Adamus, Tomasz; Lukasiewicz, Ewa; Miekus, Katarzyna; Majka, Marcin

    2015-01-01

    Rhabdomyosarcoma (RMS) is a soft tissue sarcoma, which may originate from impaired differentiation of mesenchymal stem cells (MSC). Expression of MET receptor is elevated in alveolar RMS subtype (ARMS) which is associated with worse prognosis, compared to embryonal RMS (ERMS). Forced differentiation of ARMS cells diminishes MET level and, as shown previously, MET silencing induces differentiation of ARMS. In ERMS cells introduction of TPR-MET oncogene leads to an uncontrolled overstimulation of the MET receptor downstream signaling pathways. In vivo, tumors formed by those cells in NOD-SCID mice display inhibited differentiation, enhanced proliferation, diminished apoptosis and increased infiltration of neutrophils. Consequently, tumors grow significantly faster and they display enhanced ability to metastasize to lungs and to vascularize due to elevated VEGF, MMP9 and miR-378 expression. In vitro, TPR-MET ERMS cells display enhanced migration, chemotaxis and invasion toward HGF and SDF-1. Introduction of TPR-MET into MSC increases survival and may induce expression of early myogenic factors depending on the genetic background, and it blocks terminal differentiation of skeletal myoblasts. To conclude, our results suggest that activation of MET signaling may cause defects in myogenic differentiation leading to rhabdomyosarcoma development and progression. PMID:26384300

  9. DAX-1 Expression in Pediatric Rhabdomyosarcomas: Another Immunohistochemical Marker Useful in the Diagnosis of Translocation Positive Alveolar Rhabdomyosarcoma

    PubMed Central

    Virgone, Calogero; Lalli, Enzo; Bisogno, Gianni; Lazzari, Elena; Roma, Josep; Zin, Angelica; Poli, Elena; Cecchetto, Giovanni; Dall’Igna, Patrizia; Alaggio, Rita

    2015-01-01

    Objectives The aim of this study was to investigate the expression of DAX-1 in a series of pediatric rhabdomyosarcomas (RMS) with known translocation and compare it to Ap2β, known to be selectively expressed in ARMS. Design We revised a series of 71 alveolar rhabdomyosarcomas (ARMS), enrolled in the Italian Protocols RMS 79 and 96, and 23 embryonal rhabdomyosarcomas (ERMS) as controls. Before investigating Ap2β and DAX-1, ARMS were reviewed and reclassified as 48 ARMS and 23 non-ARMS. Results Translocation positive ARMS showed a characteristic Ap2β/DAX-1+ staining pattern in 78% of cases, while 76% of classic ERMS were negative for both. Ap2β alone was positive in 3.9% of RMS lacking translocation, whereas DAX-1 alone was positive in 25.4%. Conversely, 9% and 6% of translocation positive ARMS were positive only for DAX-1 or Ap2β, respectively. The 23 non-ARMS shared the same phenotype as ERMS but had a higher frequency of DAX-1 expression. Conclusions DAX-1 is less specific than Ap2β, however it is a sensitive marker for translocation positive ARMS and can be helpful in their diagnosis if used in combination with Ap2β. PMID:26168243

  10. Constitutive activation of MET signaling impairs myogenic differentiation of rhabdomyosarcoma and promotes its development and progression.

    PubMed

    Skrzypek, Klaudia; Kusienicka, Anna; Szewczyk, Barbara; Adamus, Tomasz; Lukasiewicz, Ewa; Miekus, Katarzyna; Majka, Marcin

    2015-10-13

    Rhabdomyosarcoma (RMS) is a soft tissue sarcoma, which may originate from impaired differentiation of mesenchymal stem cells (MSC). Expression of MET receptor is elevated in alveolar RMS subtype (ARMS) which is associated with worse prognosis, compared to embryonal RMS (ERMS). Forced differentiation of ARMS cells diminishes MET level and, as shown previously, MET silencing induces differentiation of ARMS. In ERMS cells introduction of TPR-MET oncogene leads to an uncontrolled overstimulation of the MET receptor downstream signaling pathways. In vivo, tumors formed by those cells in NOD-SCID mice display inhibited differentiation, enhanced proliferation, diminished apoptosis and increased infiltration of neutrophils. Consequently, tumors grow significantly faster and they display enhanced ability to metastasize to lungs and to vascularize due to elevated VEGF, MMP9 and miR-378 expression. In vitro, TPR-MET ERMS cells display enhanced migration, chemotaxis and invasion toward HGF and SDF-1. Introduction of TPR-MET into MSC increases survival and may induce expression of early myogenic factors depending on the genetic background, and it blocks terminal differentiation of skeletal myoblasts. To conclude, our results suggest that activation of MET signaling may cause defects in myogenic differentiation leading to rhabdomyosarcoma development and progression. PMID:26384300

  11. Role of the YAP Oncoprotein in Priming Ras-Driven Rhabdomyosarcoma

    PubMed Central

    Slemmons, Katherine K.; Crose, Lisa E. S.; Rudzinski, Erin; Bentley, Rex C.; Linardic, Corinne M.

    2015-01-01

    Rhabdomyosarcoma (RMS), a cancer characterized by features of skeletal muscle histogenesis, is the most common soft tissue sarcoma of childhood and adolescence. Survival for high-risk groups is less than 30% at 5 years. RMS also occurs during adulthood, with a lower incidence but higher mortality. Recently, mutational profiling has revealed a correlation between activating Ras mutations in the embryonal (eRMS) and pleomorphic (pRMS) histologic variants of RMS, and a poorer outcome for those patients. Independently, the YAP transcriptional coactivator, an oncoprotein kept in check by the Hippo tumor suppressor pathway, is upregulated in eRMS. Here we show that YAP promotes cell proliferation and antagonizes apoptosis and myogenic differentiation of human RMS cells bearing oncogenic Ras mutations in cell culture studies in vitro and in murine xenografts in vivo. Pharmacologic inhibition of YAP by the benzoporphyrin derivative verteporfin decreased cell proliferation and tumor growth in vivo. To interrogate the temporal contribution of YAP in eRMS tumorigenesis, we used a primary human cell-based genetic model of Ras-driven RMS. Constitutively active YAP functioned as an early genetic lesion, permitting bypass of senescence and priming myoblasts to tolerate subsequent expression of hTERT and oncogenic Ras, which were necessary and sufficient to generate murine xenograft tumors mimicking RMS in vivo. This work provides evidence for cooperation between YAP and oncogenic Ras in RMS tumorigenesis, laying the foundation for preclinical co-targeting of these pathways. PMID:26496700

  12. Multi-Year Elevation Changes Near the West Margin of the Greenland Ice Sheet from Satellite Radar Altimetry

    NASA Technical Reports Server (NTRS)

    Lingle, Craig S.; Brenner, Anita C.; Zwally, H. Jay; DiMarzio, John P.

    1991-01-01

    Mean changes in the surface elevation near the west margin of the Greenland ice sheet are measured using Seasat altimetry and altimetry from the Geosat Exact Repeat Mission (ERM). The Seasat data extend from early July through early October 1978. The ERM data extend from winter 1986-87 through fall 1988. Both seasonal and multi-year changes are measured using altimetry referenced to GEM T2 orbits. The possible effects of orbit error are minimized by adjusting the orbits into a common ocean surface. Seasonal mean changes in the surface height are recognizable during the Geosat ERM. The multi-year measurements indicate the surface was lower by 0.4 +/- 0.4 m on average in late summer 1987 than in late summer 1978. The surface was lower by 0.2 +/- 0.5 m on average in late summer 1988 than in late summer 1978. As a control case, the computations art also carried out using altimetry referenced to orbits not adjusted into a common ocean surface.

  13. The RhoE/ROCK/ARHGAP25 signaling pathway controls cell invasion by inhibition of Rac activity.

    PubMed

    Thuault, Sylvie; Comunale, Franck; Hasna, Jessy; Fortier, Mathieu; Planchon, Damien; Elarouci, Nabila; De Reynies, Aurélien; Bodin, Stéphane; Blangy, Anne; Gauthier-Rouvière, Cécile

    2016-09-01

    Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma of skeletal muscle origin in children and adolescents. Among RMS subtypes, alveolar rhabdomyosarcoma (ARMS), which is characterized by the presence of the PAX3-FOXO1A or PAX7-FOXO1A chimeric oncogenic transcription factor, is associated with poor prognosis and a strong risk of metastasis compared with the embryonal subtype (ERMS). To identify molecular pathways involved in ARMS aggressiveness, we first characterized the migratory behavior of cell lines derived from ARMS and ERMS biopsies using a three-dimensional spheroid cell invasion assay. ARMS cells were more invasive than ERMS cells and adopted an ellipsoidal morphology to efficiently invade the extracellular matrix. Moreover, the invasive potential of ARMS cells depended on ROCK activity, which is regulated by the GTPase RhoE. Specifically, RhoE expression was low in ARMS biopsies, and its overexpression in ARMS cells reduced their invasion potential. Conversely, ARHGAP25, a GTPase-activating protein for Rac, was up-regulated in ARMS biopsies. Moreover, we found that ARHGAP25 inhibits Rac activity downstream of ROCKII and is required for ARMS cell invasion. Our results indicate that the RhoE/ROCK/ARHGAP25 signaling pathway promotes ARMS invasive potential and identify these proteins as potential therapeutic targets for ARMS treatment. PMID:27413008

  14. Yeast Vps13 promotes mitochondrial function and is localized at membrane contact sites.

    PubMed

    Park, Jae-Sook; Thorsness, Mary K; Policastro, Robert; McGoldrick, Luke L; Hollingsworth, Nancy M; Thorsness, Peter E; Neiman, Aaron M

    2016-08-01

    The Vps13 protein family is highly conserved in eukaryotic cells. Mutations in human VPS13 genes result in a variety of diseases, such as chorea acanthocytosis (ChAc), but the cellular functions of Vps13 proteins are not well defined. In yeast, there is a single VPS13 orthologue, which is required for at least two different processes: protein sorting to the vacuole and sporulation. This study demonstrates that VPS13 is also important for mitochondrial integrity. In addition to preventing transfer of DNA from the mitochondrion to the nucleus, VPS13 suppresses mitophagy and functions in parallel with the endoplasmic reticulum-mitochondrion encounter structure (ERMES). In different growth conditions, Vps13 localizes to endosome-mitochondrion contacts and to the nuclear-vacuole junctions, indicating that Vps13 may function at membrane contact sites. The ability of VPS13 to compensate for the absence of ERMES correlates with its intracellular distribution. We propose that Vps13 is present at multiple membrane contact sites and that separation-of-function mutants are due to loss of Vps13 at specific junctions. Introduction of VPS13A mutations identified in ChAc patients at cognate sites in yeast VPS13 are specifically defective in compensating for the lack of ERMES, suggesting that mitochondrial dysfunction might be the basis for ChAc. PMID:27280386

  15. Hepatocyte Growth Factor-mediated satellite cells niche perturbation promotes development of distinct sarcoma subtypes

    PubMed Central

    Morena, Deborah; Maestro, Nicola; Bersani, Francesca; Forni, Paolo Emanuele; Lingua, Marcello Francesco; Foglizzo, Valentina; Šćepanović, Petar; Miretti, Silvia; Morotti, Alessandro; Shern, Jack F; Khan, Javed; Ala, Ugo; Provero, Paolo; Sala, Valentina; Crepaldi, Tiziana; Gasparini, Patrizia; Casanova, Michela; Ferrari, Andrea; Sozzi, Gabriella; Chiarle, Roberto; Ponzetto, Carola; Taulli, Riccardo

    2016-01-01

    Embryonal Rhabdomyosarcoma (ERMS) and Undifferentiated Pleomorphic Sarcoma (UPS) are distinct sarcoma subtypes. Here we investigate the relevance of the satellite cell (SC) niche in sarcoma development by using Hepatocyte Growth Factor (HGF) to perturb the niche microenvironment. In a Pax7 wild type background, HGF stimulation mainly causes ERMS that originate from satellite cells following a process of multistep progression. Conversely, in a Pax7 null genotype ERMS incidence drops, while UPS becomes the most frequent subtype. Murine EfRMS display genetic heterogeneity similar to their human counterpart. Altogether, our data demonstrate that selective perturbation of the SC niche results in distinct sarcoma subtypes in a Pax7 lineage-dependent manner, and define a critical role for the Met axis in sarcoma initiation. Finally, our results provide a rationale for the use of combination therapy, tailored on specific amplifications and activated signaling pathways, to minimize resistance emerging from sarcomas heterogeneity. DOI: http://dx.doi.org/10.7554/eLife.12116.001 PMID:26987019

  16. Molecular identification and quantification of tetracycline and erythromycin resistance genes in Spanish and Italian retail cheeses.

    PubMed

    Belén Flórez, Ana; Alegría, Ángel; Rossi, Franca; Delgado, Susana; Felis, Giovanna E; Torriani, Sandra; Mayo, Baltasar

    2014-01-01

    Large antibiotic resistance gene pools in the microbiota of foods may ultimately pose a risk for human health. This study reports the identification and quantification of tetracycline- and erythromycin-resistant populations, resistance genes, and gene diversity in traditional Spanish and Italian cheeses, via culturing, conventional PCR, real-time quantitative PCR (qPCR), and denaturing gradient gel electrophoresis (DGGE). The numbers of resistant bacteria varied widely among the antibiotics and the different cheese varieties; in some cheeses, all the bacterial populations seemed to be resistant. Up to eight antibiotic resistance genes were sought by gene-specific PCR, six with respect to tetracycline, that is, tet(K), tet(L), tet(M), tet(O), tet(S), and tet(W), and two with respect to erythromycin, that is, erm(B) and erm(F). The most common resistance genes in the analysed cheeses were tet(S), tet(W), tet(M), and erm(B). The copy numbers of these genes, as quantified by qPCR, ranged widely between cheeses (from 4.94 to 10.18log10/g). DGGE analysis revealed distinct banding profiles and two polymorphic nucleotide positions for tet(W)-carrying cheeses, though the similarity of the sequences suggests this tet(W) to have a monophyletic origin. Traditional cheeses would therefore appear to act as reservoirs for large numbers of many types of antibiotic resistance determinants. PMID:25302306

  17. Role of the YAP Oncoprotein in Priming Ras-Driven Rhabdomyosarcoma.

    PubMed

    Slemmons, Katherine K; Crose, Lisa E S; Rudzinski, Erin; Bentley, Rex C; Linardic, Corinne M

    2015-01-01

    Rhabdomyosarcoma (RMS), a cancer characterized by features of skeletal muscle histogenesis, is the most common soft tissue sarcoma of childhood and adolescence. Survival for high-risk groups is less than 30% at 5 years. RMS also occurs during adulthood, with a lower incidence but higher mortality. Recently, mutational profiling has revealed a correlation between activating Ras mutations in the embryonal (eRMS) and pleomorphic (pRMS) histologic variants of RMS, and a poorer outcome for those patients. Independently, the YAP transcriptional coactivator, an oncoprotein kept in check by the Hippo tumor suppressor pathway, is upregulated in eRMS. Here we show that YAP promotes cell proliferation and antagonizes apoptosis and myogenic differentiation of human RMS cells bearing oncogenic Ras mutations in cell culture studies in vitro and in murine xenografts in vivo. Pharmacologic inhibition of YAP by the benzoporphyrin derivative verteporfin decreased cell proliferation and tumor growth in vivo. To interrogate the temporal contribution of YAP in eRMS tumorigenesis, we used a primary human cell-based genetic model of Ras-driven RMS. Constitutively active YAP functioned as an early genetic lesion, permitting bypass of senescence and priming myoblasts to tolerate subsequent expression of hTERT and oncogenic Ras, which were necessary and sufficient to generate murine xenograft tumors mimicking RMS in vivo. This work provides evidence for cooperation between YAP and oncogenic Ras in RMS tumorigenesis, laying the foundation for preclinical co-targeting of these pathways. PMID:26496700

  18. [Studies on the possible application of molecular methods in diagnosing carriers and in similarity analysis of group B streptococci (Streptococcus agalactiae)].

    PubMed

    Brzychczy-Włoch, Monika; Strus, Magdalena; Pawlik, Dorota; Gosiewski, Tomasz; Krzysztof, Rytlewski; Drzewiecki, Artur; Lauterbach, Ryszard; Heczko, Piotr B

    2008-01-01

    The most popular method of GBS identification in Poland currently is by culturing on enriched agar and verifying the Lancefield Group using special latex agglutination kits. However, the classical methods are time-consuming and their sensitivity is insufficient therefore it is becoming more common to try and apply molecular methods which are characterized by high sensitivity and rapid results. Moreover, molecular methods give us the possibility to carry out epidemiological investigations and gene detection, for instance for antibiotic resistance. It was confirmed that PCR and FISH procedures may be effective in rapid detection of GBS. Thanks to RAPD methods we showed that newborns born to colonized mothers were colonized by GBS strains which originated from the mother, irrespective of the way and the course of labour. Additionally, we detected GBS colonization in children who were born to mothers who were not colonized by GBS. These children were probably colonized with strains coming from hospital environment. More studies are needed to elucidate the route of transmission and the role of colonization of the medical staff. Using multiplex PCR we showed the presence of ermA, ermB and ermC genes in phenotypically confirmed MLS, GBS strains. PMID:18819445

  19. Antimicrobial susceptibility and analysis of macrolide resistance genes in Streptococcus pneumoniae isolated in Hamadan

    PubMed Central

    Mosleh, Mohammad Najafi; Gharibi, Marzieh; Alikhani, Mohammad Yousef; Saidijam, Massoud; Vakhshiteh, Faezeh

    2014-01-01

    Objective(s): Macrolide resistant Streptococcus pneumoniae pose an emerging problem globally. The aim of this study was to investigate the prevalence of ermB and mefA genes (macrolide resistant genes) by polymerase chain reaction (PCR) method and to detect drug resistance patterns of S. pneumoniae isolated from clinical samples to macrolides and other antibiotic agents by E-test method. Materials and Methods: Fifty five isolates of S. pneumoniae were obtained from clinical samples with microbial tests. The antibiotic susceptibility of isolates for erythromycin, azithromycin, clarithromycin, ceftazidime, ciprofloxacin and vancomycin were determined by E-test method. Genotypic antibiotic resistance pattern was determined by PCR with primer designed for ermB and mefA genes. Results: The number of S. pneumoniae isolates resistance to erythromycin, azithromycin, clarithromycin, ceftazidim, ciprofloxacin were 25.5%, 18.2%, 16.4%, 21.8% and 10.9%, respectively while no resistance to vancomycin was observed. The macrolide resistance genes of ermB and mefA were found in 10.9% and 18.2% of the isolates, respectively. Conclusion: The result of the current study suggests the necessity of evaluation the changes in MIC (minimum inhibitory concentration) values as well as genetic mutations to estimate the prevalence of the resistance antimicrobial agents in S. pneumoniae. PMID:25422753

  20. Optical Coherence Tomography Evaluation in the Multicenter Uveitis Steroid Treatment (MUST) Trial

    PubMed Central

    Domalpally, Amitha; Altaweel, Michael M.; Kempen, John H.; Myers, Dawn; Davis, Janet L; Foster, C Stephen; Latkany, Paul; Srivastava, Sunil K.; Stawell, Richard J.; Holbrook, Janet T.

    2013-01-01

    Purpose To describe the evaluation of optical coherence tomography (OCT) scans in the Muliticenter Uveitis Steroid Treatment (MUST) trial and report baseline OCT features of enrolled participants. Methods Time domain OCTs acquired by certified photographers using a standardized scan protocol were evaluated at a Reading Center. Accuracy of retinal thickness data was confirmed with quality evaluation and caliper measurement of centerpoint thickness (CPT) was performed when unreliable. Morphological evaluation included cysts, subretinal fluid,epiretinal membranes (ERMs),and vitreomacular traction. Results Of the 453 OCTs evaluated, automated retinal thickness was accurate in 69.5% of scans, caliper measurement was performed in 26%,and 4% were ungradable. Intraclass correlation was 0.98 for reproducibility of caliper measurement. Macular edema (centerpoint thickness ≥ 240um) was present in 36%. Cysts were present in 36.6% of scans and ERMs in 27.8%, predominantly central. Intergrader agreement ranged from 78 − 82% for morphological features. Conclusion Retinal thickness data can be retrieved in a majority of OCT scans in clinical trial submissions for uveitis studies. Small cysts and ERMs involving the center are common in intermediate and posterior/panuveitis requiring systemic corticosteroid therapy. PMID:23163490

  1. Influence of two-phase anaerobic digestion on fate of selected antibiotic resistance genes and class I integrons in municipal wastewater sludge.

    PubMed

    Wu, Ying; Cui, Erping; Zuo, Yiru; Cheng, Weixiao; Rensing, Christopher; Chen, Hong

    2016-07-01

    The response of representative antibiotic resistance genes (ARGs) to lab-scale two-phase (acidogenic/methanogenic phase) anaerobic digestion processes under thermophilic and mesophilic conditions was explored. The associated microbial communities and bacterial pathogens were characterized by 16S rRNA gene sequencing. A two-phase thermophilic digestion reduced the presence of tetA, tetG, tetX, sul1, ermB, dfrA1, dfrA12 and intI1 exhibiting 0.1-0.72 log unit removal; in contrast, tetO, tetW, sul3, ermF and blaTEM even increased relative to the feed, and sul2 showed no significant decrease. The acidogenic phase of thermophilic digestion was primarily responsible for reducing the quantity of these genes, while the subsequent methanogenic phase caused a rebound in their quantity. In contrast, a two-phase mesophilic digestion process did not result in reducing the quantity of all ARGs and intI1 except for ermB and blaTEM. ARGs patterns were correlated with Proteobacteria and Actinobacteria during the two-phase anaerobic digestion. PMID:27035472

  2. First description of non-motile Yersinia ruckeri serovar I strains causing disease in rainbow trout, Oncorhynchus mykiss (Walbaum), cultured in Spain.

    PubMed

    Fouz, B; Zarza, C; Amaro, C

    2006-06-01

    Yersinia ruckeri, the causal agent of enteric redmouth (ERM) disease, was isolated from epizootics that occurred in different Spanish rainbow trout, Oncorhynchus mykiss (Walbaum), farms in which vaccination against ERM had been performed. In all episodes, the most pronounced clinical signs exhibited by affected fish were severe haemorrhages in the mouth, eyes and around the vent. The isolates were identified as Y. ruckeri serovar I by 16S rRNA sequencing together with serological tests. They lacked motility and lipase activity and thus belonged to biotype 2, and were highly virulent for juvenile rainbow trout, both by intraperitoneal injection (from 3.1 x 10(2) to 6.3 x 10(3) cfu per fish) and bath challenge (5.1-7.3 x 10(6) cfu mL(-1)). This is the first description of Y. ruckeri serovar I biotype 2 causing disease in cultured trout in Spain vaccinated with commercial ERM vaccines. The occurrence of this emergent pathogen in Spanish continental aquaculture from its first isolation in 2001 to date is also documented. PMID:16768714

  3. Protein 4.1 tumor suppressors: getting a FERM grip on growth regulation.

    PubMed

    Sun, Chun-Xiao; Robb, Victoria A; Gutmann, David H

    2002-11-01

    Members of the Protein 4.1 superfamily have highly conserved FERM domains that link cell surface glycoproteins to the actin cytoskeleton. Within this large and constantly expanding superfamily, at least five subgroups have been proposed. Two of these subgroups, the ERM and prototypic Protein 4.1 molecules, include proteins that function as tumor suppressors. The ERM subgroup member merlin/schwannomin is inactivated in the tumor-predisposition syndrome neurofibromatosis 2 (NF2), and the prototypic 4.1 subgroup member, Protein 4.1B, has been implicated in the molecular pathogenesis of breast, lung and brain cancers. This review focuses on what is known of mechanisms of action and critical protein interactions that may mediate the unique growth inhibitory signals of these two Protein 4.1 tumor suppressors. On the basis of insights derived from studying the NF2 tumor suppressor, we propose a model for merlin growth regulation in which CD44 links growth signals from plasma membrane to the nucleus by interacting with ERM proteins and merlin. PMID:12356905

  4. Observation of crack propagation in saline ice and freshwater ice with fluid inclusion

    NASA Astrophysics Data System (ADS)

    Arakawa, M.; Petrenko, V. F.

    2003-01-01

    A key process of crack propagation in saline ice is the interaction between the crack and fluid inclusions. We observed their interaction in freshwater ice using very high-speed photography (VHSP) and found that the low-density fluids (air and inert liquid, Fluorinert, 1.78 g/cm(3)) could not impede the crack effectively, interrupting the propagation for less than 10 mus. The high-density liquid mercury, (13.8 g/cm(3)) impeded the crack more effectively, stalling the development of the crack for more than 20 mus. The crack velocity in saline ice was measured using two different methods: electrical resistance method (ERM) and VHSP. These two methods returned very different mean velocities, 15 m/s for the ERM and 250 m/s for the VHSP. We found that in ice with conductive liquid inclusions, the ERM measured the time it took to break liquid bridges stretched across a crack rather than the crack velocity. Results from the VHSP show that the maximum crack velocity in saline ice was 500 m/s, which is one-half of that found in freshwater ice. From our results using freshwater ice with inclusions, we conclude that liquid inclusions in saline ice may play a role in this retardation.

  5. Constraining cosmic reionization with quasar, gamma ray burst, and Lyalpha emitter observations

    NASA Astrophysics Data System (ADS)

    Gallerani, S.; Ferrara, A.; Choudhury, T. Roy; Fan, X.; Salvaterra, R.; Dayal, P.

    We investigate the cosmic reionization history by comparing semi-analytical models of the Lyalpha forest with observations of high-z quasars and gamma ray bursts absorption spectra. In order to constrain the reionization epoch z_rei, we consider two physically motivated scenarios in which reionization ends either early (ERM, z_reigtrsim 7) or late (LRM, z_rei≈ 6). We analyze the transmitted flux in a sample of 17 QSOs spectra at 5.7< z_em< 6.4 and in the spectrum of the GRB 050904 at z=6.3, studying the wide dark portions (gaps) in the observed absorption spectra. By comparing the statistics of these spectral features with our models, we conclude that current observational data do not require any sudden change in the ionization state of the IGM at z≈ 6, favouring indeed a highly ionized Universe at these epochs, as predicted by the ERM. Moreover, we test the predictions of this model through Lyalpha emitters observations, finding that the ERM provide a good fit to the evolution of the luminosity function of Lyalpha emitting galaxies in the redshift range z=5.7-6.5. The overall result points towards an extended reionization process which starts at zgtrsim 11 and completes at z_reigtrsim 7, in agreement with the recent WMAP5 data.

  6. A phospholipid transfer function of ER-mitochondria encounter structure revealed in vitro

    PubMed Central

    Kojima, Rieko; Endo, Toshiya; Tamura, Yasushi

    2016-01-01

    As phospholipids are synthesized mainly in the endoplasmic reticulum (ER) and mitochondrial inner membranes, how cells properly distribute specific phospholipids to diverse cellular membranes is a crucial problem for maintenance of organelle-specific phospholipid compositions. Although the ER-mitochondria encounter structure (ERMES) was proposed to facilitate phospholipid transfer between the ER and mitochondria, such a role of ERMES is still controversial and awaits experimental demonstration. Here we developed a novel in vitro assay system with isolated yeast membrane fractions to monitor phospholipid exchange between the ER and mitochondria. With this system, we found that phospholipid transport between the ER and mitochondria relies on membrane intactness, but not energy sources such as ATP, GTP or the membrane potential across the mitochondrial inner membrane. We further found that lack of the ERMES component impairs the phosphatidylserine transport from the ER to mitochondria, but not the phosphatidylethanolamine transport from mitochondria to the ER. This in vitro assay system thus offers a powerful tool to analyze the non-vesicular phospholipid transport between the ER and mitochondria. PMID:27469264

  7. Differences in the composition of arbuscular mycorrhizal fungal communities promoted by different propagule forms from a Mediterranean shrubland.

    PubMed

    Varela-Cervero, Sara; López-García, Álvaro; Barea, José Miguel; Azcón-Aguilar, Concepción

    2016-07-01

    As it is well known, arbuscular mycorrhizal (AM) colonization can be initiated from the following three types of fungal propagules: spores, extraradical mycelium (ERM), and mycorrhizal root fragments harboring intraradical fungal structures. It has been shown that biomass allocation of AM fungi (AMF) among these three propagule types varies between fungal taxa, as also differs the ability of the different AMF propagule fractions to initiate new colonizations. In this study, the composition of the AMF community in the roots of rosemary (Rosmarinus officinalis L., a characteristic Mediterranean shrub), inoculated with the three different propagule types, was analyzed. Accordingly, cuttings from this species were inoculated with either AMF spores, ERM, or colonized roots extracted from a natural soil. The AMF diversity within the rosemary roots was characterized using terminal restriction fragment length polymorphism (T-RFLP) of the small subunit (SSU) rDNA region. The AMF community established in the rosemary plants was significantly different according to the type of propagule used as inoculum. AMF taxa differed in their ability to initiate new colonizations from each propagule type. Results suggest different colonization strategies for the different AMF families involved, Glomeraceae and Claroideoglomeraceae colonizing mainly from colonized roots whereas Pacisporaceae and Diversisporaceae from spores and ERM. This supports that AMF taxa show contrasting life-history strategies in terms of their ability to initiate new colonizations from the different propagule types. Further research to fully understand the colonization and dispersal abilities of AMF is essential for their rational use in ecosystem restoration programs. PMID:26883142

  8. Mcp3 is a novel mitochondrial outer membrane protein that follows a unique IMP-dependent biogenesis pathway.

    PubMed

    Sinzel, Monika; Tan, Tao; Wendling, Philipp; Kalbacher, Hubert; Özbalci, Cagakan; Chelius, Xenia; Westermann, Benedikt; Brügger, Britta; Rapaport, Doron; Dimmer, Kai Stefan

    2016-07-01

    Mitochondria are separated from the remainder of the eukaryotic cell by the mitochondrial outer membrane (MOM). The MOM plays an important role in different transport processes like lipid trafficking and protein import. In yeast, the ER-mitochondria encounter structure (ERMES) has a central, but poorly defined role in both activities. To understand the functions of the ERMES, we searched for suppressors of the deficiency of one of its components, Mdm10, and identified a novel mitochondrial protein that we named Mdm10 complementing protein 3 (Mcp3). Mcp3 partially rescues a variety of ERMES-related phenotypes. We further demonstrate that Mcp3 is an integral protein of the MOM that follows a unique import pathway. It is recognized initially by the import receptor Tom70 and then crosses the MOM via the translocase of the outer membrane. Mcp3 is next relayed to the TIM23 translocase at the inner membrane, gets processed by the inner membrane peptidase (IMP) and finally integrates into the MOM. Hence, Mcp3 follows a novel biogenesis route where a MOM protein is processed by a peptidase of the inner membrane. PMID:27226123

  9. Ensemble Empirical Mode Decomposition Analysis of EEG Data Collected during a Contour Integration Task

    PubMed Central

    Al-Subari, Karema; Al-Baddai, Saad; Tomé, Ana Maria; Volberg, Gregor; Hammwöhner, Rainer; Lang, Elmar W.

    2015-01-01

    We discuss a data-driven analysis of EEG data recorded during a combined EEG/fMRI study of visual processing during a contour integration task. The analysis is based on an ensemble empirical mode decomposition (EEMD) and discusses characteristic features of event related modes (ERMs) resulting from the decomposition. We identify clear differences in certain ERMs in response to contour vs noncontour Gabor stimuli mainly for response amplitudes peaking around 100 [ms] (called P100) and 200 [ms] (called N200) after stimulus onset, respectively. We observe early P100 and N200 responses at electrodes located in the occipital area of the brain, while late P100 and N200 responses appear at electrodes located in frontal brain areas. Signals at electrodes in central brain areas show bimodal early/late response signatures in certain ERMs. Head topographies clearly localize statistically significant response differences to both stimulus conditions. Our findings provide an independent proof of recent models which suggest that contour integration depends on distributed network activity within the brain. PMID:25910061

  10. Inter- and intraspecies transfer of a Clostridium difficile conjugative transposon conferring resistance to MLSB.

    PubMed

    Wasels, François; Monot, Marc; Spigaglia, Patrizia; Barbanti, Fabrizio; Ma, Laurence; Bouchier, Christiane; Dupuy, Bruno; Mastrantonio, Paola

    2014-12-01

    Resistance to the macrolide-lincosamide-streptogramin B group of antibiotics in Clostridium difficile is generally due to erm(B) genes. Tn6194, a conjugative transposon initially detected in PCR-ribotype 027 isolates, is an erm(B)-containing element also detected in other relevant C. difficile PCR-ribotypes. In this study, the genome of a C. difficile PCR-ribotype 001 strain was sequenced, and an element with two nucleotidic changes compared to Tn6194 was detected. This element was transferred by filter mating assays to recipient strains of C. difficile belonging to PCR-ribotype 009 and 027 and to a recipient strain of Enterococcus faecalis. Transconjugants were characterized by Southern blotting and genome sequencing, and integration sites in all transconjugants were identified. The element integrated the genome of C. difficile at different sites and the genome of E. faecalis at a unique site. This study is the first molecular characterization of an erm(B)-containing conjugative transposon in C. difficile and provides additional evidence of the antibiotic resistance transmission risk among pathogenic bacteria occupying the same human intestinal niche. PMID:25055190

  11. Induction of ribosome methylation in MLS-resistant Streptococcus pneumoniae by macrolides and ketolides.

    PubMed

    Zhong, P; Cao, Z; Hammond, R; Chen, Y; Beyer, J; Shortridge, V D; Phan, L Y; Pratt, S; Capobianco, J; Reich, K A; Flamm, R K; Or, Y S; Katz, L

    1999-01-01

    One major mechanism for resistance to macrolide antibiotics in Streptococcus pneumoniae is MLS (macrolide, lincosamide, and streptogramin B) resistance, manifested when the 23S rRNA is methylated by the product of an erm gene. This modification results in the decreased binding of all known macrolide, lincosamide, and streptogramin B antibiotics to the ribosome. More than 30 ermAM-containing clinical isolates of S. pneumoniae were examined in our lab and showed high-level resistance (MIC > or =128 microg/ml) to erythromycin, azithromycin, tylosin, clindamycin, and ketolide (macrolides that lack the cladinose sugar) TE-802. We found that the new generation of ketolides A965 and A088 displayed variable activity against the same group of resistant S. pneumoniae strains. To understand the basis of variability of the minimal inhibitory concentration (MIC) values of A965 and A088, we examined the effects of a series of macrolides and ketolides on the level of 23S rRNA methylation in five ermAM-containing resistant S. pneumoniae isolates. We show here that the basal levels of ribosomal methylation vary from strain to strain. The level of rRNA methylation can be strongly induced by erythromycin, azithromycin, and TE-802, resulting in high-level of resistance to these compounds. Ketolide A965 and A088, however, are weak inducers at sub-MIC drug concentrations, therefore showing variable activities in strains with differential methylation levels. PMID:10566867

  12. Molecular Identification and Quantification of Tetracycline and Erythromycin Resistance Genes in Spanish and Italian Retail Cheeses

    PubMed Central

    Flórez, Ana Belén; Alegría, Ángel; Delgado, Susana

    2014-01-01

    Large antibiotic resistance gene pools in the microbiota of foods may ultimately pose a risk for human health. This study reports the identification and quantification of tetracycline- and erythromycin-resistant populations, resistance genes, and gene diversity in traditional Spanish and Italian cheeses, via culturing, conventional PCR, real-time quantitative PCR (qPCR), and denaturing gradient gel electrophoresis (DGGE). The numbers of resistant bacteria varied widely among the antibiotics and the different cheese varieties; in some cheeses, all the bacterial populations seemed to be resistant. Up to eight antibiotic resistance genes were sought by gene-specific PCR, six with respect to tetracycline, that is, tet(K), tet(L), tet(M), tet(O), tet(S), and tet(W), and two with respect to erythromycin, that is, erm(B) and erm(F). The most common resistance genes in the analysed cheeses were tet(S), tet(W), tet(M), and erm(B). The copy numbers of these genes, as quantified by qPCR, ranged widely between cheeses (from 4.94 to 10.18log⁡10/g). DGGE analysis revealed distinct banding profiles and two polymorphic nucleotide positions for tet(W)-carrying cheeses, though the similarity of the sequences suggests this tet(W) to have a monophyletic origin. Traditional cheeses would therefore appear to act as reservoirs for large numbers of many types of antibiotic resistance determinants. PMID:25302306

  13. Open Conformation of Ezrin Bound to Phosphatidylinositol 4,5-Bisphosphate and to F-actin Revealed by Neutron Scattering*

    PubMed Central

    Jayasundar, Jayant James; Ju, Jeong Ho; He, Lilin; Liu, Dazhi; Meilleur, Flora; Zhao, Jinkui; Callaway, David J. E.; Bu, Zimei

    2012-01-01

    Ezrin is a member of the ezrin-radixin-moesin family (ERM) of adapter proteins that are localized at the interface between the cell membrane and the cortical actin cytoskeleton, and they regulate a variety of cellular functions. The structure representing a dormant and closed conformation of an ERM protein has previously been determined by x-ray crystallography. Here, using contrast variation small angle neutron scattering, we reveal the structural changes of the full-length ezrin upon binding to the signaling lipid phosphatidylinositol 4,5-bisphosphate (PIP2) and to F-actin. Ezrin binding to F-actin requires the simultaneous binding of ezrin to PIP2. Once bound to F-actin, the opened ezrin forms more extensive contacts with F-actin than generally depicted, suggesting a possible role of ezrin in regulating the interfacial structure and dynamics between the cell membrane and the underlying actin cytoskeleton. In addition, using gel filtration, we find that the conformational opening of ezrin in response to PIP2 binding is cooperative, but the cooperativity is disrupted by a phospho-mimic mutation S249D in the 4.1-ezrin/radixin/moesin (FERM) domain of ezrin. Using surface plasmon resonance, we show that the S249D mutation weakens the binding affinity and changes the kinetics of 4.1-ERM to PIP2 binding. The study provides the first structural view of the activated ezrin bound to PIP2 and to F-actin. PMID:22927432

  14. Unfurling of the band 4.1, ezrin, radixin, moesin (FERM) domain of the merlin tumor suppressor

    SciTech Connect

    Yogesha, S.D.; Sharff, Andrew J.; Giovannini, Marco; Bricogne, Gerard; Izard, Tina

    2014-10-02

    The merlin-1 tumor suppressor is encoded by the Neurofibromatosis-2 (Nf2) gene and loss-of-function Nf2 mutations lead to nervous system tumors in man and to several tumor types in mice. Merlin is an ERM (ezrin, radixin, moesin) family cytoskeletal protein that interacts with other ERM proteins and with components of cell-cell adherens junctions (AJs). Merlin stabilizes the links of AJs to the actin cytoskeleton. Thus, its loss destabilizes AJs, promoting cell migration and invasion, which in Nf2{sup +/-} mice leads to highly metastatic tumors. Paradoxically, the 'closed' conformation of merlin-1, where its N-terminal four-point-one, ezrin, radixin, moesin (FERM) domain binds to its C-terminal tail domain, directs its tumor suppressor functions. Here we report the crystal structure of the human merlin-1 head domain when crystallized in the presence of its tail domain. Remarkably, unlike other ERM head-tail interactions, this structure suggests that binding of the tail provokes dimerization and dynamic movement and unfurling of the F2 motif of the FERM domain. We conclude the 'closed' tumor suppressor conformer of merlin-1 is in fact an 'open' dimer whose functions are disabled by Nf2 mutations that disrupt this architecture.

  15. Anthropogenic contamination of metals in sediments of the Santa Rosalía harbor, Baja California peninsula.

    PubMed

    Shumilin, Evgueni; Jiménez-Illescas, Ángel Rafael; López-López, Silverio

    2013-03-01

    To know the environment impact on a harbor of the Santa Rosalía port on the Baja California peninsula, the concentrations of metals (Cd, Co, Cu, Mn, Pb, U and Zn) in harbor sediments were determined for 13 stations and compared with their average upper Earth´s crust abundance. The mean enrichment factors, calculated using Al as a normalizer, were higher than the unity for Ag, Ba, Bi, Cd, Co, Cu, Mn, Ni, Pb, Sb, U, V and Zn. Concentrations of slightly enriched Cd in the sediments are below the effect range low (ERL) sediment quality guidelines value only at three stations. The levels of Pb in the harbor sediments at four stations (1, 6, 9 and 11) are between the ERL (46.7 mg kg(-1)) and the effect range medium (ERM) (218 mg kg(-1)), and Pb content in the rest of the sediment samples is higher than the ERM, demonstrating the high extent of the anthropogenic impact of this metal on the sedimentary environment. The total concentrations of the potentially toxic elements Cu (3,390 ± 804 mg kg(-1)) and Zn (1,916 ± 749 mg kg(-1)) very strongly exceed their ERM, showing a high possibility of toxicological danger for marine biota, living inside or entering the harbor. PMID:23277367

  16. JNK1 determines the oncogenic or tumor-suppressive activity of the integrin-linked kinase in human rhabdomyosarcoma.

    PubMed

    Durbin, Adam D; Somers, Gino R; Forrester, Michael; Pienkowska, Malgorzata; Hannigan, Gregory E; Malkin, David

    2009-06-01

    Although most reports describe the protein kinase integrin-linked kinase (ILK) as a proto-oncogene, occasional studies detail opposing functions in the regulation of normal and transformed cell proliferation, differentiation, and apoptosis. Here, we demonstrated that ILK functions as an oncogene in the highly aggressive pediatric sarcoma alveolar rhabdomyosarcoma (ARMS) and as a tumor suppressor in the related embryonal rhabdomyosarcoma (ERMS). These opposing functions hinge on signaling through a noncanonical ILK target, JNK1, to the proto-oncogene c-Jun. RNAi-mediated depletion of ILK induced activation of JNK and its target, c-Jun, resulting in growth of ERMS cells, whereas in ARMS cells, it led to loss of JNK/c-Jun signaling and suppression of growth both in vitro and in vivo. Ectopic expression of the fusion gene characteristic of ARMS (paired box 3-forkhead homolog in rhabdomyosarcoma [PAX3-FKHR]) in ERMS cells was sufficient to convert them to an ARMS signaling phenotype and render ILK activity oncogenic. Furthermore, restoration of JNK1 in ARMS reestablished a tumor-suppressive function for ILK. These findings indicate what we believe to be a novel effector pathway regulated by ILK, provide a mechanism for interconversion of oncogenic and tumor-suppressor functions of a single regulatory protein based on the genetic background of the tumor cells, and suggest a rationale for tailored therapy of rhabdomyosarcoma based on the different activities of ILK. PMID:19478459

  17. A phospholipid transfer function of ER-mitochondria encounter structure revealed in vitro.

    PubMed

    Kojima, Rieko; Endo, Toshiya; Tamura, Yasushi

    2016-01-01

    As phospholipids are synthesized mainly in the endoplasmic reticulum (ER) and mitochondrial inner membranes, how cells properly distribute specific phospholipids to diverse cellular membranes is a crucial problem for maintenance of organelle-specific phospholipid compositions. Although the ER-mitochondria encounter structure (ERMES) was proposed to facilitate phospholipid transfer between the ER and mitochondria, such a role of ERMES is still controversial and awaits experimental demonstration. Here we developed a novel in vitro assay system with isolated yeast membrane fractions to monitor phospholipid exchange between the ER and mitochondria. With this system, we found that phospholipid transport between the ER and mitochondria relies on membrane intactness, but not energy sources such as ATP, GTP or the membrane potential across the mitochondrial inner membrane. We further found that lack of the ERMES component impairs the phosphatidylserine transport from the ER to mitochondria, but not the phosphatidylethanolamine transport from mitochondria to the ER. This in vitro assay system thus offers a powerful tool to analyze the non-vesicular phospholipid transport between the ER and mitochondria. PMID:27469264

  18. Multiplex PCR To Identify Macrolide Resistance Determinants in Mannheimia haemolytica and Pasteurella multocida

    PubMed Central

    Rose, Simon; Desmolaize, Benoit; Jaju, Puneet; Wilhelm, Cornelia; Warrass, Ralf

    2012-01-01

    The bacterial pathogens Mannheimia haemolytica and Pasteurella multocida are major etiological agents in respiratory tract infections of cattle. Although these infections can generally be successfully treated with veterinary macrolide antibiotics, a few recent isolates have shown resistance to these drugs. Macrolide resistance in members of the family Pasteurellaceae is conferred by combinations of at least three genes: erm(42), which encodes a monomethyltransferase and confers a type I MLSB (macrolide, lincosamide, and streptogramin B) phenotype; msr(E), which encodes a macrolide efflux pump; and mph(E), which encodes a macrolide-inactivating phosphotransferase. Here, we describe a multiplex PCR assay that detects the presence of erm(42), msr(E), and mph(E) and differentiates between these genes. In addition, the assay distinguishes P. multocida from M. haemolytica by amplifying distinctive fragments of the 23S rRNA (rrl) genes. One rrl fragment acts as a general indicator of gammaproteobacterial species and confirms whether the PCR assay has functioned as intended on strains that are negative for erm(42), msr(E), and mph(E). The multiplex system has been tested on more than 40 selected isolates of P. multocida and M. haemolytica and correlated with MICs for the veterinary macrolides tulathromycin and tilmicosin, and the newer compounds gamithromycin and tildipirosin. The multiplex PCR system gives a rapid and robustly accurate determination of macrolide resistance genotypes and bacterial genus, matching results from microbiological methods and whole-genome sequencing. PMID:22564832

  19. Frequent HRAS Mutations in Malignant Ectomesenchymoma: Overlapping Genetic Abnormalities With Embryonal Rhabdomyosarcoma.

    PubMed

    Huang, Shih-Chiang; Alaggio, Rita; Sung, Yun-Shao; Chen, Chun-Liang; Zhang, Lei; Kao, Yu-Chien; Agaram, Narasimhan P; Wexler, Leonard H; Antonescu, Cristina R

    2016-07-01

    Malignant ectomesenchymoma (MEM) is an exceedingly rare pediatric sarcoma with a predilection for infants and young children and is composed of dual malignant mesenchymal and neuroectodermal components. Microscopically, MEM displays areas of rhabdomyosarcoma (RMS) with intermixed neuronal/neuroblastic foci. The molecular alterations associated with MEM and its relationship with embryonal RMS (ERMS) and malignant peripheral nerve sheath tumor (MPNST) have not yet been elucidated. In this study we used whole-transcriptome sequencing in 2 MEM index cases with available frozen tissue, followed by screening of the identified genetic abnormalities in 5 additional cases. No candidate fusion genes were detected by FusionSeq analysis; however, the mutation detection algorithms revealed HRAS and PTPRD hotspot mutations in both index cases, with 1 case harboring an additional FBXW7 mutation. As these mutation profiles have been previously described in ERMS we have tested their incidence in a control group of 7 age-matched ERMS. In addition, the gene signature of MEM was compared with that of RMS, MPNST, and neuronal lineage. All 7 MEM patients were male, with a mean age of 7.5 months (range, 0.6 to 17 mo). All except 1 occurred in the pelvic/urogenital region. Most cases showed ERMS elements, with occasional spindle or undifferentiated/round cell areas. The intermixed neuroectodermal components were mostly scattered ganglion cells, ganglioneuroma, or ganglioneuroblastoma. By Sanger sequencing, 6 of 7 (86%) MEMs had HRAS mutations, with no additional case harboring PTPRD or FBXW7 mutations. The only case lacking HRAS mutation showed neuroblastic micronodules without ganglion cells. The trimethylation at lysine 27 of histone H3 (H3K27me3) expression, typically lost in MPNST, was retained in all cases. In the control ERMS group, 5 of 7 (71%) showed RAS mutations, equally distributed among NRAS, KRAS, and HRAS genes. The expression profiling of MEM showed upregulation of skeletal

  20. Clinical outcomes of double membrane peeling with or without simultaneous phacoemulsification/gas tamponade for vitreoretinal-interface-associated (VRI) disorders.

    PubMed

    Kumar, Kshitiz; Chandnani, Nisha; Raj, Pallavi; Agarwal, Amar

    2016-08-01

    The purpose of this study is to evaluate the clinical outcomes of double membrane (ERM & ILM) peeling and the effect of combined cataract surgery and SF6 gas injection in vitreoretinal interface (VRI) disorders. This is a retrospective interventional study. Seventy-two eyes with idiopathic vitreoretinal interface abnormalities that underwent 23 gauge pars plana vitrectomy with "double stain and double peel" technique were reviewed. SD-OCT was used to classify VRI disorders into following 4 groups: 44 in ERM type, 17 in VMTS type, 7 in macular pseudohole (MPH) type, and 4 in lamellar macular hole (LMH) type. ERM was a common association in all types. Mean preoperative BCVA improved from 0.58 ± 0.14 logMAR to 0.27 ± 0.16 logMAR units (p = 0.001). Mean CFT reduced from 409.17 ± 122.31 µm preoperatively to 277.28 ± 0.16 µm postoperatively (p < 0.0001). Among the VRI subtypes, visual improvement was significant except in LMH variety (ERM type, p = 0.0029; VMTS type, p = 0.0281; MPH type, p = 0.05; and LMH type, p = 0.7926). Mean change in CFT from baseline was least in LMH cases (p = 0.0093). There was no significant difference in BCVA and CFT in the group who had combined phacovitrectomy versus pseudophakic group (p > 0.05). Use of intraocular SF6 gas tamponade did not show any added benefits among the groups (p > 0.05). Improvement in foveal contour was seen in all groups. Simultaneous removal of ILM along with ERM during surgery for VRI disorders helps in restoring normal foveal contour with a favorable visual outcome. Combined cataract extraction or use of intraocular SF6 gas injection does not affect the surgical results. PMID:26659009

  1. The roles of RgpB and Kgp in late onset gingipain activity in the vimA-defective mutant of Porphyromonas gingivalis W83.

    PubMed

    Dou, Y; Robles, A; Roy, F; Aruni, A W; Sandberg, L; Nothnagel, E; Fletcher, H M

    2015-10-01

    Previous studies have shown that VimA, an acetyltransferase, can modulate gingipain biogenesis in Porphyromonas gingivalis. Inactivation of the vimA gene resulted in isogenic mutants that showed a late onset of gingipain activity that only occurred during the stationary growth phase. To further elucidate the role and contribution of the gingipains in this VimA-dependent process, isogenic mutants defective in the gingipain genes in the vimA-deficient genetic background were evaluated. In contrast with the wild-type strain, RgpB and Kgp gingipain activities were absent in exponential phase in the ∆rgpA::tetQ-vimA::ermF mutant. However, these activities increased to 31 and 53%, respectively, of that of the wild-type during stationary phase. In the ∆rgpA::cat-∆kgp::tetQ-vimA::ermF mutant, the RgpB protein was observed in the extracellular fraction but no activity was present even at the stationary growth phase. There was no gingipain activity observed in the ∆rgpB::cat-∆kgp::tetQ-vimA::ermF mutant whereas Kgp activity in ∆rgpA::cat-∆rgpB::tetQ-vimA::ermF mutant was 24% of the wild-type at late stationary phase. In contrast to RgpA, the glycosylation profile of the RgpB catalytic domain from both W83 and P. gingivalis FLL92 (vimA::ermF) showed similarity. Taken together, the results suggest multiple gingipain activation pathways in P. gingivalis. Whereas the maturation pathways for RgpA and RgpB are different, the late-onset gingipain activity in the vimA-defective mutant was due to activation/maturation of RgpB and Kgp. Moreover, unlike RgpA, which is VimA-dependent, the maturation/activation pathways for RgpB and Kgp are interdependent in the absence VimA. PMID:25858089

  2. Assessment of geochemical mobility of metals in surface sediments of the Santa Rosalia mining region, Western Gulf of California.

    PubMed

    Shumilin, Evgueni; Gordeev, Vyacheslav; Figueroa, Griselda Rodríguez; Demina, Liudmila; Choumiline, Konstantin

    2011-01-01

    To asses the geomobility of cadmium, copper, iron, manganese, nickel, lead, and zinc in marine sediments near the Santa Rosalía copper smelter, which is located on the eastern coast of the Baja California Peninsula, sequential leaching was applied to sediment samples containing different levels of Cu: (1) uncontaminated or slightly contaminated (<55 mg kg⁻¹ Cu); (2) moderately contaminated (55-500 mg kg⁻¹ Cu); and (3) heavily contaminated (>500 mg kg⁻¹ Cu). Concentrations of Cd, Cu, Fe, Mn, Ni, Pb, and Zn in four fractions of the leachate (mobile fraction F1, relatively mobile fraction F2, associated with organic matter/sulphides fraction F3, and residual fraction F4) were measured by atomic absorption spectrophotometry (AAS). The sediments with Cu concentration <500 mg kg⁻¹ displayed prevalence of mobile acid-leachable fraction F1 and reducible fraction F2 for Cd, Cu, Mn, and Pb, whereas the relative contribution of fraction F3 was relatively low for all of the examined metals. Residual fraction F4 was highest (>65%) for Fe and Ni because both metals are associated with the crystalline matrix of natural sediments. The sediments heavily contaminated with Cu (>500 mg kg⁻¹) had dramatically increased percentages of Cu, Mn, Pb, and Zn, ranging on average from 63 to 81%, in the residual fraction. In the case of Cu, for example, the relative abundances of this element in the different fractions of such sediments followed this sequence: residual fraction F4 (76 ± 5%) >absorbed form and carbonates fraction F1 (15 ± 5%) >Fe and Mn oxyhydroxides fraction F2 (5 ± 2%) >fraction associated with organic matter and sulphides F3 (4.5 ± 3.9%). Copper, Pb, and Zn contents in each geochemical fraction of all samples were compared with sediment-quality guideline values ("effects range low" [ERL] and "effects range medium" [ERM]) to assess their possible negative effects on biota. Copper contents in mobile fractions F1 and F2, which were moderately contaminated

  3. Non-linear dynamics of operant behavior: a new approach via the extended return map.

    PubMed

    Li, Jay-Shake; Huston, Joseph P

    2002-01-01

    Previous efforts to apply non-linear dynamic tools to the analysis of operant behavior revealed some promise for this kind of approach, but also some doubts, since the complexity of animal behavior seemed to be beyond the analyzing ability of the available tools. We here outline a series of studies based on a novel approach. We modified the so-called 'return map' and developed a new method, the 'extended return map' (ERM) to extract information from the highly irregular time series data, the inter-response time (IRT) generated by Skinner-box experiments. We applied the ERM to operant lever pressing data from rats using the four fundamental reinforcement schedules: fixed interval (FI), fixed ratio (FR), variable interval (VI) and variable ratio (VR). Our results revealed interesting patterns in all experiment groups. In particular, the FI and VI groups exhibited well-organized clusters of data points. We calculated the fractal dimension out of these patterns and compared experimental data with surrogate data sets, that were generated by randomly shuffling the sequential order of original IRTs. This comparison supported the finding that patterns in ERM reflect the dynamics of the operant behaviors under study. We then built two models to simulate the functional mechanisms of the FI schedule. Both models can produce similar distributions of IRTs and the stereotypical 'scalloped' curve characteristic of FI responding. However, they differ in one important feature in their formulation: while one model uses a continuous function to describe the probability of occurrence of an operant behavior, the other one employs an abrupt switch of behavioral state. Comparison of ERMs showed that only the latter was able to produce patterns similar to the experimental results, indicative of the operation of an abrupt switch from one behavioral state to another over the course of the inter-reinforcement period. This example demonstrated the ERM to be a useful tool for the analysis of

  4. Evaluation of plasmid and genomic DNA calibrants used for the quantification of genetically modified organisms.

    PubMed

    Caprioara-Buda, M; Meyer, W; Jeynov, B; Corbisier, P; Trapmann, S; Emons, H

    2012-07-01

    The reliable quantification of genetically modified organisms (GMOs) by real-time PCR requires, besides thoroughly validated quantitative detection methods, sustainable calibration systems. The latter establishes the anchor points for the measured value and the measurement unit, respectively. In this paper, the suitability of two types of DNA calibrants, i.e. plasmid DNA and genomic DNA extracted from plant leaves, for the certification of the GMO content in reference materials as copy number ratio between two targeted DNA sequences was investigated. The PCR efficiencies and coefficients of determination of the calibration curves as well as the measured copy number ratios for three powder certified reference materials (CRMs), namely ERM-BF415e (NK603 maize), ERM-BF425c (356043 soya), and ERM-BF427c (98140 maize), originally certified for their mass fraction of GMO, were compared for both types of calibrants. In all three systems investigated, the PCR efficiencies of plasmid DNA were slightly closer to the PCR efficiencies observed for the genomic DNA extracted from seed powders rather than those of the genomic DNA extracted from leaves. Although the mean DNA copy number ratios for each CRM overlapped within their uncertainties, the DNA copy number ratios were significantly different using the two types of calibrants. Based on these observations, both plasmid and leaf genomic DNA calibrants would be technically suitable as anchor points for the calibration of the real-time PCR methods applied in this study. However, the most suitable approach to establish a sustainable traceability chain is to fix a reference system based on plasmid DNA. PMID:22638881

  5. Resistance Genes and Genetic Elements Associated with Antibiotic Resistance in Clinical and Commensal Isolates of Streptococcus salivarius.

    PubMed

    Chaffanel, Fanny; Charron-Bourgoin, Florence; Libante, Virginie; Leblond-Bourget, Nathalie; Payot, Sophie

    2015-06-15

    The diversity of clinical (n = 92) and oral and digestive commensal (n = 120) isolates of Streptococcus salivarius was analyzed by multilocus sequence typing (MLST). No clustering of clinical or commensal strains can be observed in the phylogenetic tree. Selected strains (92 clinical and 46 commensal strains) were then examined for their susceptibilities to tetracyclines, macrolides, lincosamides, aminoglycosides, and phenicol antibiotics. The presence of resistance genes tet(M), tet(O), erm(A), erm(B), mef(A/E), and catQ and associated genetic elements was investigated by PCR, as was the genetic linkage of resistance genes. High rates of erythromycin and tetracycline resistance were observed among the strains. Clinical strains displayed either the erm(B) (macrolide-lincosamide-streptogramin B [MLSB] phenotype) or mef(A/E) (M phenotype) resistance determinant, whereas almost all the commensal strains harbored the mef(A/E) resistance gene, carried by a macrolide efflux genetic assembly (MEGA) element. A genetic linkage between a macrolide resistance gene and genes of Tn916 was detected in 23 clinical strains and 5 commensal strains, with a predominance of Tn3872 elements (n = 13), followed by Tn6002 (n = 11) and Tn2009 (n = 4) elements. Four strains harboring a mef(A/E) gene were also resistant to chloramphenicol and carried a catQ gene. Sequencing of the genome of one of these strains revealed that these genes colocalized on an IQ-like element, as already described for other viridans group streptococci. ICESt3-related elements were also detected in half of the isolates. This work highlights the potential role of S. salivarius in the spread of antibiotic resistance genes both in the oral sphere and in the gut. PMID:25862227

  6. Characterization of Three New Glutaredoxin Genes in the Arbuscular Mycorrhizal Fungus Rhizophagus irregularis: Putative Role of RiGRX4 and RiGRX5 in Iron Homeostasis.

    PubMed

    Tamayo, Elisabeth; Benabdellah, Karim; Ferrol, Nuria

    2016-01-01

    Glutaredoxins (GRXs) are small ubiquitous oxidoreductases involved in the regulation of the redox state in living cells. In an attempt to identify the full complement of GRXs in the arbuscular mycorrhizal (AM) fungus Rhizophagus irregularis, three additional GRX homologs, besides the formerly characterized GintGRX1 (renamed here as RiGRX1), were identified. The three new GRXs (RiGRX4, RiGRX5 and RiGRX6) contain the CXXS domain of monothiol GRXs, but whereas RiGRX4 and RiGRX5 belong to class II GRXs, RiGRX6 belongs to class I together with RiGRX1. By using a yeast expression system, we observed that the newly identified homologs partially reverted sensitivity of the GRX deletion yeast strains to external oxidants. Furthermore, our results indicated that RiGRX4 and RiGRX5 play a role in iron homeostasis in yeast. Gene expression analyses revealed that RiGRX1 and RiGRX6 were more highly expressed in the intraradical (IRM) than in the extraradical mycelium (ERM). Exposure of the ERM to hydrogen peroxide induced up-regulation of RiGRX1, RiGRX4 and RiGRX5 gene expression. RiGRX4 expression was also up-regulated in the ERM when the fungus was grown in media supplemented with a high iron concentration. These data indicate the two monothiol class II GRXs, RiGRX4 and RiGRX5, might be involved in oxidative stress protection and in the regulation of fungal iron homeostasis. Increased expression of RiGRX1 and RiGRX6 in the IRM suggests that these GRXs should play a key role in oxidative stress protection of R. irregularis during its in planta phase. PMID:26900849

  7. Detection of Multiple Resistances, Biofilm Formation and Conjugative Transfer of Bacillus cereus from Contaminated Soils.

    PubMed

    Anjum, Reshma; Krakat, Niclas

    2016-03-01

    The purpose of this study was to detect microbial resistances to a set of antibiotics/pesticides (multi-resistance) within pesticide and antibiotic-contaminated alluvial soils and to identify the corresponding antibiotic resistance genes (ARGs). To assess whether identified multi-resistant isolates are able to construct biofilms, several biofilm formation and conjugation experiments were conducted. Out of 35 isolates, six strains were used for filter mating experiments. Nine strains were identified by 16S rDNA gene sequence analyses and those were closely related to Pseudomonas sp., Citrobacter sp., Acinetobacter sp., Enterobacter sp., and in addition, Bacillus cereus was chosen for multi-resistant and pesticide-tolerant studies. Antibiotic-resistant and pesticide-tolerant bacterial strains were tested for the presence of ARGs. All nine strains were containing multiple ARGs (ampC, ermB, ermD, ermG, mecA, tetM) in different combinations. Interestingly, only strain WR34 (strongly related to Bacillus cereus) exhibited a high biofilm forming capacity on glass beads. Results obtained by filter mating experiments demonstrated gene transfer frequencies from 10(-5) to 10(-8). This study provides evidence that alluvial soils are hot spots for the accumulation of antibiotics, pesticides and biofilm formation. Particularly high resistances to tetracycline, ampicillin, amoxicillin and methicillin were proved. Apparently, isolate WR34 strongly correlated to a pathogenic organism had high potential to deploy biofilms in alluvial soils. Thus, we assume that loosened and unconsolidated soils investigated pose a high risk of an enhanced ARG prevalence. PMID:26650381

  8. Genetic detection and multilocus sequence typing of vanA-containing Enterococcus strains from mullets fish (Liza ramada).

    PubMed

    Araújo, Carlos; Torres, Carmen; Gonçalves, Alexandre; Carneiro, Catarina; López, Maria; Radhouani, Hajer; Pardal, Miguel; Igrejas, Gilberto; Poeta, Patrícia

    2011-09-01

    Enterococci have emerged as important nosocomial and community-acquired pathogens in humans. The presence of vanA-enterococci was investigated in 103 fecal samples recovered from mullets fish (Liza ramada). All fecal samples were inoculated in Slanetz-Bartley agar plates supplemented with 4 mg/L of vancomycin for vancomycin-resistant enterococci (VRE) recovery and two isolates/sample were characterized. Antibiotic susceptibility was tested for 11 antibiotics by disk diffusion and agar dilution methods. VRE identification was performed by biochemical and molecular methods. Additionally, the mechanisms of resistance to glycopeptides (vanA, vanB, vanC1, vanC2, and vanD) and other antibiotics [erm(A), erm(B), tet(L), tet(M), aph(2'')-aac(6'), aph(3')-IIIa, ant(6'), vat(D), vat(E)] as well as the presence of enterococcal surface protein (esp) and hyl virulence factors were investigated. vanA-Enterococcus faecium isolates were recovered from 4 of 103 tested samples, and they showed glycopeptide and erythromycin resistances. Three of them were also ampicillin resistant, two showed resistance to tetracycline, ciprofloxacin, and kanamycin, and one showed resistance to gentamicin. The tet(M) and erm(B) genes were found in all tetracycline- and erythromycin-resistant strains, respectively. The aph(3')-III and aph(2'')-aac(6') genes were identified in the kanamycin- and gentamicin-resistant isolates, respectively. The IS1216 element was identified within vanX-vanY region of Tn1546 in two vanA isolates. The hyl and esp virulence genes were found in four and two isolates, respectively. vanA-strains were ascribed to sequence types ST280 (two isolates) and ST273 (two isolates), including both lineages into the clonal complex CC17. Mullets fish can excrete VRE in their feces and may be a reservoir for such resistant bacteria that can be transmitted to other animals including humans. PMID:21563953

  9. Unraveling antimicrobial resistance genes and phenotype patterns among Enterococcus faecalis isolated from retail chicken products in Japan.

    PubMed

    Hidano, Arata; Yamamoto, Takehisa; Hayama, Yoko; Muroga, Norihiko; Kobayashi, Sota; Nishida, Takeshi; Tsutsui, Toshiyuki

    2015-01-01

    Multidrug-resistant enterococci are considered crucial drivers for the dissemination of antimicrobial resistance determinants within and beyond a genus. These organisms may pass numerous resistance determinants to other harmful pathogens, whose multiple resistances would cause adverse consequences. Therefore, an understanding of the coexistence epidemiology of resistance genes is critical, but such information remains limited. In this study, our first objective was to determine the prevalence of principal resistance phenotypes and genes among Enterococcus faecalis isolated from retail chicken domestic products collected throughout Japan. Subsequent analysis of these data by using an additive Bayesian network (ABN) model revealed the co-appearance patterns of resistance genes and identified the associations between resistance genes and phenotypes. The common phenotypes observed among E. faecalis isolated from the domestic products were the resistances to oxytetracycline (58.4%), dihydrostreptomycin (50.4%), and erythromycin (37.2%), and the gene tet(L) was detected in 46.0% of the isolates. The ABN model identified statistically significant associations between tet(L) and erm(B), tet(L) and ant(6)-Ia, ant(6)-Ia and aph(3')-IIIa, and aph(3')-IIIa and erm(B), which indicated that a multiple-resistance profile of tetracycline, erythromycin, streptomycin, and kanamycin is systematic rather than random. Conversely, the presence of tet(O) was only negatively associated with that of erm(B) and tet(M), which suggested that in the presence of tet(O), the aforementioned multiple resistance is unlikely to be observed. Such heterogeneity in linkages among genes that confer the same phenotypic resistance highlights the importance of incorporating genetic information when investigating the risk factors for the spread of resistance. The epidemiological factors that underlie the persistence of systematic multiple-resistance patterns warrant further investigations with appropriate

  10. In Vitro Activity of the New Ketolide Telithromycin Compared with Those of Macrolides against Streptococcus pyogenes: Influences of Resistance Mechanisms and Methodological Factors

    PubMed Central

    Bemer-Melchior, Pascale; Juvin, Marie-Emmanuelle; Tassin, Sandrine; Bryskier, Andre; Schito, Gian Carlo; Drugeon, Henri-B.

    2000-01-01

    One hundred and seven clinical isolates of Streptococcus pyogenes, 80 susceptible to macrolides and 27 resistant to erythromycin A (MIC >0.5 μg/ml), were examined. The erythromycin A-lincomycin double-disk test assigned 7 resistant strains to the M-phenotype, 8 to the inducible macrolide, lincosamide, and streptogramin B resistance (iMLSB) phenotype, and 12 to the constitutive MLSB resistance (cMLSB) phenotype. MICs of erythromycin A, clarithromycin, azithromycin, roxithromycin, and clindamycin were determined by a broth microdilution method. MICs of telithromycin were determined by three different methods (broth microdilution, agar dilution, and E-test methods) in an ambient air atmosphere and in a 5 to 6% CO2 atmosphere. Erythromycin A resistance genes were investigated by PCR in the 27 erythromycin A-resistant isolates. MICs of erythromycin A and clindamycin showed six groups of resistant strains, groups A to F. iMLSB strains (A, B, and D groups) are characterized by two distinct patterns of resistance correlated with genotypic results. A- and B-group strains were moderately resistant to 14- and 15-membered ring macrolides and highly susceptible to telithromycin. All A- and B-group isolates harbored erm TR gene, D-group strains, highly resistant to macrolides and intermediately resistant to telithromycin (MICs, 1 to 16 μg/ml), were all characterized by having the ermB gene. All M-phenotype isolates (C group), resistant to 14- and 15-membered ring macrolides and susceptible to clindamycin and telithromycin, harbored the mefA gene. All cMLSB strains (E and F groups) with high level of resistance to macrolides, lincosamide, and telithromycin had the ermB gene. The effect of 5 to 6% CO2 was remarkable on resistant strains, by increasing MICs of telithromycin from 1 to 6 twofold dilutions against D-E- and F-group isolates. PMID:11036012

  11. Impact of Ribosomal Modification on the Binding of the Antibiotic Telithromycin Using a Combined Grand Canonical Monte Carlo/Molecular Dynamics Simulation Approach

    PubMed Central

    Small, Meagan C.; Lopes, Pedro; Andrade, Rodrigo B.; MacKerell, Alexander D.

    2013-01-01

    Resistance to macrolide antibiotics is conferred by mutation of A2058 to G or methylation by Erm methyltransferases of the exocyclic N6 of A2058 (E. coli numbering) that forms the macrolide binding site in the 50S subunit of the ribosome. Ketolides such as telithromycin mitigate A2058G resistance yet remain susceptible to Erm-based resistance. Molecular details associated with macrolide resistance due to the A2058G mutation and methylation at N6 of A2058 by Erm methyltransferases were investigated using empirical force field-based simulations. To address the buried nature of the macrolide binding site, the number of waters within the pocket was allowed to fluctuate via the use of a Grand Canonical Monte Carlo (GCMC) methodology. The GCMC water insertion/deletion steps were alternated with Molecular Dynamics (MD) simulations to allow for relaxation of the entire system. From this GCMC/MD approach information on the interactions between telithromycin and the 50S ribosome was obtained. In the wild-type (WT) ribosome, the 2′-OH to A2058 N1 hydrogen bond samples short distances with a higher probability, while the effectiveness of telithromycin against the A2058G mutation is explained by a rearrangement of the hydrogen bonding pattern of the 2′-OH to 2058 that maintains the overall antibiotic-ribosome interactions. In both the WT and A2058G mutation there is significant flexibility in telithromycin's imidazole-pyridine side chain (ARM), indicating that entropic effects contribute to the binding affinity. Methylated ribosomes show lower sampling of short 2′-OH to 2058 distances and also demonstrate enhanced G2057-A2058 stacking leading to disrupted A752-U2609 Watson-Crick (WC) interactions as well as hydrogen bonding between telithromycin's ARM and U2609. This information will be of utility in the rational design of novel macrolide analogs with improved activity against methylated A2058 ribosomes. PMID:23785274

  12. Microperimetric Assessment after Epiretinal Membrane Surgery: 4-Year Follow-Up

    PubMed Central

    Dal Vecchio, Marco; Lavia, Carlo; Nassisi, Marco; Grignolo, Federico M.; Fea, Antonio M.

    2016-01-01

    Purpose. To investigate retinal function using microperimetry in patients affected by idiopathic epiretinal membrane (iERM) and cataract who underwent combined surgery: 4-year follow-up. Design. Prospective, interventional case series. Methods. 30 eyes of 30 consecutive patients with iERM and age-related cataract underwent 25-gauge vitrectomy and cataract surgery. At baseline, 90 and 180 days, and 1 and 4 years, we examined retinal mean sensitivity (MS), retinal mean defect (MD), fixation stability, and frequency of microscotomas using MP1 microperimetry. Best-corrected visual acuity (BCVA) and central retinal thickness (CRT) using a spectral domain optical coherence tomography (SD-OCT) were also performed. Results. All patients completed 1-year follow-up, while 23 patients reached last follow-up. Baseline MS and MD (10.48 ± 4.17 and −9.18 ± 4.40 dB) significantly changed at one year (12.33 ± 3.66 and −7.49 ± 3.31 dB, p < 0.01), at four years (14.18 ± 3.46 and −4.66 ± 2.85, p < 0.01), and between one and four years (p < 0.01) after surgery. Compared to baseline, CRT and BCVA significantly changed at one year and remained stable at four years. No variations were observed in fixation stability and frequency of microscotomas compared to baseline. Conclusions. Long-term follow-up using microperimetry seems useful to evaluate patients after iERM surgery: retinal sensitivity changes even when BCVA and CRT remain stable. PMID:27088008

  13. Antistreptococcal Activity of Telithromycin Compared with Seven Other Drugs in Relation to Macrolide Resistance Mechanisms in Russia

    PubMed Central

    Kozlov, Roman S.; Bogdanovitch, Tatiana M.; Appelbaum, Peter C.; Ednie, Lois; Stratchounski, Leonid S.; Jacobs, Michael R.; Bozdogan, Bülent

    2002-01-01

    The susceptibilities of 468 recent Russian clinical Streptococcus pneumoniae isolates and 600 Streptococcus pyogenes isolates, from 14 centers in Russia, to telithromycin, erythromycin, azithromycin, clarithromycin, clindamycin, levofloxacin, quinupristin-dalfopristin, and penicillin G were tested. Penicillin-nonsusceptible S. pneumoniae strains were rare except in Siberia, where their prevalence rate was 13.5%: most were penicillin intermediate, but for three strains (two from Smolensk and one from Novosibirsk) the MICs of penicillin G were 4 or 8 μg/ml. Overall, 2.5% of S. pneumoniae isolates were resistant to erythromycin. Efflux was the prevalent resistance mechanism (five strains; 41.7%), followed by ribosomal methylation encoded by constitutive erm(B), which was found in four isolates. Ribosomal mutation was the mechanism of macrolide resistance in three isolates; one erythromycin-resistant S. pneumoniae isolate had an A2059G mutation in 23S rRNA, and two isolates had substitution of GTG by TPS at positions 69 to 71 in ribosomal protein L4. All S. pyogenes isolates were susceptible to penicillin, and 11% were erythromycin resistant. Ribosomal methylation was the most common resistance mechanism for S. pyogenes (89.4%). These methylases were encoded by erm(A) [subclass erm(TR)] genes, and their expression was inducible in 96.6% of isolates. The rest of the erythromycin-resistant Russian S. pyogenes isolates (7.6%) had an efflux resistance mechanism. Telithromycin was active against 100% of pneumococci and 99.2% of S. pyogenes, and levofloxacin and quinupristin-dalfopristin were active against all isolates of both species. PMID:12183254

  14. Combining multiple altimeter missions

    NASA Astrophysics Data System (ADS)

    Jacobs, G. A.; Mitchell, J. L.

    1997-10-01

    Viewing altimeter data only at the points where separate altimeter missions' ground tracks cross provides a method to observe long time period sea surface height (SSH) variations and avoids many of the problems inherent in combining separate altimeter data sets through an independently determined geoid. TOPEX/POSEIDON (T/P) data over the time period from January 1, 1993, to December 31, 1995, form a mean SSH that is used as a reference by other altimeter data sets. A least squares analysis of the mean T/P SSH determines the portion of the Geographically Correlated Orbit Error (GCOE) that may be observed through crossover differences and removes this portion of the GCOE. The analysis removes errors of 0.86 cm RMS at 1 cycle per orbit revolution (cpr) and indicates negligible errors at higher frequencies. After the GCOE removal, the accuracy of the T/P reference mean is better than 1 cm RMS as measured by crossover differences. The GCOE contained in the Geosat-Exact Repeat Mission (ERM) and ERS 1 data with orbit solutions using the Joint Gravity Model (JGM) 3 is evaluated through an adjustment to the T/P reference mean surface. The Geosat-ERM data indicate a bias of about 28 cm averaged over the globe, and the ERS 1 bias is 44 cm. The T/P data used here is not corrected for the oscillator drift correction error so that the actual bias is less by about 13 cm. Both the Geosat-ERM and ERS 1 GCOE are mainly 1 cpr. GCOE estimates at frequencies above 1 cpr indicate little actual orbit error but are more correlated to instrument correction errors (particularly water vapor). Simultaneous T/P and ERS 1 SSH anomalies to the T/P mean indicate good correlation.

  15. Radiative Impacts on the Growth of a Population of Drops within Simulated Summertime Arctic Stratus.

    NASA Astrophysics Data System (ADS)

    Harrington, Jerry Y.; Feingold, Graham; Cotton, William R.

    2000-03-01

    The impact of solar heating and infrared cooling on the growth of a population of drops is studied with two numerical modeling frameworks. An eddy-resolving model (ERM) simulation of Arctic stratus clouds is used to generate a dataset of 500 parcel trajectories that follow the mean dynamic motions of the simulated cloud. The 500-parcel dataset is used to drive a trajectory ensemble model (TEM) coupled to an explicit microphysical model that includes the radiative term in the vapor growth equation. The second framework is that of the ERM itself.Results from the TEM show that the production of drizzle-sized drops is strongly dependent upon parcel cloud-top residence time for both radiative- and nonradiative-influenced growth. Drizzle-sized drops can be produced between 20 and 50 min earlier through the inclusion of the radiative term, which corroborates earlier results. The radiative effect may also cause drops with r < 10 m to evaporate, producing a bimodal size spectrum. Parcel cloud-top residence times as short as 12 min can initiate this bimodal spectrum. TEM results show that the radiative effect increases drizzle drop mass predominately in parcels that tend to contribute to drizzle even in the absence of the radiative term. Activation of large cloud condensation nuclei appears to have a larger effect on drizzle production than does the radiative term. ERM simulations show a weak overall influence of the radiative term. Drizzle onset occurs earlier when the radiative term is included (about 20 min), but there is no strong change in the overall structure or evolution of the cloud.

  16. Resistance Genes and Genetic Elements Associated with Antibiotic Resistance in Clinical and Commensal Isolates of Streptococcus salivarius

    PubMed Central

    Chaffanel, Fanny; Charron-Bourgoin, Florence; Libante, Virginie; Leblond-Bourget, Nathalie

    2015-01-01

    The diversity of clinical (n = 92) and oral and digestive commensal (n = 120) isolates of Streptococcus salivarius was analyzed by multilocus sequence typing (MLST). No clustering of clinical or commensal strains can be observed in the phylogenetic tree. Selected strains (92 clinical and 46 commensal strains) were then examined for their susceptibilities to tetracyclines, macrolides, lincosamides, aminoglycosides, and phenicol antibiotics. The presence of resistance genes tet(M), tet(O), erm(A), erm(B), mef(A/E), and catQ and associated genetic elements was investigated by PCR, as was the genetic linkage of resistance genes. High rates of erythromycin and tetracycline resistance were observed among the strains. Clinical strains displayed either the erm(B) (macrolide-lincosamide-streptogramin B [MLSB] phenotype) or mef(A/E) (M phenotype) resistance determinant, whereas almost all the commensal strains harbored the mef(A/E) resistance gene, carried by a macrolide efflux genetic assembly (MEGA) element. A genetic linkage between a macrolide resistance gene and genes of Tn916 was detected in 23 clinical strains and 5 commensal strains, with a predominance of Tn3872 elements (n = 13), followed by Tn6002 (n = 11) and Tn2009 (n = 4) elements. Four strains harboring a mef(A/E) gene were also resistant to chloramphenicol and carried a catQ gene. Sequencing of the genome of one of these strains revealed that these genes colocalized on an IQ-like element, as already described for other viridans group streptococci. ICESt3-related elements were also detected in half of the isolates. This work highlights the potential role of S. salivarius in the spread of antibiotic resistance genes both in the oral sphere and in the gut. PMID:25862227

  17. Epidemiological and molecular characteristics of clinical isolates of Streptococcus pyogenes collected between 2005 and 2008 from Chinese children.

    PubMed

    Liang, Yunmei; Liu, Xiaorong; Chang, Hesheng; Ji, Lili; Huang, Guoying; Fu, Zhou; Zheng, Yuejie; Wang, Libo; Li, Chengrong; Shen, Ying; Yu, Sangjie; Yao, Kaihu; Ma, Lin; Shen, Xuzhuang; Yang, Yonghong

    2012-07-01

    The aim of this study was to explore the epidemiological and molecular characteristics of Streptococcus pyogenes in children from different cities in mainland China who were diagnosed with scarlet fever, impetigo and pharyngitis, as well as to detect asymptomatic carriers, between 2005 and 2008, and to compare the results with isolates from rural Chinese children with acute glomerulonephritis in 2005 and in the 1990s. Susceptibility tests to determine MICs and analysis of the presence of erythromycin-resistant genes (mefA, ermB and ermA) and emm gene typing were performed on 466 S. pyogenes isolates from Beijing, Shanghai, Chongqing and Shenzhen. Superantigen genes (speA and speC) were examined by performing PCR on isolates with the most prevalent emm genotype. All isolates were sensitive to penicillin, cefradine and ofloxacin. The highest rate of resistance was against clarithromycin (98.1 %), followed by erythromycin (97.6 %), azithromycin and clindamycin (both 97.2 %), and tetracycline (94.0 %). Among the 466 isolates, 421 (90.3 %) harboured the ermB gene, 145 (31.1 %) were speA-positive and 273 (58.6 %) were speC-positive. The speA gene was common in emm1.0 (88.8 %) and emm6.5 (83.3 %) genotypes. The speC gene was frequently observed in emm4.0 (90.0 %), emm12.0 (69.6 %), emm18.0 (66.7 %), emm22.0 (75.9 %) and emm80.0 (80.0 %) genotypes. The most prevalent emm genotypes in mainland China in recent years were emm1.0 and emm12.0. All isolates remained sensitive to β-lactams and quinolone. PMID:22442290

  18. Changes in metamorphopsia in daily life after successful epiretinal membrane surgery and correlation with M-CHARTS score

    PubMed Central

    Kinoshita, Takamasa; Imaizumi, Hiroko; Miyamoto, Hirotomo; Okushiba, Utako; Hayashi, Yuki; Katome, Takashi; Mitamura, Yoshinori

    2015-01-01

    Purpose To determine the correlation between the changes in metamorphopsia in daily life environment and the M-CHARTS scores after epiretinal membrane (ERM) removal, and to determine the criterion for determining whether clinically significant changes in the metamorphopsia score have occurred in M-CHARTS. Methods We studied 65 eyes undergoing vitrectomy for unilateral ERM. Self-administered questionnaires were used to examine the metamorphopsia in their daily life. The degree of metamorphopsia was determined by M-CHARTS. The receiver operating characteristic curve was used to determine the best predictor of the changes in metamorphopsia in daily life. To determine the reproducibility of the M-CHARTS score, another set of 56 eyes with ERM was tested twice on two different days. Results The postoperative changes in the logarithm of the M-CHARTS score was defined as M2-value. The area under the receiver operating characteristic curve for the M2-value as a predictor of the changes in metamorphopsia in daily life was larger than area under the receiver operating characteristic curve obtained for any other parameter. The optimal cutoff value was −0.4. The 95% limits of agreement between test and retest measurements had a reproducibility of ±0.3 logarithm of the M-CHARTS score. Taking into account not only the reproducibility but also the consistency with the subjective changes, we determined the criterion for clinically significant changes in the M-CHARTS scores as a change of the M2-value by ≥0.4. Conclusion Evaluating the changes in the M-CHARTS scores in logarithmic form is favorable not only theoretically but also from the perspective of consistency with the subjective changes. PMID:25678770

  19. Characterization of Three New Glutaredoxin Genes in the Arbuscular Mycorrhizal Fungus Rhizophagus irregularis: Putative Role of RiGRX4 and RiGRX5 in Iron Homeostasis

    PubMed Central

    Tamayo, Elisabeth; Benabdellah, Karim; Ferrol, Nuria

    2016-01-01

    Glutaredoxins (GRXs) are small ubiquitous oxidoreductases involved in the regulation of the redox state in living cells. In an attempt to identify the full complement of GRXs in the arbuscular mycorrhizal (AM) fungus Rhizophagus irregularis, three additional GRX homologs, besides the formerly characterized GintGRX1 (renamed here as RiGRX1), were identified. The three new GRXs (RiGRX4, RiGRX5 and RiGRX6) contain the CXXS domain of monothiol GRXs, but whereas RiGRX4 and RiGRX5 belong to class II GRXs, RiGRX6 belongs to class I together with RiGRX1. By using a yeast expression system, we observed that the newly identified homologs partially reverted sensitivity of the GRX deletion yeast strains to external oxidants. Furthermore, our results indicated that RiGRX4 and RiGRX5 play a role in iron homeostasis in yeast. Gene expression analyses revealed that RiGRX1 and RiGRX6 were more highly expressed in the intraradical (IRM) than in the extraradical mycelium (ERM). Exposure of the ERM to hydrogen peroxide induced up-regulation of RiGRX1, RiGRX4 and RiGRX5 gene expression. RiGRX4 expression was also up-regulated in the ERM when the fungus was grown in media supplemented with a high iron concentration. These data indicate the two monothiol class II GRXs, RiGRX4 and RiGRX5, might be involved in oxidative stress protection and in the regulation of fungal iron homeostasis. Increased expression of RiGRX1 and RiGRX6 in the IRM suggests that these GRXs should play a key role in oxidative stress protection of R. irregularis during its in planta phase. PMID:26900849

  20. Quantitative analysis of macular contraction in idiopathic epiretinal membrane

    PubMed Central

    2014-01-01

    Background We aimed to quantify the displacement of macular capillaries using infrared fundus photographs and image processing software (ImageJ) in patients with idiopathic epiretinal membrane (ERM) who have undergone vitrectomy and to analyze the correlation between vessel displacement and retinal thickness. Methods This prospective study included 16 patients who underwent vitrectomy for idiopathic ERM. Ophthalmic examination and optical coherence tomography (OCT) were performed before and 3 months after surgery. The length of radial vessel segment included in each area (VLA) and the length from the foveola to the vessel branching point (FBL) depending on the superior, inferior, nasal, and temporal areas of the macula were measured using infrared fundus images and image processing software (ImageJ). Preoperative and postoperative parameters were compared and correlations between VLA, FBL, macular thickness, and visual acuity were assessed. Results The VLA of superior, inferior, and temporal areas showed a significant postoperative reduction. VLA differences showed a positive correlation with differences in macular thickness, which corresponded to the superior, inferior, and temporal areas; however, no correlation was observed in the nasal area. The FBL of the superior and inferior areas was significantly increased postoperatively. A positive correlation was observed between FBL differences and macular thickness differences in the superior area. Postoperative change in VLA and FBL did not show a significant correlation with postoperative best corrected visual acuity (BCVA) and BCVA differences. Conclusions Infrared fundus photographs and image processing software can be useful for quantifying progressive changes in retinal surface distortion after surgical removal of ERM. Macular edema and vascular distortion showed significant improvement after surgery. Furthermore, a correlation was observed between topographic and tomographic changes. PMID:24735324

  1. Periostin, discovered by nano-flow liquid chromatography and mass spectrometry, is a novel marker of diabetic retinopathy

    SciTech Connect

    Takada, Michiya; Ban, Yoshiyuki; Yamamoto, Gou; Ueda, Toshihiko; Saito, Yuta; Nishimura, Eiichi; Fujisawa, Kunimi; Koide, Ryohei; Mizutani, Masakazu; Kozawa, Tadahiko; Shiraishi, Yuji; Bando, Yasuhiko; Tachikawa, Tetsuhiko; Hirano, Tsutomu

    2010-08-20

    Research highlights: {yields} In proliferative membrane and epiretinal membrane specimens, the numbers of proteins are 225 and 154, respectively, and 123 proteins are common to both. {yields} Periostin and thrombospondin-1 proteins are unique to the proliferative membrane specimens. {yields} The expression of periostin is significantly up-regulated in proliferative membrane specimens. -- Abstract: Diabetes can lead to serious microvascular complications including proliferative diabetic retinopathy (PDR), the leading cause of blindness in adults. Recent studies using gene array technology have attempted to apply a hypothesis-generating approach to elucidate the pathogenesis of PDR, but these studies rely on mRNA differences, which may or may not be related to significant biological processes. To better understand the basic mechanisms of PDR and to identify potential new biomarkers, we performed shotgun liquid chromatography (LC)/tandem mass spectrometry (MS/MS) analysis on pooled protein extracts from neovascular membranes obtained from PDR specimens and compared the results with those from non-vascular epiretinal membrane (ERM) specimens. We detected 226 distinct proteins in neovascular membranes and 154 in ERM. Among these proteins, 102 were specific to neovascular membranes and 30 were specific to ERM. We identified a candidate marker, periostin, as well as several known PDR markers such as pigment epithelium-derived factor (PEDF). We then performed RT-PCR using these markers. The expression of periostin was significantly up-regulated in proliferative membrane specimens. Periostin induces cell attachment and spreading and plays a role in cell adhesion. Proteomic analysis by LC/MS/MS, which permits accurate quantitative comparison, was useful in identifying new candidates such as periostin potentially involved in the pathogenesis of PDR.

  2. Emerging non-PCV13 serotypes of noninvasive Streptococcus pneumoniae with macrolide resistance genes in northern Japan

    PubMed Central

    Kawaguchiya, M.; Urushibara, N.; Aung, M.S.; Morimoto, S.; Ito, M.; Kudo, K.; Sumi, A.; Kobayashi, N.

    2015-01-01

    In Japan, the 7-valent pneumococcal conjugate vaccine (PCV7) was introduced to the nation's routine immunization program in April 2013 and was replaced by the 13-valent pneumococcal conjugate vaccine (PCV13) in November 2013. Distribution of serotypes and macrolide resistance genotypes was investigated for a total of 1097 (975 children, 122 adults) and 960 (873 children, 87 adults) clinical isolates of Streptococcus pneumoniae from noninvasive infections in Hokkaido (northern main island of Japan) in the routine immunization periods for PCV7 and PCV13 (April–October 2013 and November 2013–November 2014, respectively). Serotype was determined by sequential multiplex PCR and additional genetic analyses. Macrolide resistance genes erm(B) and mef(A/E) were detected by multiplex PCR. Although the most prevalent serotypes in children were 23A and 6C in the PCV7 period, after replacement with PCV13, 19A became the most common, followed by 6C, 15A and 23A. Among adults, serotype 3 was consistently the most frequent throughout the study periods. Compared with values from the pre-PCV7 routine immunization period, PCV7 serotypes decreased from 48.3 to 3.3% in the PCV13 period among children, while the rates of non-PCV13 serotypes (particularly 15A, 23A, 11A, 10A and 35B) increased from 39.7 to 75.1% (p < 0.001). In the PCV13 period, erm(B), mef(A/E) and both of these genes were detected in 75.8, 31.6 and 11.3% of all isolates, respectively. Serotype 19A accounted for 76.9% of the isolates with both the macrolide resistance genes, and emerging non-PCV13 serotypes 15A, 15C and 23A mostly harboured erm(B). PMID:26909157

  3. Emerging non-PCV13 serotypes of noninvasive Streptococcus pneumoniae with macrolide resistance genes in northern Japan.

    PubMed

    Kawaguchiya, M; Urushibara, N; Aung, M S; Morimoto, S; Ito, M; Kudo, K; Sumi, A; Kobayashi, N

    2016-01-01

    In Japan, the 7-valent pneumococcal conjugate vaccine (PCV7) was introduced to the nation's routine immunization program in April 2013 and was replaced by the 13-valent pneumococcal conjugate vaccine (PCV13) in November 2013. Distribution of serotypes and macrolide resistance genotypes was investigated for a total of 1097 (975 children, 122 adults) and 960 (873 children, 87 adults) clinical isolates of Streptococcus pneumoniae from noninvasive infections in Hokkaido (northern main island of Japan) in the routine immunization periods for PCV7 and PCV13 (April-October 2013 and November 2013-November 2014, respectively). Serotype was determined by sequential multiplex PCR and additional genetic analyses. Macrolide resistance genes erm(B) and mef(A/E) were detected by multiplex PCR. Although the most prevalent serotypes in children were 23A and 6C in the PCV7 period, after replacement with PCV13, 19A became the most common, followed by 6C, 15A and 23A. Among adults, serotype 3 was consistently the most frequent throughout the study periods. Compared with values from the pre-PCV7 routine immunization period, PCV7 serotypes decreased from 48.3 to 3.3% in the PCV13 period among children, while the rates of non-PCV13 serotypes (particularly 15A, 23A, 11A, 10A and 35B) increased from 39.7 to 75.1% (p < 0.001). In the PCV13 period, erm(B), mef(A/E) and both of these genes were detected in 75.8, 31.6 and 11.3% of all isolates, respectively. Serotype 19A accounted for 76.9% of the isolates with both the macrolide resistance genes, and emerging non-PCV13 serotypes 15A, 15C and 23A mostly harboured erm(B). PMID:26909157

  4. Development of a rule-based algorithm for rice cultivation mapping using Landsat 8 time series

    NASA Astrophysics Data System (ADS)

    Karydas, Christos G.; Toukiloglou, Pericles; Minakou, Chara; Gitas, Ioannis Z.

    2015-06-01

    In the framework of ERMES project (FP7 66983), an algorithm for mapping rice cultivation extents using mediumhigh resolution satellite data was developed. ERMES (An Earth obseRvation Model based RicE information Service) aims to develop a prototype of downstream service for rice yield modelling based on a combination of Earth Observation and in situ data. The algorithm was designed as a set of rules applied on a time series of Landsat 8 images, acquired throughout the rice cultivation season of 2014 from the plain of Thessaloniki, Greece. The rules rely on the use of spectral indices, such as the Normalized Difference Vegetation Index (NDVI), the Normalized Difference Water Index (NDWI), and the Normalized Seasonal Wetness Index (NSWI), extracted from the Landsat 8 dataset. The algorithm is subdivided into two phases: a) a hard classification phase, resulting in a binary map (rice/no-rice), where pixels are judged according to their performance in all the images of the time series, while index thresholds were defined after a trial and error approach; b) a soft classification phase, resulting in a fuzzy map, by assigning scores to the pixels which passed (as `rice') the first phase. Finally, a user-defined threshold of the fuzzy score will discriminate rice from no-rice pixels in the output map. The algorithm was tested in a subset of Thessaloniki plain against a set of selected field data. The results indicated an overall accuracy of the algorithm higher than 97%. The algorithm was also applied in a study are in Spain (Valencia) and a preliminary test indicated a similar performance, i.e. about 98%. Currently, the algorithm is being modified, so as to map rice extents early in the cultivation season (by the end of June), with a view to contribute more substantially to the rice yield prediction service of ERMES. Both algorithm modes (late and early) are planned to be tested in extra Mediterranean study areas, in Greece, Italy, and Spain.

  5. Characterization of the effect of TIMAP phosphorylation on its interaction with protein phosphatase 1.

    PubMed

    Czikora, István; Kim, Kyung-mi; Kása, Anita; Bécsi, Bálint; Verin, Alexander D; Gergely, Pál; Erdodi, Ferenc; Csortos, Csilla

    2011-07-01

    TIMAP, TGF-β inhibited, membrane-associated protein, is highly abundant in endothelial cells (EC). We have shown earlier the involvement of TIMAP in PKA-mediated ERM (ezrin-radixin-moesin) dephosphorylation as part of EC barrier protection by TIMAP (Csortos et al., 2008). Emerging data demonstrate the regulatory role of TIMAP on protein phosphatase 1 (PP1) activity. We provide here evidence for specific interaction (K(a) = 1.80 × 10(6) M(-1)) between non-phosphorylated TIMAP and the catalytic subunit of PP1 (PP1c) by surface plasmon resonance based binding studies. Thiophosphorylation of TIMAP by PKA, or sequential thiophosphorylation by PKA and GSK3β slightly modifies the association constant for the interaction of TIMAP with PP1c and decreases the rate of dissociation. However, dephosphorylation of phospho-moesin substrate by PP1cβ is inhibited to different extent in the presence of non- (~60% inhibition), mono- (~50% inhibition) or double-thiophosphorylated (<10% inhibition) form of TIMAP. Our data suggest that double-thiophosphorylation of TIMAP has minor effect on its binding ability to PP1c, but considerably attenuates its inhibitory effect on the activity of PP1c. PKA activation by forskolin treatment of EC prevented thrombin evoked barrier dysfunction and ERM phosphorylation at the cell membrane (Csortos et al., 2008). With the employment of specific GSK3β inhibitor it is shown here that PKA activation is followed by GSK3β activation in bovine pulmonary EC and both of these activations are required for the rescuing effect of forskolin in thrombin treated EC. Our results suggest that the forskolin induced PKA/GSK3β activation protects the EC barrier via TIMAP-mediated decreasing of the ERM phosphorylation level. PMID:21466834

  6. Macular Surgery Using Intraoperative Spectral Domain Optical Coherence Tomography

    PubMed Central

    Riazi-Esfahani, Mohammad; Khademi, Mohammad Reza; Mazloumi, Mehdi; Khodabandeh, Alireza; Riazi-Esfahani, Hamid

    2015-01-01

    Purpose: To report the use of intraoperative spectral domain optical coherence tomography (SD-OCT) for detecting anatomical changes during macular surgery. Methods: In a consecutive case series, 32 eyes of 32 patients undergoing concurrent pars plana vitrectomy and intraoperative SD-OCT for macular hole (MH), epiretinal membrane (ERM) and vitreomacular traction (VMT) were enrolled. Intraoperative changes in retinal thickness and dimensions of the macular hole were measured in patients with ERM and VMT following surgical manipulation using a hand-held SD-OCT device (iVue, Optovue Inc., Fremont, CA, USA). Results: SD-OCT images of sixteen eyes with macular hole were subjected to quantitative and qualitative analysis. All MH dimensions remained stable during consecutive stages of surgery except for MH apex diameter, which showed a significant decrease after internal limiting membrane (ILM) peeling (P=0.025). Quantitative analysis of ten patients with ERM showed a significant decrease in retinal thickness after membrane removal (P=0.018) which did not remain significant until the end of the procedure (P=0.8). In three cases, subretinal fluid was formed after ILM peeling. Quantitative analysis of five patients with VMT showed a decrease in retinal thickness during consecutive steps of the surgery, although these changes were not significant. In two cases, subretinal fluid was formed after ILM peeling. Conclusion: Intraoperative SD-OCT is a useful imaging technique which provides vitreoretinal surgeons with rapid awareness of changes in macular anatomy during surgery and may therefore result in better anatomical and visual outcomes. PMID:26730318

  7. Radiative impacts on the growth of a population of drops within simulated summertime Arctic stratus

    SciTech Connect

    Harrington, J.Y.; Feingold, G.; Cotton, W.R.

    2000-03-01

    The impact of solar heating and infrared cooling on the growth of a population of drops is studied with two numerical modeling frameworks. An eddy-resolving model (ERM) simulation of Arctic stratus clouds is used to generate a dataset of 500 parcel trajectories that follow the mean dynamic motions of the simulated cloud. The 500-parcel dataset is used to drive a trajectory ensemble model (TEM) coupled to an explicit microphysical model that includes the radiative term in the vapor growth equation. The second framework is that of the ERM itself. Results from the TEM show that the production of drizzle-sized drops is strongly dependent upon parcel cloud-top resistance time for both radiative- and nonradiative-influenced growth. drizzle-sized drops can be produced between 20 and 50 min earlier through the inclusion of the radiative terms, which corroborates earlier results. The radiative effect may also cause drops with r < 10 {mu}m to evaporate, producing a bimodal size spectrum. Parcel cloud-top residence times as short as 12 min can initiate this bimodal spectrum. TEM results show that the radiative effect increases drizzle drop mass predominantly in parcels that tend to contribute to drizzle even in the absence of the radiative term. Activation of large cloud condensation nuclei appears to have a larger effect on drizzle production than does the radiative term. ERM simulations show a weak overall influence of the radiative term. Drizzle onset occurs earlier when the radiative term is included (about 20 min), but there is no strong change in the overall structure or evolution of the cloud.

  8. Chemometric tools to evaluate the spatial distribution of trace metals in surface sediments of two Spanish rías.

    PubMed

    Quelle, Cristina; Besada, Victoria; Andrade, José Manuel; Gutiérrez, Noemí; Schultze, Fernando; Gago, Jesús; González, Juan José

    2011-12-15

    A suite of relevant trace metals (Hg, Pb, Cd, Cu, Zn and Ni) was measured in surface sediment samples to assess the environmental situation of the largest two Atlantic Spanish 'rías' (a form of estuaries, ría of Pontevedra, ROP, and ría of Vigo, ROV). The level of contamination originated by these metals was assessed against international guidelines, the threshold effect, ERL, and the midrange effect, ERM. Six unsupervised and supervised multivariate chemometric techniques were applied to model each ría, compare them and select those metals that characterize the samples. This is first time that such a study is performed for these two important seafood-producing areas. Maximum concentrations at ROP occurred in the vicinities of an inner island, where Cu, Zn, Ni and Pb presented concentrations over the ERL and Hg over the ERM. Highest concentrations of metals in ROV were observed in the proximities of Vigo shipyards and port, except for Pb, with peak values in San Simon Bay. ERL limits were exceeded in the inner part of this ría for Cu, Zn and Hg and in a wider area for Pb and Ni. Levels for Pb went beyond the ERM boundary in the axial part of San Simon Bay. In general, the distribution of the metals was more homogeneous in ría of Pontevedra than in ría of Vigo (where three morphological zones were characterized). Both rías could be differentiated using only two metals: Ni and Hg, as deduced from the multivariate techniques. PMID:22099668

  9. Novel insights of the gastric gland organization revealed by chief cell specific expression of moesin.

    PubMed

    Zhu, Lixin; Hatakeyama, Jason; Zhang, Bing; Makdisi, Joy; Ender, Cody; Forte, John G

    2009-02-01

    ERM (ezrin, radixin, and moesin) proteins play critical roles in epithelial and endothelial cell polarity, among other functions. In gastric glands, ezrin is mainly expressed in acid-secreting parietal cells, but not in mucous neck cells or zymogenic chief cells. In looking for other ERM proteins, moesin was found lining the lumen of much of the gastric gland, but it was not expressed in parietal cells. No significant radixin expression was detected in the gastric glands. Moesin showed an increased gradient of expression from the neck to the base of the glands. In addition, the staining pattern of moesin revealed a branched morphology for the gastric lumen. This pattern of short branches extending from the glandular lumen was confirmed by using antibody against zonula occludens-1 (ZO-1) to stain tight junctions. With a mucous neck cell probe (lectin GSII, from Griffonia simplicifolia) and a chief cell marker (pepsinogen C), immunohistochemistry revealed that the mucous neck cells at the top of the glands do not express moesin, but, progressing toward the base, mucous cells showing decreased GSII staining had low or moderate level of moesin expression. The level of moesin expression continued to increase toward the base of the glands and reached a plateau in the base where chief cells and parietal cells abound. The level of pepsinogen expression also increased toward the base. Pepsinogen C was located on cytoplasmic granules and/or more generally distributed in chief cells, whereas moesin was exclusively expressed on the apical membrane. This is a clear demonstration of distinctive cellular expression of two ERM family members in the same tissue. The results provide the first evidence that moesin is involved in the cell biology of chief cells. Novel insights on gastric gland morphology revealed by the moesin and ZO-1 staining provide the basis for a model of cell maturation and migration within the gland. PMID:19074636

  10. Novel insights of the gastric gland organization revealed by chief cell specific expression of moesin

    PubMed Central

    Zhu, Lixin; Hatakeyama, Jason; Zhang, Bing; Makdisi, Joy; Ender, Cody; Forte, John G.

    2009-01-01

    ERM (ezrin, radixin, and moesin) proteins play critical roles in epithelial and endothelial cell polarity, among other functions. In gastric glands, ezrin is mainly expressed in acid-secreting parietal cells, but not in mucous neck cells or zymogenic chief cells. In looking for other ERM proteins, moesin was found lining the lumen of much of the gastric gland, but it was not expressed in parietal cells. No significant radixin expression was detected in the gastric glands. Moesin showed an increased gradient of expression from the neck to the base of the glands. In addition, the staining pattern of moesin revealed a branched morphology for the gastric lumen. This pattern of short branches extending from the glandular lumen was confirmed by using antibody against zonula occludens-1 (ZO-1) to stain tight junctions. With a mucous neck cell probe (lectin GSII, from Griffonia simplicifolia) and a chief cell marker (pepsinogen C), immunohistochemistry revealed that the mucous neck cells at the top of the glands do not express moesin, but, progressing toward the base, mucous cells showing decreased GSII staining had low or moderate level of moesin expression. The level of moesin expression continued to increase toward the base of the glands and reached a plateau in the base where chief cells and parietal cells abound. The level of pepsinogen expression also increased toward the base. Pepsinogen C was located on cytoplasmic granules and/or more generally distributed in chief cells, whereas moesin was exclusively expressed on the apical membrane. This is a clear demonstration of distinctive cellular expression of two ERM family members in the same tissue. The results provide the first evidence that moesin is involved in the cell biology of chief cells. Novel insights on gastric gland morphology revealed by the moesin and ZO-1 staining provide the basis for a model of cell maturation and migration within the gland. PMID:19074636

  11. Ezrin: a regulator of actin microfilaments in cell junctions of the rat testis.

    PubMed

    Gungor-Ordueri, N Ece; Celik-Ozenci, Ciler; Cheng, C Yan

    2015-01-01

    Ezrin, radixin, moesin and merlin (ERM) proteins are highly homologous actin-binding proteins that share extensive sequence similarity with each other. These proteins tether integral membrane proteins and their cytoplasmic peripheral proteins (e.g., adaptors, nonreceptor protein kinases and phosphatases) to the microfilaments of actin-based cytoskeleton. Thus, these proteins are crucial to confer integrity of the apical membrane domain and its associated junctional complex, namely the tight junction and the adherens junction. Since ectoplasmic specialization (ES) is an F-actin-rich testis-specific anchoring junction-a highly dynamic ultrastructure in the seminiferous epithelium due to continuous transport of germ cells, in particular spermatids, across the epithelium during the epithelial cycle-it is conceivable that ERM proteins are playing an active role in these events. Although these proteins were first reported almost 25 years and have since been extensively studied in multiple epithelia/endothelia, few reports are found in the literature to examine their role in the actin filament bundles at the ES. Studies have shown that ezrin is also a constituent protein of the actin-based tunneling nanotubes (TNT) also known as intercellular bridges, which are transient cytoplasmic tubular ultrastructures that transport signals, molecules and even organelles between adjacent and distant cells in an epithelium to coordinate cell events that occur across an epithelium. Herein, we critically evaluate recent data on ERM in light of recent findings in the field in particular ezrin regarding its role in actin dynamics at the ES in the testis, illustrating additional studies are warranted to examine its physiological significance in spermatogenesis. PMID:25652626

  12. Stolt's f-k migration for plane wave ultrasound imaging.

    PubMed

    Garcia, Damien; Le Tarnec, Louis; Muth, Stéphan; Montagnon, Emmanuel; Porée, Jonathan; Cloutier, Guy

    2013-09-01

    Ultrafast ultrasound is an emerging modality that offers new perspectives and opportunities in medical imaging. Plane wave imaging (PWI) allows one to attain very high frame rates by transmission of planar ultrasound wave-fronts. As a plane wave reaches a given scatterer, the latter becomes a secondary source emitting upward spherical waves and creating a diffraction hyperbola in the received RF signals. To produce an image of the scatterers, all the hyperbolas must be migrated back to their apexes. To perform beamforming of plane wave echo RFs and return high-quality images at high frame rates, we propose a new migration method carried out in the frequency-wavenumber (f-k) domain. The f-k migration for PWI has been adapted from the Stolt migration for seismic imaging. This migration technique is based on the exploding reflector model (ERM), which consists in assuming that all the scatterers explode in concert and become acoustic sources. The classical ERM model, however, is not appropriate for PWI. We showed that the ERM can be made suitable for PWI by a spatial transformation of the hyperbolic traces present in the RF data. In vitro experiments were performed to outline the advantages of PWI with Stolt's f-k migration over the conventional delay-and-sum (DAS) approach. The Stolt's f-k migration was also compared with the Fourier-based method developed by J.-Y. Lu. Our findings show that multi-angle compounded f-k migrated images are of quality similar to those obtained with a stateof- the-art dynamic focusing mode. This remained true even with a very small number of steering angles, thus ensuring a highly competitive frame rate. In addition, the new FFT-based f-k migration provides comparable or better contrast-to-noise ratio and lateral resolution than the Lu's and DAS migration schemes. Matlab codes for the Stolt's f-k migration for PWI are provided. PMID:24626107

  13. Co-addition of manure increases the dissipation rates of tylosin A and the numbers of resistance genes in laboratory incubation experiments.

    PubMed

    Li, Qian; Wang, Yan; Zou, Yong-De; Liao, Xin-Di; Liang, Juan-Boo; Xin, Wen; Wu, Yin-Bao

    2015-09-15

    The behavior of veterinary antibiotics in the soil is commonly studied using the following methods to add antibiotics to the soil: (A) adding manure collected from animals fed a diet that includes antibiotics; (B) adding antibiotic-free animal manure spiked with antibiotics; and (C) the direct addition of antibiotics. However, most studies have only used methods (B) and (C) in their research, and few studies have simultaneously compared the different antibiotic addition methods. This study used tylosin A (TYLA) as a model antibiotic to compare the effects of these three commonly used antibiotic addition methods on the dissipation rates of TYLA and the numbers of resistance genes in laboratory incubation experiments. The results showed that the three treatment methods produced similar TYLA degradation trends; however, there were significant differences (P<0.05) in the TYLA degradation half-life (t1/2) among the three methods. The half-life of TYLA degradation in treatments A, B and C was 2.44 ± 0.04, 1.21 ± 0.03 and 5.13 ± 0.11 days, respectively. The presence of manure resulted in a higher electrical conductivity (EC), higher relative abundance of Citrobacter amalonaticus, higher macrolide resistant gene (ermB, ermF and ermT) count and lower ecological toxicity in the soil, which could partially explain the higher TYLA degradation rate in the treatments containing manure. The higher degradation rate of TYLA in treatment B when compared to treatment A could be due to the lower concentrations of tylosin B (TYLB) and tylosin D (TYLD). The main route for veterinary antibiotics to enter the soil is via the manure of animals that have been administered antibiotics. Therefore, the more appropriate method to study the degradation and ecotoxicity of antibiotic residues in the soil is by using manure from animals fed/administered the particular antibiotic rather than by adding the antibiotic directly to the soil. PMID:25958362

  14. Presence of superantigen genes and antimicrobial resistance in Staphylococcus isolates obtained from the uteri of dairy cows with clinical endometritis.

    PubMed

    Zhao, J-L; Ding, Y-X; Zhao, H-X; He, X-L; Li, P-F; Li, Z-F; Guan, H; Guo, X

    2014-10-11

    Clinical endometritis is an important disease of dairy cattle and results in decreased reproductive performance. This disease is caused by contamination of the uterus with a broad spectrum of microorganisms after calving. In this study, staphylococcal isolates from the uterus of dairy cows with clinical endometritis were tested for their distribution of superantigen (SAg) genes and antimicrobial resistance. Between the 127 staphylococcal isolates collected in this study, 10 species were identified. The predominant strain identified was Staphylococcus aureus (n=53), followed by Staphylococcus saprophyticus (n=38) and Staphylococcus chromogenes (n=22). PCR analysis demonstrated that most isolates (63.0 per cent) harboured at least one SAg gene. The most commonly observed SAg gene and genotype was selj (38.6 per cent) and sec-selj-seln (24.0 per cent), respectively. Most isolates were resistant to penicillin (79.5 per cent), ampicillin (71.7 per cent), erythromycin (56.7 per cent), and tetracycline (52.0 per cent). PCR analysis demonstrated that the antimicrobial resistance determinants ermA, ermB, ermC, tetK, tetM and blaZ were detected in 0 per cent, 44.4 per cent, 51.4 per cent, 68.2 per cent, 13.6 per cent and 86.1 per cent of the erythromycin, tetracycline and β-lactam resistant isolates, respectively. There were 22 (17.3 per cent of all isolates) coagulase-negative staphylococci shown to be methicillin resistant. In the methicillin-resistant isolates, significant resistances to ampicillin, erythromycin and penicillin were observed (P<0.01). The results of this study demonstrate that staphylococci recovered from dairy cows with clinical endometritis contain an extensive and complex prevalence of SAg genes. Significant resistances to antibiotics were also seen, highlighting the need for the rational appliance of antibiotics in veterinary medicine. PMID:24989035

  15. Species Diversity and Pheno- and Genotypic Antibiotic Resistance Patterns of Staphylococci Isolated from Retail Ground Meats.

    PubMed

    Guran, Husnu Sahan; Kahya, Serpil

    2015-06-01

    The presence and species diversity of staphylococci in 250 ground beef and lamb meat samples obtained from Diyarbakir, Turkey were investigated. The presence of the 16S rRNA gene, mecA, nuc, pvl, and femA was analyzed by multiplex PCR. Pheno- and genotypic antibiotic resistance profiles of 208 staphylococci isolates were established. Of the ground beef and ground lamb samples, 86.4% and 62.4% were positive for staphylococci, respectively. Staphylococcus aureus, S. saprophyticus, S. hominis, S. lentus, S. pasteuri, S. warneri, S. intermedius, and S. vitulinus made up 40.8%, 28.8%, 11%, 3.8%, 3.8%, 2.4%, 2.4%, and 2.4% of isolates, respectively. Of the 85 S. aureus isolates, 40%, 47%, and 5.8% carried femA, mecA, and pvl, respectively, whereas the corresponding rates for the 118 coagulase-negative staphylococci (CoNS) were 0%, 10.1%, and 0%, respectively. We determined from the 208 isolates, the highest antibiotic resistances were to tetracycline and oxytetracycline (85.5%), followed by penicillin (51.4%), novobiocin (45.6%), ampicillin (39.9%), and doxycycline (31.7%), using the Clinical and Laboratory Standards Inst. (CLSI) method. All isolates were sensitive to gentamycin, ofloxacin, and tobramycin, but 2.3% of the S. aureus isolates had resistance to vancomycin. The staphylococci isolates carried tet(K), blaZ, tet(L), tet(W), cat, tet(S), tet(M), ermB, ermA, and ermC antibiotic resistance genes at rates of 59%, 51.7%, 36.9%, 31.8%, 27.2%, 27.2%, 24.4%, 18.1%, 7.9%, and 3.9%, respectively. PMID:25944650

  16. Environmental radiation monitoring plan for depleted uranium and beryllium areas, Yuma Proving Ground

    SciTech Connect

    Ebinger, M.H.; Hansen, W.R.

    1994-05-11

    This Environmental Radiation Monitoring Plan (ERM) discusses sampling soils, vegetation, and biota for depleted uranium (DU) and beryllium (Be) at Yuma Proving Ground (YPG). The existing ERM plan was used and modified to more adequately assess the potential of DU and Be migration through the YPG ecosystem. The potential pathways for DU and Be migration are discussed and include soil to vegetation, soil to animals, vegetation to animals, animals to animals, and animals to man. Sample collection will show DU deposition and will be used to estimate DU migration. The number of samples from each area varies and depends on if the firing range of interest is currently used for DU testing (GP 17A) or if the range is not used currently for DU testing (GP 20). Twenty to thirty-five individual mammals or lizards will be sampled from each transect. Air samples and samples of dust in the air fall will be collected in three locations in the active ranges. Thirty to forty-five sediment samples will be collected from different locations in the arroys near the impact areas. DU and Be sampling in the Hard Impact and Soft Impact areas changed only slightly from the existing ERM. The modifications are changes in sample locations, addition of two sediment transport locations, addition of vegetation samples, mammal samples, and air sampling from three to five positions on the impact areas. Analysis of samples for DU or total U by inductively-coupled mass spectroscopy (ICP/MS), cc spectroscopy, neutron activation analysis (NAA), and kinetic phosphorimetric analysis (KPA) are discussed, and analysis for Be by ICP/MS are recommended. Acquiring total U (no isotope data) from a large number of samples and analysis of those samples with relatively high total U concentrations results in fewer isotopic identifications but more information on U distribution. From previous studies, total U concentrations greater than about 3 times natural background are usually DU by isotopic confirmation.

  17. GHG emissions inventory for on-road transportation in the town of Sassari (Sardinia, Italy)

    NASA Astrophysics Data System (ADS)

    Sanna, Laura; Ferrara, Roberto; Zara, Pierpaolo; Duce, Pierpaolo

    2016-04-01

    The IPCC Fifth Assessment Report (AR5) accounts an increase of the total annual anthropogenic GHG emissions between 2000 and 2010 that directly came from the transport sector. In 2010, 14% of GHG emissions were released by transport and fossil-fuel-related CO2 emissions reached about 32 GtCO2 per year. The report also considers adaptation and mitigation as complementary strategies for reducing the risks of climate change for sustainable development of urban areas. This paper describes the on-road traffic emission estimated in the framework of a Sardinian regional project [1] for the town of Sassari (Sardinia, Italy), one of the Sardinian areas where the fuel consumption for on-road transportation purposes is higher [2]. The GHG emissions have been accounted (a) by a calculation-based methodology founded on a linear relationship between source activity and emission, and (b) by the COPERT IV methodology through the EMITRA (EMIssions from road TRAnsport) software tool [3]. Inventory data for annual fossil fuel consumption associated with on-road transportation (diesel, gasoline, gas) have been collected through the Dogane service, the ATP and ARST public transport services and vehicle fleet data are available from the Public Vehicle Database (PRA), using 2010 as baseline year. During this period, the estimated CO2 emissions accounts for more than 180,000 tCO2. The calculation of emissions due to on-road transport quantitatively estimates CO2 and other GHG emissions and represents a useful baseline to identify possible adaptation and mitigation strategies to face the climate change risks at municipal level. Acknowledgements This research was funded by the Sardinian Regional Project "Development, functional checking and setup of an integrated system for the quantification of CO2 net exchange and for the evaluation of mitigation strategies at urban and territorial scale", (Legge Regionale 7 agosto 2007, No. 7). References [1] Sanna L., Ferrara R., Zara P. & Duce P. (2014

  18. Readout Techniques for Drift and Low Frequency Noise Rejection in Infrared Arrays

    NASA Astrophysics Data System (ADS)

    Finger, G.; Dorn, R. J.; Hoffman, A. W.; Mehrgan, H.; Meyer, M.; Moorwood, A. F. M.; Steigmeier, J.

    Three different methods are presented to subtract thermal drifts and low-frequency noise from the signal of infrared array. The first is dead pixels with open Indium bumps, the second is reference output as implemented on the Hawaii2 multiplexer, and the third is dark pixels to emulate reference cells having a capacity connected to the gate of the unit cell field-effect transistor (FET). The third method is the most effective and yields a reduction in readout noise from 15.4-9.4 erms. A novel method will be described to extend this readout technique to the Aladdin 1Kx1K InSb array.

  19. Readout Techniques for Drift and Low Frequency Noise Rejection in Infrared Arrays

    NASA Astrophysics Data System (ADS)

    Finger, Gert; Dorn, Reinhold J.; Hoffman, Alan W.; Mehrgan, Hamid; Meyer, Manfred; Moorwood, Alan F. M.; Stegmeier, Joerg

    2002-10-01

    Three different methods are presentedto subtract thermal drifts and low-frequency noise from thesignal of infrared array. The first is dead pixels with openIndium bumps, the second is reference output as implemented onthe Hawaii2 multiplexer, and the third is dark pixels to emulatereference cells having a capacity connected to the gate of theunit cell field-effect transistor (FET). The third method is themost effective and yields a reduction in readout noise from15.4 9.4 erms. A novel method will be described to extend this readouttechnique to the Aladdin 1 K × 1 K InSb array.

  20. Detection and genetic characterisation of vanA-containing Enterococcus strains in healthy Lusitano horses.

    PubMed

    Moura, I; Radhouani, H; Torres, C; Poeta, P; Igrejas, G

    2010-03-01

    Lusitano horses were investigated in order to detect the presence of vancomycin-resistant enterococci. vanA isolates showed high level vancomycin (Minimum inhibitory concentration; MIC > or = 128 mg/l) and teicoplanin resistance (MIC 64 mg/l), as well as resistance to ciprofloxacin, erythromycin and tetracycline. The tet(L) and erm(B) genes, associated with tetracycline and erythromycin resistance, respectively, were found in all vanA isolates. The intestinal tract of Lusitano horses can be a potential reservoir for vanA-containing enterococci. PMID:20156257

  1. Vernetzung im Kfz

    NASA Astrophysics Data System (ADS)

    Reif, Konrad

    Elektrische und elektronische Systeme im Kfz sind vielfach nicht voneinander unabhängig, sondern beeinflussen und ergänzen sich gegenseitig. Deshalb wurden schon bei den frühen Einspritz- und Zündsystemen Signalleitungen eingesetzt, um eine einfache Kommunikation zwischen diesen beiden Systemen zu ermöglichen. Die zunehmende Anzahl elektronischer Systeme erhöhte jedoch rasch den Bedarf und die Vielfalt an auszutauschenden Informationen. Die Anzahl der hierzu erforderlichen Signalleitungen und Stecker anschlüsse stiegen gleichermaßen, so dass die bis dahin angewandte Technik an ihre Grenzen stieß.

  2. Echtzeitfähigkeit von Satellitenkompassen in der Binnenschifffahrt

    NASA Astrophysics Data System (ADS)

    Haberkamp, G.; Zöbel, D.

    In der maritimen Schifffahrt finden Satellitenkompasse schon seit längerem Verwendung. Nachdem diese Geräte immer erschwinglicher werden, stellt sich die Frage, ob sie auch als Multisensoren für Telematikanwendungen in der Binnenschifffahrt geeignet sind. Im Rahmen einer umfassenden Untersuchung wurde auch die Fragestellung untersucht, wie zeitnah die Sensorinformationen verfügbar werden. Es wurde ein Versuchsaufbau entwickelt, der es ermöglicht, Satellitenkompasse auf ihre Echtzeitfähigkeit hin zu überprüfen. Das vorliegende Papier beschreibt den entwickelten Versuchsaufbau, dokumentiert die Ergebnisse und gibt einen Ausblick auf Verbesserungspotenziale.

  3. Excretion of Antibiotic Resistance Genes by Dairy Calves Fed Milk Replacers with Varying Doses of Antibiotics

    PubMed Central

    Thames, Callie H.; Pruden, Amy; James, Robert E.; Ray, Partha P.; Knowlton, Katharine F.

    2012-01-01

    Elevated levels of antibiotic resistance genes (ARGs) in soil and water have been linked to livestock farms and in some cases feed antibiotics may select for antibiotic resistant gut microbiota. The purpose of this study was to examine the establishment of ARGs in the feces of calves receiving milk replacer containing no antibiotics versus subtherapeutic or therapeutic doses of tetracycline and neomycin. The effect of antibiotics on calf health was also of interest. Twenty-eight male and female dairy calves were assigned to one of the three antibiotic treatment groups at birth and fecal samples were collected at weeks 6, 7 (prior to weaning), and 12 (5 weeks after weaning). ARGs corresponding to the tetracycline (tetC, tetG, tetO, tetW, and tetX), macrolide (ermB, ermF), and sulfonamide (sul1, sul2) classes of antibiotics along with the class I integron gene, intI1, were monitored by quantitative polymerase chain reaction as potential indicators of direct selection, co-selection, or horizontal gene transfer of ARGs. Surprisingly, there was no significant effect of antibiotic treatment on the absolute abundance (gene copies per gram wet manure) of any of the ARGs except ermF, which was lower in the antibiotic-treated calf manure, presumably because a significant portion of host bacterial cells carrying ermF were not resistant to tetracycline or neomycin. However, relative abundance (gene copies normalized to 16S rRNA genes) of tetO was higher in calves fed the highest dose of antibiotic than in the other treatments. All genes, except tetC and intI1, were detectable in feces from 6 weeks onward, and tetW and tetG significantly increased (P < 0.10), even in control calves. Overall, the results provide new insight into the colonization of calf gut flora with ARGs in the early weeks. Although feed antibiotics exerted little effect on the ARGs monitored in this study, the fact that they also provided no health benefit suggests that the greater than conventional

  4. Comparison of different multiple flow algorithms for topographic RUSLE factor (LS) calculation in Switzerland

    NASA Astrophysics Data System (ADS)

    Bircher, Pascal; Liniger, Hanspeter; Prasuhn, Volker

    2016-04-01

    Soil erosion is a well-known challenge both from a global perspective and in Switzerland, and it is assessed and discussed in many projects (e.g. national or European erosion risk maps). Meaningful assessment of soil erosion requires models that adequately reflect surface water flows. Various studies have attempted to achieve better modelling results by including multiple flow algorithms in the topographic length and slope factor (LS-factor) of the Revised Universal Soil Loss Equation (RUSLE). The choice of multiple flow algorithms is wide, and many of them have been implemented in programs or tools like Saga-Gis, GrassGis, ArcGIS, ArcView, Taudem, and others. This study compares six different multiple flow algorithms with the aim of identifying a suitable approach to calculating the LS factor for a new soil erosion risk map of Switzerland. The comparison of multiple flow algorithms is part of a broader project to model soil erosion for the entire agriculturally used area in Switzerland and to renew and optimize the current erosion risk map of Switzerland (ERM2). The ERM2 was calculated in 2009, using a high resolution digital elevation model (2 m) and a multiple flow algorithm in ArcView. This map has provided the basis for enforcing soil protection regulations since 2010 and has proved its worth in practice, but it has become outdated (new basic data are now available, e.g. data on land use change, a new rainfall erosivity map, a new digital elevation model, etc.) and is no longer user friendly (ArcView). In a first step towards its renewal, a new data set from the Swiss Federal Office of Topography (Swisstopo) was used to generate the agricultural area based on the existing field block map. A field block is an area consisting of farmland, pastures, and meadows which is bounded by hydrological borders such as streets, forests, villages, surface waters, etc. In our study, we compared the six multiple flow algorithms with the LS factor calculation approach used in

  5. Phage Transduction.

    PubMed

    Goh, Shan

    2016-01-01

    Bacteriophages mediate horizontal gene transfer through a mechanism known as transduction. Phage transduction carried out in the laboratory involves a bacterial donor and a recipient, both of which are susceptible to infection by the phage of interest. Phage is propagated in the donor, concentrated, and exposed transiently to recipient at different multiplicity of infection ratios. Transductants are selected for the desired phenotype by culture on selective medium. Here we describe transduction of ermB conferring resistance to erythromycin by the C. difficile phage ϕC2. PMID:27507341

  6. Antibiotic Susceptibility Profiles of Dairy Leuconostoc, Analysis of the Genetic Basis of Atypical Resistances and Transfer of Genes In Vitro and in a Food Matrix

    PubMed Central

    Delgado, Susana; Alegría, Ángel; Salvetti, Elisa; Felis, Giovanna E.; Mayo, Baltasar; Torriani, Sandra

    2016-01-01

    In spite of a global concern on the transfer of antibiotic resistances (AR) via the food chain, limited information exists on this issue in species of Leuconostoc and Weissella, adjunct cultures used as aroma producers in fermented foods. In this work, the minimum inhibitory concentration was determined for 16 antibiotics in 34 strains of dairy origin, belonging to Leuconostoc mesenteroides (18), Leuconostoc citreum (11), Leuconostoc lactis (2), Weissella hellenica (2), and Leuconostoc carnosum (1). Atypical resistances were found for kanamycin (17 strains), tetracycline and chloramphenicol (two strains each), and erythromycin, clindamycin, virginiamycin, ciprofloxacin, and rifampicin (one strain each). Surprisingly, L. mesenteroides subsp. mesenteroides LbE16, showed resistance to four antibiotics, kanamycin, streptomycin, tetracycline and virginiamycin. PCR analysis identified tet(S) as responsible for tetracycline resistance in LbE16, but no gene was detected in a second tetracycline-resistant strain, L. mesenteroides subsp. cremoris LbT16. In Leuconostoc mesenteroides subsp. dextranicum LbE15, erythromycin and clindamycin resistant, an erm(B) gene was amplified. Hybridization experiments proved erm(B) and tet(S) to be associated to a plasmid of ≈35 kbp and to the chromosome of LbE15 and LbE16, respectively. The complete genome sequence of LbE15 and LbE16 was used to get further insights on the makeup and genetic organization of AR genes. Genome analysis confirmed the presence and location of erm(B) and tet(S), but genes providing tetracycline resistance in LbT16 were again not identified. In the genome of the multi-resistant strain LbE16, genes that might be involved in aminoglycoside (aadE, aphA-3, sat4) and virginiamycin [vat(E)] resistance were further found. The erm(B) gene but not tet(S) was transferred from Leuconostoc to Enterococcus faecalis both under laboratory conditions and in cheese. This study contributes to the characterization of AR in the

  7. Comparison of methods for marine gravity determination from satellite altimetry data in the Labrador Sea

    NASA Astrophysics Data System (ADS)

    Zhang, Changyou; Blais, J. A. R.

    1995-09-01

    One-year average satellite altimetry data from the Exact Repeat Missions (ERM) of GEOSAT have been used to determine marine gravity disturbances in the Labrador Sea region using the inverse Hotine approach with FFT techniques. The derived satellite gravity information has been compared to shipboard gravity as well as gravity information derived by least-squares collocation (LSC), GEMT3 and OSU91A geopotential models in the Orphan Knoll area. The RMS and mean differences between satellite and shipboard gravity disturbances are about 8.0 and 2.8 mGal, respectively. There is no significantly difference between the results obtained using FFT and LSC.

  8. SURF_ER—surface electron spin resonance (ESR) of the surface domain of large objects

    NASA Astrophysics Data System (ADS)

    Herrling, Th.; Rehberg, J.; Jung, K.; Groth, N.

    2002-04-01

    SURF_ER is a method for spectral and spatial electron spin resonance measurements on the surface of large objects which extension is only restricted by the width of the pole gap of the magnet and the homogeneity of the magnetic field and not by the cavity dimensions. The application of several techniques like SURF_ER for spectroscopic measurements, SURF_ERM for spatial scanning and SURF_ERI for spatial measurements of the depth of the surface region are discussed and represented for the skin of a human being as an example.

  9. Antibiotic resistance genes detected in the marine sponge Petromica citrina from Brazilian coast.

    PubMed

    Laport, Marinella Silva; Pontes, Paula Veronesi Marinho; Dos Santos, Daniela Silva; Santos-Gandelman, Juliana de Fátima; Muricy, Guilherme; Bauwens, Mathieu; Giambiagi-deMarval, Marcia; George, Isabelle

    2016-01-01

    Although antibiotic-resistant pathogens pose a significant threat to human health, the environmental reservoirs of the resistance determinants are still poorly understood. This study reports the detection of resistance genes (ermB, mecA, mupA, qnrA, qnrB and tetL) to antibiotics among certain culturable and unculturable bacteria associated with the marine sponge Petromica citrina. The antimicrobial activities elicited by P. citrina and its associated bacteria are also described. The results indicate that the marine environment could play an important role in the development of antibiotic resistance and the dissemination of resistance genes among bacteria. PMID:27287338

  10. Das Neutron, der Kosmos und die Kräfte: Neutronen in der Teilchenphysik

    NASA Astrophysics Data System (ADS)

    Soldner, Torsten

    2003-05-01

    Das Neutron besitzt eine einzigartige Kombination von Eigenschaften. Sie ermöglicht die Untersuchung aller vier elementaren Kräfte. Dabei wurden beeindruckende Resultate erzielt, wie die Präzisionsmessungen der elektrischen Ladung des Neutrons oder der Feinstrukturkonstante zeigen. Die genaue Bestimmung der schwachen Wechselwirkungsstärke der Nukleonen liefert der Astrophysik wichtige Daten. Die Messung eines von Null verschiedenen elektrischen Dipolmoments des Neutrons könnte einen entscheidenden Hinweis über die Physik jenseits des Standardmodells der Teilchenphysik geben. Doch auch zur Aufklärung von Unsicherheiten innerhalb des Standardmodells selbst tragen Neutronen bei.

  11. [Epidemiological analysis of Streptococcus pneumoniae in Gifu prefecture and the northern district of Aichi prefecture--2009].

    PubMed

    Yamagishi, Yuka; Mikamo, Hiroshige; Sawamura, Haruki; Suematsu, Hiroyuki; Asano, Yuko; Ishigo, Shiomi; Hatano, Masakazu; Matsubara, Shigenori; Ohta, Hirotoshi; Matsukawa, Yoko; Saeki, Hiroikazu; Mutou, Toshihiro; Teraji, Mayumi; Mouri, Tetsuo; Kawahara, Yuki; Akita, Shigeki; Miyabe, Takanori; Okada, Masako; Terada, Hiroshi; Sakuma, Takashi; Morita, Eri; Miyamoto, Naoya; Tuchiya, Yoko; Yamada, Yukiji; Yamaoka, Kazukiyo; Miyaki, Yuki; Tanaka, Kaori; Watanabe, Kunitomo

    2012-02-01

    High pathogenicity and drug resistance of Streptococcus pneumoniae are serious problem in clinical practice. Since 1999, we have conducted epidemiologic analyses of S. pneumoniae in Chubu district. We report the results of the analysis conducted in 2009. Three hundred and eight (308) S. pneumoniae isolates with a gene coding for autolysin lyt-A, which had been isolated from patients at 21 medical institutions in Gifu prefecture and the northern part of Aichi prefecture in 2009, were enrolled in this study. The strains were classified according to their drug resistance based on the presence of the pbp mutation, and examined for the presence of the two macrolide-resistance genes, ermB and mefA. Moreover, they were serotyped using type-specific antisera. The mean age of the patients from whom these S. pneumoniae strains were isolated, was 23.4 +/- 30.1 years old, and children aged 15 years old or less accounted for 66% of all the patients. Genotype penicillin-susceptible S. pneumoniae (gPSSP), genotype penicillin-intermediate S. pneumoniae (gPISP) and genotype penicillin-resistant S. pneumoniae (gPRSP) were 22 (7.1%), 131 (42.5%) and 155 (50.3%), respectively. The strains with mefA positive and ermB negative, mefA negative and ermB positive, and mefA positive and ermB positive were 80 (26.0%), 153 (49.7%), and 47 (15.3%), respectively. The MIC90 values of tebipenem (TBPM) and faropenem were 0.06 microg/mL and 0.5 microg/mL, respectively. TBPM showed the high bactericidal activity against gPRSP. In carbapenems, panipenem and biapenem exhibited higher bactericidal activities. Quinolone-resistant S. pneumoniae (QRSP) were isolated from 10 (3.2%). QRSP dominated 5 (7.9%) and 3 (1.5%) among the elderly (over 65 years old) and children, respectively. (As for the serotype, serotypes 6, 19 and 23 were 60 (19.5%), 62 (20.1%), and 44 (14.3%), respectively. Further epidemiologic studies on S. pneumoniae might be required also in the future, including the relationship between the

  12. Fourier-domain optical coherence tomography of eyes with idiopathic epiretinal membrane: correlation between macular morphology and visual function

    PubMed Central

    Pilli, S; Lim, P; Zawadzki, R J; Choi, S S; Werner, J S; Park, S S

    2011-01-01

    Purpose The purpose of this study is to evaluate the macular morphological changes associated with idiopathic epiretinal membrane (iERM) using high-resolution Fourier-domain optical coherence tomography (FD-OCT), as they correlate with visual acuity and microperimetry (MP-1). Methods In all, 24 eyes (19 subjects) with iERM were imaged prospectively using FD-OCT with axial resolution of 4.5 μm and transverse resolution of 10 to 15 μm. MP-1 and Stratus OCT were carried out in a subset of eyes. Results The mean log of the minimum angle of resolution best-corrected visual acuity (BCVA) was 0.18±0.16 (range: −0.08 to 0.48, Snellen equivalent 20/15−1 to 20/60). ERM was visualized in all 24 eyes with FD-OCT and in 17 eyes (85%) of 20 eyes imaged with Stratus OCT. Although BCVA correlated with macular thickening in the central 1 mm sub-field of the Stratus ETDRS (P=0.0005) and macular volume (central 3 mm area) on FD-OCT (P<0.0001), macular thickening on thickness map and volume correlated poorly with decrease in macular sensitivity on MP-1 (P=0.16). On FD-OCT, foveal morphological changes correlated best with decrease in BCVA, the strongest being central foveal thickness (P<0.0001). Other significant changes included blurring of the foveal inner segment–outer segment (IS–OS) junction and/or Verhoeff's membrane, vitreal displacement of foveal outer nuclear layer and foveal detachment (P<0.05). Foveal IS–OS junction disruption was seen in 25% of eyes on Stratus OCT but in none of the eyes on FD-OCT. Conclusion FD-OCT allowed improved visualization of ERM and associated foveal morphological changes that correlated best with BCVA. Macular thickening correlated weakly with decreased macular function as assessed by MP-1. PMID:21436847

  13. Automatische Segmentierung der Gewebegrenzen eines schlagenden embryonalen Hühnerherzens im 2D-Videobild

    NASA Astrophysics Data System (ADS)

    Thommes, Jan; Yelbuz, Talât Mesud

    Eine Klimakammer ermöglicht es, die Herzentwicklung bei Hühnerembryonen in einer schalenlosen Kultur über einen Zeitraum von etwa 50 Stunden visuell zu beobachten. Eine Hochgeschwindigkeits-Mikroskopkamera liefert eine Aufsicht auf das schlagende embryonale Herz. Da die für eine Analyse der Herzfunktion notwendige Segmentierung der Myokardgrenzen und des blutgefüllte Cavums manuell sehr zeitaufwändig ist, wird ein Ansatz zur kontinuierlichen, automatischen Segmentierung des schlagenden embryonalen Herzens mit Active Appearance Modellen präsentiert.

  14. Comparative study of 27-gauge vs 25-gauge vitrectomy for epiretinal membrane.

    PubMed

    Mitsui, K; Kogo, J; Takeda, H; Shiono, A; Sasaki, H; Munemasa, Y; Kitaoka, Y; Takagi, H

    2016-04-01

    PurposeThe purpose of this study was to compare 27-gauge (27G) with 25-gauge (25G) microincision vitrectomy in patients with epiretinal membrane (ERM).ParticipantsSeventy-four eyes of 66 patients undergoing 3-port pars plana vitrectomy using 27G or 25G instrumentation.MethodsSeventy-four eyes of 66 patients with ERM, who underwent 27G or 25G microincision vitrectomy were prospectively evaluated.ResultsThe mean operation time for vitrectomy was significantly longer in the 27G group than in the 25G group (9.9±3.5 vs 6.2±2.7 min, respectively, P<0.0001). No statistically significant difference was found between the two groups in terms of the mean operation time for ERM-inner limiting membrane peeling (27G vs 25G: 20.2±9.9 vs 16.1±9.3 min, P=0.14), although the time for vitreous cutting was longer in the 27G group (9.9±3.5 vs 6.2±2.7 min, respectively, P<0.0001). The flare value, intraocular pressure (IOP), and rate of hypotony 1 day after surgery did not differ between the 27G and 25G groups (flare value: 18.7 vs 17.2; IOP: 8.8 vs 9.7 mm Hg; rate of hypotony: 30 vs 35%, respectively). There was no significant difference in the surgically induced astigmatism between the two groups in the follow-up period. The mean time required for wound closure did not show a significant difference between the 27G and 25G groups (7.7 vs 8.6 weeks, respectively).ConclusionThe 27G system is as safe and useful for ERM vitrectomy as the 25G system. Based on its potential, further improvement of 27G instruments could result in greater efficiency. PMID:26742862

  15. Antibiotic Susceptibility Profiles of Dairy Leuconostoc, Analysis of the Genetic Basis of Atypical Resistances and Transfer of Genes In Vitro and in a Food Matrix.

    PubMed

    Flórez, Ana Belén; Campedelli, Ilenia; Delgado, Susana; Alegría, Ángel; Salvetti, Elisa; Felis, Giovanna E; Mayo, Baltasar; Torriani, Sandra

    2016-01-01

    In spite of a global concern on the transfer of antibiotic resistances (AR) via the food chain, limited information exists on this issue in species of Leuconostoc and Weissella, adjunct cultures used as aroma producers in fermented foods. In this work, the minimum inhibitory concentration was determined for 16 antibiotics in 34 strains of dairy origin, belonging to Leuconostoc mesenteroides (18), Leuconostoc citreum (11), Leuconostoc lactis (2), Weissella hellenica (2), and Leuconostoc carnosum (1). Atypical resistances were found for kanamycin (17 strains), tetracycline and chloramphenicol (two strains each), and erythromycin, clindamycin, virginiamycin, ciprofloxacin, and rifampicin (one strain each). Surprisingly, L. mesenteroides subsp. mesenteroides LbE16, showed resistance to four antibiotics, kanamycin, streptomycin, tetracycline and virginiamycin. PCR analysis identified tet(S) as responsible for tetracycline resistance in LbE16, but no gene was detected in a second tetracycline-resistant strain, L. mesenteroides subsp. cremoris LbT16. In Leuconostoc mesenteroides subsp. dextranicum LbE15, erythromycin and clindamycin resistant, an erm(B) gene was amplified. Hybridization experiments proved erm(B) and tet(S) to be associated to a plasmid of ≈35 kbp and to the chromosome of LbE15 and LbE16, respectively. The complete genome sequence of LbE15 and LbE16 was used to get further insights on the makeup and genetic organization of AR genes. Genome analysis confirmed the presence and location of erm(B) and tet(S), but genes providing tetracycline resistance in LbT16 were again not identified. In the genome of the multi-resistant strain LbE16, genes that might be involved in aminoglycoside (aadE, aphA-3, sat4) and virginiamycin [vat(E)] resistance were further found. The erm(B) gene but not tet(S) was transferred from Leuconostoc to Enterococcus faecalis both under laboratory conditions and in cheese. This study contributes to the characterization of AR in the

  16. Von Donuts und Zucker: Mit Neutronen biologische Makromoleküle erforschen

    NASA Astrophysics Data System (ADS)

    May, Roland P.

    2003-05-01

    Für die Erforschung von Biomolekülen bieten Neutronen einzigartige Eigenschaften. Vor allem ihre unterschiedliche Wechselwirkung mit dem natürlichen Wasserstoff und seinem schweren Isotop Deuterium ermöglicht tiefe Einblicke in Struktur, Funktion und Dynamik von Proteinen, Nukleinsäuren und Biomembranen. Bei vielen Fragestellungen zur Strukturaufklärung gibt es kaum oder keine Alternative zum Neutron. Das Institut Laue-Langevin trägt Bahnbrechendes zum Erfolg der Neutronen-Methoden in der Biologie bei.

  17. "Agnatha", Kieferlose

    NASA Astrophysics Data System (ADS)

    Rieger, Gunde; Maier, Wolfgang

    An der Basis der Wirbeltiere standen kieferlose Organismen, vermutlich mit Kiemendarm und noch filtrierender Lebensweise. Ihre wenigen heute noch lebenden Nachkommen sind die Myxinoida (Schleimaale) und die Petromyzontida (Neunaugen). Beide Taxa sind nur Relikte einer Vielzahl und Vielfalt im frühen Paläozoikum (Ordovizium bis ins Devon: vor 470-355 Mio. Jahren). Fossil traten die beiden heute lebenden Gruppen bereits im Karbon bzw. im Oberdevon auf. Obwohl diese rezenten Formen hoch spezialisiert sind, ermöglichen sie — zusammen mit den fossilen kieferlosen Formen (früher "†Ostracodermata") — Struktur und Lebensweise der frühen Craniota in Umrissen zu erkennen.

  18. Relationship between variations in posterior vitreous detachment and visual prognosis in idiopathic epiretinal membranes

    PubMed Central

    Ota, Ayumi; Tanaka, Yoshiaki; Toyoda, Fumihiko; Shimmura, Machiko; Kinoshita, Nozomi; Takano, Hiroko; Kakehashi, Akihiro

    2016-01-01

    Purpose To clarify the relationship between variations in posterior vitreous detachments (PVDs) and visual prognoses in idiopathic epiretinal membranes (ERMs). Methods In this retrospective, observational, and consecutive case series, we observed variations in PVDs in 37 patients (mean age, 65.7±11.0 years) with ERMs and followed them for 2 years. Three PVD types were found biomicroscopically: no PVD, complete PVD with collapse (C-PVD with collapse), and partial PVD without shrinkage, with persistent vitreous attachment to the macula through the premacular hole of the posterior hyaloid membrane (P-PVD without shrinkage [M]). The best-corrected visual acuity (BCVA) was measured and converted to the logarithm of the minimum angle of resolution (logMAR) BCVA at the first visit and 2 years later. Results No PVD was observed in 16 of the 37 eyes (mean age, 61.3±11.3 years), C-PVD with collapse in 11 of the 37 eyes (mean age, 69.1±9.9 years), and P-PVD without shrinkage (M) in 10 of the 37 eyes (mean age, 69.3±10.9 years). The logMAR BCVA at the first visit was the worst in the P-PVD without shrinkage (M) group (0.22±0.35) compared with the no-PVD group (−0.019±0.07; P<0.01) and the C-PVD group (0.029±0.08; P<0.05). The logMAR BCVA 2 years later was also worst in the P-PVD without shrinkage (M) group (0.39±0.35) compared with the no-PVD group (0.04±0.13) and the C-PVD with collapse group (0.03±0.09; P<0.05 for both comparisons). The change in the logMAR BCVA over the 2-year follow-up period was worst in the P-PVD without shrinkage (M) group (0.17±0.23) compared with the no-PVD group (0.06±0.14) and the C-PVD with collapse group (0.0009±0.09; P<0.05 for both comparisons). Conclusion Cases with an ERM with a P-PVD without shrinkage (M) had a worse visual prognosis than those with an ERM with no PVD and C-PVD with collapse. PMID:26730176

  19. Antibiotic resistance of Streptococcus agalactiae from cows with mastitis.

    PubMed

    Gao, Jian; Yu, Fu-Qing; Luo, Li-Ping; He, Jian-Zhong; Hou, Rong-Guang; Zhang, Han-Qi; Li, Shu-Mei; Su, Jing-Liang; Han, Bo

    2012-12-01

    The aim of this study was to characterise the phenotypic and genotypic antibiotic resistance patterns of Streptococcus agalactiae isolated from cows with mastitis in China. Antibiotic resistance was based on minimum inhibitory concentrations and detection of resistance genes by PCR. S. agalactiae isolates most frequently exhibited phenotypic resistance to tetracycline, while the resistance genes most frequently detected were ermB, tetL and tetM. Resistance genes were detected in some susceptible isolates, whereas no resistance genes could be detected in some resistant isolates, indicating that the resistance genotype does not accurately predict phenotypic resistance. PMID:22627045

  20. Development of antibiotic resistance genes in microbial communities during long-term operation of anaerobic reactors in the treatment of pharmaceutical wastewater.

    PubMed

    Aydin, Sevcan; Ince, Bahar; Ince, Orhan

    2015-10-15

    Biological treatment processes offer the ideal conditions in which a high diversity of microorganisms can grow and develop. The wastewater produced during these processes is contaminated with antibiotics and, as such, they provide the ideal setting for the acquisition and proliferation of antibiotic resistance genes (ARGs). This research investigated the occurrence and variation in the ARGs found during the one-year operation of the anaerobic sequencing batch reactors (SBRs) used to treat pharmaceutical wastewater that contained combinations of sulfamethoxazole-tetracycline-erythromycin (STE) and sulfamethoxazole-tetracycline (ST). The existence of eighteen ARGs encoding resistance to sulfamethoxazole (sul1, sul2, sul3), erythromycin (ermA, ermF, ermB, msrA, ereA), tetracycline (tetA, tetB, tetC, tetD, tetE, tetM, tetS, tetQ, tetW, tetX) and class Ι integron gene (intΙ 1) in the STE and ST reactors was investigated by quantitative real-time PCR. Due to the limited availability of primers to detect ARGs, Illumina sequencing was also performed on the sludge and effluent of the STE and ST reactors. Although there was good reactor performance in the SBRs, which corresponds to min 80% COD removal efficiency, tetA, tetB, sul1, sul2 and ermB genes were among those ARGs detected in the effluent from STE and ST reactors. A comparison of the ARGs acquired from the STE and ST reactors revealed that the effluent from the STE reactor had a higher number of ARGs than that from the ST reactor; this could be due to the synergistic effects of erythromycin. According to the expression of genes results, microorganisms achieve tetracycline and erythromycin resistance through a combination of three mechanisms: efflux pumping protein, modification of the antibiotic target and modifying enzymes. There was also a significant association between the presence of the class 1 integron and sulfamethoxazole resistance genes. PMID:26188597

  1. Improvement of chloramphenicol production in Streptomyces venezuelae ATCC 10712 by overexpression of the aroB and aroK genes catalysing steps in the shikimate pathway.

    PubMed

    Vitayakritsirikul, Vipawan; Jaemsaeng, Ratchaniwan; Lohmaneeratana, Karan; Thanapipatsiri, Anyarat; Daduang, Ratama; Chuawong, Pitak; Thamchaipenet, Arinthip

    2016-03-01

    Streptomyces venezuelae ATCC 10712 produces chloramphenicol in small amounts. To enhance chloramphenicol production, two genes, aroB and aroK, encoding rate-limiting enzymes of the shikimate pathway were overexpressed using the expression vector pIJ86 under the control of the strong constitutive ermE* promoter. The recombinant strains, S. venezuelae/pIJ86-aroB and S. venezuelae/pIJ86-aroK, produced 2.5- and 4.3-fold greater amounts respectively of chloramphenicol than wild type at early stationary phase of growth. High transcriptional levels of aroB and aroK genes were detected at the early exponential growth of both recombinant strains and consistent with the enhanced expression of pabB gene encoding an early enzyme in chloramphenicol biosynthesis. The results suggested that the increment of carbon flux was directed towards intermediates in the shikimate pathway required for the production of chorismic acid, and consequently resulted in the enhancement of chloramphenicol production. This work is the first report of a convenient genetic approach to manipulate primary metabolite genes in S. venezuelae in order to increase chloramphenicol production. PMID:26715388

  2. Determination of inorganic contaminants in polyamide textiles used for manufacturing sport T-shirts.

    PubMed

    Matoso, Erika; Cadore, Solange

    2012-01-15

    An acid microwave closed vessel digestion method was used for the determination of inorganic contaminants (Sb, As, Pb, Cd, Cr, Co, Cu, Ni and Hg) in polyamide raw materials (pellets) and textiles by inductively coupled plasma optical emission spectrometry (ICP OES). The initial tests were carried out with samples of polyamide pellets, which is the main raw material used to manufacture sport textiles. The recovery factors obtained were 94.4-105.7% with relative standard deviation (RSD) of 0.5-2.2%. The proposed method was evaluated by addition and recovery tests and also using certified reference materials (ERM-BCR680 and ERM-BCR681) showing good accuracy. The residual acidity was about 4% HNO(3) (w/w) and the quantification limits were from 0.1 to 6.6 mg kg(-1). After the development of these parameters for the raw material, the method was applied to textile samples from different sport fabrics obtained from three different brands. The residual carbon after sample digestion was 0.2% (w/w) and the most significant result was obtained for chromium, 901 mg kg(-1), in black fabric. Lixiviation tests using synthetic sweat and temperature were carried out on two black samples, showing that only 0.3% of the initial concentration migrated to the solution. PMID:22265532

  3. The Prevalence and Distribution of Vitreoretinal Interface Abnormalities among Urban Community Population in China

    PubMed Central

    Liu, Lei; Yue, Song; Wu, Jingyang; Zhang, Jiahua; Lian, Jie; Huang, Desheng; Teng, Weiping; Chen, Lei

    2015-01-01

    The aim of this research was to identify the prevalence and distribution of vitreoretinal interface abnormalities (VIAs) among urban community population in Shenyang, China. According to the WHO criteria, a cross-sectional study was carried out among 304 Type 2 diabetes (T2D) patients and 304 people without diabetes as control over 45 years old. The presence of VIAs was determined by standardized grading of macular optical coherence tomography (Optovue OCT; Optovue, Inc., Fremont, CA) scans and two-field fundus photographs in at least one eye. For both men and women, high prevalence of VIAs (70.79%) was observed among over 65-years-old T2D patients. Prevalence of VIAs was observed to be high among T2D patients in all age groups compared to normal subjects. Prevalence of VIAs increased with age in all subjects. Prevalence of components of VIAs was epiretinal membrane (ERM) 11.43%, posterior vitreous detachment (PVD) 17.76%, vitreomacular traction syndrome (VMT) 5.67%, macular cysts/macular edema (MC/ME) 4.61%, full-thickness macular hole (FTMH) 0.82%, and partial thickness macular hole (PTMH) 0.74% in any eye, respectively. ERM and MC/ME were more prevalent in T2D in both males and females. The results highlight the need for early detection using OCT and approaches for the prevention of VIAs of diabetes in urban community. PMID:26759726

  4. Clonal dissemination of macrolide-resistant and penicillin-susceptible serotype 3 and penicillin-resistant Taiwan 19F-14 and 23F-15 Streptococcus pneumoniae isolates in Japan: a pilot surveillance study.

    PubMed

    Kasahara, Kei; Maeda, Koichi; Mikasa, Keiichi; Uno, Kenji; Takahashi, Ken; Konishi, Mitsuru; Yoshimoto, Eiichiro; Murakawa, Koichi; Kita, Eiji; Kimura, Hiroshi

    2005-04-01

    Large-scale surveillance studies using molecular techniques such as pulsed-field gel electrophoresis (PFGE) have revealed that the spread of antibiotic-resistant pneumococci is due to clonal spread. However, in Japan, surveillance studies using such molecular techniques have never been done. Therefore, we conducted a pilot surveillance study to elucidate the present situation in Japan. Among the 145 isolates examined, the most prevalent serotype was type 19F (20%), for which most isolates were not susceptible to penicillin (86.2%) but were positive for the mef(A)/mef(E) gene (89.7%). The secondmost prevalent was serotype 3 (16.6%), for which most isolates were susceptible to penicillin (87.5%) and positive for the erm(B) gene (91.7%). PFGE analysis showed that both serotypes consisted mainly of clonally identical or related isolates and, in particular, 38% of the type 19F isolates were indistinguishable from or closely related to the Taiwan 19F-14 clone. In addition, some of the Japanese type 23F isolates with the erm(B) gene were indistinguishable from or related to the Taiwan 23F-15 clone as analyzed by PFGE. Based on the results of our pilot study performed in a single institution, it is likely that international antibiotic-resistant clones have already spread in Japan; therefore, a nationwide surveillance study should be urgently conducted. PMID:15814978

  5. Antimicrobial resistance of invasive Streptococcus pneumoniae isolates in a British district general hospital: the international connection.

    PubMed

    Birtles, Andrew; Virgincar, Nilangi; Sheppard, Carmen L; Walker, Rachel A; Johnson, Alan P; Warner, Marina; Edwards-Jones, Valerie; George, Robert C

    2004-12-01

    Between January 2000 and March 2001, Streptococcus pneumoniae were isolated from the blood of 56 patients admitted to a single district general hospital in the South-East of England. The serotype and antibiotic susceptibility were determined for all isolates and, for those resistant to erythromycin, the presence or absence of the mef(A) and erm(B) genes was determined by PCR. Multi-locus sequence typing, along with PFGE, was undertaken on all isolates resistant to penicillin or erythromycin and a group of antibiotic-susceptible isolates, to identify whether globally distributed pneumococcal clones, as described by the Pneumococcal Molecular Epidemiology Network (PMEN), were present in the study population. Three serotype 9V penicillin-resistant isolates were identified as belonging to the Spain9V-3 clone, while 14 erythromycin-resistant isolates of serotype 14 belonged to the England14-9 clone. A single multi-resistant isolate of serotype 6B, was found to be a single-locus variant of the Spain6B-2 clone. All 14 erythromycin-resistant serotype 14 isolates possessed the mef(A) gene, while the single multi-resistant isolate possessed the erm(B) gene. These findings confirm the wide distribution and clinical impact of PMEN clones, which accounted for all of the penicillin and erythromycin resistance observed amongst invasive isolates in a district general hospital over a 15-month period. PMID:15585504

  6. Macrolones Are a Novel Class of Macrolide Antibiotics Active against Key Resistant Respiratory Pathogens In Vitro and In Vivo.

    PubMed

    Čipčić Paljetak, Hana; Verbanac, Donatella; Padovan, Jasna; Dominis-Kramarić, Miroslava; Kelnerić, Željko; Perić, Mihaela; Banjanac, Mihailo; Ergović, Gabrijela; Simon, Nerrisa; Broskey, John; Holmes, David J; Eraković Haber, Vesna

    2016-09-01

    As we face an alarming increase in bacterial resistance to current antibacterial chemotherapeutics, expanding the available therapeutic arsenal in the fight against resistant bacterial pathogens causing respiratory tract infections is of high importance. The antibacterial potency of macrolones, a novel class of macrolide antibiotics, against key respiratory pathogens was evaluated in vitro and in vivo MIC values against Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus aureus, and Haemophilus influenzae strains sensitive to macrolide antibiotics and with defined macrolide resistance mechanisms were determined. The propensity of macrolones to induce the expression of inducible erm genes was tested by the triple-disk method and incubation in the presence of subinhibitory concentrations of compounds. In vivo efficacy was assessed in a murine model of S. pneumoniae-induced pneumonia, and pharmacokinetic (PK) profiles in mice were determined. The in vitro antibacterial profiles of macrolones were superior to those of marketed macrolide antibiotics, including the ketolide telithromycin, and the compounds did not induce the expression of inducible erm genes. They acted as typical protein synthesis inhibitors in an Escherichia coli transcription/translation assay. Macrolones were characterized by low to moderate systemic clearance, a large volume of distribution, a long half-life, and low oral bioavailability. They were highly efficacious in a murine model of pneumonia after intraperitoneal application even against an S. pneumoniae strain with constitutive resistance to macrolide-lincosamide-streptogramin B antibiotics. Macrolones are the class of macrolide antibiotics with an outstanding antibacterial profile and reasonable PK parameters resulting in good in vivo efficacy. PMID:27353268

  7. Occurrence of sulfonamide-, tetracycline-, plasmid-mediated quinolone- and macrolide-resistance genes in livestock feedlots in Northern China.

    PubMed

    Mu, Quanhua; Li, Jin; Sun, Yingxue; Mao, Daqing; Wang, Qing; Luo, Yi

    2015-05-01

    Antibiotic resistance genes (ARGs) in livestock feedlots deserve attention because they are prone to transfer to human pathogens and thus pose threats to human health. In this study, the occurrence of 21 ARGs, including tetracycline (tet)-, sulfonamide (sul)-, plasmid-mediated quinolone (PMQR)- and macrolide-resistance (erm) genes were investigated in feces and adjacent soils from chicken, swine, and cattle feedlots in Northern China. PMQR and sul ARGs were the most prevalent and account for over 90.0 % of the total ARGs in fecal samples. Specifically, PMQR genes were the most prevalent, accounting for 59.6 % of the total ARGs, followed by sul ARGs (34.2 %). The percentage of tet ARGs was 3.4 %, and erm ARGs accounted for only 1.9 %. Prevalence of PMQR and sul ARGs was also found in swine and cattle feces. The overall trend of ARG concentrations in feces of different feeding animals was chicken > swine > beef cattle in the studied area. In soils, sul ARGs had the highest concentration and account for 71.1 to 80.2 % of the total ARGs, which is possibly due to the widely distributed molecular carriers (i.e., class one integrons), facilitating sul ARG propagation. Overall, this study provides integrated profiles of various types of ARGs in livestock feedlots and thus provides a reference for the management of antibiotic use in livestock farming. PMID:25475616

  8. MEKs/ERKs inhibitor U0126 increases the radiosensitivity of rhabdomyosarcoma cells in vitro and in vivo by down regulating growth and DNA repair signals

    PubMed Central

    Marampon, Francesco; Gravina, Giovanni Luca; Di Rocco, Agnese; Bonfili, Pierluigi; Di Staso, Mario; Fardella, Caterina; Polidoro, Lorella; Ciccarelli, Carmela; Festuccia, Claudio; Popov, Vladimir M.; Pestell, Richard G.; Tombolini, Vincenzo; Zani, Bianca Maria

    2010-01-01

    Multimodal treatment has improved the outcome of many solid tumors, and in some cases the use of radiosensitizers has significantly contributed to this gain. Activation of the extracellular signaling kinase pathway (MEK/ERK) generally results in stimulation of cell growth and confers a survival advantage playing the major role in human cancer. The potential involvement of this pathway in cellular radiosensitivity remains unclear. We previously reported that the disruption of c-Myc through MEK/ERK inhibition blocks the expression of the transformed phenotype, affects in vitro and in vivo growth, angiogenic signaling, and induces myogenic differentiation in the embryonal rhabdomyosarcoma (ERMS) cell lines (RD). The present study was designed to examine whether the ERK pathway affects intrinsic radiosensitivity of rhabdomyosarcoma cancer cells. Exponentially growing human ERMS, RD, xenograft-derived RD-M1 and TE671 cell lines were used. The specific MEK/ERK inhibitor, U0126, reduced the clonogenic potential of the three cell lines, and was effected by radiation. U0126 inhibited phospho/active ERK1/2 and reduced DNAPKcs suggesting that ERKs and DNA-PKcs cooperate in radioprotection of rhabdomyosarcoma cells. The TE671 cell line-xenotransplanted in mice showed a reduction in tumor mass and increase in the time of tumor progression with U0126 treatment associated with reduced DNAPKcs, an effect enhanced by radiotherapy. Thus, our results show that MEK/ERK inhibition enhances radiosensitivity of rhabdomyosarcoma cells suggesting a rational approach in combination with radiotherapy. PMID:21220498

  9. The Ohio State 1991 geopotential and sea surface topography harmonic coefficient models

    NASA Technical Reports Server (NTRS)

    Rapp, Richard H.; Wang, Yan Ming; Pavlis, Nikolaos K.

    1991-01-01

    The computation is described of a geopotential model to deg 360, a sea surface topography model to deg 10/15, and adjusted Geosat orbits for the first year of the exact repeat mission (ERM). This study started from the GEM-T2 potential coefficient model and it's error covariance matrix and Geosat orbits (for 22 ERMs) computed by Haines et al. using the GEM-T2 model. The first step followed the general procedures which use a radial orbit error theory originally developed by English. The Geosat data was processed to find corrections to the a priori geopotential model, corrections to a radial orbit error model for 76 Geosat arcs, and coefficients of a harmonic representation of the sea surface topography. The second stage of the analysis took place by doing a combination of the GEM-T2 coefficients with 30 deg gravity data derived from surface gravity data and anomalies obtained from altimeter data. The analysis has shown how a high degree spherical harmonic model can be determined combining the best aspects of two different analysis techniques. The error analysis was described that has led to the accuracy estimates for all the coefficients to deg 360. Significant work is needed to improve the modeling effort.

  10. Chirurgie angeborener Herzfehler

    NASA Astrophysics Data System (ADS)

    Schreiber, Christian; Libera, Paul; Lange, Rüdiger

    Störungen der embryonalen Entwicklung in der frühen Phase der Schwangerschaft können zu Fehlbildungen am Herz- und Gefäßsystem führen. Die Häufigkeit liegt bei 0.8-1 % aller lebend geborenen Kinder. In Deutschland werden jedes Jahr etwa 6.000 Kinder mit einem Herzfehler geboren (Quelle: http://www.kompetenznetzahf.de). Das Spektrum reicht von einfachen Fehlern, die das Herz-Kreislauf-System wenig beeinträchtigen, bis zu sehr schweren Herzerkrankungen, die unbehandelt zum Tode führen. Fortschritte der Kinderkardiologie, Herzchirurgie und Anästhesie ermöglichen heute ein Überleben bei über 90 % der Patienten. Auch die spezialisierte Pränataldiagnostik (vorgeburtliche Diagnostik) ermöglicht schon die frühe Weichenstellung für mögliche Therapieoptionen. Bei der chirurgischen Therapie ist jedoch festzuhalten, dass ein Herzfehler entweder korrigierend behandelt wird oder nur "palliiert“ werden kann. Bei letzterer Therapie wird bei einem Patienten eine medizinische Maßnahme durchgeführt, die nicht die Herstellung normaler Körperfunktionen zum Ziel hat, sondern in Anpassung an die physiologischen Besonderheiten des Patienten dessen Zustand lediglich stabilisiert und optimiert. Dies kann beispielsweise bei einer nicht korrigierbaren angeborenen Fehlbildung notwendig sein, bei der lediglich eine funktionelle Herzkammer vorhanden ist (z. B. hypoplastisches Linksherz). Hierbei muss eine prothetische Verbindung zur Lungenstrombahn in der Folgezeit entfernt werden.

  11. NHERF1/EBP50 and NF2 as diagnostic markers for choroid plexus tumors.

    PubMed

    Georgescu, Maria-Magdalena; Mobley, Bret C; Orr, Brent A; Shang, Ping; Lehman, Norman L; Zhu, Xiaoping; O'Neill, Thomas J; Rajaram, Veena; Hatanpaa, Kimmo J; Timmons, Charles F; Raisanen, Jack M

    2016-01-01

    The adaptor protein NHERF1 (Na/H exchanger-3 regulatory factor-1) and its associated ezrin-radixin-moesin-merlin/neurofibromin-2 (ERM-NF2) family proteins are required for epithelial morphogenesis and have been implicated in cancer progression. NHERF1 is expressed in ependymal cells and constitutes a highly sensitive diagnostic marker for ependymoma, where it labels membrane polarity structures. Since NHERF1 and ERM-NF2 proteins show polarized expression in choroid plexus (CP) cells, we tested their diagnostic utility in CP neoplasms. NHERF1 immunohistochemistry in 43 adult and pediatric tumors with papillary morphology revealed strong apical plasma membrane staining in CP papilloma (WHO grade I) and cytoplasmic expression in CP carcinoma (WHO grade III). Ezrin and moesin showed similar but less distinctive staining. NHERF1 also labeled papillary tumors of the pineal region in a microlumen and focal apical membrane pattern, suggestive of a transitional morphology between CP papilloma and ependymoma. CP tumors of all grades could be differentiated from metastatic carcinomas with papillary architecture by NF2, which showed polarized membranous staining in CP tumors. NHERF1 and NF2 immunohistochemistry showed enhanced sensitivity and specificity for CP tumors compared to commonly used markers, including cytokeratins and Kir7.1, emerging as reliable diagnostic tools for the differential diagnosis of papillary tumors of the central nervous system. PMID:27229317

  12. The evolution of ERMIONE in mitochondrial biogenesis and lipid homeostasis: An evolutionary view from comparative cell biology.

    PubMed

    Wideman, Jeremy G; Muñoz-Gómez, Sergio A

    2016-08-01

    The ER-mitochondria organizing network (ERMIONE) in Saccharomyces cerevisiae is involved in maintaining mitochondrial morphology and lipid homeostasis. ERMES and MICOS are two scaffolding complexes of ERMIONE that contribute to these processes. ERMES is ancient but has been lost in several lineages including animals, plants, and SAR (stramenopiles, alveolates and rhizaria). On the other hand, MICOS is ancient and has remained present in all organisms bearing mitochondrial cristae. The ERMIONE precursor evolved in the α-proteobacterial ancestor of mitochondria which had the central subunit of MICOS, Mic60. The subsequent evolution of ERMIONE and its interactors in eukaryotes reflects the integrative co-evolution of mitochondria and their hosts and the adaptive paths that some lineages have followed in their specialization to certain environments. By approaching the ERMIONE from a perspective of comparative evolutionary cell biology, we hope to shed light on not only its evolutionary history, but also how ERMIONE components may function in organisms other than S. cerevisiae. This article is part of a Special Issue entitled: The cellular lipid landscape edited by Tim P. Levine and Anant K. Menon. PMID:26825688

  13. Tailor-Made Ezrin Actin Binding Domain to Probe Its Interaction with Actin In-Vitro

    PubMed Central

    Shrivastava, Rohini; Köster, Darius; Kalme, Sheetal; Mayor, Satyajit; Neerathilingam, Muniasamy

    2015-01-01

    Ezrin, a member of the ERM (Ezrin/Radixin/Moesin) protein family, is an Actin-plasma membrane linker protein mediating cellular integrity and function. In-vivo study of such interactions is a complex task due to the presence of a large number of endogenous binding partners for both Ezrin and Actin. Further, C-terminal actin binding capacity of the full length Ezrin is naturally shielded by its N-terminal, and only rendered active in the presence of Phosphatidylinositol bisphosphate (PIP2) or phosphorylation at the C-terminal threonine. Here, we demonstrate a strategy for the design, expression and purification of constructs, combining the Ezrin C-terminal actin binding domain, with functional elements such as fusion tags and fluorescence tags to facilitate purification and fluorescence microscopy based studies. For the first time, internal His tag was employed for purification of Ezrin actin binding domain based on in-silico modeling. The functionality (Ezrin-actin interaction) of these constructs was successfully demonstrated by using Total Internal Reflection Fluorescence Microscopy. This design can be extended to other members of the ERM family as well. PMID:25860910

  14. Enviromental sampling at remote sites based on radiological screening assessments

    SciTech Connect

    Ebinger, M.H.; Hansen, W.R.; Wenz, G.; Oxenberg, T.P.

    1996-06-01

    Environmental radiation monitoring (ERM) data from remote sites on the White Sands Missile Range, New Mexico, were used to estimate doses to humans and terrestrial mammals from residual radiation deposited during testing of components containing depleted uranium (DU) and thorium (Th). ERM data were used with the DOE code RESRAD and a simple steady-state pathway code to estimate the potential adverse effects from DU and Th to workers in the contaminated zones, to hunters consuming animals from the contaminated zones, and to terrestrial mammals that inhabit the contaminated zones. Assessments of zones contaminated with DU and Th and DU alone were conducted. Radiological doses from Th and DU in soils were largest with a maximum of about 3.5 mrem y{sup -1} in humans and maximum of about 0.1 mrad d{sup -1} in deer. Dose estimates from DU alone in soils were significantly less with a maximum of about 1 mrem y{sup -1} in humans and about 0.04 mrad d{sup -1} in deer. The results of the dose estimates suggest strongly that environmental sampling in these affected areas can be infrequent and still provide adequate assessments of radiological doses to workers, hunters, and terrestrial mammals.

  15. Ecological risk screen for PAHs in sediments near two produced water discharges at coastal production platforms in the Gulf of Mexico

    SciTech Connect

    Holtzman, S.; Meinhold, A.F.; DePhillips, M.P.

    1995-12-01

    Preliminary screens for risks to biota, were done on PAHs in sediments associated with produced waters from platforms at Delacroix Island and at Bay de Chene, in open bays of the Louisiana coast. Sediment samples were taken in Spring 1993 at the discharge sites, along three transacts at Delacroix Island and along four transacts at Bay de Chene (at intervals of 100, 300, 500 and 1000 ft), and at two reference locations for each discharge site. A screen for deleterious effects on biota was done by comparing concentrations to the Effects Range-Median (ERM) and Effects Range-Low (ERL) criteria of Long et al. 1995. Only sediment samples from the discharge site at Bay de Chene exceeded ERM concentrations for either total PAH, or individual and total high molecular weight PAHS. The ERL criteria for total and individual PAH concentrations were exceeded at, and 100 m from the discharge at Delacroix Island. At Bay de Chene the ERL criteria for total and individual PAH concentrations were exceeded at the discharge, as well as at 100 and 300 m stations.

  16. Determining the optimal number of individual samples to pool for quantification of average herd levels of antimicrobial resistance genes in Danish pig herds using high-throughput qPCR.

    PubMed

    Clasen, Julie; Mellerup, Anders; Olsen, John Elmerdahl; Angen, Øystein; Folkesson, Anders; Halasa, Tariq; Toft, Nils; Birkegård, Anna Camilla

    2016-06-30

    The primary objective of this study was to determine the minimum number of individual fecal samples to pool together in order to obtain a representative sample for herd level quantification of antimicrobial resistance (AMR) genes in a Danish pig herd, using a novel high-throughput qPCR assay. The secondary objective was to assess the agreement between different methods of sample pooling. Quantification of AMR was achieved using a high-throughput qPCR method to quantify the levels of seven AMR genes (ermB, ermF, sulI, sulII, tet(M), tet(O) and tet(W)). A large variation in the levels of AMR genes was found between individual samples. As the number of samples in a pool increased, a decrease in sample variation was observed. It was concluded that the optimal pooling size is five samples, as an almost steady state in the variation was observed when pooling this number of samples. Good agreement between different pooling methods was found and the least time-consuming method of pooling, by transferring feces from each individual sample to a tube using a 10μl inoculation loop and adding 3.5ml of PBS, approximating a 10% solution, can therefore be used in future studies. PMID:27259826

  17. Self-similar rupture implied by scaling properties of volcanic earthquakes occurring during the 2004-2008 eruption of Mount St. Helens, Washington

    NASA Astrophysics Data System (ADS)

    Harrington, Rebecca M.; Kwiatek, Grzegorz; Moran, Seth C.

    2015-07-01

    We analyze a group of 6073 low-frequency earthquakes recorded during a week-long temporary deployment of broadband seismometers at distances of less than 3 km from the crater at Mount St. Helens in September of 2006. We estimate the seismic moment (M0) and spectral corner frequency (f0) using a spectral ratio approach for events with a high signal-to-noise (SNR) ratio that have a cross-correlation coefficient of 0.8 or greater with at least five other events. A cluster analysis of cross-correlation values indicates that the group of 421 events meeting the SNR and cross-correlation criteria forms eight event families that exhibit largely self-similar scaling. We estimate the M0 and f0 values of the 421 events and calculate their static stress drop and scaled energy (ER/M0) values. The estimated values suggest self-similar scaling within families, as well as between five of eight families (i.e., M0∝f0-3 and ER/M0∝ constant). We speculate that differences in scaled energy values for the two families with variable scaling may result from a lack of resolution in the velocity model. The observation of self-similar scaling is the first of its kind for such a large group of low-frequency volcanic tectonic events occurring during a single active dome extrusion eruption.

  18. The Ca2+-activated Cl- channel Ano1 controls microvilli length and membrane surface area in the oocyte.

    PubMed

    Courjaret, Raphael; Hodeify, Rawad; Hubrack, Satanay; Ibrahim, Awab; Dib, Maya; Daas, Sahar; Machaca, Khaled

    2016-07-01

    Ca(2+)-activated Cl(-) channels (CaCCs) play important physiological functions in epithelia and other tissues. In frog oocytes the CaCC Ano1 regulates resting membrane potential and the block to polyspermy. Here, we show that Ano1 expression increases the oocyte surface, revealing a novel function for Ano1 in regulating cell morphology. Confocal imaging shows that Ano1 increases microvilli length, which requires ERM-protein-dependent linkage to the cytoskeleton. A dominant-negative form of the ERM protein moesin precludes the Ano1-dependent increase in membrane area. Furthermore, both full-length and the truncated dominant-negative forms of moesin co-localize with Ano1 to the microvilli, and the two proteins co-immunoprecipitate. The Ano1-moesin interaction limits Ano1 lateral membrane mobility and contributes to microvilli scaffolding, therefore stabilizing larger membrane structures. Collectively, these results reveal a newly identified role for Ano1 in shaping the plasma membrane during oogenesis, with broad implications for the regulation of microvilli in epithelia. PMID:27173493

  19. CLIC5 Stabilizes Membrane-Actin Filament Linkages at the Base of Hair Cell Stereocilia in a Molecular Complex with Radixin, Taperin, and Myosin VI

    PubMed Central

    Salles, Felipe T.; Andrade, Leonardo R.; Tanda, Soichi; Grati, M’hamed; Plona, Kathleen L.; Gagnon, Leona H.; Johnson, Kenneth R.; Kachar, Bechara; Berryman, Mark A.

    2015-01-01

    Chloride intracellular channel 5 protein (CLIC5) was originally isolated from microvilli in complex with actin binding proteins including ezrin, a member of the Ezrin-Radixin-Moesin (ERM) family of membrane-cytoskeletal linkers. CLIC5 concentrates at the base of hair cell stereocilia and is required for normal hearing and balance in mice, but its functional significance is poorly understood. This study investigated the role of CLIC5 in postnatal development and maintenance of hair bundles. Confocal and scanning electron microscopy of CLIC5-deficient jitterbug (jbg) mice revealed progressive fusion of stereocilia as early as postnatal day 10. Radixin (RDX), protein tyrosine phosphatase receptor Q (PTPRQ), and taperin (TPRN), deafness-associated proteins that also concentrate at the base of stereocilia, were mislocalized in fused stereocilia of jbg mice. TPRQ and RDX were dispersed even prior to stereocilia fusion. Biochemical assays showed interaction of CLIC5 with ERM proteins, TPRN, and possibly myosin VI (MYO6). In addition, CLIC5 and RDX failed to localize normally in fused stereocilia of MYO6 mutant mice. Based on these findings, we propose a model in which these proteins work together as a complex to stabilize linkages between the plasma membrane and subjacent actin cytoskeleton at the base of stereocilia. PMID:24285636

  20. Heavy metal pollution in sediments and mussels: assessment by using pollution indices and metallothionein levels.

    PubMed

    Okay, Oya S; Ozmen, Murat; Güngördü, Abbas; Yılmaz, Atilla; Yakan, Sevil D; Karacık, Burak; Tutak, Bilge; Schramm, Karl-Werner

    2016-06-01

    In the present work, the concentration of eight metals (Cd, Cr, Cu, Fe, Mn, Ni, Pb, Zn) was determined in the sediments and transplanted and native mussels (Mytilus galloprovincialis). The study was conducted in Turkish marinas, shipyards, and shipbreaking yards. The effect of metal pollution was evaluated by determining the levels of metallothionein (MT) in the mussels. The extent of contamination for each single metal was assessed by using the geoaccumulation index (I geo) and enrichment factor (EF). Whereas, to evaluate the overall metal pollution and effect, the pollution load index (PLI), modified contamination degree (mC d), potential toxicity response index (RI), mean effects range median (ERM) quotient (m-ERM-Q), and mean PEL quotient (m-PEL-Q) were calculated. The influence of different background values on the calculations was discussed. The results indicated a significant metal pollution caused by Cu, Pb, and Zn especially in shipyard and shipbreaking sites. Higher concentrations of MT were observed in the ship/breaking yard samples after the transplantation. PMID:27188302

  1. [Pollution characteristics and sources of polycyclic aromatic hydrocarbons in urban rivers of Wenzhou city].

    PubMed

    Zhou, Jie-Cheng; Chen, Zhen-Lou; Bi, Chun-Juan; Lü, Jin-Gang; Xu, Shi-Yuan; Pan, Qi

    2012-12-01

    Concentrations of 18 polycyclic aromatic hydrocarbons (PAHs) in water and surface sediments collected from the urban rivers of Wenzhou city in spring and summer were measured by GC-MS. The results showed that the total PAHs concentrations in water and sediments of the studied rivers varied in ranges of 146.74-3 047.89 ng x L(-1) and 21.01-11 990.48 ng x g(-1), respectively. Higher concentrations occurred in spring. The low and middle rings of 2-4-ring were dominant in both water and sediments, but the concentrations of 5-ring and 6-ring PAHs in sediments were relatively higher than those in water. The EBaP values of PAHs in water of the studied rivers in spring and summer were 1.69-51.95 ng x L(-1) and 0-3.03 ng x L(-1), respectively. Eighty percent of water samples in spring surpassed the limits of BaP in surface water of China. The concentrations of sigma PAHs in the sediments both in spring and summer were lower than the ERM value, but part of the components of PAHs had values higher than the ERM, suggesting possible toxic effect on living organisms. Based on the PAHs molecule ratios and principal component analysis, a mixed PAHs source of petroleum and combustion in water and sediments was diagnosed, while sediments showed a greater proportion of combustion sources. PMID:23379146

  2. Identification of position isomers by energy-resolved mass spectrometry.

    PubMed

    Menachery, Sunil Paul M; Laprévote, Olivier; Nguyen, Thao P; Aravind, Usha K; Gopinathan, Pramod; Aravindakumar, Charuvila T

    2015-07-01

    This study reports an energy-resolved mass spectrometric (ERMS) strategy for the characterization of position isomers derived from the reaction of hydroxyl radicals ((●)OH) with diphenhydramine (DPH) that are usually hard to differentiate by other methods. The isomer analogues formed by (●)OH attack on the side chain of DPH are identified with the help of a specific fragment ion peak (m/z 88) in the collision-induced dissociation (CID) spectrum of the protonated molecule. In the negative ion mode, the breakdown curves of the deprotonated molecules show an order of stability (supported by density functional theory (DFT) calculations) ortho > meta > para of the positional isomers formed by the hydroxylation of the aromatic ring. The gas phase stability of the deprotonated molecules [M - H](-) towards the benzylic cleavage depends mainly on the formation of intramolecular hydrogen bonds and of the mesomeric effect of the phenol hydroxyl. The [M - H](-) molecules of ortho and meta isomers result a peak at m/z 183 with notably different intensities because of the presence/absence of an intramolecular hydrogen bonding between the OH group and C9 protons. The ERMS approach discussed in this report might be an effective replacement for the conventional methods that requires very costly and time-consuming separation/purification methods along with the use of multi-spectroscopic methods. PMID:26349650

  3. Distribution, sources and ecological risk assessment of heavy metals in surface sediments from Lake Taihu, China

    NASA Astrophysics Data System (ADS)

    Yin, Hongbin; Gao, Yongnian; Fan, Chengxin

    2011-10-01

    The distribution, sources and ecological risk of heavy metals in surface sediments from Lake Taihu were studied. Results showed that the measured heavy metals had varied spatial distribution patterns, indicating that they had complex origins and controlling factors. Pearson's correlation analysis revealed that the total phosphorus and the loss on ignition were positively correlated with the measured metals except Cd. Principal component analysis and cluster analysis demonstrated that Hg, Cu, Cr, Cd and Pb might originate from domestic sewage and industrial wastewater, whereas As predominantly originated from natural processes. Potential ecological risk indices indicated that sediment from Wuli Lake, Gonghu Bay and the Northwest Area suffered high pollution, whereas other areas of Lake Taihu were moderately polluted. A comparison of metal levels with the effects range low (ERL) and effects range median (ERM) showed that metals exceeded their corresponding ERL limit at 13.6-72.3% (72.3% for As, 52.4% for Pb, 27.7% for Cu, 22.8% for Cd, 16.0 for Hg and 13.6% for Cr) of the sites investigated. Moreover, 3.90% and 0.50% of the sites sampled exceeded the ERM thresholds for Hg and Pb, respectively.

  4. Polycyclic aromatic hydrocarbons (PAHs) in coastal sediments from urban and industrial areas of Asaluyeh Harbor, Iran: distribution, potential source and ecological risk assessment.

    PubMed

    Raeisi, Alireza; Arfaeinia, Hossein; Seifi, Morteza; Shirzad-Siboni, Mehdi; Keshtkar, Mozhgan; Dobaradaran, Sina

    2016-01-01

    The distribution and toxicity levels of 16 EPA priority pollutant polycyclic aromatic hydrocarbons (PAHs) in the sediments of Asaluyeh shore, Iran were investigated. The total concentrations of the PAHs in surface sediments ranged from 1,054 to 17,448 ng/g dry weights with a mean concentration of 8,067 ng/g. The spatial distribution of PAHs showed that PAH levels are much higher in the industrial areas in comparison with urban areas. Based on diagnostic ratios, pyrogenic activities were dominant sources of PAHs pollution in sediments comparing petroleum sources. The toxic equivalent concentrations (TEQ Carc) of PAHs ranged from 172 to 2,235 ng TEQ/g with mean value of 997.9. Toxicity levels were evaluated using sediment quality guidelines (SQGs) and toxic equivalent factors. Samples were collected from industrial and urban stations in Asaluyeh shores. According to SQGs, ΣPAHs concentrations in sediments of urban areas were below the ERL (effects range low), but the industrial samples had ΣPAHs concentrations between ERL and ERM (effects range median). Furthermore, ΣHPAHs (heavy PAHs) and some individual PAHs in some industrial stations exceeded ERM, indicating adverse ecological risk effects frequently occur. Findings demonstrate that the surface sediment from Asaluyeh shore is highly to very highly contaminated with PAHs. PMID:27533870

  5. Molecular conformation of the full-length tumor suppressor NF2/Merlin—a small angle neutron scattering study

    PubMed Central

    Khajeh, Jahan Ali; Ju, Jeong Ho; Atchiba, Moussoubaou; Allaire, Marc; Stanley, Christopher; Heller, William T.; Callaway, David J.E.; Bu, Zimei

    2014-01-01

    Summary The tumor suppressor protein Merlin inhibits cell proliferation upon establishing cell-cell contacts. Because Merlin has high sequence similarity to the Ezrin-Radixin-Moesin (ERM) family of proteins, the structural model of ERM protein autoinhibition and cycling between closed/resting and open/active conformational states is often employed to explain Merlin function. However, recent biochemical studies suggest alternative molecular models of Merlin function. Here, we have determined the low resolution molecular structure and binding activity of Merlin and a Merlin(S518D) mutant that mimics the inactivating phosphorylation at S518 using small angle neutron scattering (SANS) and binding experiments. SANS shows that in solution both Merlin and Merlin(S518D) adopt a closed conformation, but binding experiments indicate that a significant fraction of either Merlin or Merlin(S518D) is capable of binding to the target protein NHERF1. Upon binding to the phosphatidylinositol 4,5-bisphosphate lipid, the wild-type Merlin adopts a more open conformation than in solution, but Merlin(S518D) remains in a closed conformation. This study supports a rheostat model of Merlin in NHERF1 binding, and contributes to resolve a controversy about the molecular conformation and binding activity of Merlin. PMID:24882693

  6. Vertical geochemical distribution of some contaminants in estuarine sediments from the Jaboatão River, Pernambuco, Northeastern Brazil.

    NASA Astrophysics Data System (ADS)

    Lima, Marta; Neumann, Virgínio; Castro, Maria Teresa; Lima, Enjôlras; Lima, Edmilson; Silva, Ricardo

    2014-05-01

    Sediment core samples with length ranging from 50 cm was collected in the Jaboatão river estuary and analyzed to ( As, Cr, Pb, Hg, Ni and Zn) level concentrations using instrumental neutron activation analysis (INAA). The studied area is located in the northeastern coast of Pernambuco State and is comprised of a densely urbanized region, the largest industrial complex of the state, with a predominance of fertilizer plants. These elements have a consistent vertical evolution compatible with the increase of the pelitic-organic material. There is an increase from bottom to top of the core but this may not represent a real increase in the input, but rather a concentration associated to the fine particle fraction and organic matter that increases to the top of the core profile. These conditions of clay predominance in estuarine sediments on the top indicate a decrease of the energy condition in the system and not necessarily an increase in the supply of contaminating metals. The majority of the chemical species, the concentration did not reach toxicity levels, when considering the values (ERL and ERM) established by USEPA (1998). The only exception is Cr, which has already surpassed the ERM value, and suggests that this increase is related to industrial contribution. Chromium concentration reached preliminary alert levels according to international environmental agencies, whereas the other species analyzed do not show toxicity. Key-words: environmental geochemistry, estuarine sediments

  7. iGen 0.1: the automated generation of a parameterisation of entrainment in marine stratocumulus

    NASA Astrophysics Data System (ADS)

    Tang, D. F.; Dobbie, S.

    2011-09-01

    In a previous paper we described a new technique for automatically generating parameterisations using a program called iGen. iGen generates parameterisations by analysing the source code of a~high resolution model that resolves the physics to be parameterised. In order to demonstrate that this technique scales up to deal with models of realistic complexity we have used iGen to generate a parameterisation of entrainment in marine stratocumulus. We describe how iGen was used to analyse the source code of an eddy resolving model (ERM) and generate a parameterisation of entrainment velocity in marine stratocumulus in terms of the large-scale state of the boundary layer. The parameterisation was tested against results from the DYCOMS-II intercomparison of ERM models and iGen's parameterisation of mean entrainment velocity was found to be 5.27 × 10-3 ± 0.62 × 10-3 m s-1 compared to 5.2 × 10-3 ± 0.8 × 10-3 m s-1 for the DYCOMS-II ensemble of large eddy simulation (LES) models.

  8. Electronic radon monitoring with the CMOS System-on-Chip AlphaRad

    NASA Astrophysics Data System (ADS)

    Higueret, S.; Husson, D.; Le, T. D.; Nourreddine, A.; Michielsen, N.

    2008-01-01

    The development of the integrated circuit AlphaRad as a new System-on-Chip for detection of α-particles has already been reported. This paper deals with electronic monitoring of atmospheric radon, which is one of the promising applications of the chip. The future electronic radon monitor (ERM) is designed to be compact, inexpensive, operating at low voltage and fully stand-alone. We present here the complete electronic board of the future ERM: it is made of three independent AlphaRad chips running in parallel, mounted on a small printed-circuit board which includes a numeric block for data treatment based on a Xilinx programmable gate array. The maximal counting rate of the AlphaRad chip has been pushed to at least 3×10 6 α-particles cm -2. The complete system for detection of the solid aerosols will be published separately, and this paper will focus on the electronic board alone. Already 20 times faster than our first measurement with a CMOS pixel sensor, the system was tested at low and high activities, showing an excellent linearity for 222Rn levels up to 80 kBq m -3.

  9. Project Columbiad: Mission to the Moon. Book 2, volume 3: Stage configuration designs; volume 4: Program plan

    NASA Astrophysics Data System (ADS)

    The Earth Orbital Rendezvous (EOR) configuration for the piloted mission is composed of three propulsive elements in addition to the Crew Module (CM): Primary Trans-Lunar Injection (PTLI), Lunar Braking Module (LBM), and Earth Return Module (ERM). The precursor mission is also composed of three propulsive elements in addition to its surface payloads: PTLI, LBM and the Payload Landing Module (PLM). Refer to Volume 1, Section 5.1 and 5.2 for a break-up of the different stages into the four launches. A quick summary is as follows: PTLI is on Launch 1 and 3 while the LBM, PLM, and surface payloads are on Launch 2 and another LBM, ERM, and CM on Launch 4. The precursor mission is designed to be as modular as possible with the piloted mission for developmental cost considerations. The following topics are discussed: launch vehicle description; primary trans-lunar injection stage; lunar braking module; earth return module; crew module; payload landing module; and surface payload description.

  10. Combining topological analysis matrices-based active learning on networked data classification

    NASA Astrophysics Data System (ADS)

    He, Xiaoqi; Liu, Yangguang; Jin, Xiaogang

    2010-12-01

    Active learning is an important technique to improve the learned model using unlabeled data, when labeled data is difficult to obtain, and unlabeled data is available in large quantity and easy to collect. Several instance querying strategies have been suggested recently. These works show that empirical risk minimization (ERM) can find the next instance to label effectively, but the computation time consumption is large. This paper introduces a new approach to select the best instance with less time consumption. In the case where the data is graphical in nature, we can implement the graph topological analysis to rapidly select instances that are likely to be good candidates for labeling. This paper describes an approach of using degree of a node metric to identify the best instance next to label. We experiment on Zachary's Karate Club dataset and 20 newsgroups dataset with four binary classification tasks, the results show that the strategy of degree of a node has similar performance to ERM with less time consumption.

  11. Heavy metals in coastal sediments of the Ligurian sea off Vado Ligure

    NASA Astrophysics Data System (ADS)

    Bertolotto, R. M.; Tortarolo, B.; Frignani, M.; Bellucci, L. G.; Albanese, S.; Cuneo, C.

    2003-05-01

    This paper combines data from a survey on heavy metal contamination of surficial sediments and the analysis of a short sediment core (30 cm) carried out in 1999 and 2001, respectively. Heavy metals (As, Cd, Cr, Cu, Hg, Ni, Pb, Zn) and Al were analysed by AAS after complete dissolution of the samples. Surficial sediments are particularly rich in Cr, Cu, Hg and Pb, which reach concentrations of 322, 47.0, 5.85 and 145 mg kg^{-1}, respectively. While most metals show high concentrations close to the built-up area, the maximum values of Cr characterise offshore samples, thus suggesting a different origin. Sediment accumulation rates and chronologies were calculated on the basis of both ^{137}Cs and ^{210}Pb activity-depth profiles. The depth distribution of Al is peculiar, showing several peaks (from ca. 17 to ca. 106 mg g^{-1}) that are not justified by changes in sediment grain size and mineralogical composition. Very recent inputs of Cr and Ni are accounted for by surficial peak values, whereas Hg and Cu reach high concentrations at depth in core (before the early 1960s) and then decrease. Cu shows also a recent peak. Sediment grain size as well as organic carbon content do not seem to be correlated and significantly influence the metal distributions. Hg concentrations exceed the ERM guidelines all over the study area, whereas Cr is higher than the ERM at the top of the core. Only Cd is always lower than the ERL guidelines.

  12. CLIC5 stabilizes membrane-actin filament linkages at the base of hair cell stereocilia in a molecular complex with radixin, taperin, and myosin VI.

    PubMed

    Salles, Felipe T; Andrade, Leonardo R; Tanda, Soichi; Grati, M'hamed; Plona, Kathleen L; Gagnon, Leona H; Johnson, Kenneth R; Kachar, Bechara; Berryman, Mark A

    2014-01-01

    Chloride intracellular channel 5 protein (CLIC5) was originally isolated from microvilli in complex with actin binding proteins including ezrin, a member of the Ezrin-Radixin-Moesin (ERM) family of membrane-cytoskeletal linkers. CLIC5 concentrates at the base of hair cell stereocilia and is required for normal hearing and balance in mice, but its functional significance is poorly understood. This study investigated the role of CLIC5 in postnatal development and maintenance of hair bundles. Confocal and scanning electron microscopy of CLIC5-deficient jitterbug (jbg) mice revealed progressive fusion of stereocilia as early as postnatal day 10. Radixin (RDX), protein tyrosine phosphatase receptor Q (PTPRQ), and taperin (TPRN), deafness-associated proteins that also concentrate at the base of stereocilia, were mislocalized in fused stereocilia of jbg mice. TPRQ and RDX were dispersed even prior to stereocilia fusion. Biochemical assays showed interaction of CLIC5 with ERM proteins, TPRN, and possibly myosin VI (MYO6). In addition, CLIC5 and RDX failed to localize normally in fused stereocilia of MYO6 mutant mice. Based on these findings, we propose a model in which these proteins work together as a complex to stabilize linkages between the plasma membrane and subjacent actin cytoskeleton at the base of stereocilia. PMID:24285636

  13. Conjugal mobilization of the mega element carrying mef(E) from Streptococcus salivarius to Streptococcus pneumoniae.

    PubMed

    Santagati, Maria; Lupo, Agnese; Scillato, Marina; Di Martino, Andrea; Stefani, Stefania

    2009-01-01

    We report the isolation and characterization of an unusual strain of Streptococcus salivarius, 3C30, displaying both the macrolide-lincosamide-streptogramin B and the tetracycline resistance phenotypes. It harbours the mef(E), erm(B), and tet(M) genes carried by different genetic elements. The genetic element carrying mef(E), named mega, was investigated by long PCR and sequencing, while the presence of the Tn3872-like element, carrying tet(M) and erm(B), was demonstrated by sequencing of both the int-xis-Tn and the fragment between the two resistance genes. In strain 3C30 the mega element is 5388 bp in size and its nucleotide sequence is identical to that of the element described previously in S. salivarius, with the exception of a 912 bp deletion at the left end. The composite Tn3872-like element appeared to be nonconjugative while the mega element was transferred by conjugation to Streptococcus pneumoniae. It was, however, impossible to transfer it again from these transconjugants to other strains. In addition, only in the 3C30 strain did mega form circular structures, as identified by real-time PCR. In conclusion, we found a clinical strain of S. salivarius carrying both mega and Tn3872-like genetic elements. Mega is transferable by conjugation to S. pneumoniae but it is not transferable again from the transconjugants, suggesting a possible mobilization by recombinases of the coresident Tn3872-like transposon. PMID:19025575

  14. Heterologous coexpression of Vitreoscilla hemoglobin and Bacillus megaterium glucanase in Streptomyces lydicus A02 enhanced its production of antifungal metabolites.

    PubMed

    Wu, Huiling; Li, Jinjin; Dong, Dan; Liu, Ting; Zhang, Taotao; Zhang, Dianpeng; Liu, Weicheng

    2015-12-01

    Streptomyces lydicus A02 is a novel producer of commercially important polyene macrocyclic antibiotic natamycin and a potential biocontrol agent to several plant fungal diseases, including wilt caused by Fusarium oxysporum f. spp. To improve the natamycin production and the antifungal activity of S. lydicus A02, we coexpressed gene vgb encoding Vitreoscilla hemoglobin (VHb) and bglC encoding Bacillus megaterium L103 glucanase, both under the control of the strong constitutive ermE* promoter, in S. lydicus A02. Our results showed that coexpressing VHb and glucanase improved cell growth, and the engineered strain produced 26.90% more biomass than the wild-type strain after 72h fermentation in YSG medium. In addition, coexpressing genes encoding VHb and glucanase led to increased natamycin production, higher endogenous chitinase activity and exogenous glucanase activity, as well as enhanced antifungal activity in the engineered S. lydicus AVG02 and AGV02, regardless of the position of the two genes on the plasmids. Compared with model strains, few reports have successfully coexpressed VHb and other foreign proteins in industrial strains. Our results illustrated an effective approach for improving antifungal activity in an industrial strain by the rational engineering of combined favorable factors. PMID:26453475

  15. Heterogeneity in resistant fecal Bacteroides fragilis group collected from healthy people.

    PubMed

    Narimani, T; Douraghi, M; Owlia, P; Rastegar, A; Esghaei, M; Nasr, B; Talebi, M

    2016-06-01

    Normal nonpathogenic flora would represent a constant lake of resistance genes potentially transferable to human pathogens. To assess the prevalence of resistance genes and genetic variability of Bacteroides fragilis group (BFG) from normal flora, 177 Bacteroides isolates obtained from the fecal samples of healthy individuals. These isolates were subjected to antibiotic susceptibility testing and pulsed field gel electrophoresis (PFGE). The isolates were further tested for the presence of ermF, tetQ and bft genes by PCR. Our results indicated the presence of different clonal strains (1 common type and 57 single types) among the resistant isolates. The resistance rate for the six antibiotics in this study was between 1% and 95%. Most of the isolates (99%) were susceptible to metronidazole. ermF and tetQ were detected in all erythromycin and tetracycline resistant isolates. None of the isolates were carried bft gene. These data suggest dissemination of heterogenic clonal groups in healthy persons and resistance to 5 high commonly used antibiotics. PMID:26945562

  16. Differentially Private Empirical Risk Minimization

    PubMed Central

    Chaudhuri, Kamalika; Monteleoni, Claire; Sarwate, Anand D.

    2011-01-01

    Privacy-preserving machine learning algorithms are crucial for the increasingly common setting in which personal data, such as medical or financial records, are analyzed. We provide general techniques to produce privacy-preserving approximations of classifiers learned via (regularized) empirical risk minimization (ERM). These algorithms are private under the ε-differential privacy definition due to Dwork et al. (2006). First we apply the output perturbation ideas of Dwork et al. (2006), to ERM classification. Then we propose a new method, objective perturbation, for privacy-preserving machine learning algorithm design. This method entails perturbing the objective function before optimizing over classifiers. If the loss and regularizer satisfy certain convexity and differentiability criteria, we prove theoretical results showing that our algorithms preserve privacy, and provide generalization bounds for linear and nonlinear kernels. We further present a privacy-preserving technique for tuning the parameters in general machine learning algorithms, thereby providing end-to-end privacy guarantees for the training process. We apply these results to produce privacy-preserving analogues of regularized logistic regression and support vector machines. We obtain encouraging results from evaluating their performance on real demographic and benchmark data sets. Our results show that both theoretically and empirically, objective perturbation is superior to the previous state-of-the-art, output perturbation, in managing the inherent tradeoff between privacy and learning performance. PMID:21892342

  17. Multigen-2 Pre-Flight Testing: Science Testing Unit (STU) and Stowage Conditions

    NASA Astrophysics Data System (ADS)

    Kittang, A.-I.; Kvaloy, B.; Berg, C.; Rakvaag, G.; Iversen, T.-H.

    2008-06-01

    The Multigen-2 experiment Science Testing Unit (STU) proved to be a useful tool in optimizing experiment environment settings for cultivation of Arabidopsis thaliana (Col-0) in the European Modular Cultivation System (EMCS). By using the EMCS Experiment Reference Model (ERM); light, temperature and air flow regimes for optimal growth could be tested. Healthy seedlings were obtained using the STU#2 and STU#3 in the EMCS ERM. It was concluded that the Experiment Container Development Kit (ECDK) is unsuitable for the Multigen-2 testing due to limitation in the ECDK temperature control. The results from the stowage condition tests showed that the selected growth medium (agar) can be used after 3 months at +4°C. The seeds show a germination rate of ≥80% after sterilisation and stowed for 5 months. The Multigen-2 plant samples will be fixed in RNA later and stored at - 80 °C. Three methods with different RNA isolation kits showed that the Qiagen kit (#74904) gave the highest amount and the best quality of Total RNA from RNA Later and frozen samples. The amount of plant material from one cultivation chamber gives two RNA isolations. Each of the isolations gives Total RNA sufficient for at least two microarray analyses.

  18. Dasatinib affects focal adhesion and myosin regulation to inhibit matrix contraction by Müller cells.

    PubMed

    Tsukahara, Rintaro; Umazume, Kazuhiko; Yamakawa, Naoyuki; McDonald, Kevin; Kaplan, Henry J; Tamiya, Shigeo

    2015-10-01

    Epiretinal membrane (ERM) contraction is associated with a variety of ocular diseases that cause macular dysfunction. Trans-differentiated Müller cells have been identified in ERMs, and have been implicated to be involved in the contractile process. In this study, we tested the effect of dasatinib, an FDA-approved tyrosine kinase inhibitor, on matrix contraction caused by Müller cells, and examined molecular mechanism of action. Type I collagen matrix contraction assays were used to examine the effect of drugs on matrix contraction by trans-differentiated Müller cells. Fluophore-conjugated phalloidin was used for the detection of actin cytoskeleton, and Western-blot analyses were carried out to examine protein expression and phosphorylation status. Dasatinib inhibited collagen matrix contraction by trans-differentiated Müller cells that was associated with decreased cell spreading and reduction of actomyosin stress fibers. Concomitantly, dasatinib-treated Müller cells had reduced phosphorylation of Src family kinase, paxillin, as well as myosin II light chain. Specific inhibitors of Rho/ROCK and myosin II confirmed the critical role played by this pathway in Müller cell contraction. Our data demonstrate that dasatinib significantly reduced matrix contraction by Müller cells via inhibition of focal adhesion, as well as actomyosin contraction. PMID:26240967

  19. Specific requirements of nonbilayer phospholipids in mitochondrial respiratory chain function and formation.

    PubMed

    Baker, Charli D; Basu Ball, Writoban; Pryce, Erin N; Gohil, Vishal M

    2016-07-15

    Mitochondrial membrane phospholipid composition affects mitochondrial function by influencing the assembly of the mitochondrial respiratory chain (MRC) complexes into supercomplexes. For example, the loss of cardiolipin (CL), a signature non-bilayer-forming phospholipid of mitochondria, results in disruption of MRC supercomplexes. However, the functions of the most abundant mitochondrial phospholipids, bilayer-forming phosphatidylcholine (PC) and non-bilayer-forming phosphatidylethanolamine (PE), are not clearly defined. Using yeast mutants of PE and PC biosynthetic pathways, we show a specific requirement for mitochondrial PE in MRC complex III and IV activities but not for their formation, whereas loss of PC does not affect MRC function or formation. Unlike CL, mitochondrial PE or PC is not required for MRC supercomplex formation, emphasizing the specific requirement of CL in supercomplex assembly. Of interest, PE biosynthesized in the endoplasmic reticulum (ER) can functionally substitute for the lack of mitochondrial PE biosynthesis, suggesting the existence of PE transport pathway from ER to mitochondria. To understand the mechanism of PE transport, we disrupted ER-mitochondrial contact sites formed by the ERMES complex and found that, although not essential for PE transport, ERMES facilitates the efficient rescue of mitochondrial PE deficiency. Our work highlights specific roles of non-bilayer-forming phospholipids in MRC function and formation. PMID:27226479

  20. 20 g PPV with indocyanine green-assisted ILM peeling versus 23 g PPV with brilliant blue G-assisted ILM peeling for epiretinal membrane.

    PubMed

    Manousaridis, Kleanthis; Peter, Silvia; Mennel, Stefan

    2016-06-01

    To compare the anatomical and visual outcomes of 20 gauge (g) pars plana vitrectomy (PPV) with indocyanine green (ICG)-assisted internal limiting membrane (ILM) peeling and 23 g PPV with brilliant blue G (BBG)-assisted ILM peeling for idiopathic epiretinal membrane (ERM). 38 eyes of 38 patients with idiopathic ERM were included. They were divided in two groups: group 1 (18 eyes) underwent 20 g PPV with ICG-assisted ILM peeling and group 2 (20 eyes) 23 g PPV with BBG-assisted ILM peeling. Postoperative best-corrected visual acuity (BCVA) and central macular thickness (CMT) were compared. Average BCVA in group 1 improved significantly from 0.60 logarithm of the minimal angle of resolution (log MAR) at baseline to 0.3 log MAR postoperatively. Average BCVA in group 2 improved significantly from 0.60 log MAR at baseline to 0.3 log MAR postoperatively. Mean CMT reduced significantly from 473 to 375 μm in group 1 and from 486 to 396 μm in group 2. There were no significant differences in the BCVA and CMT between the groups. Both surgical methods appeared to be safe and provided similar anatomical and visual outcomes. PMID:26499510

  1. Clinical and Microbiological Aspects of Linezolid Resistance Mediated by the cfr Gene Encoding a 23S rRNA Methyltransferase▿

    PubMed Central

    Arias, Cesar A.; Vallejo, Martha; Reyes, Jinnethe; Panesso, Diana; Moreno, Jaime; Castañeda, Elizabeth; Villegas, Maria V.; Murray, Barbara E.; Quinn, John P.

    2008-01-01

    The cfr (chloramphenicol-florfenicol resistance) gene encodes a 23S rRNA methyltransferase that confers resistance to linezolid. Detection of linezolid resistance was evaluated in the first cfr-carrying human hospital isolate of linezolid and methicillin-resistant Staphylococcus aureus (designated MRSA CM-05) by dilution and diffusion methods (including Etest). The presence of cfr was investigated in isolates of staphylococci colonizing the patient's household contacts and clinical isolates recovered from patients in the same unit where MRSA CM-05 was isolated. Additionally, 68 chloramphenicol-resistant Colombian MRSA isolates recovered from hospitals between 2001 and 2004 were screened for the presence of the cfr gene. In addition to erm(B), the erm(A) gene was also detected in CM-05. The isolate belonged to sequence type 5 and carried staphylococcal chromosomal cassette mec type I. We were unable to detect the cfr gene in any of the human staphylococci screened (either clinical or colonizing isolates). Agar and broth dilution methods detected linezolid resistance in CM-05. However, the Etest and disk diffusion methods failed to detect resistance after 24 h of incubation. Oxazolidinone resistance mediated by the cfr gene is rare, and acquisition by a human isolate appears to be a recent event in Colombia. The detection of cfr-mediated linezolid resistance might be compromised by the use of the disk diffusion or Etest method. PMID:18174304

  2. Molecular characterization of SCO0765 as a cellotriose releasing endo-β-1,4-cellulase from Streptomyces coelicolor A(3).

    PubMed

    Hong, Joo-Bin; Dhakshnamoorthy, Vijayalakshmi; Lee, Chang-Ro

    2016-09-01

    The sco0765 gene was annotated as a glycosyl hydrolase family 5 endoglucanase from the genomic sequence of Streptomyces coelicolor A3(2) and consisted of 2,241 bp encoding a polypeptide of 747 amino acids (molecular weight of 80.5 kDa) with a 29-amino acid signal peptide for secretion. The SCO0765 recombinant protein was heterogeneously over-expressed in Streptomyces lividans TK24 under the control of a strong ermE* promoter. The purified SCO0765 protein showed the expected molecular weight of the mature form (718 aa, 77.6 kDa) on sodium dodecyl sulfate-polyacryl amide gel electrophoresis. SCO0765 showed high activity toward β-glucan and carboxymethyl cellulose (CMC) and negligible activity to Avicel, xylan, and xyloglucan. The SCO0765 cellulase had a maximum activity at pH 6.0 and 40°C toward CMC and at pH 9.0 and 50-60°C toward β-glucan. Thin layer chromatography of the hydrolyzed products of CMC and β-glucan by SCO0765 gave cellotriose as the major product and cellotetraose, cellopentaose, and longer oligosaccharides as the minor products. These results clearly demonstrate that SCO0765 is an endo-β-1,4-cellulase, hydrolyzing the β-1,4 glycosidic bond of cellulose into cellotriose. PMID:27572512

  3. Identification and characterization of the afsR homologue regulatory gene from Streptomyces peucetius ATCC 27952.

    PubMed

    Parajuli, Niranjan; Viet, Hung Trinh; Ishida, Kenji; Tong, Hang Thi; Lee, Hei Chan; Liou, Kwangkyoung; Sohng, Jae Kyung

    2005-01-01

    We have isolated an afsR homologue, called afsR-p, through genome analysis of Streptomyces peucetius ATCC 27952. AfsR-p shares 60% sequence identity with AfsR from Streptomyces coelicolor A3 (2). afsR-p was expressed under the control of the ermE* promoter in its hosts S. peucetius, Streptomyces lividans TK 24, Streptomyces clavuligerus and Streptomyces griseus. We observed overproduction of doxorubicin (4-fold) in S. peucetius, gamma-actinorhodin (2.6-fold) in S. lividans, clavulanic acid (1.5-fold) in S. clavuligerus and streptomycin (slight) in S. griseus. Overproduction was due to expression of the gene in these strains as compared to the wild-type strains harboring the vector only. Comparative study of the expression of afsR-p revealed that regulatory networking in Streptomyces is not uniform. We speculate that phosphorylated AfsR-p becomes bound to the promoter region of afsS. The latter activates other regulatory genes, including pathway regulatory genes, and induces the production of secondary metabolites including antibiotics. We identified specific conserved amino acids and exploited them for the isolation of the partial sequence of the afsR homologue from S. clavuligerus and Streptomyces achromogens (rubradirin producer). Such findings provide additional evidence for the presence of a serine/threonine and tyrosine kinase-dependent global regulatory network in Streptomyces. PMID:15921897

  4. Characterization and antibiotic susceptibility of Streptococcus agalactiae isolates causing urinary tract infections.

    PubMed

    Piccinelli, Giorgio; Biscaro, Valeria; Gargiulo, Franco; Caruso, Arnaldo; De Francesco, Maria Antonia

    2015-08-01

    Streptococcus agalactiae (GBS) has been implicated in urinary tract infections but the microbiological characteristics and antimicrobial susceptibility of these strains are poorly investigated. In this study, 87 isolates recovered from urine samples of patients who had attended the Spedali Civili of Brescia (Italy) and had single organism GBS cultured were submitted to antimicrobial susceptibility testing, molecular characterization of macrolide and levofloxacin resistance, PCR-based capsular typing and analysis of surface protein genes. By automated broth microdilution method, all isolates were susceptible to penicillin, cefuroxime, cefaclor, and ceftriaxone; 80%, 19.5% and 3.4% of isolates were non-susceptible to tetracycline, erythromycin, and levofloxacin, respectively. Macrolide resistance determinants were iMLS(B) (n=1), cMLS(B) (n=10) and M (n=5), associated with ermTR, ermB and mefA/E. Levofloxacin resistance was linked to mutations in gyrA and parC genes. Predominant capsular types were III, Ia, V, Ib and IX. Type III was associated with tetracycline resistance, while type Ib was associated with levofloxacin resistance. Different capsular type-surface protein gene combinations (serotype V-alp2, 3; serotype III-rib; serotype Ia-epsilon) were detected. A variety of capsular types are involved in significant bacteriuria. The emergence of multidrug resistant GBS may become a significant public health concern and highlights the importance of careful surveillance to prevent the emergence of these virulent GBS. PMID:26144658

  5. Spatial and temporal analysis of the risks posed by polycyclic aromatic hydrocarbon, polychlorinated biphenyl and metal contaminants in sediments in UK estuaries and coastal waters.

    PubMed

    Manuel Nicolaus, E E; Law, Robin J; Wright, Serena R; Lyons, Brett P

    2015-06-15

    The environmental risks of 22 contaminants, comprising 6 metals, 10 PAHs and 6 PCB congeners occurring in UK estuaries and coastal waters were assessed as single substances. Sediment samples were taken within 12 nautical miles of the English and Welsh coastlines between 1999 and 2011. The measured environmental concentrations were compared to quality standards including ERL, ERM and EAC, all of which have been established internationally. Out of a total of 38,031 individual samples analysed, 42.6% and 7.7% exceeded the ERL/EAC and ERM values, respectively. The highest Risk Characterisation Ratios (RCRs) for metals, PAHs and PCBs were observed for copper, fluorene and CB118 (2,3',4,4',5-pentachlorobiphenyl). In general, the highest concentrations of PAHs and PCBs were observed in 2011 in the Lower Medway indicating a potential risk to the aquatic environment. This study suggests that re-suspension of contaminants banned over 20years ago is still an ongoing issue. PMID:25813718

  6. Distribution of streptogramin resistance genes and genetic relatedness among quinupristin/dalfopristin-resistant Enterococcus faecium recovered from pigs and chickens in Korea.

    PubMed

    Hwang, I Y; Ku, H O; Lim, S K; Lee, K J; Park, C K; Jung, G S; Jung, S C; Park, Y H; Nam, H M

    2010-08-01

    Fifty-four quinupristin/dalfopristin-resistant Enterococcus faecium (QDREF) isolated from chickens and pigs during 2002-2003 in Korea were screened by PCR for the presence of streptogramin resistance genes vatD, vatE, and vgbA, and macrolide resistance gene ermB. None of the QDREF isolates carried vgbA and vatD genes, while vatE and ermB were detected in 9.2% and 74% of the isolates, respectively. Twenty-six percent (14/54) of the QDREF isolates contained none of the resistance determinants tested. Pulsed-field gel electrophoresis (PFGE) patterns revealed high heterogeneity: 47 different patterns for 54 QDREF evaluated. Identical PFGE types were observed in two pairs of chicken isolates and a pair of pig isolates, respectively, but chicken isolates did not share PFGE pattern with pig isolates, suggesting clonal spread of QDREF strain between the same species of animals but not between different species of animals. This is the first report, to our knowledge, of vatE-positive E. faecium isolates and also the first evidence of clonal spread of QDREF strain between animals in Korea. PMID:20206952

  7. Echinoderms from Azores islands: an unexpected source of antibiotic resistant Enterococcus spp. and Escherichia coli isolates.

    PubMed

    Marinho, Catarina; Silva, Nuno; Pombo, Sofia; Santos, Tiago; Monteiro, Ricardo; Gonçalves, Alexandre; Micael, Joana; Rodrigues, Pedro; Costa, Ana Cristina; Igrejas, Gilberto; Poeta, Patrícia

    2013-04-15

    The prevalence of antibiotic resistance and the implicated mechanisms of resistance were evaluated in Enterococcus spp. and Escherichia coli, isolated from a total of 250 faecal samples of echinoderms collected from Azorean waters (Portugal). A total of 144 enterococci (120 Enterococcus faecium, 14 E. hirae, 8 E. faecalis, 2 E. gallinarum) and 10 E. coli were recovered. High percentages of resistance in enterococci were found for erythromycin, ampicillin, tetracyclin and ciprofloxacin. The erm(A) or erm(B), tet(M) and/or tet(L), vat(D), aac(6')-aph(2″) and aph(3')-IIIa genes were found in isolates resistant to erythromycin, tetracycline, quinupristin/dalfopristin, high-level gentamicin and high-level kanamycin, respectively. Resistance in E. coli isolates was detected for streptomycin, amikacin, tetracycline and tobramycin. The aadA gene was found in streptomycin-resistant isolates and tet(A)+tet(B) genes in tetracycline-resistant isolates. The data recovered are essential to improve knowledge about the dissemination of resistant strains through marine ecosystems and the possible implications involved in transferring these resistances either to other animals or to humans. PMID:23419753

  8. Technological characterisation, antibiotic susceptibility and antimicrobial activity of wild-type Leuconostoc strains isolated from North Italian traditional cheeses.

    PubMed

    Morandi, Stefano; Cremonesi, Paola; Silvetti, Tiziana; Brasca, Milena

    2013-11-01

    Genotypic and technological properties, antibiotic susceptibility and antimicrobial activity of 35 Leuconostoc strains, isolated from different Italian raw milk cheeses, were investigated. RAPD-PCR was used to study genetic variability and to distinguish closely related strains. The results showed a high degree of heterogeneity among isolates. All the strains had weak acidifying activity and showed low proteolytic and lipolytic activities. Reduction activity, was generally low. All the Leuconostoc were susceptible to ampicillin, mupirocin, erythromycin, quinupristin/dalfopristin and tetracycline. Many strains were classified as resistant to oxacillin, ciprofloxacin and nitrofurantonin, while all isolates were found resistant to vancomycin. PCR-based detection did not identify any of the common genetic determinants for vancomycin (vanA, vanB, vanC1, vanC2, vanC3, vanD, vanE, vanG) or erythromycin (ermB and ermC). Tetracycline resistance genes were detected in 25 tetracycline susceptible strains, the most frequent one being tetM. One strain, belonging to Ln. pseudomesenteroides species, was positive for the presence of the int gene of the Tn916/Tn1545 trasposon family. This is the first time the conjugative transposon Tn916 has been detected inside the Leuconostoc species. All strains showed antimicrobial activity against Enterococcus faecalis and Ent. faecium. The presence of genes encoding amino-acid decarboxylases (hdc and tdc) was not detected. Some strains are interesting in view of their use in cheese production as starter and non starter cultures. PMID:24067095

  9. Eubacterial Diterpene Cyclase Genes Essential for Production of the Isoprenoid Antibiotic Terpentecin

    PubMed Central

    Dairi, Tohru; Hamano, Yoshimitsu; Kuzuyama, Tomohisa; Itoh, Nobuya; Furihata, Kazuo; Seto, Haruo

    2001-01-01

    A gene cluster containing the mevalonate pathway genes (open reading frame 2 [ORF2] to ORF7) for the formation of isopentenyl diphosphate and a geranylgeranyl diphosphate (GGDP) synthase gene (ORF1) had previously been cloned from Streptomyces griseolosporeus strain MF730-N6, a diterpenoid antibiotic, terpentecin (TP) producer (Y. Hamano, T. Dairi, M. Yamamoto, T. Kawasaki, K Kaneda, T. Kuzuyama, N. Itoh, and H. Seto, Biosci. Biotech. Biochem. 65:1627–1635, 2001). Sequence analysis in the upstream region of the cluster revealed seven new ORFs, ORF8 to ORF14, which were suggested to encode TP biosynthetic genes. We constructed two mutants, in which ORF11 and ORF12, which encode a protein showing similarities to eukaryotic diterpene cyclases (DCs) and a eubacterial pentalenene synthase, respectively, were inactivated by gene disruptions. The mutants produced no TP, confirming that these cyclase genes are essential for the production of TP. The two cyclase genes were also expressed in Streptomyces lividans together with the GGDP synthase gene under the control of the ermE* constitutive promoter. The transformant produced a novel cyclic diterpenoid, ent-clerod-3,13(16),14-triene (terpentetriene), which has the same basic skeleton as TP. The two enzymes, each of which was overproduced in Escherichia coli and purified to homogeneity, converted GGDP into terpentetriene. To the best of our knowledge, this is the first report of a eubacterial DC. PMID:11567009

  10. Molecular dissection of radixin: distinct and interdependent functions of the amino- and carboxy-terminal domains

    PubMed Central

    1995-01-01

    The ERM proteins--ezrin, radixin, and moesin--occur in particular cortical cytoskeletal structures. Several lines of evidence suggest that they interact with both cytoskeletal elements and plasma membrane components. Here we described the properties of full-length and truncated radixin polypeptides expressed in transfected cells. In stable transfectants, exogenous full-length radixin behaves much like endogenous ERM proteins, localizing to the same cortical structures. However, the presence of full-length radixin or its carboxy-terminal domain in cortical structures correlates with greatly diminished staining of endogenous moesin in those structures, suggesting that radixin and moesin compete for a limiting factor required for normal associations in the cell. The results also reveal distinct roles for the amino- and carboxy-terminal domains. At low levels relative to endogenous radixin, the carboxy-terminal polypeptide is associated with most of the correct cortical targets except cleavage furrows. In contrast, the amino-terminal polypeptide is diffusely localized throughout the cell. Low level expression of full-length radixin or either of the truncated polypeptides has no detectable effect on cell physiology. However, high level expression of the carboxy-terminal domain dramatically disrupts normal cytoskeletal structures and functions. At these high levels, the amino-terminal polypeptide does localize to cortical structures, but does not affect the cells. We conclude that the behavior of radixin in cells depends upon activities contributed by separate domains of the protein, but also requires modulating interactions between those domains. PMID:7744951

  11. Characterization of wild-type and deltaF508 cystic fibrosis transmembrane regulator in human respiratory epithelia.

    PubMed

    Kreda, Silvia M; Mall, Marcus; Mengos, April; Rochelle, Lori; Yankaskas, James; Riordan, John R; Boucher, Richard C

    2005-05-01

    Previous studies in native tissues have produced conflicting data on the localization and metabolic fate of WT and deltaF508 cystic fibrosis transmembrane regulator (CFTR) in the lung. Combining immunocytochemical and biochemical studies utilizing new high-affinity CFTR mAbs with ion transport assays, we examined both 1) the cell type and region specific expression of CFTR in normal airways and 2) the metabolic fate of deltaF508 CFTR and associated ERM proteins in the cystic fibrosis lung. Studies of lungs from a large number of normal subjects revealed that WT CFTR protein localized to the apical membrane of ciliated cells within the superficial epithelium and gland ducts. In contrast, other cell types in the superficial, gland acinar, and alveolar epithelia expressed little WT CFTR protein. No deltaF508 CFTR mature protein or function could be detected in airway specimens freshly excised from a large number of deltaF508 homozygous subjects, despite an intact ERM complex. In sum, our data demonstrate that WT CFTR is predominantly expressed in ciliated cells, and deltaF508 CFTR pathogenesis in native tissues, like heterologous cells, reflects loss of normal protein processing. PMID:15716351

  12. Characterization of Wild-Type and ΔF508 Cystic Fibrosis Transmembrane Regulator in Human Respiratory Epithelia

    PubMed Central

    Kreda, Silvia M.; Mall, Marcus; Mengos, April; Rochelle, Lori; Yankaskas, James; Riordan, John R.; Boucher, Richard C.

    2005-01-01

    Previous studies in native tissues have produced conflicting data on the localization and metabolic fate of WT and ΔF508 cystic fibrosis transmembrane regulator (CFTR) in the lung. Combining immunocytochemical and biochemical studies utilizing new high-affinity CFTR mAbs with ion transport assays, we examined both 1) the cell type and region specific expression of CFTR in normal airways and 2) the metabolic fate of ΔF508 CFTR and associated ERM proteins in the cystic fibrosis lung. Studies of lungs from a large number of normal subjects revealed that WT CFTR protein localized to the apical membrane of ciliated cells within the superficial epithelium and gland ducts. In contrast, other cell types in the superficial, gland acinar, and alveolar epithelia expressed little WT CFTR protein. No ΔF508 CFTR mature protein or function could be detected in airway specimens freshly excised from a large number of ΔF508 homozygous subjects, despite an intact ERM complex. In sum, our data demonstrate that WT CFTR is predominantly expressed in ciliated cells, and ΔF508 CFTR pathogenesis in native tissues, like heterologous cells, reflects loss of normal protein processing. PMID:15716351

  13. Microincision Vitrectomy Surgery in Vitreomacular Traction Syndrome of Retinitis Pigmentosa Patients

    PubMed Central

    Vingolo, Enzo Maria; Gerace, Emanuele; Valente, Stefano; Spadea, Leopoldo; Nebbioso, Marcella

    2014-01-01

    Purpose. To investigate long-term retinal changes after microincision pars plana vitrectomy surgery (MIVS) and internal limiting membrane (ILM) peeling outcome in retinitis pigmentosa (RP) patients affected by vitreomacular traction syndrome (VMT) with higher vitreous surface adhesion or coexisting epiretinal membrane (ERM). Methods. Eight RP patients suffering from VMT were evaluated by means of best corrected visual acuity (BCVA), anterior and posterior binocular examination, spectral-domain optical coherence tomography (SD-OCT), MP-1 microperimetry (MP-1), and full-field electroretinogram (ERG), before MIVS and ILM peeling and during the 36-month follow-up. Patients were hospitalized for two days after the surgery. Surgical procedure was performed following this schedule: surgical removal of crystalline lens (MICS), MIVS with 23-gauge sutureless system trocars, core vitreous body removal, and balanced-sterile-salin-solution- (BSS-) air-gas (SF6) exchange. Results. All patients presented visual acuity (VA) increase after MIVS. None of the patients developed ocular hypertension or vitreomacular adhesions during the 3-year follow-up. MP-1 bivariate contour ellipse area (BCEA) was reduced in its dimensions and improved in all patients demonstrating a better fixation. Conclusions. MIVS could be the gold standard therapy in RP patients with VMT and higher vitreous surface adhesion or coexisting ERM if medical therapy is not applicable or not effective. PMID:25009817

  14. Mycobacterium abscessus. "Pleased to meet you, hope you guess my name...".

    PubMed

    Griffith, David E; Brown-Elliott, Barbara A; Benwill, Jeana L; Wallace, Richard J

    2015-03-01

    Mycobacterium abscessus is a formidable and difficult-to-treat mycobacterial pathogen with multiple drug-resistance mechanisms. The most important of these mechanisms is the presence of an inducible erythromycin methylase (erm) gene, because it confers macrolide resistance. It has recently been found that "M. abscessus" can be split into three species or subspecies based on gene sequence analysis other than the 16S rRNA gene and the presence or absence of a functional erm(41) gene. Several names have been applied to these three organisms, including M. abscessus or M. abscessus subsp. abscessus, Mycobacterium massiliense or M. abscessus subsp. massiliense, and Mycobacterium bolletii or M. abscessus subsp. bolletii. No universally accepted or recognized species or subspecies designations have emerged, and no names have been universally adopted for these organisms. This uncertainty has led to inconsistencies in the medical literature and understandable confusion by clinicians about the appropriate labels for "M. abscessus" isolates. We discuss the complexities involved in mycobacterial species/subspecies identification and taxonomy and suggest possible ways to improve the present state of uncertainty surrounding the labels for "M. abscessus" clinical isolates. We also suggest necessary changes in mycobacterial laboratory processing and reporting procedures for mycobacterial isolates. PMID:25643064

  15. IMEP-115: determination of methylmercury in seafood by elemental mercury analysis: collaborative study.

    PubMed

    Cordeiro, Fernando; Calderón, Josep; Gonçalves, Susana; Lourenço, Maria Helena; Robouch, Piotr; Emteborg, Hakan; Conneely, Patrick; Tumba-Tshilumba, Marie-France; de la Calle, Maria Beatriz

    2014-01-01

    A collaborative study IMEP-115 was organized by the European Union Reference Laboratory for Heavy Metals in Feed and Food (EURL-HM) to validate a method for the determination of methylmercury in seafood. The method was based on a liquid-liquid extraction with an organic solvent and with an aqueous cysteine solution. The final quantitation was done with an elemental mercury analyzer. Fifteen laboratories experienced in elemental mercury analyses, from 10 European countries, took part in the exercise. Five test items were selected to cover the concentration range from 0.013 to 5.12 mg/kg. All test items were reference materials certified for the methylmercury mass fraction: DOLT-4 (dogfish liver), TORT-2 (lobster hepatopancreas), SRM 2974a (mussel), SRM 1566b (oyster), and ERM CE-464 (tuna). Participants also received a bottle of ERM CE-463 (tuna) to test their analytical method before starting the collaborative study. Method validation showed adequate accuracy and acceptable precision for all test items, thus fitting its intended analytical purpose. The repeatability RSD ranged from 3.9 to 12.3%, while the reproducibility RSD ranged from 8.4 to 24.8%. PMID:24830172

  16. Project Columbiad: Mission to the Moon. Book 2, volume 3: Stage configuration designs; volume 4: Program plan

    NASA Technical Reports Server (NTRS)

    1992-01-01

    The Earth Orbital Rendezvous (EOR) configuration for the piloted mission is composed of three propulsive elements in addition to the Crew Module (CM): Primary Trans-Lunar Injection (PTLI), Lunar Braking Module (LBM), and Earth Return Module (ERM). The precursor mission is also composed of three propulsive elements in addition to its surface payloads: PTLI, LBM and the Payload Landing Module (PLM). Refer to Volume 1, Section 5.1 and 5.2 for a break-up of the different stages into the four launches. A quick summary is as follows: PTLI is on Launch 1 and 3 while the LBM, PLM, and surface payloads are on Launch 2 and another LBM, ERM, and CM on Launch 4. The precursor mission is designed to be as modular as possible with the piloted mission for developmental cost considerations. The following topics are discussed: launch vehicle description; primary trans-lunar injection stage; lunar braking module; earth return module; crew module; payload landing module; and surface payload description.

  17. Antibiotic resistance among cultured bacterial isolates from bioethanol fermentation facilities across the United States.

    PubMed

    Murphree, Colin A; Heist, E Patrick; Moe, Luke A

    2014-09-01

    Bacterial contamination of fuel ethanol fermentations by lactic acid bacteria (LAB) can have crippling effects on bioethanol production. Producers have had success controlling bacterial growth through prophylactic addition of antibiotics to fermentors, yet concerns have arisen about antibiotic resistance among the LAB. Here, we report on mechanisms used by 32 LAB isolates from eight different US bioethanol facilities to persist under conditions of antibiotic stress. Minimum inhibitory concentration assays with penicillin, erythromycin, and virginiamycin revealed broad resistance to each of the antibiotics as well as high levels of resistance to individual antibiotics. Phenotypic assays revealed that antibiotic inactivation mechanisms contributed to the high levels of individual resistances among the isolates, especially to erythromycin and virginiamycin, yet none of the isolates appeared to use a β-lactamase. Biofilm formation was noted among the majority of the isolates and may contribute to persistence under low levels of antibiotics. Nearly all of the isolates carried at least one canonical antibiotic resistance gene and many carried more than one. The erythromycin ribosomal methyltransferase (erm) gene class was found in 19 of 32 isolates, yet a number of these isolates exhibit little to no resistance to erythromycin. The erm genes were present in 15 isolates that encoded more than one antibiotic resistance mechanism, suggestive of potential genetic linkages. PMID:24748439

  18. Root-associated fungi of Vaccinium carlesii in subtropical forests of China: intra- and inter-annual variability and impacts of human disturbances

    PubMed Central

    Zhang, Yanhua; Ni, Jian; Tang, Fangping; Pei, Kequan; Luo, Yiqi; Jiang, Lifen; Sun, Lifu; Liang, Yu

    2016-01-01

    Ericoid mycorrhiza (ERM) are expected to facilitate establishment of ericaceous plants in harsh habitats. However, diversity and driving factors of the root-associated fungi of ericaceous plants are poorly understood. In this study, hair-root samples of Vaccinium carlesii were taken from four forest types: old growth forests (OGF), secondary forests with once or twice cutting (SEC I and SEC II), and Cunninghamia lanceolata plantation (PLF). Fungal communities were determined using high-throughput sequencing, and impacts of human disturbances and the intra- and inter-annual variability of root-associated fungal community were evaluated. Diverse fungal taxa were observed and our results showed that (1) Intra- and inter-annual changes in root-associated fungal community were found, and the Basidiomycota to Ascomycota ratio was related to mean temperature of the sampling month; (2) Human disturbances significantly affected structure of root-associated fungal community of V. carlesii, and two secondary forest types were similar in root-associated fungal community and were closer to that of the old growth forest; (3) Plant community composition, edaphic parameters, and geographic factors significantly affected root-associated fungal communities of V. carlesii. These results may be helpful in better understanding the maintenance mechanisms of fungal diversity associated with hair roots of ERM plants under human disturbances. PMID:26928608

  19. Myogenin, AP2β, NOS-1 and HMGA2 are surrogate markers of fusion status in rhabdomyosarcoma

    PubMed Central

    Rudzinski, Erin R.; Anderson, James R.; Lyden, Elizabeth R.; Bridge, Julia A.; Barr, Frederic G.; Gastier-Foster, Julie M.; Bachmeyer, Karen; Skapek, Stephen X.; Hawkins, Douglas S.; Teot, Lisa A.; Parham, David M.

    2014-01-01

    Pediatric rhabdomyosarcoma (RMS) is traditionally classified by histologic appearance into alveolar (ARMS) and embryonal (ERMS) subtypes. The majority of ARMS contain a PAX3-FOXO1 or PAX7-FOXO1 gene fusion, but about 20% do not. Intergroup Rhabdomyosarcoma Study (IRS) Stage and group-matched ARMS typically behaves more aggressively than the embryonal subtype, but recent studies have shown that it is, in fact, fusion status that drives outcome for RMS. Gene expression microarray data indicate that several genes discriminate between fusion positive and fusion negative RMS with high specificity. Using tissue microarrays containing a series of both ARMS and ERMS, we identified a panel of four immunohistochemical markers, myogenin, AP2β, NOS-1 and HMGA2, which can be used as surrogate markers of fusion status in RMS. These antibodies provide an alternative to molecular methods for identification of fusion positive RMS, particularly in cases where there is scant or poor quality material. Additionally, these antibodies, may be useful in fusion negative ARMS as an indicator that a variant gene fusion may be present. PMID:24618610

  20. Influence of different mineral nitrogen sources (NO3(-)-N vs. NH4(+)-N) on arbuscular mycorrhiza development and N transfer in a Glomus intraradices-cowpea symbiosis.

    PubMed

    Ngwene, Benard; Gabriel, Elke; George, Eckhard

    2013-02-01

    Labeled nitrogen ((15)N) was applied to a soil-based substrate in order to study the uptake of N by Glomus intraradices extraradical mycelium (ERM) from different mineral N (NO(3)(-) vs. NH(4)(+)) sources and the subsequent transfer to cowpea plants. Fungal compartments (FCs) were placed within the plant growth substrate to simulate soil patches containing root-inaccessible, but mycorrhiza-accessible, N. The fungus was able to take up both N-forms, NO(3)(-) and NH(4)(+). However, the amount of N transferred from the FC to the plant was higher when NO(3)(-) was applied to the FC. In contrast, analysis of ERM harvested from the FC showed a higher (15)N enrichment when the FC was supplied with (15)NH(4)(+) compared with (15)NO(3)(-). The (15)N shoot/root ratio of plants supplied with (15)NO(3)(-) was much higher than that of plants supplied with (15)NH(4)(+), indicative of a faster transfer of (15)NO(3)(-) from the root to the shoot and a higher accumulation of (15)NH (4)(+) in the root and/or intraradical mycelium. It is concluded that hyphae of the arbuscular mycorrhizal fungus may absorb NH(4)(+) preferentially over NO(3)(-) but that export of N from the hyphae to the root and shoot may be greater following NO(3)(-) uptake. The need for NH(4)(+) to be assimilated into organically bound N prior to transport into the plant is discussed. PMID:22810583

  1. Prevalence and Molecular Genetics of Macrolide Resistance among Streptococcus pneumoniae Isolates Collected in Finland in 2002

    PubMed Central

    Rantala, M.; Huikko, S.; Huovinen, P.; Jalava, J.

    2005-01-01

    The prevalence and mechanisms of macrolide resistance among 1,007 clinical pneumococcal isolates collected in Finland were investigated. Of these, 217 (21.5%) were resistant to erythromycin and 11% to clindamycin. Among the erythromycin-resistant isolates, mef(E) was present in 95 isolates (44%), mef(A) was present in 12 isolates (6%), and erm(B) was present in 90 isolates (41%). A double mechanism, mef(E) and erm(B), was detected in five isolates (2%). Ribosomal mutation was detected in 14 (6%) macrolide-resistant isolates in which no other determinant was found. Based on the telithromycin MICs, two groups of isolates were formed: 83.3% of the isolates belonged to a major group for which the telithromycin MIC range was ≤0.008 to 0.063 μg/ml, and 16.7% belonged to a minor group for which the telithromycin MIC range was 0.125 to 8 μg/ml. All except three isolates in the minor population carried a macrolide resistance gene. PMID:16189096

  2. Immediate and long-term posttherapy neuropsychologic performance in children with acute lymphoblastic leukemia treated without central nervous system radiation.

    PubMed

    Tamaroff, M; Miller, D R; Murphy, M L; Salwen, R; Ghavimi, F; Nir, Y

    1982-10-01

    Intellectual and neuropsychologic test performance was studied in 41 children with acute lymphoblastic leukemia within one year of completing multidrug therapy in which intrathecal methotrexate was the sole agent of central nervous system prophylaxis. To control for possible effects of intensive systemic chemotherapy and the disruptive psychosocial effects of treatment on psychometric test function, 33 children with embryonal rhabdomyosarcoma who had no CNS disease or treatment were similarly studied. All patients studied had been in continuous complete remission since diagnosis. Within each diagnostic group, patients were placed in a younger (less than 8 years) or an older (more than 8 years) group depending on age at the time of testing. No significant difference was found in intellectual function between the young ALL and ERMS groups or between the total ALL and ERMS groups. Further five-year follow-up of 12 of the youngest ALL patients showed no significant long-term change in intellectual or other specific neuropsychologic functions. Children with ALL who remained in continuous complete remission from diagnosis and who were treated with prophylactic intrathecal methotrexate and no CNS radiation did not have global or specific neuropsychologic impairment. PMID:6956704

  3. Staphylococcus saprophyticus surface-associated protein (Ssp) is associated with lifespan reduction in Caenorhabditis elegans

    PubMed Central

    Szabados, Florian; Mohner, Amelie; Kleine, Britta; Gatermann, Sören G

    2013-01-01

    Staphylococcal lipases have been proposed as pathogenicity factors. In Staphylococcus saprophyticus the surface-associated protein (Ssp) has been previously characterized as a cell wall-associated true lipase. A S. saprophyticus Δssp::ermB mutant has been described as less virulent in an in vivo model of urinary tract infection compared with its wild-type. This is the first report showing that S. saprophyticus induced a lifespan reduction in Caenorhabditis elegans similar to that of S. aureus RN4220. In two S. saprophyticus Δssp::ermB mutants lifespan reduction in C. elegans was partly abolished. In order to attribute virulence to the lipase activity itself and distinguish this phenomenon from the presence of the Ssp-protein, the conserved active site of the lipase was modified by site-directed ligase-independent mutagenesis and lipase activity-deficient mutants were constructed. These results indicate that the Ssp is associated with pathogenicity in C. elegans and one could speculate that the lipase activity itself is responsible for this virulence. PMID:23959029

  4. Assimilation of organic and inorganic nutrients by Erica root fungi from the fynbos ecosystem.

    PubMed

    Bizabani, Christine; Dames, Joanna Felicity

    2016-03-01

    Erica dominate the fynbos ecosystem, which is characterized by acidic soils that are rich in organic matter. The ericaceae associate with ericoid mycorrhizal (ERM) fungi for survival. In this study fungal biomass accumulation in vitro was used to determine nutrient utilisation of various inorganic and organic substrates. This is an initial step towards establishment of the ecological roles of typical ERM fungi and other root fungi associated with Erica plants, with regard to host nutrition. Meliniomyces sp., Acremonium implicatum, Leohumicola sp., Cryptosporiopsis erica, Oidiodendron maius and an unidentified Helotiales fungus were selected from fungi previously isolated and identified from Erica roots. Sole nitrogen sources ammonium, nitrate, arginine and Bovine Serum Albumin (BSA) were tested. Meliniomyces and Leohumicola species were able to utilise BSA effectively. Phosphorus nutrition was tested using orthophosphate, sodium inositol hexaphosphate and DNA. Most isolates preferred orthophosphate. Meliniomyces sp. and A. implicatum were able to accumulate significant biomass using DNA. Carbon utilisation was tested using glucose, cellobiose, carboxymethylcellulose, pectin and tannic acid substrates. All fungal isolates produced high biomass on glucose and cellobiose. The ability to utilize organic nutrient sources in culture, illustrates their potential role of these fungi in host nutrition in the fynbos ecosystem. PMID:26895865

  5. Glycogen synthase kinase 3 regulates PAX3-FKHR-mediated cell proliferation in human alveolar rhabdomyosarcoma cells

    SciTech Connect

    Zeng, Fu-Yue; Dong, Hanqing; Cui, Jimmy; Liu, Lingling; Chen, Taosheng

    2010-01-01

    Patients with alveolar rhabdomyosarcoma (ARMS) have poorer response to conventional chemotherapy and lower survival rates than those with embryonal RMS (ERMS). To identify compounds that preferentially block the growth of ARMS, we conducted a small-scale screen of 160 kinase inhibitors against the ARMS cell line Rh30 and ERMS cell line RD and identified inhibitors of glycogen synthase kinase 3 (GSK3), including TWS119 as ARMS-selective inhibitors. GSK3 inhibitors inhibited cell proliferation and induced apoptosis more effectively in Rh30 than RD cells. Ectopic expression of fusion protein PAX3-FKHR in RD cells significantly increased their sensitivity to TWS119. Down-regulation of GSK3 by GSK3 inhibitors or siRNA significantly reduced the transcriptional activity of PAX3-FKHR. These results suggest that GSK3 is directly involved in regulating the transcriptional activity of PAX3-FKHR. Also, GSK3 phosphorylated PAX3-FKHR in vitro, suggesting that GSK3 might regulate PAX3-FKHR activity via phosphorylation. These findings support a novel mechanism of PAX3-FKHR regulation by GSK3 and provide a novel strategy to develop GSK inhibitors as anti-ARMS therapies.

  6. Glycogen synthase kinase 3 regulates PAX3-FKHR-mediated cell proliferation in human alveolar rhabdomyosarcoma cells.

    PubMed

    Zeng, Fu-Yue; Dong, Hanqing; Cui, Jimmy; Liu, Lingling; Chen, Taosheng

    2010-01-01

    Patients with alveolar rhabdomyosarcoma (ARMS) have poorer response to conventional chemotherapy and lower survival rates than those with embryonal RMS (ERMS). To identify compounds that preferentially block the growth of ARMS, we conducted a small-scale screen of 160 kinase inhibitors against the ARMS cell line Rh30 and ERMS cell line RD and identified inhibitors of glycogen synthase kinase 3 (GSK3), including TWS119 as ARMS-selective inhibitors. GSK3 inhibitors inhibited cell proliferation and induced apoptosis more effectively in Rh30 than RD cells. Ectopic expression of fusion protein PAX3-FKHR in RD cells significantly increased their sensitivity to TWS119. Down-regulation of GSK3 by GSK3 inhibitors or siRNA significantly reduced the transcriptional activity of PAX3-FKHR. These results suggest that GSK3 is directly involved in regulating the transcriptional activity of PAX3-FKHR. Also, GSK3 phosphorylated PAX3-FKHR in vitro, suggesting that GSK3 might regulate PAX3-FKHR activity via phosphorylation. These findings support a novel mechanism of PAX3-FKHR regulation by GSK3 and provide a novel strategy to develop GSK inhibitors as anti-ARMS therapies. PMID:19995556

  7. Occurrence, distribution, and ecological risk assessment of potentially toxic elements in surface sediments of Lake Awassa and Lake Ziway, Ethiopia.

    PubMed

    Mekonnen, Kebede N; Ambushe, Abayneh A; Chandravanshi, Bhagwan S; Redi-Abshiro, Mesfin; McCrindle, Robert I

    2015-01-01

    Microwave-assisted acid digestion and modified aqua regia (HNO3:HCl:HF:H3BO3) leaching techniques were used for the determination of 15 potentially toxic elements (V, Cr, Fe, Mn, Co, Ni, Cu, Zn, As, Se, Ag, Cd, Sn, Hg and Pb) in sediment samples from Lake Awassa and Lake Ziway, Ethiopia. The digests were subsequently analyzed by inductively coupled plasma-mass spectrometry (ICP-MS) and inductively coupled plasma-optical emission spectrometry (ICP-OES). Mercury was directly determined in the solid samples using an elemental mercury analyzer. The precision and accuracy of the digestion procedures were verified using certified reference materials. The experimental results were in good agreement with the certified values (P < 0.05) and the recoveries were quantitative (>90%). The average relative standard deviations were below 10%. There is significant correlation between the two lakes at the 0.01 level (2-tailed). Using the sediment quality guidelines, both lakes are heavily polluted with Zn and some of the sites are heavily polluted with Cu, Ni and Pb. Based on effect range low (ERL) - effect range medium (ERM), in both lakes for Ag were greater than the ERM, indicating that the areas could be toxic to aquatic organisms, while for Cr, Cu, As and Hg the values were less than ERL. PMID:25438135

  8. Clinical and microbiological aspects of linezolid resistance mediated by the cfr gene encoding a 23S rRNA methyltransferase.

    PubMed

    Arias, Cesar A; Vallejo, Martha; Reyes, Jinnethe; Panesso, Diana; Moreno, Jaime; Castañeda, Elizabeth; Villegas, Maria V; Murray, Barbara E; Quinn, John P

    2008-03-01

    The cfr (chloramphenicol-florfenicol resistance) gene encodes a 23S rRNA methyltransferase that confers resistance to linezolid. Detection of linezolid resistance was evaluated in the first cfr-carrying human hospital isolate of linezolid and methicillin-resistant Staphylococcus aureus (designated MRSA CM-05) by dilution and diffusion methods (including Etest). The presence of cfr was investigated in isolates of staphylococci colonizing the patient's household contacts and clinical isolates recovered from patients in the same unit where MRSA CM-05 was isolated. Additionally, 68 chloramphenicol-resistant Colombian MRSA isolates recovered from hospitals between 2001 and 2004 were screened for the presence of the cfr gene. In addition to erm(B), the erm(A) gene was also detected in CM-05. The isolate belonged to sequence type 5 and carried staphylococcal chromosomal cassette mec type I. We were unable to detect the cfr gene in any of the human staphylococci screened (either clinical or colonizing isolates). Agar and broth dilution methods detected linezolid resistance in CM-05. However, the Etest and disk diffusion methods failed to detect resistance after 24 h of incubation. Oxazolidinone resistance mediated by the cfr gene is rare, and acquisition by a human isolate appears to be a recent event in Colombia. The detection of cfr-mediated linezolid resistance might be compromised by the use of the disk diffusion or Etest method. PMID:18174304

  9. Conjugative transfer of resistance determinants among human and bovine Streptococcus agalactiae.

    PubMed

    Pinto, Tatiana Castro Abreu; Costa, Natália Silva; Corrêa, Ana Beatriz de Almeida; de Oliveira, Ivi Cristina Menezes; de Mattos, Marcos Correa; Rosado, Alexandre Soares; Benchetrit, Leslie Claude

    2014-01-01

    Streptococcus agalactiae (GBS) is a major source of human perinatal diseases and bovine mastitis. Erythromycin (Ery) and tetracycline (Tet) are usually employed for preventing human and bovine infections although resistance to such agents has become common among GBS strains. Ery and Tet resistance genes are usually carried by conjugative transposons (CTns) belonging to the Tn916 family, but their presence and transferability among GBS strains have not been totally explored. Here we evaluated the presence of Tet resistance genes (tetM and tetO) and CTns among Ery-resistant (Ery-R) and Ery-susceptible (Ery-S) GBS strains isolated from human and bovine sources; and analyzed the ability for transferring resistance determinants between strains from both origins. Tet resistance and int-Tn genes were more common among Ery-R when compared to Ery-S isolates. Conjugative transfer of all resistance genes detected among the GBS strains included in this study (ermA, ermB, mef, tetM and tetO), in frequencies between 1.10(-7) and 9.10(-7), was possible from bovine donor strains to human recipient strain, but not the other way around. This is, to our knowledge, the first report of in vitro conjugation of Ery and Tet resistance genes among GBS strains recovered from different hosts. PMID:25477908

  10. Distribution of serotypes and evaluation of antimicrobial susceptibility among human and bovine Streptococcus agalactiae strains isolated in Brazil between 1980 and 2006.

    PubMed

    Pinto, Tatiana Castro Abreu; Costa, Natália Silva; Vianna Souza, Aline Rosa; Silva, Ligia Guedes da; Corrêa, Ana Beatriz de Almeida; Fernandes, Flavio Gimenis; Oliveira, Ivi Cristina Menezes; Mattos, Marcos Corrêa de; Rosado, Alexandre Soares; Benchetrit, Leslie Claude

    2013-01-01

    Streptococcus agalactiae is a common agent of clinical and subclinical bovine mastitis and an important cause of human infections, mainly among pregnant women, neonates and nonpregnant adults with underlying diseases. The present study describes the genetic and phenotypic diversity among 392 S. agalactiae human and bovine strains isolated between 1980 and 2006 in Brazil. The most prevalent serotypes were Ia, II, III and V and all the strains were susceptible to penicillin, vancomycin and levofloxacin. Resistance to clindamycin, chloramphenicol, erythromycin, rifampicin and tetracycline was observed. Among the erythromycin resistant strains, mefA/E, ermA and, mainly, ermB gene were detected, and a shift of prevalence from the macrolide resistance phenotype to the macrolide-lincosamide-streptogramin B resistance phenotype over the years was observed. The 23 macrolide-resistant strains showed 19 different pulsed-field gel electrophoresis profiles. Regarding macrolide resistance, a major concern in S. agalactiae epidemiology, the present study describes an increase in erythromycin resistance from the 80s to the 90s followed by a decrease in the 2000-2006 period. Also, the genetic heterogeneity described points out that erythromycin resistance in Brazil is rather due to horizontal gene transmission than to spreading of specific macrolide-resistant clones. PMID:23453948

  11. Development of a Synthetic Oxytetracycline-Inducible Expression System for Streptomycetes Using de Novo Characterized Genetic Parts.

    PubMed

    Wang, Weishan; Yang, Tongjian; Li, Yihong; Li, Shanshan; Yin, Shouliang; Styles, Kathryn; Corre, Christophe; Yang, Keqian

    2016-07-15

    Precise control of gene expression using exogenous factors is of great significance. To develop ideal inducible expression systems for streptomycetes, new genetic parts, oxytetracycline responsive repressor OtrR, operator otrO, and promoter otrBp from Streptomyces rimosus, were selected de novo and characterized in vivo and in vitro. OtrR showed strong affinity to otrO (KD = 1.7 × 10(-10) M) and oxytetracycline induced dissociation of the OtrR/DNA complex in a concentration-dependent manner. On the basis of these genetic parts, a synthetic inducible expression system Potr* was optimized. Induction of Potr* with 0.01-4 μM of oxytetracycline triggered a wide-range expression level of gfp reporter gene in different Streptomyces species. Benchmarking Potr* against the widely used constitutive promoters ermE* and kasOp* revealed greatly enhanced levels of expression when Potr* was fully induced. Finally, Potr* was used as a tool to activate and optimize the expression of the silent jadomycin biosynthetic gene cluster in Streptomyces venezuelae. Altogether, the synthetic Potr* presents a new versatile tool for fine-tuning gene expression in streptomycetes. PMID:27100123

  12. The Na+/H+ exchanger NHE1, but not the Na+, HCO3(-) cotransporter NBCn1, regulates motility of MCF7 breast cancer cells expressing constitutively active ErbB2.

    PubMed

    Lauritzen, Gitte; Stock, Christian-Martin; Lemaire, Justine; Lund, Stine F; Jensen, Mie Frid; Damsgaard, Britt; Petersen, Katrine Seide; Wiwel, Maria; Rønnov-Jessen, Lone; Schwab, Albrecht; Pedersen, Stine Falsig

    2012-04-28

    We and others have shown central roles of the Na(+)/H(+) exchanger NHE1 in cell motility. The aim of this study was to determine the roles of NHE1 and of the Na(+), HCO(3)(-) cotransporter NBCn1 in motility of serum-starved MCF-7 breast cancer cells expressing constitutively active ErbB2 (ΔNErbB2). ΔNErbB2 expression elicited NBCn1 upregulation, Ser(703)-phosphorylation of NHE1, and NHE1-inhibitor (EIPA)-sensitive pericellular acidification, in conjunction with increased expression of β1 integrin and ERM proteins. Active ERM proteins and NHE1 colocalized strongly to invadopodial rosettes, the diameter of which was increased by ΔNErbB2. Adhesion and migration on collagen-I were augmented by ΔNErbB2, unaffected by the NBC inhibitor S0859, and further stimulated by EIPA in a manner potentiated by PI3K-Akt-inhibition. These findings demonstrate that NHE1 inhibition can enhance cancer cell motility, adding an important facet to the understanding of NHE1 in cancer. PMID:22120673

  13. Epidemiological and Inducible Resistance in Coagulase Negative Staphylococci

    PubMed Central

    Abdollahi, Shadieh; Ramazanzadeh, Rashid; Khiabani, Zahra Delami; Kalantar, Enayat

    2016-01-01

    Background and Objectives: Coagulase negative staphylococci (CNS) are potential pathogens with the increased use of implants in hospitals. Macrolide, lincosamide and streptogramin B (MLSB) are used in the treatment of staphylococcal infections. The aim of this study was to molecular detection of inducible clindamycin resistance and genetic pattern in CNS isolates and their transmission between hospitals. Materials and Methods: 110 CNS strains, isolated from hospitalized patients in the intensive care unit and infectious wards of Besat and Toohid hospitals, Sanandaj. Methicillin resistance was done by agar screen test and the resistance inducible Clindamycin by the D-Test. Multiplex PCR was performed, using primers specific for erm (A, B, C, and TR) genes. Diversity of strains was determined by ERIC–PCR technique based on the similarities between DNA fingerprints by using Jaccards coefficient in the SAHN program of the NTSYS-pc software. Results: Of the 110 isolates, 64(58.2%) were methicillin -resistant CNS (MRCNS), 48(43.6%) were resistant to erythromycin (ERCNS). Out of 48 Erythromycin-resistant strains 5 (10.4%) were iMLSB phenotypes that 4 isolates showed genes erm by Multiplex PCR. The ERIC–PCR profiles allowed typing of the 110 isolates into 90 ERIC-types which were grouped into fourteen main clusters (C1–C14). Conclusion: The results of this study also showed that most of CNS isolated produced different genomic fingerprint patterns, therefore, source of infection is differen t. PMID:26573051

  14. Cost-effectiveness of ocriplasmin for the treatment of vitreomacular traction and macular hole

    PubMed Central

    Bennison, Craig; Stephens, Stephanie; Lescrauwaet, Benedicte; Van Hout, Ben; Jackson, Timothy L.

    2016-01-01

    Background If left untreated, vitreomacular traction (VMT) will infrequently improve through spontaneous resolution of vitreomacular adhesion (VMA), and patients remain at risk of further deterioration in vision. The mainstay of treatment for VMT is vitrectomy, an invasive procedure that carries the risk of rare but serious complications and further vision loss. As such, a ‘watch and wait’ approach is often adopted before this surgical intervention is performed. Ocriplasmin (microplasmin) is a potential alternative treatment for patients with symptomatic VMA/VMT that may remove the requirement for vitrectomy. Objective The purpose of this study was to evaluate the cost-effectiveness of ocriplasmin for the treatment of VMT in comparison to standard of care. Study design A cohort-based computer simulation model was developed, capturing three mutually exclusive subgroups: 1) VMT without epiretinal membrane (ERM) or full thickness macular hole (FTMH), 2) VMT with ERM but no FTMH, and 3) VMT with FTMH. Transition probabilities between health states, utilities, and resource utilisation were estimated based on clinical trial results, the literature, and expert opinion. The cost per quality-adjusted life year (QALY) gained was estimated over a lifetime, using UK unit costs and utilities associated with visual acuity, adverse events, metamorphopsia, and surgical interventions. Setting Analyses were conducted from a UK payer perspective. Population Transition probabilities for the model were primarily estimated from patient-level data from the combined Phase 3 MIVI-TRUST trials in patients with symptomatic VMA/VMT, including when associated with a FTMH ≤400 µm. Intervention Ocriplasmin (microplasmin) is a one-time intravitreal injection designed specifically to release the abnormal traction between the macula and the vitreous and thereby treat VMT, as well as macular hole with persistent vitreous attachment. Main outcome measure The main outcome measure of the

  15. Distribution of Antimicrobial Resistance and Virulence Genes in Enterococcus spp. and Characterization of Isolates from Broiler Chickens ▿ † ‡

    PubMed Central

    Diarra, Moussa S.; Rempel, Heidi; Champagne, Julie; Masson, Luke; Pritchard, Jane; Topp, Edward

    2010-01-01

    Enterococci are now frequent causative agents of nosocomial infections. In this study, we analyzed the frequency and distribution of antibiotic resistance and virulence genotypes of Enterococcus isolates from broiler chickens. Fecal and cecal samples from nine commercial poultry farms were collected to quantify total enterococci. Sixty-nine presumptive enterococci were isolated and identified by API 20 Strep, and their susceptibilities to antibiotics were determined. Genotypes were assessed through the use of a novel DNA microarray carrying 70 taxonomic, 17 virulence, and 174 antibiotic resistance gene probes. Total enterococcal counts were different from farm to farm and between sample sources (P < 0.01). Fifty-one (74%) of the isolates were identified as E. faecium, whereas nine (13%), seven (10%), and two (3%) isolates were identified as E. hirae, E. faecalis, and E. gallinarum, respectively. Multiple-antibiotic resistance was evident in E. faecium and E. faecalis isolates. The most common multiple-antibiotic resistance phenotype was Bac Ery Tyl Lin Str Gen Tet Cip. Genes conferring resistance to aminoglycoside (aac, aacA-aphD, aadB, aphA, sat4), macrolide (ermA, ermB, ermAM, msrC), tetracycline (tetL, tetM, tetO), streptogramin (satG_vatE8), bacitracin (bcrR), and lincosamide (linB) antibiotics were detected in corresponding phenotypes. A range of 9 to 12 different virulence genes was found in E. faecalis, including ace, agg, agrBEfs (agrB gene of E. faecalis), cad1, the cAM373 and cCF10 genes, cob, cpd1, cylAB, efaAEfs, and gelE. All seven E. faecalis isolates were found to carry the gelE gene and to hydrolize gelatin and bile salts. Results from this study showed the presence of enterococci of public and environmental health concerns in broiler chicken farms and demonstrated the utility of a microarray to quickly and reliably analyze resistance and virulence genotypes of Enterococcus spp. PMID:20971861

  16. Resistance gene pool to co-trimoxazole in non-susceptible Nocardia strains

    PubMed Central

    Valdezate, Sylvia; Garrido, Noelia; Carrasco, Gema; Villalón, Pilar; Medina-Pascual, María J.; Saéz-Nieto, Juan A.

    2015-01-01

    The soil-borne pathogen Nocardia sp. causes severe cutaneous, pulmonary, and central nervous system infections. Against them, co-trimoxazole (SXT) constitutes the mainstay of antimicrobial therapy. However, some Nocardia strains show resistance to SXT, but the underlying genetic basis is unknown. We investigated the presence of genetic resistance determinants and class 1–3 integrons in 76 SXT-resistant Nocardia strains by PCR and sequencing. By E test, these clinical strains showed SXT minimum inhibitory concentrations of ≥32:608 mg/L (ratio of 1:19 for trimethoprim: sulfamethoxazole). They belonged to 12 species, being the main representatives Nocardia farcinica (32%), followed by N. flavorosea (6.5%), N. nova (11.8%), N. carnea (10.5%), N. transvalensis (10.5%), and Nocardia sp. (6.5%). The prevalence of resistance genes in the SXT-resistant strains was as follows: sul1 and sul2 93.4 and 78.9%, respectively, dfrA(S1) 14.7%, blaTEM-1 and blaZ 2.6 and 2.6%, respectively, VIM-2 1.3%, aph(3′)-IIIa 40.8%, ermA, ermB, mefA, and msrD 2.6, 77.6, 14.4, and 5.2%, respectively, and tet(O), tet(M), and tet(L) 48.6, 25.0, and 3.9%, respectively. Detected amino acid changes in GyrA were not related to fluoroquinolone resistance, but probably linked to species polymorphism. Class 1 and 3 integrons were found in 93.42 and 56.57% strains, respectively. Class 2 integrons and sul3 genes were not detected. Other mechanisms, different than dfrA(S1), dfrD, dfrF, dfrG, and dfrK, could explain the strong trimethoprim resistance shown by the other 64 strains. For first time, resistance determinants commonly found in clinically important bacteria were detected in Nocardia sp. sul1, sul2, erm(B), and tet(O) were the most prevalent in the SXT-resistant strains. The similarity in their resistome could be due to a common genetic platform, in which these determinants are co-transferred. PMID:25972856

  17. Comparison of national and regional sediment quality guidelines for classifying sediment toxicity in California.

    PubMed

    Bay, Steven M; Ritter, Kerry J; Vidal-Dorsch, Doris E; Field, L Jay

    2012-10-01

    A number of sediment quality guidelines (SQGs) have been developed for relating chemical concentrations in sediment to their potential for effects on benthic macroinvertebrates, but there have been few studies evaluating the relative effectiveness of different SQG approaches. Here we apply 6 empirical SQG approaches to assess how well they predict toxicity in California sediments. Four of the SQG approaches were nationally derived indices that were established in previous studies: effects range median (ERM), logistic regression model (LRM), sediment quality guideline quotient 1 (SQGQ1), and Consensus. Two approaches were variations of nationally derived approaches that were recalibrated to California-specific data (CA LRM and CA ERM). Each SQG approach was applied to a standardized set of matched chemistry and toxicity data for California and an index of the aggregate magnitude of contamination (e.g., mean SQG quotient or maximum probability of toxicity) was calculated. A set of 3 thresholds for classification of the results into 4 categories of predicted toxicity was established for each SQG approach using a statistical optimization procedure. The performance of each SQG approach was evaluated in terms of correlation and categorical classification accuracy. Each SQG index had a significant, but low, correlation with toxicity and was able to correctly classify the level of toxicity for up to 40% of samples. The CA LRM had the best overall performance, but the magnitude of differences in classification accuracy among the SQG approaches was relatively small. Recalibration of the indices using California data improved performance of the LRM, but not the ERM. The LRM approach is more amenable to revision than other national SQGs, which is a desirable attribute for use in programs where the ability to incorporate new information or chemicals of concern is important. The use of a consistent threshold development approach appeared to be a more important factor than type

  18. Support vector regression for porosity prediction in a heterogeneous reservoir: A comparative study

    NASA Astrophysics Data System (ADS)

    Al-Anazi, A. F.; Gates, I. D.

    2010-12-01

    In wells with limited log and core data, porosity, a fundamental and essential property to characterize reservoirs, is challenging to estimate by conventional statistical methods from offset well log and core data in heterogeneous formations. Beyond simple regression, neural networks have been used to develop more accurate porosity correlations. Unfortunately, neural network-based correlations have limited generalization ability and global correlations for a field are usually less accurate compared to local correlations for a sub-region of the reservoir. In this paper, support vector machines are explored as an intelligent technique to correlate porosity to well log data. Recently, support vector regression (SVR), based on the statistical learning theory, have been proposed as a new intelligence technique for both prediction and classification tasks. The underlying formulation of support vector machines embodies the structural risk minimization (SRM) principle which has been shown to be superior to the traditional empirical risk minimization (ERM) principle employed by conventional neural networks and classical statistical methods. This new formulation uses margin-based loss functions to control model complexity independently of the dimensionality of the input space, and kernel functions to project the estimation problem to a higher dimensional space, which enables the solution of more complex nonlinear problem optimization methods to exist for a globally optimal solution. SRM minimizes an upper bound on the expected risk using a margin-based loss function ( ɛ-insensitivity loss function for regression) in contrast to ERM which minimizes the error on the training data. Unlike classical learning methods, SRM, indexed by margin-based loss function, can also control model complexity independent of dimensionality. The SRM inductive principle is designed for statistical estimation with finite data where the ERM inductive principle provides the optimal solution (the

  19. [Sensitivity surveillance of Streptococcus pneumoniae isolates for several antibacterial agents in Gifu and Aichi prefectures (2011-2012)].

    PubMed

    Funatsu, Tori; Mizunaga, Shingo; Fukuda, Yoshiko; Nomura, Nobuhiko; Hashido, Hikonori; Mitsuyama, Junichi; Hatano, Masakazu; Yamaoka, Kazukiyo; Watanabe, Kunitomo; Asano, Yuko; Suematsu, Hiroyuki; Sawamura, Haruki; Matsukawa, Yoko; Ohta, Hirotoshi; Yamagishi, Yuka; Mikamo, Hiroshige; Matsubara, Shigenori; Shibata, Naohiro

    2015-08-01

    We investigated the susceptibility to antibacterial agents, genotype of penicillin-binding protein (PBP) genes and macrolide resistant genes, and the serotypes against 270 strains of Streptococcus pneumoniae isolated from medical facilities in Gifu and Aichi prefectures between October 2011 and April 2012. These results were compared with those against S. pneumoniae isolated in 2008-2009 and 2010-2011. The number of gPSSP with 3 normal PBP genes, gPISP with 1 or 2 normal PBP genes and gPRSP with 3 abnormal genes isolated in 2011-2012 was 15 (5.6%), 162 (60.0%) and 93 (34.4%) strains, respectively. Compared with those isolated in 2008-2009 and 2010-2011, the numbers of gPRSP were decreasing. On the other hand, the isolates with no macrolide-resistant gene, only mefA, only ermB, and both mefA and ermB were 16 (5.9%), 75 (27.8%), 153 (56.7%) and 26 (9.6%). Compared with those isolated in 2008-2009 and 2010-2011, the numbers of isolates with ermB, which was usually associated with high-level resistance, were increasing. The prevalent pneumococcal serotypes in children were type 3 (14.4%), following by type 15 and 19F (9.3%). The coverages of 7-valent pneumococcal conjugate vaccine (PCV7) and 13-valent pneumococcal conjugate vaccine (PCV13) were calculated as 22.9% and 49.2%, respectively. The coverages of PCV7 and PCV13 in gPRSP isolated from children were 47.7% (21/44 strains) and 72.7% (32/44 strains). The MIC90 of each antibacterial agent was as follows; 0.125pg/mL for imipenem, panipenem and garenoxacin, 0.25 μg/mL for meropenem and doripenem, 0.5 μg/mL for cefditoren, moxifloxacin and tosufloxacin, 1 μg/mL for amoxicillin, clavulanic acid/amoxicillin, cefteram, cefcapene and ceftriaxone, 2 μg/mL for benzylpenicillin, ampicillin, sulbactam/ampicillin, piperacillin, tazobactam/piperacillin and levofloxacin, 4 μg/mL for cefdinir, flomoxef and pazufloxacin, 16 μg/mL for minocycline, > 64 μg/mL for clarithromycin and azithromycin, and these MIC90s were about the

  20. Prevalence and mechanism of resistance against macrolides, lincosamides, and streptogramins among Enterococcus faecium isolates from food-producing animals and hospital patients in Belgium.

    PubMed

    De Graef, E M; Decostere, A; De Leener, E; Goossens, H; Baele, M; Haesebrouck, F

    2007-01-01

    The prevalence of acquired resistance to streptogramins, macrolides, and lincosamides and the genetic background of this resistance was investigated in Enterococcus faecium strains isolated from food-producing animals and hospital patients 4-5 years after the ban of streptogramins as growth promoters. The minimum inhibitory concentrations (MICs) of quinupristin/dalfopristin (Q/D), virginiamycin M1 (virgM1), erythromycin (ery), tylosin (tyl), and lincomycin (lin) were determined by the agar dilution method for E. faecium isolates derived from pigs (80), broilers (45), and hospitalized patients (103). Resistance or susceptibility was interpreted using a microbiological criterion and breakpoints recommended by the Clinical Laboratory Standards Institute (CLSI), if available. The isolates were also screened by PCR for erm(B), lnu(A), lnu(B), mef(A/E), vat(D), vat(E), vga(A), vga(B), and vgb(A) genes. Acquired resistance to Q/D, virgM1, ery, tyl, and lin was detected in 34%, 96%, 46%, 46%, and 69% of the porcine strains, respectively. For broiler strains this was 15% (Q/D), 98% (virgM1), 69% (ery), 71% (tyl), and 89% (lin) and for human strains 23% (Q/D), 65% (virgM1), 54% (ery), 52% (tyl), and 60% (lin). Strains showing cross-resistance against macrolides and lincosamides almost always carried the erm(B) gene. This gene was present in 64% of the Q/D-resistant isolates. Only in two human and three broiler Q/D- and virgM1-resistant isolates, a combination of the erm(B) and vat(D) or vat(E) genes was found. The genetic background of resistance could not be determined in the other Q/D- or virgM1-resistant strains. This study demonstrates that streptogramin resistance is frequently present in strains from hospitalized patients and food-producing animals, but the genetic basis hitherto mostly remains obscure. PMID:17650967

  1. Molecular Epidemiologic Analysis of Enterococcus faecalis Isolates in Cuba by Multilocus Sequence Typing

    PubMed Central

    Kobayashi, Nobumichi; Nagashima, Shigeo

    2009-01-01

    We carried out the first study of Enterococcus faecalis clinical isolates in Cuba by multilocus sequence typing linking the molecular typing data with the presence of virulence determinants and the antibiotic resistance genes. A total of 23 E. faecalis isolates recovered from several clinic sources and geographic areas of Cuba during a period between 2000 and 2005 were typed by multilocus sequence typing. Thirteen sequence types (STs) including five novel STs were identified, and the ST 64 (clonal complex [CC] 8), ST 6 (CC2), ST 21(CC21), and ST 16 (CC58) were found in more than one strain. Sixty-seven percent of STs corresponded to STs reported previously in Spain, Poland, and The Netherlands, and other STs (ST115, ST64, ST6, and ST40) were genetically close to those detected in the United States. Prevalence of both antimicrobial resistance genes [aac(6′)-aph(2″), aph(3′), ant(6), ant(3″)(9), aph(2″)-Id, aph(2″)-Ic, erm(B), erm(A), erm(C), mef(A), tet(M), and tet(L)] and virulence genes (agg, gelE, cylA, esp, ccf, and efaAfs) were examined by polymerase chain reaction. Aminoglycoside resistance genes aac(6′)-Ie-aph(2″)-Ia, aph(3′), ant(6), ant(3″)(9) were more frequently detected in ST6, ST16, ST23, ST64, and ST115. The multidrug resistance was distributed to all STs detected, except for ST117 and singleton ST225. The presence of cyl gene was specifically linked to the ST64 and ST16. Presence of the esp, gel, and agg genes was not specific to any particular ST. This research provided the first insight into the population structure of E. faecalis in Cuba, that is, most Cuban strains were related to European strains, whereas others to U.S. strains. The CC2, CC21, and CC8, three of the biggest CCs in the world, were evidently circulating in Cuba, associated with multidrug resistance and virulence traits. PMID:19857135

  2. Longitudinal characterization of antimicrobial resistance genes in feces shed from cattle fed different subtherapeutic antibiotics

    PubMed Central

    2011-01-01

    Background Environmental transmission of antimicrobial-resistant bacteria and resistance gene determinants originating from livestock is affected by their persistence in agricultural-related matrices. This study investigated the effects of administering subtherapeutic concentrations of antimicrobials to beef cattle on the abundance and persistence of resistance genes within the microbial community of fecal deposits. Cattle (three pens per treatment, 10 steers per pen) were administered chlortetracycline, chlortetracycline plus sulfamethazine, tylosin, or no antimicrobials (control). Model fecal deposits (n = 3) were prepared by mixing fresh feces from each pen into a single composite sample. Real-time PCR was used to measure concentrations of tet, sul and erm resistance genes in DNA extracted from composites over 175 days of environmental exposure in the field. The microbial communities were analyzed by quantification and denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S-rRNA. Results The concentrations of 16S-rRNA in feces were similar across treatments and increased by day 56, declining thereafter. DGGE profiles of 16S-rRNA differed amongst treatments and with time, illustrating temporal shifts in microbial communities. All measured resistance gene determinants were quantifiable in feces after 175 days. Antimicrobial treatment differentially affected the abundance of certain resistance genes but generally not their persistence. In the first 56 days, concentrations of tet(B), tet(C), sul1, sul2, erm(A) tended to increase, and decline thereafter, whereas tet(M) and tet(W) gradually declined over 175 days. At day 7, the concentration of erm(X) was greatest in feces from cattle fed tylosin, compared to all other treatments. Conclusion The abundance of genes coding for antimicrobial resistance in bovine feces can be affected by inclusion of antibiotics in the feed. Resistance genes can persist in feces from cattle beyond 175 days with concentrations

  3. Stolt’s f-k migration for plane wave ultrasound imaging

    PubMed Central

    Garcia, Damien; Le Tarnec, Louis; Muth, Stéphan; Montagnon, Emmanuel; Porée, Jonathan; Cloutier, Guy

    2013-01-01

    Ultrafast ultrasound is an emerging modality that offers new perspectives and opportunities in medical imaging. Plane wave imaging (PWI) allows one to attain very high frame rates by transmission of planar ultrasound wavefronts. As a plane wave reaches a given scatterer, the latter becomes a secondary source emitting upward spherical waves and creating a diffraction hyperbola in the received RF (radio-frequency) signals. To produce an image of the scatterers, all the hyperbolas must be migrated back to their apexes. In order to perform beamforming of plane wave echo RFs and return high-quality images at high frame rates, we propose a new migration method carried out in the frequency-wavenumber (f-k) domain. The f-k migration for PWI has been adapted from the Stolt migration for seismic imaging. This migration technique is based on the exploding reflector model (ERM), which consists in assuming that all the scatterers explode in concert and become acoustic sources. The classical ERM model, however, is not appropriate for PWI. We showed that the ERM can be made suitable for PWI by a spatial transformation of the hyperbolic traces present in the RF data. In vitro experiments were performed to sketch the advantages of PWI with Stolt’s f-k migration over the conventional delay-and-sum (DAS) approach. The Stolt’s f-k migration was also compared with the Fourier-based method developed by J-Y Lu. Our findings show that multi-angle compounded f-k migrated images are of quality similar to those obtained with a state-of-the-art dynamic focusing mode. This remained true even with a very small number of steering angles thus ensuring a highly competitive frame rate. In addition, the new FFT-based f-k migration provides comparable or better contrast-to-noise ratio and lateral resolution than the Lu’s and DAS migration schemes. Matlab codes of the Stolt’s f-k migration for PWI are provided. PMID:24626107

  4. Ezrin/Radixin/Moesin Proteins and Flotillins Cooperate to Promote Uropod Formation in T Cells

    PubMed Central

    Martinelli, Sibylla; Chen, Emily J. H.; Clarke, Fiona; Lyck, Ruth; Affentranger, Sarah; Burkhardt, Janis K.; Niggli, Verena

    2013-01-01

    T cell uropods are enriched in specific proteins including adhesion receptors such as P-selectin glycoprotein ligand-1 (PSGL-1), lipid raft-associated proteins such as flotillins and ezrin/radixin/moesin (ERM) proteins which associate with cholesterol-rich raft domains and anchor adhesion receptors to the actin cytoskeleton. Using dominant mutants and siRNA technology we have tested the interactions among these proteins and their role in shaping the T cell uropod. Expression of wild type (WT) ezrin-EGFP failed to affect the morphology of human T cells or chemokine-induced uropod recruitment of PSGL-1 and flotillin-1 and -2. In contrast, expression of constitutively active T567D ezrin-EGFP induced a motile, polarized phenotype in some of the transfected T cells, even in the absence of chemokine. These cells featured F-actin-rich ruffles in the front and uropod enrichment of PSGL-1 and flotillins. T567D ezrin-EGFP was itself strongly enriched in the rear of the polarized T cells. Uropod formation induced by T567D ezrin-EGFP was actin-dependent as it was attenuated by inhibition of Rho-kinase or myosin II, and abolished by disruption of actin filaments. While expression of constitutively active ezrin enhanced cell polarity, expression of a dominant-negative deletion mutant of ezrin, 1–310 ezrin-EGFP, markedly reduced uropod formation induced by the chemokine SDF-1, T cell front-tail polarity, and capping of PSGL-1 and flotillins. Transfection of T cells with WT or T567D ezrin did not affect chemokine-mediated chemotaxis whereas 1–310 ezrin significantly impaired spontaneous 2D migration and chemotaxis. siRNA-mediated downregulation of flotillins in murine T cells attenuated moesin capping and uropod formation, indicating that ERM proteins and flotillins cooperate in uropod formation. In summary, our results indicate that activated ERM proteins function together with flotillins to promote efficient chemotaxis of T cells by structuring the uropod of migrating T

  5. Metrology and Characterization Challenges for Emerging Research Materials and Devices

    SciTech Connect

    Garner, C. Michael; Herr, Dan; Obeng, Yaw

    2011-11-10

    The International Technology Roadmap for Semiconductors (ITRS) Emerging Research Materials (ERM) and Emerging Research Devices (ERD) Technology Workgroups have identified materials and devices that could enable continued increases in the density and performance of future integrated circuit (IC) technologies and the challenges that must be overcome; however, this will require significant advances in metrology and characterization to enable progress. New memory devices and beyond CMOS logic devices operate with new state variables (e.g., spin, redox state, etc.) and metrology and characterization techniques are needed to verify their switching mechanisms and scalability, and enable improvement of operation of these devices. Similarly, new materials and processes are needed to enable these new devices. Additionally, characterization is needed to verify that the materials and their interfaces have been fabricated with required quality and performance.

  6. Ein mobiler und offener Kernspintomograph: Kernspintomographie für Medizin und Materialforschung

    NASA Astrophysics Data System (ADS)

    Blümich, Bernhard; Kölker, Christian; Casanova, Federico; Perlo, Juan; Felder, Jörg

    2005-09-01

    An der RWTH Aachen wurde der erste offene und mobile Kernspintomograph entwickelt. Anders als die normalerweise riesigen und unbeweglichen Geräte besitzt er keine enge Magnetröhre. So kann er auch in große Untersuchungsobjekte hinein schauen, ohne dass diese zerstört werden müssen. Wie eine Lupe erfasst er dabei ein begrenztes Volumen um den Aufsatzpunkt herum. Neben der Medizin ermöglicht der offene Tomograph viele neue Anwendungsgebiete, vor allem in der Materialprüfung und Qualitätskontrolle. Der Aachener Prototyp kann auch das Fließprofil und die Geschwindigkeitsverteilung strömender Flüssigkeiten sichtbar machen. Sein offenes Prinzip erforderte neue technische Komponenten und modifizierte bildgebende Verfahren. Die Messzeit pro Bild kann heute schon unter einer Viertelstunde liegen.

  7. Das AL-Konzept: Ein Konzept für Bearbeitungszentren zur Großserienfertigung

    NASA Astrophysics Data System (ADS)

    Müllner, Ralf

    In der Großserienfertigung hält der Trend weg von Transferstraßen und Rundtaktmaschinen nun schon mehrere Jahre ungebrochen an. Vielfach wurden diese klassischen Einrichtungen zur Produktion hoher Stückzahlen inzwischen durch Fertigungsanlagen, wie z.B. Fertigungsmodule oder mehrspindlige Bearbeitungszentren ersetzt. Die Motivation hierzu ist vielschichtig. Eine ständig zunehmende Variantenanzahl der zu fertigenden Teile neben einem oft sehr unterschiedlichen Stückzahlbedarf begründet den Wunsch nach kleineren flexiblen Einheiten für die Fertigung. Um während der Produktionsphase eines Bauteils auf Stückzahlschwankungen reagieren zu können wird zudem eine bessere Skalierbarkeit gefordert, als es Transferstraßen und Rundtaktmaschinen ermöglichen.

  8. Das mechatronische Fahrwerk der Zukunft

    NASA Astrophysics Data System (ADS)

    Rieth, Peter

    Technologiesprünge gab es immer wieder in der Geschichte der Automobiltechnologie, so auch in der Kraftfahrzeugbremse, die seit ihren Anfängen eine stete Weiterentwicklung erfahren hat (Bild 40-1). So erlangten schon mechanische Bremsen durchaus ein hohes Niveau, ehe die Hydraulik Mitte der zwanziger Jahre für Komfort und Sicherheit vollkommen neue Perspektiven eröffnete, welche durch Einführung der Hilfskraft(servo)bremsanlagen Mitte des letzten Jahrhunderts noch erweitert wurden. Den wichtigsten Technologiesprung, den der Kunde als Fortschritt erkennt und dementsprechend honoriert, ermöglichte die Elektronik Mitte der siebziger Jahre. ABS, ASR, EBV und nicht zuletzt ESC wären ohne sie nicht vorstellbar.

  9. Development of Final A-Fault Rupture Models for WGCEP/ NSHMP Earthquake Rate Model 2

    USGS Publications Warehouse

    Field, Edward H.; Weldon, Ray J., II; Parsons, Thomas; Wills, Chris J.; Dawson, Timothy E.; Stein, Ross S.; Petersen, Mark D.

    2008-01-01

    This appendix discusses how we compute the magnitude and rate of earthquake ruptures for the seven Type-A faults (Elsinore, Garlock, San Jacinto, S. San Andreas, N. San Andreas, Hayward-Rodgers Creek, and Calaveras) in the WGCEP/NSHMP Earthquake Rate Model 2 (referred to as ERM 2. hereafter). By definition, Type-A faults are those that have relatively abundant paleoseismic information (e.g., mean recurrence-interval estimates). The first section below discusses segmentation-based models, where ruptures are assumed be confined to one or more identifiable segments. The second section discusses an un-segmented-model option, the third section discusses results and implications, and we end with a discussion of possible future improvements. General background information can be found in the main report.

  10. Umsetzung von CE-Prozessen durch neue Methoden und interaktive 3-D-Modelle

    NASA Astrophysics Data System (ADS)

    Lange, Andrea; Schulz, Torsten; Szymanski, Hans

    Innerhalb der EU gelten einheitliche Anforderungen an die Gestaltung von Maschinen, die in Form von grundlegenden Sicherheits- und Gesundheitsschutzanforderungen in der Maschinenrichtlinie (MaschRL) formuliert sind. Ihre Einhaltung dokumentiert der Hersteller mit der Konformitätserklärung und der Vergabe des CE-Zeichens. Die Maschinenrichtlinie ist ein Element des 1985 von der Europäischen Gemeinschaft (EG) beschriebenen "New Approach“ Rechtsetzungsmodells, das technologische Innovation ermöglicht und die Wettbewerbsfähigkeit der europäischen Industrie regelt. Es wurde erschaffen, um technische Handelshemmnisse im europäischen Binnenmarkt zu beseitigen und ein einheitliches Niveau hinsichtlich der Sicherheit der Produkte und dem Gesundheits- bzw. Verbraucherschutz zu gewährleisten. Alle Produkt-Richtlinien der EG, die später auf Grundlage des New Approach entstanden sind, weisen eine einheitliche Vorgehensweise auf, mit der diese Ziele erreicht werden sollen (VDI Verlag 11/2008).

  11. Seneca Compressed Air Energy Storage (CAES) Project

    SciTech Connect

    2012-11-30

    This report provides a review and an analysis of potential environmental justice areas that could be affected by the New York State Electric & Gas (NYSEG) compress air energy storage (CAES) project and identifies existing environmental burden conditions on the area and evaluates additional burden of any significant adverse environmental impact. The review assesses the socioeconomic and demographic conditions of the area surrounding the proposed CAES facility in Schuyler County, New York. Schuyler County is one of 62 counties in New York. Schuyler County’s 2010 population of 18,343 makes it one of the least populated counties in the State (U.S. Census Bureau, 2010). This report was prepared for WorleyParsons by ERM and describes the study area investigated, methods and criteria used to evaluate this area, and the findings and conclusions from the evaluation.

  12. Convergent losses of decay mechanisms and rapid turnover of symbiosis genes in mycorrhizal mutualists

    SciTech Connect

    Kohler, Annegret; Kuo, Alan; Nagy, Laszlo G.; Morin, Emmanuelle; Barry, Kerrie W.; Buscot, Francois; Canbäck, Björn; Choi, Cindy; Cichocki, Nicolas; Clum, Alicia; Colpaert, Jan; Copeland, Alex; Costa, Mauricio D.; Doré, Jeanne; Floudas, Dimitrios; Gay, Gilles; Girlanda, Mariangela; Henrissat, Bernard; Herrmann, Sylvie; Hess, Jaqueline; Högberg, Nils; Johansson, Tomas; Khouja, Hassine-Radhouane; LaButti, Kurt; Lahrmann, Urs; Levasseur, Anthony; Lindquist, Erika A.; Lipzen, Anna; Marmeisse, Roland; Martino, Elena; Murat, Claude; Ngan, Chew Y.; Nehls, Uwe; Plett, Jonathan M.; Pringle, Anne; Ohm, Robin A.; Perotto, Silvia; Peter, Martina; Riley, Robert; Rineau, Francois; Ruytinx, Joske; Salamov, Asaf; Shah, Firoz; Sun, Hui; Tarkka, Mika; Tritt, Andrew; Veneault-Fourrey, Claire; Zuccaro, Alga; Tunlid, Anders; Grigoriev, Igor V.; Hibbett, David S.; Martin, Francis

    2015-02-23

    To elucidate the genetic bases of mycorrhizal lifestyle evolution, we sequenced new fungal genomes, including 13 ectomycorrhizal (ECM), orchid (ORM) and ericoid (ERM) species, and five saprotrophs, which we analyzed along with other fungal genomes. Ectomycorrhizal fungi have a reduced complement of genes encoding plant cell wall-degrading enzymes (PCWDEs), as compared to their ancestral wood decayers. Nevertheless, they have retained a unique array of PCWDEs, thus suggesting that they possess diverse abilities to decompose lignocellulose. Similar functional categories of nonorthologous genes are induced in symbiosis. Of induced genes, 7-38% are orphan genes, including genes that encode secreted effector-like proteins. Convergent evolution of the mycorrhizal habit in fungi occurred via the repeated evolution of a 'symbiosis toolkit', with reduced numbers of PCWDEs and lineage-specific suites of mycorrhiza-induced genes.

  13. Phenotypic and molecular characterization of Yersinia ruckeri isolates from rainbow trout (Oncorhynchus mykiss, Walbaum, 1792) in Turkey.

    PubMed

    Onuk, Ertan Emek; Ciftci, Alper; Findik, Arzu; Ciftci, Gülay; Altun, Soner; Balta, Fikri; Ozer, Selmin; Coban, Ahmet Yilmaz

    2011-01-01

    The aim of the study was the phenotypic and molecular characterization of Yersinia (Y) ruckeri strains, the causative agent of Enteric Redmouth Disease (ERM), by antibiotyping, random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) and sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE) patterns of whole cell proteins. For this aim, a total of 97 Y ruckeri isolates were analyzed. The isolates were distinguished into ten antibiotypes and six phenotypes according to their resistance properties and whole cell protein profiles, respectively. Also, a glycoprotein band of approximately 25.5 kDa was observed in all Y ruckeri strains tested. In all strains, six different RAPD types were observed. In conclusion, Y ruckeri strains isolated from rainbow trout of fish farms in Turkey showed variation according to their phenotypic and genotypic characteristics, and the use of these three typing techniques in double and triple combinations could be more useful for discriminating the strains. PMID:21848040

  14. Cyber-Physical Systems - Wissenschaftliche Herausforderungen Bei Der Entwicklung

    NASA Astrophysics Data System (ADS)

    Broy, Manfred

    Cyber-Physical Systems adressieren die enge Verbindung eingebetteter Systeme zur Überwachung und Steuerung physikalischer Vorgänge mittels Sensoren und Aktuatoren über Kommunikationseinrichtungen mit den globalen digitalen Netzen (dem Cyberspace"). Dieser Typus von Systemen ermöglicht über Wirkketten eine Verbindung zwischen Vorgängen der physischen Realität und den heute verfügbaren digitalen Netzinfrastrukturen. Dies erlaubt vielfältige Applikationen mit hohem wirtschaftlichen Potential, und mit starker Innovationskraft. Die vollständige Ausschöpfung des Potentials erfordert aber gezielte wissenschaftliche Anstrengungen bei der Entwicklung solcher Systeme im Hinblick auf Methodik, Technologie, Kostenbeherrschung und funktionale Angemessenheit.

  15. Mycobacterium abscessus Complex Infections in Humans

    PubMed Central

    Lee, Meng-Rui; Sheng, Wang-Huei; Hung, Chien-Ching; Yu, Chong-Jen; Lee, Li-Na

    2015-01-01

    Mycobacterium abscessus complex comprises a group of rapidly growing, multidrug-resistant, nontuberculous mycobacteria that are responsible for a wide spectrum of skin and soft tissue diseases, central nervous system infections, bacteremia, and ocular and other infections. M. abscessus complex is differentiated into 3 subspecies: M. abscessus subsp. abscessus, M. abscessus subsp. massiliense, and M. abscessus subsp. bolletii. The 2 major subspecies, M. abscessus subsp. abscessus and M. abscessus subsp. massiliense, have different erm(41) gene patterns. This gene provides intrinsic resistance to macrolides, so the different patterns lead to different treatment outcomes. M. abscessus complex outbreaks associated with cosmetic procedures and nosocomial transmissions are not uncommon. Clarithromycin, amikacin, and cefoxitin are the current antimicrobial drugs of choice for treatment. However, new treatment regimens are urgently needed, as are rapid and inexpensive identification methods and measures to contain nosocomial transmission and outbreaks. PMID:26295364

  16. Potentials and limits for the use of ozone as a fish disease control agent

    USGS Publications Warehouse

    Wedemeyer, Gary A.; Nelson, Nancy C.; Yasutake, Wm. T.

    1979-01-01

    Ozone and chlorine inactivation curves were determined in three types of freshwater at 20 C for the destruction of the fish pathogens Aeromonas salmonicida the etiologic agent of furunculosis, and Yersinia ruckeri the enteric redmouth bacterium (ERM). Ozone and chlorine inactivation curves were also obtained in the same water types at 10 C for the fish pathogenic viruses infectious hematopoietic necrosis (IHNV), and infectious pancreatic necrosis (IPNV). Acute toxicity tests using the rainbow trout as a representative salmonid revealed that ozone was highly toxic at the dose levels used. Partial chronic (3. mo.) testing revealed that ozone exposure at 2 μg/L causes only minimal physiological changes, none of which would be expected to compromise biological function.

  17. 1995 Department of Energy Records Management Conference

    SciTech Connect

    1995-07-01

    The Department of Energy (DOE) Records Management Group (RMG) provides a forum for DOE and its contractor personnel to review and discuss subjects, issues, and concerns of common interest. This forum will include the exchange of information, and interpretation of requirements, and a dialog to aid in cost-effective management of the DOE Records Management program. Issues addressed by the RMG may result in recommendations for DOE-wide initiatives. Proposed DOE-wide initiatives shall be, provided in writing by the RMG Steering Committee to the DOE Records Management Committee and to DOE`s Office of ERM Policy, Records, and Reports Management for appropriate action. The membership of the RMG is composed of personnel engaged in Records Management from DOE Headquarters, Field sites, contractors, and other organizations, as appropriate. Selected papers are indexed separately for inclusion in the Energy Science and Technology Database.

  18. Calculation of geoid undulations and gravity anomalies in the South China Sea by using the TOPEX/Poseidon and Geosat altimeter data

    NASA Astrophysics Data System (ADS)

    Zhang, Youguang; Zhang, Jie; Ji, Yonggang; Zhang, Huiqin

    2003-05-01

    In this paper, using TOPEX/Poseidon (9~346cycle) and Geosat/ERM(1~60cycle) altimeter data, the author applies combined adjustment model for calculating the South China Sea geoid undulations after data preprocessing. The difference between calculation result and OSU91A model is 30cm (spatial resolution is 22km). In addition, 12"x12" South China Sea gravity anomalies are calculated by using above geoid data and improved Stokes inverse formula. Contrasted with Scripps Institution of Oceanography gravity anomalies data, the accuracy of computation of gravity anomalies is 12´x10-5m/s2. These computations show that calculation speed is fast and calculation efficiency is high, so the method can calculate rapidly gravity anomalies in special sea area.

  19. On the high-density expansion for Euclidean random matrices

    NASA Astrophysics Data System (ADS)

    Grigera, T. S.; Martin-Mayor, V.; Parisi, G.; Urbani, P.; Verrocchio, P.

    2011-02-01

    Diagrammatic techniques to compute perturbatively the spectral properties of Euclidean random matrices (ERM) in the high-density regime are introduced and discussed in detail. Such techniques are developed in two alternative and very different formulations of the mathematical problem and are shown to give identical results up to second order in the perturbative expansion. One method, based on writing the so-called resolvent function as a Taylor series, allows us to group the diagrams into a small number of topological classes, providing a simple way to determine the infrared (small momenta) behaviour of the theory up to third order, which is of interest for the comparison with experiments. The other method, which reformulates the problem as a field theory, can instead be used to study the infrared behaviour at any perturbative order.

  20. Report on the 10th European Fusion Physics Workshop (Vaals, The Netherlands, 9-11 December 2002)

    NASA Astrophysics Data System (ADS)

    Campbell, D. J.; Borba, D.; Bucalossi, J.; Moreau, D.; Sauter, O.; Stober, J.; Vayakis, G.

    2003-06-01

    The 10th European Fusion Physics Workshop took place in December 2002 at Vaals in The Netherlands, hosted by the Trilateral Euregio Cluster (TEC: Associations EURATOM-ERM/KMS, FZJ and FOM), and sponsored by the European Commission and the Foundation SOFT. Within an overall theme of `Operational limits in toroidal devices, with particular reference to steady-state operation', four topics of importance to the future development of magnetically confined fusion were discussed in detail. In addition, a review of the JET scientific and technical programme under EFDA and an assessment of ITER's measurement requirements and diagnostic development programme were presented. The main issues discussed and the areas identified as requiring further study are summarized here.