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Sample records for erm tarmo soomere

  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 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,"…

  3. Intrinsic Macrolide Resistance in Mycobacterium smegmatis Is Conferred by a Novel erm Gene, erm(38)

    PubMed Central

    Nash, Kevin A.

    2003-01-01

    High-level, acquired macrolide resistance in mycobacteria is conferred by mutation within the 23S rRNA gene. However, several mycobacteria are naturally resistant to macrolides, including the Mycobacterium smegmatis group and Mycobacterium tuberculosis complex. Thus, the aim of this study was to characterize this resistance. Intrinsic macrolide resistance in M. smegmatis was inducible and showed cross-resistance to lincosamides but not to streptogramin B (i.e., ML resistance). A similar phenotype was found with Mycobacterium microti and macrolide-resistant Mycobacterium fortuitum. A search of the DNA sequence data for M. smegmatis strain mc2155 identified a novel erm gene, erm(38), and expression analysis showed that erm(38) RNA levels increased >10-fold after a 2-h incubation with macrolide. Inducible ML resistance was not expressed by an erm(38) knockout mutant, and complementation of this mutant with intact erm(38) in trans resulted in high-level ML resistance (e.g., clarithromycin MIC of >512 μg/ml). Thus, the results indicate that erm(38) confers the intrinsic ML resistance of M. smegmatis. Southern blot analysis with an erm(38)-specific probe indicated that a similar gene may be present in macrolide-resistant M. fortuitum. This finding, with the presence of the erm(37) gene (Rv1988) in the M. tuberculosis complex, suggests that such genes are widespread in mycobacteria with intrinsic macrolide resistance. PMID:14506008

  4. Ermüdungs- und Wachsamkeitsmanagement

    NASA Astrophysics Data System (ADS)

    Ebermann, Hans-Joachim; Murtha, Maria-Pascaline

    1994 wurde erstmals Ermüdung (Fatigue) durch die amerikanische Unfalluntersuchungsbehörde NTSB (National Transportation Safety Board) offiziell als Hauptursache für einen Unfall in der Luftfahrt erklärt. Seitdem wird Müdigkeit bei der Untersuchung von Unfällen systematisch abgefragt.

  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. Mycobacterium smegmatis Erm(38) Is a Reluctant Dimethyltransferase

    PubMed Central

    Madsen, Christian Toft; Jakobsen, Lene; Douthwaite, Stephen

    2005-01-01

    The waxy cell walls of mycobacteria provide intrinsic tolerance to a broad range of antibiotics, and this effect is augmented by specific resistance determinants. The inducible determinant erm(38) in the nontuberculous species Mycobacterium smegmatis confers high resistance to lincosamides and some macrolides, without increasing resistance to streptogramin B antibiotics. This is an uncharacteristic resistance pattern falling between the type I and type II macrolide, lincosamide, and streptogramin B (MLSB) phenotypes that are conferred, respectively, by Erm monomethyltransferases and dimethyltransferases. Erm dimethyltransferases are typically found in pathogenic bacteria and confer resistance to all MLSB drugs by addition of two methyl groups to nucleotide A2058 in 23S rRNA. We show here by mass spectrometry analysis of the mycobacterial rRNA that Erm(38) is indeed an A2058-specific dimethyltransferase. The activity of Erm(38) is lethargic, however, and only a meager proportion of the rRNA molecules become dimethylated in M. smegmatis, while most of the rRNAs are either monomethylated or remain unmethylated. The methylation pattern produced by Erm(38) clarifies the phenotype of M. smegmatis, as it is adequate to confer resistance to lincosamides and 14-member ring macrolides such as erythromycin, but it is insufficient to raise the level of resistance to streptogramin B drugs above the already high intrinsic tolerance displayed by this species. PMID:16127056

  7. Methyltransferase Erm(37) slips on rRNA to confer atypical resistance in Mycobacterium tuberculosis.

    PubMed

    Madsen, Christian Toft; Jakobsen, Lene; Buriánková, Karolina; Doucet-Populaire, Florence; Pernodet, Jean-Luc; Douthwaite, Stephen

    2005-11-25

    Members of the Mycobacterium tuberculosis complex possess a resistance determinant, erm(37) (also termed ermMT), which is a truncated homologue of the erm genes found in a diverse range of drug-producing and pathogenic bacteria. All erm genes examined thus far encode N(6)-monomethyltransferases or N(6),N(6)-dimethyltransferases that show absolute specificity for nucleotide A2058 in 23 S rRNA. Monomethylation at A2058 confers resistance to a subset of the macrolide, lincosamide, and streptogramin B (MLS(B)) group of antibiotics and no resistance to the latest macrolide derivatives, the ketolides. Dimethylation at A2058 confers high resistance to all MLS(B) and ketolide drugs. The erm(37) phenotype fits into neither category. We show here by tandem mass spectrometry that Erm(37) initially adds a single methyl group to its primary target at A2058 but then proceeds to attach additional methyl groups to the neighboring nucleotides A2057 and A2059. Other methyltransferases, Erm(E) and Erm(O), maintain their specificity for A2058 on mycobacterial rRNA. Erm(E) and Erm(O) have a full-length C-terminal domain, which appears to be important for stabilizing the methyltransferases at their rRNA target, and this domain is truncated in Erm(37). The lax interaction of the M. tuberculosis Erm(37) with its rRNA produces a unique methylation pattern and confers resistance to the ketolide telithromycin.

  8. 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.

  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. Induction of ermSV by 16-membered-ring macrolide antibiotics.

    PubMed Central

    Kamimiya, S; Weisblum, B

    1997-01-01

    The erm family of 23S rRNA adenine-N6-methyltransferases confers resistance to all macrolide-lincosamide-streptograminB (MLS) antibiotics, but not all MLS antibiotics induce synthesis of Erm methyltransferase with equal efficiency in a given organism. The induction efficiency of a test panel of MLS antibiotics was studied by using two translational attenuator-lac reporter gene fusion constructs, one based on ermSV from Streptomyces viridochromogenes NRRL 2860 and the other based on ermC from Staphylococcus aureus RN2442. Four types of responses which were correlated with the macrolide ring size were seen, as follows: group 1, both ermSV and ermC were induced by the 14-membered-ring macrolides erythromycin, lankamycin, and matromycin, as well as by the lincosamide celesticetin; group 2, neither ermSV nor ermC was induced by the 12-membered-ring macrolide methymycin or by the lincosamide lincomycin or the streptogramin type B antibiotic ostreogrycin B; group 3, ermSV was selectively induced over ermC by the 16-membered-ring macrolides carbomycin, chalcomycin, cirramycin, kitasamycin, maridomycin, and tylosin; and group 4, ermC was selectively induced over ermSV by the 14-membered-ring macrolide megalomicin. These data suggest that the leader peptide determines the specificity of induction by different classes of MLS antibiotics and that for a given attenuator, a major factor which determines whether a given macrolide induces resistance is its size. PMID:9055987

  11. e-Learning system ERM for medical radiation physics education.

    PubMed

    Stoeva, Magdalena; Cvetkov, Asen

    2005-09-01

    The objective of this paper is to present the Education for Radiation in Medicine (ERM) e-Learning System. The system was developed, tested and piloted in the Inter-University Medical Physics Centre, Plovdiv, Bulgaria. It was based on the results of EU Project TEMPUS S-JEP 09826. The ERM e-Learning System is an integrated on-line system for remote education covering aspects of Medical Radiation Physics education (M.Sc. level). It provides user-friendly interface and optimised functionality with three different access levels: trainee, professor and administrator. The minimum server requirements and the standard client side working environment turn the system into a good, cost effective and easy to support solution for remote education.

  12. Merlin/ERM proteins establish cortical asymmetry and centrosome position

    PubMed Central

    Hebert, Alan M.; DuBoff, Brian; Casaletto, Jessica B.; Gladden, Andrew B.; McClatchey, Andrea I.

    2012-01-01

    The ability to generate asymmetry at the cell cortex underlies cell polarization and asymmetric cell division. Here we demonstrate a novel role for the tumor suppressor Merlin and closely related ERM proteins (Ezrin, Radixin, and Moesin) in generating cortical asymmetry in the absence of external cues. Our data reveal that Merlin functions to restrict the cortical distribution of the actin regulator Ezrin, which in turn positions the interphase centrosome in single epithelial cells and three-dimensional organotypic cultures. In the absence of Merlin, ectopic cortical Ezrin yields mispositioned centrosomes, misoriented spindles, and aberrant epithelial architecture. Furthermore, in tumor cells with centrosome amplification, the failure to restrict cortical Ezrin abolishes centrosome clustering, yielding multipolar mitoses. These data uncover fundamental roles for Merlin/ERM proteins in spatiotemporally organizing the cell cortex and suggest that Merlin's role in restricting cortical Ezrin may contribute to tumorigenesis by disrupting cell polarity, spindle orientation, and, potentially, genome stability. PMID:23249734

  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. Essential role of endogenously synthesized tylosin for induction of ermSF in Streptomyces fradiae.

    PubMed Central

    Memili, E; Weisblum, B

    1997-01-01

    We compared ermSF induction in wild-type Streptomyces fradiae NRRL B-2702 and that in GS-14, a tylA mutant which cannot synthesize tylosin. Our findings suggest that (i) endogenously synthesized tylosin plays an obligatory role in ermSF induction and (ii) tylosin, or a biosynthetic intermediate beyond tylactone, has an "autocrine" function that induces ErmSF synthesis, thereby enabling S. fradiae to resist higher levels of tylosin. PMID:9145902

  15. 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

  16. Inducible Expression of both ermB and ermT Conferred High Macrolide Resistance in Streptococcus gallolyticus subsp. pasteurianus Isolates in China

    PubMed Central

    Li, Meixia; Cai, Chao; Chen, Juan; Cheng, Changwei; Cheng, Guofu; Hu, Xueying; Liu, Cuiping

    2016-01-01

    Streptococcus gallolyticus subsp. pasteurianus is an under-recognized pathogen and zoonotic agent causing opportunistic infections in humans. Despite increasing recognition of this subspecies as a cause for human infectious diseases, limited information is known about its antibiotic resistance mechanism. In this study, we aim to identify the molecular mechanism underlying the high macrolide resistance of six S. gallolyticus subsp. pasteurianus isolates from dead ducklings collected in several natural outbreaks in China during 2010–2013. All isolates exhibited multi-drug resistance including high macrolide resistance (MIC ≥ 1024 mg/L for erythromycin, and 512 mg/L for clarithromycin). Efflux-encoding mefA and mefE genes were not detectable in these isolates. The presence of 23S rRNA mutations in specific isolates did not significantly change macrolide MICs. No nucleotide substitutions were found in genes encoding ribosomal proteins L4 or L22. The ermB and ermT genes were found in the genomes of all isolates. These two genes were acquired independently in one highly virulent isolate AL101002, and clustered with Tn916 and IS1216, respectively. The expression of both ermB and ermT in all isolates was erythromycin inducible and yielded comparable macrolide MICs in all six isolates. Taken together, inducible expression of both ermB and ermT conferred high macrolide resistance in these S. gallolyticus subsp. pasterianus isolates. Our findings reveal new macrolide resistance features in S. gallolyticus subsp. pasteurianus by both ermB and ermT. PMID:27669217

  17. New Genetic Element Carrying the Erythromycin Resistance Determinant erm(TR) in Streptococcus pneumoniae▿

    PubMed Central

    Camilli, Romina; Del Grosso, Maria; Iannelli, Francesco; Pantosti, Annalisa

    2008-01-01

    erm(A) subclass erm(TR), a common macrolide resistance determinant in Streptococcus pyogenes but quite rare in Streptococcus pneumoniae, was found in a clinical S. pneumoniae isolate (AP200) from Italy. In this isolate, erm(TR) was found included in a genetic element approximately 56 kb in size that did not appear to be conjugative but could be transferred by transformation. An erm(TR)-containing DNA fragment of approximately 10 kb was sequenced and 12 open reading frames (ORFs) were identified. Upstream of erm(TR), a regulatory protein of the TetR family and the two components of an efflux pump of the ABC type were found. Downstream of erm(TR), there were ORFs homologous to a spectinomycin phosphotransferase, transposases, and a relaxase. Since the genomic sequence of S. pyogenes MGAS10750 carrying erm(TR) became available, comparison between the erm(TR)-containing genetic elements in AP200 and in MGAS10750 was performed. The region flanking erm(TR) in MGAS10750 showed identity with AP200 for 10 ORFs out of 12. PCR mapping using primers designed on the sequence of MGAS10750 confirmed that AP200 carries a genetic element similar to that of MGAS10750. In AP200 the genetic element was inserted inside an ORF homologous to spr0790 of S. pneumoniae R6, coding for a type I restriction modification system. Homologies between the insertion sites in AP200 and MGAS10750 consisted of eight conserved nucleotides, of which three were duplicated, likely representing target site duplication. The structure of the erm(TR)-carrying genetic element shows characteristics of a transposon/prophage remnant chimera. In AP200 this genetic element was designated Tn1806. PMID:18070957

  18. Recombination rates of Streptococcus pneumoniae isolates with both erm(B) and mef(A) genes.

    PubMed

    Lee, Ji-Young; Song, Jae-Hoon; Ko, Kwan Soo

    2010-08-01

    Erythromycin-resistant Streptococcus pneumoniae isolates containing both erm(B) and mef(A) genes have a higher rate of multidrug resistance (MDR). We investigated the relationships between the presence of erythromycin resistance determinants and the recombination rate. We determined the mutation and recombination frequencies of 46 S. pneumoniae isolates, which included 19 with both erm(B) and mef(A), nine with only erm(B), six with only mef(A), and 11 erythromycin-susceptible isolates. Mutation frequency values were estimated as the number of rifampin-resistant colonies as a proportion of total viable count. Genotypes and serotypes of isolates with the hyper-recombination phenotype were determined. Twelve S. pneumoniae isolates were hypermutable and four isolates were determined to have hyper-recombination frequency. Streptococcus pneumoniae isolates with both erm(B) and mef(A) genes did not show a high mutation frequency. In contrast, all isolates with a hyper-recombination phenotype contained both erm(B) and mef(A) genes. In addition, the recombination rate of isolates with both erm(B) and mef(A) genes was statistically higher than the rate of other isolates. The dual presence of erm(B) and mef(A) genes in some pneumococcal isolates may be associated with high recombination frequency. This may be one of the reasons for the frequent emergence of MDR in certain pneumococcal isolates.

  19. 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.

  20. Nucleotide sequence of ermA, a macrolide-lincosamide-streptogramin B determinant in Staphylococcus aureus.

    PubMed Central

    Murphy, E

    1985-01-01

    The complete nucleotide sequence of ermA, the prototype macrolide-lincosamide-streptogramin B resistance gene from Staphylococcus aureus, has been determined. The sequence predicts a 243-amino-acid protein that is homologous to those specified by ermC, ermAM, and ermD, resistance determinants from Staphylococcus aureus, Streptococcus sanguis, and Bacillus licheniformis, respectively. The ermA transcript, identified by Northern analysis and S1 mapping, contains a 5' leader sequence of 211 bases which has the potential to encode two short peptides of 15 and 19 amino acids; the second, longer peptide has 13 amino acids in common with the putative regulatory leader peptide of ermC. The coding sequence for this peptide is deleted in several mutants in which macrolide-lincosamide-streptogramin B resistance is constitutively expressed. Potential secondary structures available to the leader sequence of the wild-type (inducible) transcript and to constitutive deletion, insertion, and point mutations provide additional support for the translational attenuation model for induction of macrolide-lincosamide-streptogramin B resistance. Images PMID:2985541

  1. Protein Phosphatase 1α Mediates Ceramide-induced ERM Protein Dephosphorylation

    PubMed Central

    Canals, Daniel; Roddy, Patrick; Hannun, Yusuf A.

    2012-01-01

    ERM (ezrin, radixin, and moesin) proteins are cytoskeletal interacting proteins that bind cortical actin, the plasma membrane, and membrane proteins, which are found in specialized plasma membrane structures such as microvilli and filopodia. ERM proteins are regulated by phosphatidylinositol 4, 5-biphosphate (PIP2) and by phosphorylation of a C-terminal threonine, and its inactivation involves PIP2 hydrolysis and/or myosin phosphatase (MP). Recently, we demonstrated that ERM proteins are also subject to counter regulation by the bioactive sphingolipids ceramide and sphingosine 1-phosphate. Plasma membrane ceramide induces ERM dephosphorylation whereas sphingosine 1-phosphate induces their phosphorylation. In this work, we pursue the mechanisms by which ceramide regulates dephosphorylation. We found that this dephosphorylation was independent of hydrolysis and localization of PIP2 and MP. However, the results show that ERM dephosphorylation was blocked by treatment with protein phosphatase 1 (PP1) pharmacological inhibitors and specifically by siRNA to PP1α, whereas okadaic acid, a PP2A inhibitor, failed. Moreover, a catalytic inactive mutant of PP1α acted as dominant negative of the endogenous PP1α. Additional results showed that the ceramide mechanism of PP1α activation is largely independent of PIP2 hydrolysis and MP. Taken together, these results demonstrate a novel, acute mechanism of ERM regulation dependent on PP1α and plasma membrane ceramide. PMID:22311981

  2. Differential expression of ezrin/radixin/moesin (ERM) and ERM-associated adhesion molecules in the blastocyst and uterus suggests their functions during implantation.

    PubMed

    Matsumoto, Hiromichi; Daikoku, Takiko; Wang, Haibin; Sato, Eimei; Dey, S K

    2004-03-01

    Development of the blastocyst to implantation competency, differentiation of the uterus to the receptive state, and a cross talk between the implantation-competent blastocyst and the uterine luminal epithelium are all essential to the process of implantation. In the present investigation, we examined the possibility for a potential cross talk between the blastocyst and uterus involving the ezrin/radixin/moesin (ERM) proteins and ERM-associated cytoskeletal cross-linker proteins CD43, CD44, ICAM-1, and ICAM-2. In normal Day 4 blastocysts and after rendering dormant blastocysts to implantation-competent by estrogen in vivo (activated), the outer surface of mural trophectoderm cells showed much higher levels of radixin as compared to those in the polar trophectoderm cells, inner cell mass (ICM), and primitive endoderm. In contrast, ezrin was present on both the mural and the polar trophectoderm cell surfaces of normal Day 4 and activated blastocysts at higher intensity than dormant blastocysts. A distinct localization was noted in the primitive endoderm of dormant blastocysts that was not apparent in activated or normal Day 4 blastocysts. The expression of moesin was modestly higher at the mural trophectoderm of implantation-competent blastocysts, while the localization appeared to be present primarily on the polar trophectoderm cell surface of Day 4 blastocysts. The localization of ERM-associated adhesion molecules CD43, CD44, and ICAM-2 was more intense in the implantation-competent blastocysts compared with the dormant blastocysts. However, while CD44 was present both in the trophectoderm and in ICM, CD43 and ICAM-2 were localized primarily to the trophectoderm. The signal for ICAM-1 was very intense in the ICM but was modest in the trophectoderm. No significant changes in fluorescence intensity were noted between activated and dormant blastocysts. In the receptive uterus on Day 4 of pregnancy, ERM proteins were localized to the uterine epithelium, while on Day 5

  3. Gem1 and ERMES do not directly affect phosphatidylserine transport from ER to mitochondria or mitochondrial inheritance.

    PubMed

    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

    2012-06-01

    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.

  4. Ubiquitination of ERMES components by the E3 ligase Rsp5 is involved in mitophagy

    PubMed Central

    Belgareh-Touzé, Naïma; Cavellini, Laetitia; Cohen, Mickael M.

    2017-01-01

    ABSTRACT Mitochondria are dynamic organelles that undergo permanent fission and fusion events. These processes play an essential role in maintaining normal cellular function. In the yeast Saccharomyces cerevisiae, the endoplasmic reticulum-mitochondrial encounter structure (ERMES) is a marker of sites of mitochondrial division, but it is also involved in a plethora of other mitochondrial functions. However, it remains unclear how these different functions are regulated. We show here that Mdm34 and Mdm12, 2 components of ERMES, are ubiquitinated by the E3 ligase Rsp5. This ubiquitination is not involved in mitochondrial dynamics or in the distribution and turnover of ERMES. Nevertheless, the ubiquitination of Mdm34 and Mdm12 was required for efficient mitophagy. We thus report here the first identification of ubiquitinated substrates participating in yeast mitophagy. PMID:27846375

  5. Tylosin Resistance in Arcanobacterium pyogenes Is Encoded by an Erm X Determinant

    PubMed Central

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

    2003-01-01

    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 ≥128 μg/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

  6. Scarce detection of mobile erm genes associated with tetQ in Bacteroides and Parabacteroides from Costa Rica.

    PubMed

    Quesada-Gómez, Carlos; Rodríguez-Cavallini, Evelyn; Rodríguez, César

    2013-06-01

    The frequency of finding of clindamycin-resistant anaerobic bacteria in clinical samples has doubled from 2008 to 2010 in Costa Rica. To determine whether this increase is due to dissemination of erm genes aided by tetQ elements, we analyzed 100 isolates of Bacteroides or Parabacteroides from a regional hospital, a national hospital, and the community. Antimicrobial susceptibilities were recorded with a broth micro-dilution method and erm genes were detected by PCR and Southern blotting. In addition, plasmid isolation and mating experiments were performed to clarify the location and mobility of the detected erm genes. Resistance to clindamycin was by far more frequent in the regional hospital (72%) than in the national hospital (29%) and the community (26%). Resistance to tetracycline was even more common, with the community (85%) outweighing the hospitals (71-72%). While MIC of clindamycin were higher in the hospitals than in the community (P < 0.05), the opposite was seen for tetracycline (P < 0.0001). Of the sought-after genes, only ermG (n = 2), ermA (n = 1), and ermF (n = 1) were detected in the hospitals and ermF in the community (n = 2). In opposition to the low frequency of finding of erm genes, 71% of the isolates were positive for tetQ. None of the detected genes were encoded on plasmids. Only three isolates from the hospitals transferred their erm genes laterally. By contrast, 13 hospital isolates and two community isolates transferred tetQ. Despite the widespread finding of tetracycline-resistant tetQ-positive bacteria, mobile erm genes were rare in our bacterial collection. We conclude that the detected erm genes are likely not included in typical conjugative transposons of Bacteroides and Parabacteroides.

  7. Characterization of a novel type of MLSB resistance plasmid from Staphylococcus saprophyticus carrying a constitutively expressed erm(C) gene.

    PubMed

    Hauschild, Tomasz; Lüthje, Petra; Schwarz, Stefan

    2006-06-15

    An erm(C)-carrying plasmid of unusual size and restriction map, designated pSES22, was identified in a Staphylococcus saprophyticus strain and sequenced completely. Constitutive expression of the erm(C) gene from pSES22 is based on a novel 22-bp tandem duplication in the erm(C) translational attenuator. Comparative analysis of the deduced Erm(C) amino acid sequence revealed that Erm(C) from pSES22 - together with an Erm(C) methylase from S. hyicus - represented a separate branch in the homology tree of Erm(C) methylases. Structural comparisons showed that plasmid pSES22 differed distinctly from all other completely sequenced erm(C)-carrying resistance plasmids. However, pSES22 was similar to several members of a diverse group of small plasmids, all of which carried closely related plasmid backbones consisting of the genes repU and pre/mob, but differed in their resistance genes.

  8. 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…

  9. Emergence and spread of Streptococcus pneumoniae with erm(B) and mef(A) resistance.

    PubMed

    Farrell, David J; Jenkins, Stephen G; Brown, Steven D; Patel, Manish; Lavin, Bruce S; Klugman, Keith P

    2005-06-01

    Streptococcus pneumoniae isolates (N = 31,001) were collected from patients with community-acquired respiratory tract infections during the PROTEKT US surveillance study (2000-2003). While the macrolide (erythromycin) resistance rate remained stable at approximately 29%, the prevalence of resistant isolates containing both erm(B) and mef(A) increased from 9.7% in year 1 to 16.4% in year 3, with substantial regional variability. Almost all (99.2%) dual erm(B) + mef(A) macrolide-resistant isolates exhibited multidrug resistance, whereas 98.6% and 99.0% were levofloxacin- and telithromycin-susceptible, respectively. These strains were most commonly isolated from the ear or middle-ear fluid of children. Of 152 representative erm(B)+mef(A) isolates, >90% were clonally related to the multidrug-resistant international Taiwan19F-14 clonal complex 271 (CC271). Of 366 erm(B)+mef(A) isolates from the PROTEKT global study (1999-2003), 83.3% were CC271, with the highest prevalence seen in South Africa, South Korea, and the United States. This study confirms the increasing global emergence and rapidly increasing US prevalence of this multidrug-resistant pneumococcal clone.

  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. Findability Enabled: The Rise of the Knowledgebase Puts the ERMS on a New Stage

    ERIC Educational Resources Information Center

    Ownes, Dodie

    2006-01-01

    As public and academic librarians rely more heavily on electronic resources to satisfy patrons' information needs, seamless searching across print and electronic holdings gets more and more essential. Enter the integrated library system (ILS)-independent electronic resource management system (ERMS), which promises to integrate, search, and expose…

  12. First report of macrolide resistance gene erm(T) harbored by a novel small plasmid from Erysipelothrix rhusiopathiae.

    PubMed

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

    2015-04-01

    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.

  13. 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

  14. Development of the Extendable and Retractable Mast (ERM), Design phase 2. Volume 1

    NASA Astrophysics Data System (ADS)

    Specht; Schmid; Chellingsworth; Wislez; Bhatti; Stenne

    1987-03-01

    An Extendable and Retractable Mast (ERM) for space applications such as deployment and retraction of high power rollable, foldable, or rigid solar arrays, large unfurlable antennas, and scientific payloads, was developed. The coverage of a large field of application requires a strong and stiff light-weight mast with good pointing accuracy allowing the adaptation of heavy tip masses as well as the fixation of line-loads to intermediate attachment points along the mast at the same time. The ERM payload capability is 40 kg on tip, plus 10 kg/m line load, plus 100N eccentric load. Deployed length is 40 m. The ERM is designed as a spindle-driven telescopic mast in carbon-fiber reinforced plastics (CFRP) technology. A 40 m solar array mast and a 20 m antenna mast were considered. The feasibility of a 60 m mast is also shown. Manufacturing and test of a breadboard model hardware, and the detailed design of an engineering model mast are described.

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

    PubMed

    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.

  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. 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.

  18. A Novel erm(44) Gene Variant from a Human Staphylococcus saprophyticus Isolate Confers Resistance to Macrolides and Lincosamides but Not Streptogramins.

    PubMed

    Strauss, Christian; Hu, Yanmin; Coates, Anthony; Perreten, Vincent

    2017-01-01

    A novel erm(44) gene variant, erm(44)v, has been identified by whole-genome sequencing in a Staphylococcus saprophyticus isolate from the skin of a healthy person. It has the particularity to confer resistance to macrolides and lincosamides but not to streptogramin B when expressed in S. aureus The erm(44)v gene resides on a 19,400-bp genomic island which contains phage-associated proteins and is integrated into the chromosome of S. saprophyticus.

  19. TcdC Does Not Significantly Repress Toxin Expression in Clostridium difficile 630ΔErm

    PubMed Central

    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

  20. 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...

  1. Knockdown of ERM family member moesin in host cells increases HIV type 1 replication.

    PubMed

    Capalbo, Gianni; Mueller-Kuller, Thea; Markovic, Sandra; Klein, Stefan A; Dietrich, Ursula; Hoelzer, Dieter; Ottmann, Oliver G; Scheuring, Urban J

    2011-12-01

    Moesin is a member of the ERM (ezrin, radixin, moesin) family of cytoskeleton/membrane structure organizing and signal transduction proteins. Previously, we found an increased expression of moesin during HIV-1 infection. Moesin was also reported to be incorporated into HIV-1 virions. To analyze whether moesin is a host factor affecting the replication cycle of human immunodeficiency virus type 1 (HIV-1), we used small interfering RNAs (siRNAs) to evaluate the effect of moesin knockdown on HIV-1 replication in P4-CCR5 cells. Moesin's knockdown did not affect the cell viability or cell phenotype. Interestingly, we observed a marked increase in viral replication, as demonstrated by enhanced HIV-1 RNA, p24 antigen, and ß-galactosidase reporter expression. Moesin-dependent enhancement of HIV-1 replication was confirmed in lymphocytic host cells (Jurkat). These results suggest an overall rather restrictive role of moesin for HIV-1 replication in host cells in vitro.

  2. Oral and anal vaccination confers full protection against enteric redmouth disease (ERM) in rainbow trout.

    PubMed

    Villumsen, Kasper Rømer; Neumann, Lukas; Ohtani, Maki; Strøm, Helene Kragelund; Raida, Martin Kristian

    2014-01-01

    The effect of oral vaccines against bacterial fish diseases has been a topic for debate for decades. Recently both M-like cells and dendritic cells have been discovered in the intestine of rainbow trout. It is therefore likely that antigens reaching the intestine can be taken up and thereby induce immunity in orally vaccinated fish. The objective of this project was to investigate whether oral and anal vaccination of rainbow trout induces protection against an experimental waterborne infection with the pathogenic enterobacteria Yersinia ruckeri O1 biotype 1 the causative agent of enteric redmouth disease (ERM). Rainbow trout were orally vaccinated with AquaVac ERM Oral (MERCK Animal Health) or an experimental vaccine bacterin of Y. ruckeri O1. Both vaccines were tested with and without a booster vaccination four months post the primary vaccination. Furthermore, two groups of positive controls were included, one group receiving the experimental oral vaccine in a 50 times higher dose, and the other group receiving a single dose administered anally in order to bypass the stomach. Each group was bath challenged with 6.3 × 10(8) CFU/ml Y. ruckeri, six months post the primary vaccination. The challenge induced significant mortality in all the infected groups except for the groups vaccinated anally with a single dose or orally with the high dose of bacterin. Both of these groups had 100% survival. These results show that a low dose of Y. ruckeri bacterin induces full protection when the bacterin is administered anally. Oral vaccination also induces full protection, however, at a dose 50 times higher than if the fish were to be vaccinated anally. This indicates that much of the orally fed antigen is digested in the stomach before it reaches the second segment of the intestine where it can be taken up as immunogenic antigens and presented to lymphocytes.

  3. 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.

  4. 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

  5. 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…

  6. 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

  7. Two novel functions of hyaluronidase-2 (Hyal2) are formation of the glycocalyx and control of CD44-ERM interactions.

    PubMed

    Duterme, Cecile; Mertens-Strijthagen, Jeannine; Tammi, Markku; Flamion, Bruno

    2009-11-27

    It has long been predicted that the members of the hyaluronidase enzyme family have important non-enzymatic functions. However, their nature remains a mystery. The metabolism of hyaluronan (HA), their major enzymatic substrate, is also enigmatic. To examine the function of Hyal2, a glycosylphosphatidylinositol-anchored hyaluronidase with intrinsically weak enzymatic activity, we have compared stably transfected rat fibroblastic BB16 cell lines with various levels of expression of Hyal2. These cell lines continue to express exclusively the standard form (CD44s) of the main HA receptor, CD44. Hyal2, CD44, and one of its main intracellular partners, ezrin-radixin-moesin (ERM), were found to co-immunoprecipitate. Functionally, Hyal2 overexpression was linked to loss of the glycocalyx, the HA-rich pericellular coat. This effect could be mimicked by exposure of BB16 cells either to Streptomyces hyaluronidase, to HA synthesis inhibitors, or to HA oligosaccharides. This led to shedding of CD44, separation of CD44 from ERM, reduction in baseline level of ERM activation, and markedly decreased cell motility (50% reduction in a wound healing assay). The effects of Hyal2 on the pericellular coat and on CD44-ERM interactions were inhibited by treatment with the Na(+)/H(+) exchanger-1 inhibitor ethyl-N-isopropylamiloride. We surmise that Hyal2, through direct interactions with CD44 and possibly some pericellular hyaluronidase activity requiring acidic foci, suppresses the formation or the stability of the glycocalyx, modulates ERM-related cytoskeletal interactions, and diminishes cell motility. These effects may be relevant to the purported in vivo tumor-suppressive activity of Hyal2.

  8. Polyclonal population structure of Streptococcus pneumoniae isolates in Spain carrying mef and mef plus erm(B).

    PubMed

    de la Pedrosa, Elia Gómez G; Morosini, María-Isabel; van der Linden, Mark; Ruiz-Garbajosa, Patricia; Galán, Juan Carlos; Baquero, Fernando; Reinert, Ralf René; Cantón, Rafael

    2008-06-01

    The population structure (serotypes, pulsed-field gel electrophoresis [PFGE] types, and multilocus sequencing types) of 45 mef-positive Streptococcus pneumoniae isolates [carrying mef alone (n = 17) or with the erm(B) gene n = 28)] were studied. They were selected from among all erythromycin-resistant isolates (n = 244) obtained from a collection of 712 isolates recovered from different Spanish geographic locations in the prevaccination period from 1999 to 2003. The overall rates of resistance (according to the criteria of the CLSI) among the 45 mef-positive isolates were as follows: penicillin G, 82.2%; cefotaxime, 22.2%; clindamycin, 62.2%; and tetracycline, 68.8% [mainly in isolates carrying erm(B) plus mef(E); P < 0.001]. No levofloxacin or telithromycin resistance was found. Macrolide resistance phenotypes (as determined by the disk diffusion approximation test) were 37.7% for macrolide resistance [with all but one due to mef(E)] and 62.2% for constitutive macrolide-lincosamide-streptogramin B resistance [cMLS(B); with all due to mef(E) plus erm(B)]. Serotypes 14 (22.2%), 6B (17.7%), 19A (13.3%), and 19F (11.1%) were predominant. Twenty-five different DNA patterns (PFGE types) were observed. Our mef-positive isolates were grouped (by eBURST analysis) into four clonal complexes (n = 18) and 19 singleton clones (n = 27). With the exception of clone Spain(9V)-3, all clonal complexes (clonal complexes 6B, Spain(6B)-2, and Sweden(15A)-25) and 73.6% of singleton clones carried both the erm(B) and the mef(E) genes. The international multiresistant clones Spain(23F)-1 and Poland(6B)-20 were represented as singleton clones. A high proportion of mef-positive S. pneumoniae isolates presented the erm(B) gene, with all isolates expressing the cMLS(B) phenotype. A polyclonal population structure was demonstrated within our Spanish mef-positive S. pneumoniae isolates, with few clonal complexes overrepresented within this collection.

  9. ERM protein moesin is phosphorylated by advanced glycation end products and modulates endothelial permeability.

    PubMed

    Guo, Xiaohua; Wang, Lingjun; Chen, Bo; Li, Qiang; Wang, Jiping; Zhao, Ming; Wu, Wei; Zhu, Ping; Huang, Xuliang; Huang, Qiaobing

    2009-07-01

    Advanced glycation end products (AGEs) accumulated in different pathological conditions have the potent capacity to alter cellular properties that include endothelial structural and functional regulations. The disruption of endothelial barrier integrity may contribute to AGE-induced microangiopathy and macrovasculopathy. Previous studies have shown that AGEs induced the rearrangement of actin and subsequent hyperpermeability in endothelial cells (ECs). However, the mechanisms involved in this AGE-evoked EC malfunction are not well understood. This study directly evaluated the involvement of moesin phosphorylation in AGE-induced alterations and the effects of the RhoA and p38 MAPK pathways on this process. Using immortalized human dermal microvascular ECs (HMVECs), we first confirmed that the ezrin/radixin/moesin (ERM) protein moesin is required in AGE-induced F-actin rearrangement and hyperpermeability responses in ECs by knockdown of moesin protein expression with small interfering RNA. We then detected AGE-induced moesin phosphorylation by Western blot analysis. The mechanisms involved in moesin phosphorylation were analyzed by blocking AGE receptor binding and inhibiting Rho and MAPK pathways. AGE-treated HMVECs exhibited time- and dose-dependent increases in the Thr(558) phosphorylation of moesin. The increased moesin phosphorylation was attenuated by preadministrations of AGE receptor antibody, Rho kinase (ROCK), or p38 inhibitor. Suppression of p38 activation via the expression of dominant negative mutants with Ad.MKK6b or Ad.p38alpha also decreased moesin phosphorylation. The activation of the p38 pathway by transfection of HMVECs with an adenoviral construct of dominant active MKK6b resulted in moesin phosphorylation. These results suggest a critical role of moesin phosphorylation in AGE-induced EC functional and morphological regulations. Activation of the ROCK and p38 pathways is required in moesin phosphorylation.

  10. 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.

  11. The protective role of MLCP-mediated ERM dephosphorylation in endotoxin-induced lung injury in vitro and in vivo

    PubMed Central

    Kovacs-Kasa, Anita; Gorshkov, Boris A.; Kim, Kyung-Mi; Kumar, Sanjiv; Black, Stephen M.; Fulton, David J.; Dimitropoulou, Christiana; Catravas, John D.; Verin, Alexander D.

    2016-01-01

    The goal of this study was to investigate the role of MLC phosphatase (MLCP) in a LPS model of acute lung injury (ALI). We demonstrate that ectopic expression of a constitutively-active (C/A) MLCP regulatory subunit (MYPT1) attenuates the ability of LPS to increase endothelial (EC) permeability. Down-regulation of MYPT1 exacerbates LPS-induced expression of ICAM1 suggesting an anti-inflammatory role of MLCP. To determine whether MLCP contributes to LPS-induced ALI in vivo, we utilized a nanoparticle DNA delivery method to specifically target lung EC. Expression of a C/A MYPT1 reduced LPS-induced lung inflammation and vascular permeability. Further, increased expression of the CS1β (MLCP catalytic subunit) also reduced LPS-induced lung inflammation, whereas the inactive CS1β mutant increased vascular leak. We next examined the role of the cytoskeletal targets of MLCP, the ERM proteins (Ezrin/Radixin/Moesin), in mediating barrier dysfunction. LPS-induced increase in EC permeability was accompanied by PKC-mediated increase in ERM phosphorylation, which was more prominent in CS1β-depleted cells. Depletion of Moesin and Ezrin, but not Radixin attenuated LPS-induced increases in permeability. Further, delivery of a Moesin phospho-null mutant into murine lung endothelium attenuated LPS-induced lung inflammation and vascular leak suggesting that MLCP opposes LPS-induced ALI by mediating the dephosphorylation of Moesin and Ezrin. PMID:27976727

  12. The protective role of MLCP-mediated ERM dephosphorylation in endotoxin-induced lung injury in vitro and in vivo.

    PubMed

    Kovacs-Kasa, Anita; Gorshkov, Boris A; Kim, Kyung-Mi; Kumar, Sanjiv; Black, Stephen M; Fulton, David J; Dimitropoulou, Christiana; Catravas, John D; Verin, Alexander D

    2016-12-15

    The goal of this study was to investigate the role of MLC phosphatase (MLCP) in a LPS model of acute lung injury (ALI). We demonstrate that ectopic expression of a constitutively-active (C/A) MLCP regulatory subunit (MYPT1) attenuates the ability of LPS to increase endothelial (EC) permeability. Down-regulation of MYPT1 exacerbates LPS-induced expression of ICAM1 suggesting an anti-inflammatory role of MLCP. To determine whether MLCP contributes to LPS-induced ALI in vivo, we utilized a nanoparticle DNA delivery method to specifically target lung EC. Expression of a C/A MYPT1 reduced LPS-induced lung inflammation and vascular permeability. Further, increased expression of the CS1β (MLCP catalytic subunit) also reduced LPS-induced lung inflammation, whereas the inactive CS1β mutant increased vascular leak. We next examined the role of the cytoskeletal targets of MLCP, the ERM proteins (Ezrin/Radixin/Moesin), in mediating barrier dysfunction. LPS-induced increase in EC permeability was accompanied by PKC-mediated increase in ERM phosphorylation, which was more prominent in CS1β-depleted cells. Depletion of Moesin and Ezrin, but not Radixin attenuated LPS-induced increases in permeability. Further, delivery of a Moesin phospho-null mutant into murine lung endothelium attenuated LPS-induced lung inflammation and vascular leak suggesting that MLCP opposes LPS-induced ALI by mediating the dephosphorylation of Moesin and Ezrin.

  13. Manual curation and reannotation of the genomes of Clostridium difficile 630Δerm and Clostridium difficile 630.

    PubMed

    Dannheim, Henning; Riedel, Thomas; Neumann-Schaal, Meina; Bunk, Boyke; Schober, Isabel; Spröer, Cathrin; Chibani, Cynthia Maria; Gronow, Sabine; Liesegang, Heiko; Overmann, Jörg; Schomburg, Dietmar

    2017-01-09

    We resequenced the genome of Clostridium difficile 630Δerm (DSM 28645), a model strain commonly used for the generation of insertion mutants. The genome sequence was obtained by a combination of single-molecule real-time (SMRT) and Illumina sequencing technology. Detailed manual curation and comparison to the previously published genomic sequence revealed sequence differences including inverted regions and the presence of plasmid pCD630. Manual curation of our previously deposited genome sequence of the parental strain 630 (DSM 27543) led to an improved genome sequence. In addition, the sequence of the transposon Tn5397 was completely identified. We manually revised the current manual annotation of the initial sequence of strain 630 and modified either gene names, gene product names or assigned EC numbers of 57 % of genes. The number of hypothetical and conserved hypothetical proteins was reduced by 152. This annotation was used as a template to annotate the most recent genome sequences of the strains 630Δerm and 630. Based on the genomic analysis, several new metabolic features of C. difficile are proposed and could be supported by literature and subsequent experiments.

  14. 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

  15. 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.

  16. Genotypes of macrolide-resistant pneumococci from children in southwestern Japan: raised incidence of strains that have both erm(B) and mef(A) with serotype 6B clones.

    PubMed

    Ikenaga, Masaaki; Kosowska-Shick, Klaudia; Gotoh, Kenji; Hidaka, Hidenobu; Koga, Hiroyasu; Masunaga, Kenji; Nagai, Kensuke; Tsumura, Naoki; Appelbaum, Peter C; Matsuishi, Toyojiro

    2008-09-01

    MICs of penicillin G, erythromycin, clarithromycin, clindamycin, azithromycin, and telithromycin were tested for 189 clinical isolates collected during 2002 to 2005 from children in southwestern Japan. Serotyping and polymerase chain reaction for presence of erm(B) and mef(A) were performed. All strains with erm(B) + mef(A) were analyzed by pulsed-field gel electrophoresis (PFGE) and compared to 3 global clones: Spain(23F)-1; Spain(9V)-3 and its variant -14; a South Korean strain same as Taiwan (19F)-14 clone and 5 strains with erm(B) + mef(A) from other countries. Of the 173 macrolide-resistant (erythromycin MIC > or =0.5 microg/mL) strains, 104 (60.1%) had erm(B), 47 (27.2%) had mef(A), and 22 (12.7%) had erm(B) + mef(A). Strains expressing erm(B) or both erm(B) and mef(A) had high macrolide MIC(90)s (>64 microg/mL), except telithromycin (MIC(90), 0.25 microg/mL). Of the 22 erm(B) + mef(A) strains, 10 had 4 distinct PFGE patterns and were mainly serotype 6B clones, which differed from those described in previous reports; 5 other strains had unique profiles.

  17. A Self-deleting Cre-lox-ermAM Cassette, CHESHIRE, for marker-less gene deletion in Streptococcus pneumoniae

    PubMed Central

    Weng, Liming; Biswas, Indranil; Morrison, Donald A.

    2009-01-01

    Although targeted mutagenesis of Streptococcus pneumoniae is readily accomplished with the aid of natural genetic transformation and chimeric donor DNA constructs assembled in vitro, the drug resistance markers often employed for selection of recombinant products can themselves be undesirable by-products of the genetic manipulation. A new cassette carrying the erythromycin-resistance marker ermAM is described that can be used as a temporary marker for selection of desired recombinants. The cassette may subsequently be removed at will by virtue of an embedded fucose-regulated Cre recombinase gene and terminal lox66 and lox71 Cre recognition sites, with retention of 34 bp from the cassette as an inert residual double-mutant lox72 site. PMID:19850089

  18. Internalization of Met requires the co-receptor CD44v6 and its link to ERM proteins.

    PubMed

    Hasenauer, Susanne; Malinger, Dieter; Koschut, David; Pace, Giuseppina; Matzke, Alexandra; von Au, Anja; Orian-Rousseau, Véronique

    2013-01-01

    Receptor Tyrosine Kinases (RTKs) are involved in many cellular processes and play a major role in the control of cell fate. For these reasons, RTK activation is maintained under tight control. Met is an essential RTK that induces proliferation, differentiation, migration, survival and branching morphogenesis. Deregulation of Met by overexpression, amplification or lack of effective degradation leads to cancer and metastasis. We have shown that Met relies on CD44v6 for its activation and for signaling in several cancer cell lines and also in primary cells. In this paper, we show that internalization of Met is dependent on CD44v6 and the binding of Ezrin to the CD44v6 cytoplasmic domain. Both CD44v6 and Met are co-internalized upon Hepatocyte Growth Factor induction suggesting that Met-induced signaling from the endosomes relies on its collaboration with CD44v6 and the link to the cytoskeleton provided by ERM proteins.

  19. An in vivo EGF receptor localization screen in C. elegans Identifies the Ezrin homolog ERM-1 as a temporal regulator of signaling.

    PubMed

    Haag, Andrea; Gutierrez, Peter; Bühler, Alessandra; Walser, Michael; Yang, Qiutan; Langouët, Maeva; Kradolfer, David; Fröhli, Erika; Herrmann, Christina J; Hajnal, Alex; Escobar-Restrepo, Juan M

    2014-05-01

    The subcellular localization of the epidermal growth factor receptor (EGFR) in polarized epithelial cells profoundly affects the activity of the intracellular signaling pathways activated after EGF ligand binding. Therefore, changes in EGFR localization and signaling are implicated in various human diseases, including different types of cancer. We have performed the first in vivo EGFR localization screen in an animal model by observing the expression of the EGFR ortholog LET-23 in the vulval epithelium of live C. elegans larvae. After systematically testing all genes known to produce an aberrant vulval phenotype, we have identified 81 genes regulating various aspects of EGFR localization and expression. In particular, we have found that ERM-1, the sole C. elegans Ezrin/Radixin/Moesin homolog, regulates EGFR localization and signaling in the vulval cells. ERM-1 interacts with the EGFR at the basolateral plasma membrane in a complex distinct from the previously identified LIN-2/LIN-7/LIN-10 receptor localization complex. We propose that ERM-1 binds to and sequesters basolateral LET-23 EGFR in an actin-rich inactive membrane compartment to restrict receptor mobility and signaling. In this manner, ERM-1 prevents the immediate activation of the entire pool of LET-23 EGFR and permits the generation of a long-lasting inductive signal. The regulation of receptor localization thus serves to fine-tune the temporal activation of intracellular signaling pathways.

  20. An In Vivo EGF Receptor Localization Screen in C. elegans Identifies the Ezrin Homolog ERM-1 as a Temporal Regulator of Signaling

    PubMed Central

    Walser, Michael; Yang, Qiutan; Langouët, Maeva; Kradolfer, David; Fröhli, Erika; Herrmann, Christina J.; Hajnal, Alex; Escobar-Restrepo, Juan M.

    2014-01-01

    The subcellular localization of the epidermal growth factor receptor (EGFR) in polarized epithelial cells profoundly affects the activity of the intracellular signaling pathways activated after EGF ligand binding. Therefore, changes in EGFR localization and signaling are implicated in various human diseases, including different types of cancer. We have performed the first in vivo EGFR localization screen in an animal model by observing the expression of the EGFR ortholog LET-23 in the vulval epithelium of live C. elegans larvae. After systematically testing all genes known to produce an aberrant vulval phenotype, we have identified 81 genes regulating various aspects of EGFR localization and expression. In particular, we have found that ERM-1, the sole C. elegans Ezrin/Radixin/Moesin homolog, regulates EGFR localization and signaling in the vulval cells. ERM-1 interacts with the EGFR at the basolateral plasma membrane in a complex distinct from the previously identified LIN-2/LIN-7/LIN-10 receptor localization complex. We propose that ERM-1 binds to and sequesters basolateral LET-23 EGFR in an actin-rich inactive membrane compartment to restrict receptor mobility and signaling. In this manner, ERM-1 prevents the immediate activation of the entire pool of LET-23 EGFR and permits the generation of a long-lasting inductive signal. The regulation of receptor localization thus serves to fine-tune the temporal activation of intracellular signaling pathways. PMID:24785082

  1. p38MAPK, Rho/ROCK and PKC pathways are involved in influenza-induced cytoskeletal rearrangement and hyperpermeability in PMVEC via phosphorylating ERM.

    PubMed

    Zhang, Chenyue; Wu, Ying; Xuan, Zinan; Zhang, Shujing; Wang, Xudan; Hao, Yu; Wu, Jun; Zhang, Shu

    2014-11-04

    Severe influenza infections are featured by acute lung injury, a syndrome of pulmonary microvascular leak. A growing number of evidences have shown that the pulmonary microvascular endothelial cells (PMVEC) are critical target of influenza virus, promoting microvascular leak. It is reported that there are multiple mechanisms by which influenza virus could elicit increased pulmonary endothelial permeability, in both direct and indirect manners. Ezrin/radixin/moesin family proteins, the linkers between plasma membrane and actin cytoskeleton, have been reported to be involved in cell adhesion, motility and may modulate endothelial permeability. Studies have also shown that ERM is phosphorylated in response to various stimuli via p38MAPK, Rho/ROCK or PKC pathways. However, it is unclear that whether influenza infection could induce ERM phosphorylation and its relocalization. In the present study, we have found that there are cytoskeletal reorganization and permeability increases in the course of influenza virus infection, accompanied by upregulated levels of p-ERM. p-ERM's aggregation along the periphery of PMVEC upon influenza virus infection was detected via confocal microscopy. Furthermore, we sought to determine the role of p38MAPK, Rho/ROCK and PKC pathways in ERM phosphorylation as well as their involvement in influenza virus-induced endothelial malfunction. The activation of p38MAPK, Rho/ROCK and PKC pathways upon influenza virus stimulation were observed, as evidenced by the evaluation of phosphorylated p38 (p-p38), phosphorylated MKK (p-MKK) in p38MAPK pathway, ROCK1 in Rho/ROCK pathway and phosphorylated PKC (p-PKC) in PKC pathway. We also showed that virus-induced ERM phosphorylation was reduced by using p38MAPK inhibitor, SB203580 (20 μM), Rho/ROCK inhibitor, Y27632 (20 μM), PKC inhibitor, LY317615 (10 μM). Additionally, influenza virus-induced F-actin reorganization and hyperpermeability were attenuated by pretreatment with SB203580, Y27632 and LY317615

  2. Analysis of transcriptional modulation of the presenilin 1 gene promoter by ZNF237, a candidate binding partner of the Ets transcription factor ERM

    PubMed Central

    Pastorcic, Martine; Das, Hriday K.

    2007-01-01

    DNA sequences required for the expression of the human presenilin 1 (PS1) gene have been identified between -118 and +178 flanking the major initiation site (+1) mapped in SK-N-SH cells. Several Ets sites are located both upstream as well as downstream from the +1 site, including an Ets motif present at -10 that controls 90% of transcription in SK-N-SH cells. However in SH-SY5Y cells transcription initiates further downstream and requires an alternative set of promoter elements including a +90 Ets motif. Ets2, ER81, ERM and Elk1 were identified by yeast one-hybrid selection in a human brain cDNA library using the -10 Ets motif as a bait. We have shown that ERM recognizes specifically Ets motifs on the PS1 promoter located at -10 as well as downstream at +90, +129 and +165 and activates PS1 transcription with promoter fragments whether or not they contain the -10 Ets site. We have now searched for ERM interacting proteins by yeast two-hybrid selection in a human brain cDNA library using the C-terminal 415 amino acid of ERM as a bait. One of the interacting proteins was ZNF237, a member of the MYM gene family. It is widely expressed in different tissues in eukaryotes under several forms derived by alternative splicing, including a large 382 amino acid form containing a single MYM domain, and 2 shorter forms of 208 and 213 amino acids respectively that do not. We show that both the 382 as well as the 208 amino acid forms are expressed in SK-N-SH cells but not in SH-SY5Y cells. Both forms interact with ERM and repress the transcription of PS1 in SH-SY5Y cells. The effect of both C-terminal and N-terminal deletions indicate that the N-terminal 120 amino acid region is required for interaction with ERM in yeast and furthermore single amino acid mutations show that residues 112 and 114 play an important role. The repression of transcription in SH-SY5Y cells also appears to require the N-terminal potion of ZNF237 and was affected by mutation of the amino acid 112. Data

  3. Analysis of transcriptional modulation of the presenilin 1 gene promoter by ZNF237, a candidate binding partner of the Ets transcription factor ERM.

    PubMed

    Pastorcic, Martine; Das, Hriday K

    2007-01-12

    DNA sequences required for the expression of the human presenilin 1 (PS1) gene have been identified between -118 and +178 flanking the major initiation site (+1) mapped in SK-N-SH cells. Several Ets sites are located both upstream as well as downstream from the +1 site, including an Ets motif present at -10 that controls 90% of transcription in SK-N-SH cells. However, in SH-SY5Y cells, transcription initiates further downstream and requires an alternative set of promoter elements including a +90 Ets motif. Ets2, ER81, ERM and Elk1 were identified by yeast one-hybrid selection in a human brain cDNA library using the -10 Ets motif as a bait. We have shown that ERM recognizes specifically Ets motifs on the PS1 promoter located at -10 as well as downstream at +90, +129 and +165 and activates PS1 transcription with promoter fragments whether or not they contain the -10 Ets site. We have now searched for ERM interacting proteins by yeast two-hybrid selection in a human brain cDNA library using the C-terminal 415 amino acid of ERM as a bait. One of the interacting proteins was ZNF237, a member of the MYM gene family. It is widely expressed in different tissues in eukaryotes under several forms derived by alternative splicing, including a large 382 amino acid form containing a single MYM domain, and 2 shorter forms of 208 and 213 amino acids respectively that do not. We show that both the 382 as well as the 208 amino acid forms are expressed in SK-N-SH cells but not in SH-SY5Y cells. Both forms interact with ERM and repress the transcription of PS1 in SH-SY5Y cells. The effect of both C-terminal and N-terminal deletions indicates that the N-terminal 120 amino acid region is required for interaction with ERM in yeast, and furthermore single amino acid mutations show that residues 112 and 114 play an important role. The repression of transcription in SH-SY5Y cells also appears to require the N-terminal potion of ZNF237 and was affected by mutation of the amino acid 112. Data

  4. Distribution of macrolide resistance genes erm(B) and mef(A) among 160 penicillin-intermediate clinical isolates of Streptococcus pneumoniae isolated in southern France.

    PubMed

    Marchandin, H; Jean-Pierre, H; Jumas-Bilak, E; Isson, L; Drouillard, B; Darbas, H; Carrière, C

    2001-09-01

    Two prevalent mechanisms of macrolide resistance are currently described in pneumococci: production of rRNA methylase that modify 23S ribosomal RNA resulting in MLSB phenotype, and an active efflux system resulting in M-phenotype. These two mechanisms are mediated by erm(B) and mef(A) genes respectively. Several studies reported a predominance of mef(A) gene in United-States and Canada. In European countries, erm(B) determinant is prevalent and mef(A)-mediated erythromycin resistance was recently reported in about 10% of strains in Belgium and Italy. In order to evaluate implication of mef(A) gene in pneumococci erythromycin resistance, 160 clinical isolates of S. pneumoniae with low-level of penicillin resistance and resistance to macrolides recovered between April 1999 and April 2000 were collected. These isolates were tested for their macrolide susceptibility by disc diffusion method, 155 showed the MLSB phenotype and 5 the M phenotype. Genotypic analysis was performed by erm(B) and mef(A) specific-mediated PCR: erm(B) gene was detected in 154 isolates, mef(A) gene in 5 isolates, and both genes in one strain. The phenotype seems to be well correlated to the genotyping result except for strain harboring both resistance determinants. Molecular typing of isolates harboring mef(A) gene performed by pulsed-field gel electrophoresis (PFGE) after restriction by Smal shows these strains to be epidemiologically unrelated. Our results show the predominance of the erm(B) gene in erythromycin resistant S. pneumoniae isolates. mef(A)-mediated resistance is effective in Southern France (3.7%) but this rate is the lowest published from European countries.

  5. Clostridioides difficile 630Δerm in silico and in vivo - quantitative growth and extensive polysaccharide secretion.

    PubMed

    Dannheim, Henning; Will, Sabine E; Schomburg, Dietmar; Neumann-Schaal, Meina

    2017-04-01

    Antibiotic-associated infections with Clostridioides difficile are a severe and often lethal risk for hospitalized patients, and can also affect populations without these classical risk factors. For a rational design of therapeutical concepts, a better knowledge of the metabolism of the pathogen is crucial. Metabolic modeling can provide a simulation of quantitative growth and usage of metabolic pathways, leading to a deeper understanding of the organism. Here, we present an elaborate genome-scale metabolic model of C. difficile 630Δerm. The model iHD992 includes experimentally determined product and substrate uptake rates and is able to simulate the energy metabolism and quantitative growth of C. difficile. Dynamic flux balance analysis was used for time-resolved simulations of the quantitative growth in two different media. The model predicts oxidative Stickland reactions and glucose degradation as main sources of energy, while the resulting reduction potential is mostly used for acetogenesis via the Wood-Ljungdahl pathway. Initial modeling experiments did not reproduce the observed growth behavior before the production of large quantities of a previously unknown polysaccharide was detected. Combined genome analysis and laboratory experiments indicated that the polysaccharide is an acetylated glucose polymer. Time-resolved simulations showed that polysaccharide secretion was coupled to growth even during unstable glucose uptake in minimal medium. This is accomplished by metabolic shifts between active glycolysis and gluconeogenesis which were also observed in laboratory experiments.

  6. 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

  7. 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.

  8. Anchoring of protein kinase A by ERM (ezrin-radixin-moesin) proteins is required for proper netrin signaling through DCC (deleted in colorectal cancer).

    PubMed

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

    2015-02-27

    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.

  9. Erythromycin- and copper-resistant Enterococcus hirae from marine sediment and co-transfer of erm(B) and tcrB to human Enterococcus faecalis.

    PubMed

    Pasquaroli, Sonia; Di Cesare, Andrea; Vignaroli, Carla; Conti, Giulia; Citterio, Barbara; Biavasco, Francesca

    2014-09-01

    An erythromycin-, copper- and cadmium-resistant isolate of Enterococcus hirae from marine sediment was shown to harbor the plasmid pRE25 and to co-transfer erm(B) and tcrB to Enterococcus faecalis JH2-2. These data highlight the scope for antibiotic resistance selection by the marine environment through heavy metals and its possible involvement in antibiotic-resistant enterococcal infections.

  10. 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

  11. Analysis of Mutations in Neurospora crassa ERMES Components Reveals Specific Functions Related to β-Barrel Protein Assembly and Maintenance of Mitochondrial Morphology

    PubMed Central

    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

  12. 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

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

    2013-07-01

    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.

  13. Rise of Streptococcus pneumoniae isolates containing both erm(B) and mef(E) genes from an adult tertiary care community hospital system.

    PubMed

    DiPersio, Linda P; DiPersio, Joseph R; Beach, Jacqueline A; DeFine, Linda A

    2006-08-01

    The emergence of macrolide- and lincosamide-resistant Streptococcus pneumoniae is a worldwide concern. Of particular interest is the increasing prevalence of erythromycin and clindamycin-resistant isolates containing both erm(B) and mef genes. This study determined the prevalence of erythromycin and clindamycin resistance in 596 clinical S. pneumoniae isolates from 2 adult tertiary care hospitals over a 4-year period (2001-2004). Erythromycin resistance increased from 24% to 34%, but S. pneumoniae isolates resistant to clindamycin as well as to erythromycin increased from 3% in 2001 to 15.5% in 2004 (5-fold increase). Among erythromycin-resistant isolates, those also resistant to clindamycin (MLS(B) phenotype) increased 3-fold (12.8-45%). Of forty-one erythromycin/clindamycin-resistant S. pneumoniae isolates tested, 29 (71%) contained both erm(B) and mef(E) genes. Pulsed-field gel electrophoresis performed on 28 erm(B) + mef(E) positive isolates identified 2 predominant and possibly related clones, which made up 64% of the isolates.

  14. Summary Describing Integration of ERM Methodology into Supervisory Control Framework with Software Package Documentation; Advanced Reactor Technology Milestone: M4AT-16PN2301052

    SciTech Connect

    Ramuhalli, Pradeep; Hirt, Evelyn H.; Dib, Gerges; Veeramany, Arun; Bonebrake, Christopher A.; Roy, Surajit

    2016-09-20

    This project involved the development of enhanced risk monitors (ERMs) for active components in Advanced Reactor (AdvRx) designs by integrating real-time information about equipment condition with risk monitors. Health monitoring techniques in combination with predictive estimates of component failure based on condition and risk monitors can serve to indicate the risk posed by continued operation in the presence of detected degradation. This combination of predictive health monitoring based on equipment condition assessment and risk monitors can also enable optimization of maintenance scheduling with respect to the economics of plant operation. This report summarizes PNNL’s multi-year project on the development and evaluation of an ERM concept for active components while highlighting FY2016 accomplishments. Specifically, this report provides a status summary of the integration and demonstration of the prototypic ERM framework with the plant supervisory control algorithms being developed at Oak Ridge National Laboratory (ORNL), and describes additional case studies conducted to assess sensitivity of the technology to different quantities. Supporting documentation on the software package to be provided to ONRL is incorporated in this report.

  15. MRSA Pediatric clone expressing ermC plus lnuA genes causing nosocomial transmission and healthcare workers colonization in a neonatal intensive care unit.

    PubMed

    Faccone, Diego; Togneri, Ana M; Podesta, Laura; Perez, Marcela; Gagetti, Paula; Sanchez, Susana; Romero, Graciela; Corso, Alejandra

    2014-07-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of both nosocomial and community-acquired infections. We describe an outbreak caused by the MRSA Pediatric clone expressing an unusual lincosamide resistant phenotype. Between January and May 2006, an MRSA outbreak was detected at the Neonatal Unit of Hospital Interzonal General de Agudos "Evita", Buenos Aires Province, Argentina that affected ten patients. Seven isolates from seven patients plus five MRSA recovered from health care workers (nasal carriage) were studied. Two phenotypes were observed: (i) ELCi (10), resistance to erythromycin and lincomycin and inducible resistance to clindamycin; (ii) ELiCi (2), resistance to erythromycin and inducible resistance to lincomycin and clindamycin. All 12 MRSA were resistant to oxacillin, erythromycin and gentamicin. Isolates expressing the ELCi-phenotype showed lincomycin MIC values between 16 and 32mg/L, while the remaining 2 isolates with ELiCi-phenotype presented a MIC value of 0.5mg/L. No differences were observed between the clindamycin MIC values in both phenotypes, ranging 0.25-0.5mg/L. Isolates showing ELCi-phenotype harbored ermC plus lnuA genes, and the other two only ermC gene. All 12 isolates were genetically related and belonged to the Pediatric clone (ST100) harboring a new variant of SCCmecIV. This is the first MRSA outbreak expressing an unusual ELCi phenotype due to a combination of ermC plus lnuA genes.

  16. Presence of the tet(O) gene in erythromycin- and tetracycline-resistant strains of Streptococcus pyogenes and linkage with either the mef(A) or the erm(A) gene.

    PubMed

    Giovanetti, Eleonora; Brenciani, Andrea; Lupidi, Remo; Roberts, Marilyn C; Varaldo, Pietro E

    2003-09-01

    Sixty-three recent Italian clinical isolates of Streptococcus pyogenes resistant to both erythromycin (MICs >or=1 microg/ml) and tetracycline (MICs >or= 8 microg/ml) were genotyped for macrolide and tetracycline resistance genes. We found 19 isolates carrying the mef(A) and the tet(O) genes; 25 isolates carrying the erm(A) and tet(O) genes; and 2 isolates carrying the erm(A), tet(M), and tet(O) genes. The resistance of all erm(A)-containing isolates was inducible, but the isolates could be divided into two groups on the basis of erythromycin MICs of either >128 or 1 to 4 microg/ml. The remaining 17 isolates included 15 isolates carrying the erm(B) gene and 2 isolates carrying both the erm(B) and the mef(A) genes, with all 17 carrying the tet(M) gene. Of these, 12 carried Tn916-Tn1545-like conjugative transposons. Conjugal transfer experiments demonstrated that the tet(O) gene moved with and without the erm(A) gene and with the mef(A) gene. These studies, together with the results of pulsed-field gel electrophoresis experiments and hybridization assays with DNA probes specific for the tet(O), erm(A), and mef(A) genes, suggested a linkage of tet(O) with either erm(A) or mef(A) in erythromycin- and tetracycline-resistant S. pyogenes isolates. By amplification and sequencing experiments, we detected the tet(O) gene ca. 5.5 kb upstream from the mef(A) gene. This is the first report demonstrating the presence of the tet(O) gene in S. pyogenes and showing that it may be linked with another gene and can be moved by conjugation from one chromosome to another.

  17. 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…

  18. Development and application of real-time PCR assays for quantification of erm genes conferring resistance to macrolides-lincosamides-streptogramin B in livestock manure and manure management systems.

    PubMed

    Chen, Jing; Yu, Zhongtang; Michel, Frederick C; Wittum, Thomas; Morrison, Mark

    2007-07-01

    Erythromycin and tylosin are commonly used in animal production, and such use is perceived to contribute to the overall antimicrobial resistance (AR) reservoirs. Quantitative measurements of this type of AR reservoir in microbial communities are required to understand AR ecology (e.g., emergence, persistence, and dissemination). We report here the development, validation, and use of six real-time PCR assays for quantifying six classes of erm genes (classes A through C, F, T, and X) that encode the major mechanism of resistance to macrolides-lincosamides-streptogramin B (MLS(B)). These real-time PCR assays were validated and used in quantifying the six erm classes in five types of samples, including those from bovine manure, swine manure, compost of swine manure, swine waste lagoons, and an Ekokan upflow biofilter system treating hog house effluents. The bovine manure samples were found to contain much smaller reservoirs of each of the six erm classes than the swine manure samples. Compared to the swine manure samples, the composted swine manure samples had substantially reduced erm gene abundances (by up to 7.3 logs), whereas the lagoon or the biofilter samples had similar erm gene abundances. These preliminary results suggest that the methods of manure storage and treatment probably have a substantial impact on the persistence and decline of MLS(B) resistance originating from food animals, thus likely affecting the dissemination of such resistance genes into the environment. The abundances of these erm genes appeared to be positively correlated with those of the tet genes determined previously among these samples. These real-time PCR assays provide a rapid, quantitative, and cultivation-independent measurement of six major classes of erm genes, which should be useful for ecological studies of AR.

  19. Addressing Analytical Challenges of the Environmental Monitoring for the Water Framework Directive: ERM-CE100, a New Biota Certified Reference Material.

    PubMed

    Dosis, Ioannis; Ricci, Marina; Majoros, Laszlo; Lava, Roberto; Emteborg, Håkan; Held, Andrea; Emons, Hendrik

    2017-02-21

    In the context of supporting the EU Member States in the implementation of the EU Water Framework Directive (WFD), a project for the production of a fish reference material (ERM-CE100) certified for its content of the two priority substances hexachlorobenzene (HCB) and hexachlorobutadiene (HCBD) was carried out at the Joint Research Centre (JRC) of the European Commission. The starting material was naturally contaminated Wels catfish (Silurus glanis), caught in the Ebro River (Spain). A novel approach for the processing of the fish was tested that resulted in a homogeneous and stable reference material in the form of a wet paste. The fresh-like texture of the matrix enhances the comparability of this material toward routinely analyzed environmental biota samples and facilitates its use as a quality assurance tool given that the WFD environmental quality standards (EQS) for biota are expressed as wet weight. Certified values for the mass fractions of HCB and HCBD were assigned with 120 ± 8 and 36 ± 4 μg/kg, respectively. The related interlaboratory comparison involved 13 expert laboratories applying a range of analytical methodologies. It is the first biota CRM ever available for HCBD. ERM-CE100 can be used to assess the performance of analytical methods employed in the mandatory monitoring of water bodies under the WFD, thus, providing a benchmark for establishing comparability among measurement results.

  20. The Corynebacterium xerosis composite transposon Tn5432 consists of two identical insertion sequences, designated IS1249, flanking the erythromycin resistance gene ermCX.

    PubMed

    Tauch, A; Kassing, F; Kalinowski, J; Pühler, A

    1995-09-01

    Analysis of the 50-kb R-plasmid pTP10 from the clinical isolate Corynebacterium xerosis M82B revealed that the erythromycin resistance gene, ermCX, is located on a 4524-bp composite transposable element, Tn5432. The ends of Tn5432 are identical, direct repeats of an insertion sequence, designated IS1249, encoding a putative transposase of the IS256 family. IS1249 consists of 1385 bp with 45/42 imperfect terminal inverted repeats. The nucleotide sequence of the 1754-bp Tn5432 central region is 99% identical to the previously sequenced erythromycin resistance region of the Corynebacterium diphtheriae plasmid pNG2. It encodes the erythromycin resistance gene, ermCX, and an ORF homologous to the amino-terminal end of the transposase of IS31831 from Corynebacterium glutamicum. Transposons with regions flanking the insertion sites were recovered from the C. glutamicum chromosome by a plasmid rescue technique. Insertion of Tn5432 created 8-bp target site duplications. A Tn5432-induced isoleucine/valine-auxotrophic mutant was found to carry the transposon in the 5' region of the ilvBNC cluster; in pTP10 the transposon is inserted in a region similar to replication and partitioning functions of the Enterococcus faecalis plasmid pAD1 and the Agrobacterium tumefaciens plasmid pTAR.

  1. 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

    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.

  2. 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

  3. 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.

  4. 23S rRNA domain V, a fragment that can be specifically methylated in vitro by the ErmSF (TlrA) methyltransferase.

    PubMed Central

    Kovalic, D; Giannattasio, R B; Jin, H J; Weisblum, B

    1994-01-01

    The DNA sequence that encodes 23S rRNA domain V of Bacillus subtilis, nucleotides 2036 to 2672 (C. J. Green, G. C. Stewart, M. A. Hollis, B. S. Vold, and K. F. Bott, Gene 37:261-266, 1985), was cloned and used as a template from which to transcribe defined domain V RNA in vitro. The RNA transcripts served as a substrate in vitro for specific methylation of B. subtilis adenine 2085 (adenine 2058 in Escherichia coli 23S rRNA) by the ErmSF methyltransferase, an enzyme that confers resistance to the macrolide-lincosamide-streptogramin B group of antibiotics on Streptomyces fradiae NRRL 2702, the host from which it was cloned. Thus, neither RNA sequences belonging to domains other than V nor the association of 23S rRNA with ribosomal proteins is needed for the specific methylation of adenine that confers resistance to the macrolide-lincosamide-streptogramin B group of antibiotics. Images PMID:7961463

  5. Pharmacodynamic Modeling of Clarithromycin against Macrolide-Resistant [PCR-Positive mef(A) or erm(B)] Streptococcus pneumoniae Simulating Clinically Achievable Serum and Epithelial Lining Fluid Free-Drug Concentrations

    PubMed Central

    Noreddin, Ayman M.; Roberts, Danielle; Nichol, Kim; Wierzbowski, Aleksandra; Hoban, Daryl J.; Zhanel, George G.

    2002-01-01

    The association between macrolide resistance mechanisms and clinical outcomes remains understudied. The present study, using an in vitro pharmacodynamic model, assessed clarithromycin (CLR) activity against mef(A)-positive and erm(B)-negative Streptococcus pneumoniae isolates by simulating free-drug concentrations in serum and both total (protein-bound and free) and free drug in epithelial lining fluid (ELF). Five mef(A)-positive and erm(B)-negative strains, one mef(A)-negative and erm(B)-positive strain, and a control [mef(A)-negative and erm(B)-negative] strain of S. pneumoniae were tested. CLR was modeled using a one-compartment model, simulating a dosage of 500 mg, per os, twice a day (in serum, free-drug Cp maximum of 2 μg/ml, t1/2 of 6 h; in ELF, CELF(total) maximum of 35μg/ml, t1/2 of 6 h; CELF(free) maximum of 14 μg/ml, t1/2 of 6 h). Starting inocula were 106 CFU/ml in Mueller-Hinton broth with 2% lysed horse blood. With sampling at 0, 4, 8, 12, 20, and 24 h, the extent of bacterial killing was assessed. Achieving CLR T/MIC values of ≥90% (AUC0-24/MIC ratio, ≥61) resulted in bacterial eradication, while T>MIC values of 40 to 56% (AUC0-24/MIC ratios of ≥30.5 to 38) resulted in a 1.2 to 2.0 log10 CFU/ml decrease at 24 h compared to that for the initial inoculum. CLR T/MIC values of ≤8% (AUC0-24/MIC ratio, ≤17.3) resulted in a static effect or bacterial regrowth. The high drug concentrations in ELF that were obtained clinically with CLR may explain the lack of clinical failures with mef(A)-producing S. pneumoniae strains, with MICs up to 8 μg/ml. However, mef(A) isolates for which MICs are ≥16 μg/ml along with erm(B) may result in bacteriological failures. PMID:12435719

  6. Pharmacodynamic modeling of clarithromycin against macrolide-resistant [PCR-positive mef(A) or erm(B)] Streptococcus pneumoniae simulating clinically achievable serum and epithelial lining fluid free-drug concentrations.

    PubMed

    Noreddin, Ayman M; Roberts, Danielle; Nichol, Kim; Wierzbowski, Aleksandra; Hoban, Daryl J; Zhanel, George G

    2002-12-01

    The association between macrolide resistance mechanisms and clinical outcomes remains understudied. The present study, using an in vitro pharmacodynamic model, assessed clarithromycin (CLR) activity against mef(A)-positive and erm(B)-negative Streptococcus pneumoniae isolates by simulating free-drug concentrations in serum and both total (protein-bound and free) and free drug in epithelial lining fluid (ELF). Five mef(A)-positive and erm(B)-negative strains, one mef(A)-negative and erm(B)-positive strain, and a control [mef(A)-negative and erm(B)-negative] strain of S. pneumoniae were tested. CLR was modeled using a one-compartment model, simulating a dosage of 500 mg, per os, twice a day (in serum, free-drug C(p) maximum of 2 micro g/ml, t(1/2) of 6 h; in ELF, C(ELF(total)) maximum of 35 micro g/ml, t(1/2) of 6 h; C(ELF(free)) maximum of 14 micro g/ml, t(1/2) of 6 h). Starting inocula were 10(6) CFU/ml in Mueller-Hinton broth with 2% lysed horse blood. With sampling at 0, 4, 8, 12, 20, and 24 h, the extent of bacterial killing was assessed. Achieving CLR T/MIC values of > or =90% (AUC(0-24)/MIC ratio, > or =61) resulted in bacterial eradication, while T>MIC values of 40 to 56% (AUC(0-24)/MIC ratios of > or =30.5 to 38) resulted in a 1.2 to 2.0 log(10) CFU/ml decrease at 24 h compared to that for the initial inoculum. CLR T/MIC values of < or =8% (AUC(0-24)/MIC ratio, < or =17.3) resulted in a static effect or bacterial regrowth. The high drug concentrations in ELF that were obtained clinically with CLR may explain the lack of clinical failures with mef(A)-producing S. pneumoniae strains, with MICs up to 8 micro g/ml. However, mef(A) isolates for which MICs are > or =16 micro g/ml along with erm(B) may result in bacteriological failures.

  7. 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

  8. 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

  9. Aurora-B and Rho-kinase/ROCK, the two cleavage furrow kinases, independently regulate the progression of cytokinesis: possible existence of a novel cleavage furrow kinase phosphorylates ezrin/radixin/moesin (ERM).

    PubMed

    Yokoyama, Tomoya; Goto, Hidemasa; Izawa, Ichiro; Mizutani, Hitoshi; Inagaki, Masaki

    2005-02-01

    Cytokinesis is regulated by several protein kinases, such as Aurora-B and Rho-kinase/ROCK. We have indicated that these two kinases are the cleavage furrow (CF) kinases that accumulate at the cleavage furrow and phosphorylate several intermediate filament (IF) proteins into two daughter cells. It has been reported that Aurora-B phosphorylates MgcRacGAP to functionally convert to a RhoGAP during cytokinesis. Therefore, we investigated here the relationship between Aurora-B and Rho-kinase/ROCK in cytokinesis, by using small interfering RNA (siRNA) technique. Aurora-B depletion did not alter the cleavage furrow-specific localization of Rho-kinase/ROCK and vice versa. Treatment of Aurora-B or Rho-kinase/ROCK siRNA increased multinucleate cells, and the effect of double depletion was additive. Aurora-B depletion induced the reduction of cleavage furrow-specific phosphorylation of vimentin at Ser72 but not vimentin at Ser71, myosin light chain (MLC) at Ser19, and myosin binding subunit of myosin phosphatase (MBS) at Ser852. In contrast, Rho-kinase/ROCK depletion led to the reduction of cleavage furrow-specific phosphorylation of MLC at Ser19, MBS at Ser852, and vimentin at Ser71 but not vimentin at Ser72. Cleavage furrow-specific ezrin/radixin/moesin (ERM) phosphorylation was not altered in the Aurora-B- and/or Rho-kinase/ROCK-depleted cells. In addition, C3 or toxin B treatment did not abolish ERM phosphorylation at the cleavage furrow in cells attaining cytokinesis. These results suggest that Aurora-B and Rho-kinase/ROCK regulate the progression of cytokinesis without communicating to each other, and there may exist a novel protein kinase which phosphorylates ERM at the cleavage furrow.

  10. Predictors of one-year mortality at hospital discharge after acute coronary syndromes: A new risk score from the EPICOR (long-tErm follow uP of antithrombotic management patterns In acute CORonary syndrome patients) study

    PubMed Central

    Pocock, Stuart; Bueno, Héctor; Licour, Muriel; Medina, Jesús; Zhang, Lin; Annemans, Lieven; Danchin, Nicholas; Huo, Yong; Van de Werf, Frans

    2015-01-01

    Aims: A reliable prediction tool is needed to identify acute coronary syndrome (ACS) patients with high mortality risk after their initial hospitalization. Methods: EPICOR (long-tErm follow uP of antithrombotic management patterns In acute CORonary syndrome patients: NCT01171404) is a prospective cohort study of 10,568 consecutive hospital survivors after an ACS event (4943 ST-segment elevation myocardial infarction (STEMI) and 5625 non-ST-elevation ACS (NSTE-ACS)). Of these cases, 65.1% underwent percutaneous coronary intervention (PCI) and 2.5% coronary artery bypass graft (CABG). Post-discharge mortality was recorded for up to two years. From over 50 potential predictor variables a new risk score for one-year mortality was developed using forward stepwise Cox regression, and examined for goodness-of-fit, discriminatory power, and external validation. Results: A total of 407 patients (3.9%) died within one year of discharge. We identified 12 highly significant independent predictors of mortality (in order of predictive strength): age, lower ejection fraction, poorer EQ-5D quality of life, elevated serum creatinine, in-hospital cardiac complications, chronic obstructive pulmonary disease, elevated blood glucose, male gender, no PCI/CABG after NSTE-ACS, low hemoglobin, peripheral artery disease, on diuretics at discharge. When combined into a new risk score excellent discrimination was achieved (c-statistic=0.81) and this was also validated on a large similar cohort (9907 patients) in Asia (c=0.78). For both STEMI and NSTE-ACS there was a steep gradient in one-year mortality ranging from 0.5% in the lowest quintile to 18.2% in the highest decile. NSTE-ACS contributes over twice as many high-risk patients as STEMI. Conclusions: Post-discharge mortality for ACS patients remains of concern. Our new user-friendly risk score available on www.acsrisk.org can readily identify who is at high risk. PMID:25301783

  11. 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…

  12. Barriers to Electronic Records Management (ERM): An Exploratory Case Study Investigating ERM in the Deployed Environment During Operations Enduring Freedom and Iraqi Freedom

    DTIC Science & Technology

    2005-03-01

    Patterson, G., & Sprehe, J. T. (2002). Principal challenges facing electronic records management in federal agencies today. Government Information Quarterly , 19...factors in managing U.S. federal recordkeeping. Government Information Quarterly , 19(3), 289-305. Stake, R. E. (1995). The art of case study

  13. 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 ...

  14. 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).

  15. Are ERM (ezrin/radixin/moesin) proteins targets for autoantibodies in demyelinating neuropathies?

    PubMed

    Miyaji, Kazuki; Shahrizaila, Nortina; Umapathi, Thirugnanam; Chan, Yee-Cheun; Hirata, Koichi; Yuki, Nobuhiro

    2014-11-01

    Ezrin, radixin and moesin, which are strongly expressed in the Schwann cell microvilli, are putative targets for autoantibodies in acute or chronic inflammatory demyelinating polyneuropathy (AIDP or CIDP). An association between anti-moesin IgG antibodies and cytomegalovirus-related AIDP has been postulated. None of 41 AIDP patients, including 8 cytomegalovirus-related AIDP patients, and 23 CIDP had IgG or IgM antibodies to ezrin, radixin and moesin; whereas, one patient with cytomegalovirus-related AIDP had anti-ezrin IgM antibodies. Ezrin, radixin and moesin are unlikely targets for autoantibodies in AIDP and CIDP, and the association of anti-moesin antibodies with cytomegalovirus-related AIDP was not confirmed.

  16. 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. ...

  17. An Initial Model for Generative Design Research: Bringing Together Generative Focus Group (GFG) and Experience Reflection Modelling (ERM)

    ERIC Educational Resources Information Center

    Bakirlioglu, Yekta; Ogur, Dilruba; Dogan, Cagla; Turhan, Senem

    2016-01-01

    Understanding people's experiences and the context of use of a product at the earliest stages of the design process has in the last decade become an important aspect of both the design profession and design education. Generative design research helps designers understand user experiences, while also throwing light on their current needs,…

  18. A simplified erythromycin resistance cassette for Treponema denticola mutagenesis

    PubMed Central

    Goetting-Minesky, M. Paula; Fenno, J. Christopher

    2010-01-01

    The primary selectable marker for genetic studies of Treponema denticola is a hybrid gene cassette containing both ermF and ermAM (ermB) genes. ErmB functions in Escherichia coli, while ErmF has been assumed to confer resistance in T. denticola. We demonstrate here that ErmB is sufficient for erythromycin selection in T. denticola and that the native ermB promoter drives ErmB expression. PMID:20691222

  19. 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.

  20. Increasing Range and Lethality of Extended-Range Munitions (ERMS) using Numerical Weather Prediction (NWP) and the AUV Workbench to Compute a Ballistic Correction (BALCOR)

    DTIC Science & Technology

    2006-12-01

    Environmental effects have a direct impact on the ability of weapons to reach their intended target. One example of this impact was during the dust storm that... dust storm , and were able to make changes to planned weapons loads, favoring GPS-guided munitions over laser guided munitions (Anderson, 2004). The...2004). An Analysis of a Dust Storm impacting Operation Iraqi Freedom, 25-27 March 2003. (M.S. Naval Postgraduate School). Annati, M. (2003

  1. The Role of Drosophila Merlin in the Control of Mitosis Exit and Development

    DTIC Science & Technology

    2006-07-01

    proteins, erm1a and erm1b of Caenorhabditis elegans [51], may represent different transcript variants of the same gene (also see below). Furthermore, we...merlin-like A. gambiae fragment of merlin A. mellifera similar to schwannomin D. melanogaster merlin C . elegans nfm 1a C . briggsae merlin-like B. malayi...rubripes moesin C . intestinalis erm-like B. glabrata erm-like L. variegates moesin C . elegans erm-like 1a C . briggsae erm-like D. melanogaster

  2. The Expression of Antibiotic Resistance Methyltransferase Correlates with mRNA Stability Independently of Ribosome Stalling

    PubMed Central

    Dzyubak, Ekaterina

    2016-01-01

    Members of the Erm methyltransferase family modify 23S rRNA of the bacterial ribosome and render cross-resistance to macrolides and multiple distantly related antibiotics. Previous studies have shown that the expression of erm is activated when a macrolide-bound ribosome stalls the translation of the leader peptide preceding the cotranscribed erm. Ribosome stalling is thought to destabilize the inhibitory stem-loop mRNA structure and exposes the erm Shine-Dalgarno (SD) sequence for translational initiation. Paradoxically, mutations that abolish ribosome stalling are routinely found in hyper-resistant clinical isolates; however, the significance of the stalling-dead leader sequence is largely unknown. Here, we show that nonsense mutations in the Staphylococcus aureus ErmB leader peptide (ErmBL) lead to high basal and induced expression of downstream ErmB in the absence or presence of macrolide concomitantly with elevated ribosome methylation and resistance. The overexpression of ErmB is associated with the reduced turnover of the ermBL-ermB transcript, and the macrolide appears to mitigate mRNA cleavage at a site immediately downstream of the ermBL SD sequence. The stabilizing effect of antibiotics on mRNA is not limited to ermBL-ermB; cationic antibiotics representing a ribosome-stalling inducer and a noninducer increase the half-life of specific transcripts. These data unveil a new layer of ermB regulation and imply that ErmBL translation or ribosome stalling serves as a “tuner” to suppress aberrant production of ErmB because methylated ribosome may impose a fitness cost on the bacterium as a result of misregulated translation. PMID:27645242

  3. 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...

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

    PubMed

    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.

  5. What Is Rhabdomyosarcoma?

    MedlinePlus

    ... The cells of ERMS look like the developing muscle cells of a 6- to 8-week-old embryo. ERMS tends to occur in the head and neck area, bladder, vagina, or in or around the prostate and testicles. Two subtypes of ERMS, botryoid and spindle cell rhabdomyosarcomas, tend to have a better prognosis ( ...

  6. 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,…

  7. Embryonal Rhabdomyosarcoma Occurring on Mandibular Gingiva in an Adult

    PubMed Central

    Patil, Gururaj; Halawar, Sangamesh; Sagari, Shitalkumar; Babannavar, Roopa; Purohit, Sharad

    2013-01-01

    An embryonal rhabdomyosarcoma (ERMS) is a primitive, malignant, soft tissue sarcoma that recapitulates the phenotypic and biological features of embryonic skeletal muscles. Occurrence of intraoral ERMS in adults is extremely rare. This unique case report highlights the clinical, radiographic, histopathological and immunohistochemical findings of an intraoral ERMS. PMID:24179953

  8. 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

  9. 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.

  10. Separating mitochondrial protein assembly and endoplasmic reticulum tethering by selective coupling of Mdm10.

    PubMed

    Ellenrieder, Lars; Opaliński, Łukasz; Becker, Lars; Krüger, Vivien; Mirus, Oliver; Straub, Sebastian P; Ebell, Katharina; Flinner, Nadine; Stiller, Sebastian B; Guiard, Bernard; Meisinger, Chris; Wiedemann, Nils; Schleiff, Enrico; Wagner, Richard; Pfanner, Nikolaus; Becker, Thomas

    2016-10-10

    The endoplasmic reticulum-mitochondria encounter structure (ERMES) connects the mitochondrial outer membrane with the ER. Multiple functions have been linked to ERMES, including maintenance of mitochondrial morphology, protein assembly and phospholipid homeostasis. Since the mitochondrial distribution and morphology protein Mdm10 is present in both ERMES and the mitochondrial sorting and assembly machinery (SAM), it is unknown how the ERMES functions are connected on a molecular level. Here we report that conserved surface areas on opposite sides of the Mdm10 β-barrel interact with SAM and ERMES, respectively. We generated point mutants to separate protein assembly (SAM) from morphology and phospholipid homeostasis (ERMES). Our study reveals that the β-barrel channel of Mdm10 serves different functions. Mdm10 promotes the biogenesis of α-helical and β-barrel proteins at SAM and functions as integral membrane anchor of ERMES, demonstrating that SAM-mediated protein assembly is distinct from ER-mitochondria contact sites.

  11. 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.

  12. Spontaneous release of epiretinal membrane in a young weight-lifting athlete by presumed central rupture and centrifugal pull

    PubMed Central

    Mansour, Ahmad M; Mansour, Hana A; Arevalo, J Fernando

    2014-01-01

    This patient presented for surgery at the age of 32 years, 14 months after his initial complaint of metamorphopsia and visual loss in the right eye. Past tests demonstrated a whitish epiretinal membrane (ERM) with translucent stress lines over a thickened macula. Visual acuity was found on last presentation to be normal with minimal alteration on Amsler grid testing. A torn ERM was found in the center with left-over ERM temporally and rolled-over ERM nasally at the site of the epicenter with no posterior vitreous detachment. Visual recovery occurred gradually over several days 2 months prior to presentation apparently following heavy weight-lifting with a sensation of severe eye pressure. Sequential funduscopy and optical coherence tomography scans demonstrated the peeling of an ERM accompanied by normalization of foveal thickness. Valsalva maneuver had put excessive tension on ERM which tore in its center at the weakest line with gradual contraction of the ERM away from the fovea towards the peripapillary area. This is a new mechanism of self-separation of ERM induced by Valsalva. ERM in young subjects is subject to rupture and subsequent separation by tangential traction. There are three mechanisms for spontaneous separation of ERM: 1) posterior vitreous detachment with pulling of ERM by detaching vitreous (most common in adults); 2) the contracting forces of the immature ERM become stronger than its adhesions to the retina resulting in slow tangential traction on the edges of the ERM and gradual separation from the edges towards the center (remodeling common in youngsters); and 3) acute tearing of ERM at its weakest central point and retraction of part of the membrane towards the epicenter (current case report). PMID:25484573

  13. [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.

  14. Paternal uniparental disomy with segmental loss of heterozygosity of chromosome 11 are hallmark characteristics of syndromic and sporadic embryonal rhabdomyosarcoma.

    PubMed

    Robbins, Katherine M; Stabley, Deborah L; Holbrook, Jennifer; Sahraoui, Rebecca; Sadreameli, Alexa; Conard, Katrina; Baker, Laura; Gripp, Karen W; Sol-Church, Katia

    2016-12-01

    Costello syndrome (CS) arises from a typically paternally derived germline mutation in the proto-oncogene HRAS, and is considered a rasopathy. CS results in failure-to-thrive, intellectual disabilities, short stature, coarse facial features, skeletal abnormalities, congenital heart disease, and a predisposition for cancer, most commonly embryonal rhabdomyosarcoma (ERMS). The goal of this study was to characterize CS ERMS at the molecular level and to determine how divergent it is from sporadic ERMS. We characterized eleven ERMS tumors from eight unrelated CS patients, carrying paternally derived HRAS c.34G>A (p.Gly12Ser; 6) or c.35G>C (p.Gly12Ala; 2) mutations. Loss of heterozygosity (LOH) was evaluated in all CS ERMS by microarray and/or short tandem repeat (STR) markers spanning the entire chromosome 11. Eight CS ERMS tumors displayed complete paternal uniparental disomy of chromosome 11 (pUPD11), whereas two displayed UPD only at 11p and a second primary ERMS tumor showed UPD limited to 11p15.5, the classical hallmark for ERMS. Three sporadic ERMS cell lines (RD, Rh36, Rh18) and eight formalin fixed paraffin embedded (FFPE) ERMS tumors were also analyzed for RAS mutations and LOH status. We found a higher than anticipated frequency of RAS mutations (HRAS or NRAS; 50%) in sporadic ERMS cell lines/tumors. Unexpectedly, complete uniparental disomy (UPD11) was observed in five specimens, while the other six showed LOH extending across the p and q arms of chromosome 11. In this study, we are able to clearly demonstrate complete UPD11 in both syndromic and sporadic ERMS. © 2016 Wiley Periodicals, Inc.

  15. Paternal Uniparental Disomy with Segmental Loss of Heterozygosity of Chromosome 11 are Hallmark Characteristics of Syndromic and Sporadic Embryonal Rhabdomyosarcoma

    PubMed Central

    Robbins, Katherine M.; Stabley, Deborah L.; Holbrook, Jennifer; Sahraoui, Rebecca; Sadreameli, Alexa; Conard, Katrina; Baker, Laura; Gripp, Karen W.; Sol-Church, Katia

    2016-01-01

    Costello syndrome (CS) arises from a typically paternally derived germline mutation in the proto-oncogene HRAS, and is considered a rasopathy. CS results in failure-to-thrive, intellectual disabilities, short stature, coarse facial features, skeletal abnormalities, congenital heart disease, and a predisposition for cancer, most commonly embryonal rhabdomyosarcoma (ERMS). The goal of this study was to characterize CS ERMS at the molecular level and to determine how divergent it is from sporadic ERMS. We characterized eleven ERMS tumors from eight unrelated CS patients, carrying paternally derived HRAS c.34G>A (p.Gly12Ser; 6) or c.35G>C (p.Gly12Ala; 2) mutations. Loss of heterozygosity (LOH) was evaluated in all CS ERMS by microarray and/or short tandem repeat (STR) markers spanning the entire chromosome 11. Eight CS ERMS tumors displayed complete paternal uniparental disomy of chromosome 11 (pUPD11), whereas two displayed UPD only at 11p and a second primary ERMS tumor showed UPD limited to 11p15.5, the classical hallmark for ERMS. Three sporadic ERMS cell lines (RD, Rh36, Rh18) and eight formalin fixed paraffin embedded (FFPE) ERMS tumors were also analyzed for RAS mutations and LOH status. We found a higher than anticipated frequency of RAS mutations (HRAS or NRAS; 50%) in sporadic ERMS cell lines/tumors. Unexpectedly, complete uniparental disomy (UPD11) was observed in five specimens, while the other six showed LOH extending across the p and q arms of chromosome 11. In this study, we are able to clearly demonstrate complete UPD11 in both syndromic and sporadic ERMS. PMID:27589201

  16. 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.

  17. Comparison of Gene Expression Profile of Epiretinal Membranes Obtained from Eyes with Proliferative Vitreoretinopathy to That of Secondary Epiretinal Membranes

    PubMed Central

    Asato, Ryo; Yoshida, Shigeo; Ogura, Atsushi; Nakama, Takahito; Ishikawa, Keijiro; Nakao, Shintaro; Sassa, Yukio; Enaida, Hiroshi; Oshima, Yuji; Ikeo, Kazuho; Gojobori, Takashi; Kono, Toshihiro; Ishibashi, Tatsuro

    2013-01-01

    Background Proliferative vitreoretinopathy (PVR) is a destructive complication of retinal detachment and vitreoretinal surgery which can lead to severe vision reduction by tractional retinal detachments. The purpose of this study was to determine the gene expression profile of epiretinal membranes (ERMs) associated with a PVR (PVR-ERM) and to compare it to the expression profile of less-aggressive secondary ERMs. Methodology/Principal Findings A PCR-amplified complementary DNA (cDNA) library was constructed using the RNAs isolated from ERMs obtained during vitrectomy. The sequence from the 5′ end was obtained for randomly selected clones and used to generate expressed sequence tags (ESTs). We obtained 1116 nonredundant clusters representing individual genes expressed in PVR-ERMs, and 799 clusters representing the genes expressed in secondary ERMs. The transcriptome of the PVR-ERMs was subdivided by functional subsets of genes related to metabolism, cell adhesion, cytoskeleton, signaling, and other functions, by FatiGo analysis. The genes highly expressed in PVR-ERMs were compared to those expressed in the secondary ERMs, and these were subdivided by cell adhesion, proliferation, and other functions. Querying 10 cell adhesion-related genes against the STRING database yielded 70 possible physical relationships to other genes/proteins, which included an additional 60 genes that were not detected in the PVR-ERM library. Of these, soluble CD44 and soluble vascular cellular adhesion molecule-1 were significantly increased in the vitreous of patients with PVR. Conclusions/Significance Our results support an earlier hypothesis that a PVR-ERM, even from genomic points of view, is an aberrant form of wound healing response. Genes preferentially expressed in PVR-ERMs may play an important role in the progression of PVR and could be served as therapeutic targets. PMID:23372684

  18. 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

  19. Molecular Basis of Intrinsic Macrolide Resistance in the Mycobacterium tuberculosis Complex

    PubMed Central

    Buriánková, Karolína; Doucet-Populaire, Florence; Dorson, Olivier; Gondran, Anne; Ghnassia, Jean-Claude; Weiser, Jaroslav; Pernodet, Jean-Luc

    2004-01-01

    The intrinsic resistance of the Mycobacterium tuberculosis complex (MTC) to most antibiotics, including macrolides, is generally attributed to the low permeability of the mycobacterial cell wall. However, nontuberculous mycobacteria (NTM) are much more sensitive to macrolides than members of the MTC. A search for macrolide resistance determinants within the genome of M. tuberculosis revealed the presence of a sequence encoding a putative rRNA methyltransferase. The deduced protein is similar to Erm methyltransferases, which confer macrolide-lincosamide-streptogramin (MLS) resistance by methylation of 23S rRNA, and was named ErmMT. The corresponding gene, ermMT (erm37), is present in all members of the MTC but is absent in NTM species. Part of ermMT is deleted in some vaccine strains of Mycobacterium bovis BCG, such as the Pasteur strain, which lack the RD2 region. The Pasteur strain was susceptible to MLS antibiotics, whereas MTC species harboring the RD2 region were resistant to them. The expression of ermMT in the macrolide-sensitive Mycobacterium smegmatis and BCG Pasteur conferred MLS resistance. The resistance patterns and ribosomal affinity for erythromycin of Mycobacterium host strains expressing ermMT, srmA (monomethyltransferase from Streptomyces ambofaciens), and ermE (dimethyltransferase from Saccharopolyspora erythraea) were compared, and the ones conferred by ErmMT were similar to those conferred by SrmA, corresponding to the MLS type I phenotype. These results suggest that ermMT plays a major role in the intrinsic macrolide resistance of members of the MTC and could be the first example of a gene conferring resistance by target modification in mycobacteria. PMID:14693532

  20. Molecular basis of intrinsic macrolide resistance in the Mycobacterium tuberculosis complex.

    PubMed

    Buriánková, Karolína; Doucet-Populaire, Florence; Dorson, Olivier; Gondran, Anne; Ghnassia, Jean-Claude; Weiser, Jaroslav; Pernodet, Jean-Luc

    2004-01-01

    The intrinsic resistance of the Mycobacterium tuberculosis complex (MTC) to most antibiotics, including macrolides, is generally attributed to the low permeability of the mycobacterial cell wall. However, nontuberculous mycobacteria (NTM) are much more sensitive to macrolides than members of the MTC. A search for macrolide resistance determinants within the genome of M. tuberculosis revealed the presence of a sequence encoding a putative rRNA methyltransferase. The deduced protein is similar to Erm methyltransferases, which confer macrolide-lincosamide-streptogramin (MLS) resistance by methylation of 23S rRNA, and was named ErmMT. The corresponding gene, ermMT (erm37), is present in all members of the MTC but is absent in NTM species. Part of ermMT is deleted in some vaccine strains of Mycobacterium bovis BCG, such as the Pasteur strain, which lack the RD2 region. The Pasteur strain was susceptible to MLS antibiotics, whereas MTC species harboring the RD2 region were resistant to them. The expression of ermMT in the macrolide-sensitive Mycobacterium smegmatis and BCG Pasteur conferred MLS resistance. The resistance patterns and ribosomal affinity for erythromycin of Mycobacterium host strains expressing ermMT, srmA (monomethyltransferase from Streptomyces ambofaciens), and ermE (dimethyltransferase from Saccharopolyspora erythraea) were compared, and the ones conferred by ErmMT were similar to those conferred by SrmA, corresponding to the MLS type I phenotype. These results suggest that ermMT plays a major role in the intrinsic macrolide resistance of members of the MTC and could be the first example of a gene conferring resistance by target modification in mycobacteria.

  1. Presence and distribution of Macrolides-Lincosamide-Streptogramin resistance genes and potential indicator ARGs in the university ponds in Guangzhou, China.

    PubMed

    Wang, Mianzhi; Sun, Jing; Zhong, Weixin; Xiong, Wenguang; Zeng, Zhenling; Sun, Yongxue

    2016-11-01

    This study aimed to determine the occurrence, abundance, and variation of seven Macrolides-Lincosamide-Streptogramin (MLS) resistance genes (ereB, ermA, ermB, ermF, mefA, vatB, mphA) and six potential indicator ARGs (tet (B), sul1, qnrS, fexA, IntI1, ermB) from three ponds at university by quantitative PCR and assess the impacts on the surroundings. Solid samples (fish feces, soil and sediment) and water samples were tested. All the genes were found at low levels in soil samples. For the MLS resistance genes, only two MLS genes (ermB, ermF) were detected in all samples and significant correlations between ermB and Σ MLS (R = 0.91 in solid samples; R = 0.86 in water samples, p < 0.01) were found. For the potential indicators, intl1 and sul1 were present at high levels in the three different ponds while the other genes showed varying levels. These findings show that the ermB gene can probably be served as an indicator to evaluate the overall level of MLS resistance genes. The fairly low abundance of all the tested resistance genes in soil samples and the moderate levels in other samples suggests that the university ponds kept a good state and did not have a significant impact on their surroundings.

  2. Regulation of flagellum biosynthesis within the fish pathogen Yersinia ruckeri

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Yersinia ruckeri, a Gram negative Enterobacterium, is the causative agent of enteric red mouth disease (ERM) within farmed rainbow trout (Oncorhynchus mykiss, Walbaum). There has been an increase of ERM outbreaks in previously vaccinated trout caused by a recently emerged, non-motile variant of Y. r...

  3. 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

  4. 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…

  5. 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…

  6. "They Will Post a Law about Playing Soccer" and Other Ethnic/Racial Microaggressions in Organized Activities Experienced by Mexican-Origin Families

    ERIC Educational Resources Information Center

    Lin, Alex R.; Menjívar, Cecilia; Vest Ettekal, Andrea; Simpkins, Sandra D.; Gaskin, Erin R.; Pesch, Annelise

    2016-01-01

    Organized activities have been found to provide positive experiences for Latino adolescents to develop confidence and learn critical life skills; however, these programs are sometimes a context where youth encounter negative experiences related to ethnic/racial microaggressions (ERMs). This qualitative study explores the types of ERMs that…

  7. 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…

  8. 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…

  9. Enterprise Risk Management Solutions: A Case Study

    DTIC Science & Technology

    2008-06-01

    MARKETING MIX .......................................................................................37...product, pricing, distribution, and communications, marketing mix and research [10]. B. BUSINESS OBJECTIVES The first task that faces ERMS as it...Mailers may have to be eliminated if Albert finds that the value they bring to ERMS is not significant. 37 F. MARKETING MIX After determining

  10. 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…

  11. 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

  12. [Investigation of macrolide, lincosamide and streptogramin B resistance in Staphylococcus aureus strains isolated from clinical samples by phenotypical and genotypical methods].

    PubMed

    Aydeniz Ozansoy, Fatma; Cevahir, Nural; Kaleli, İlknur

    2015-01-01

    Staphylococcus aureus is one of the most common cause of both community and healthcare-associated infections. As staphylococci have developed resistance to various antibiotics, initially to penicillins then to methicillin and glycopeptides and have the ability to cause epidemics, they continue to be a major problem from past to present. Methicillin resistance gave rise to the use of alternative antibiotics such as macrolides, however worldwide development of macrolide resistance limited the use of these antibiotics. Macrolide resistance occurs either through target site modification (MLS(B) phenotype, encoded by erm genes), efflux pumps (MS phenotype, encoded by msrA/B genes) or decreased cell wall permeability. The aim of this study was to investigate the MLS(B) resistance of clinical S.aureus strains with phenotypic and genotypic methods. A total of 404 S.aureus strains isolated from different clinical samples (50% wound, 15% tracheal aspirate and 35% other samples) of inpatients (93.3%) and outpatients (6.7%) were included in the study. Double disc synergy test (D-test) was used for the phenotypical research and PCR was used for the genotypical research of MLS(B) resistance of isolates. One hundred fifty eight (39.1%) of the S.aureus isolates were methicillin-resistant (MRSA), and 246 (60.9%) were methicillin-susceptible (MSSA). By the use of D-test, constitutive (cMLS(B)) and inducible (iMLS(B)) clindamycin resistance were detected in 19 and 111 isolates, respectively, while five isolates were MS phenotype and 268 isolates were S phenotype (susceptible to erythromycin and clindamycin). The resistance genes of 136 isolates with MLS(B) resistance phenotype were determined genotypically and among 111 isolates showing iMLS(B) phenotype ermA gene was found in 81.9% (83 MRSA, 8 MSSA), ermC gene in 10.8% (7 MRSA, 5 MSSA), msrA gene in 10.8% (11 MRSA, 1 MSSA), msrB gene in 1.8% (2 MRSA) and ermB gene in 0.9% (1 MRSA). Among 19 strains with cMLS(B) phenotype, ermA was

  13. Ezrin, radixin, and moesin are phosphorylated in response to 2-methoxyestradiol and modulate endothelial hyperpermeability.

    PubMed

    Bogatcheva, Natalia V; Zemskova, Marina A; Gorshkov, Boris A; Kim, Kyung Mi; Daglis, Gregory A; Poirier, Christophe; Verin, Alexander D

    2011-12-01

    We showed previously that microtubule disruptor 2-methoxyestradiol (2ME) induces hyperpermeability of the endothelial monolayer via mechanisms that include the activation of p38 and Rho kinase (ROCK) and rearrangement of the actin cytoskeleton. Using the protein kinase C (PKC) inhibitors Ro-31-7549 and Ro-32-0432, we show in vitro and in vivo that 2ME-induced barrier dysfunction is also PKC-dependent. The known PKC substrates ezrin, radixin, and moesin (ERM) were recently implicated in the regulation of endothelial permeability. This study tested the hypotheses that ERM proteins are phosphorylated in response to 2ME, and that this phosphorylation is involved in 2ME-induced barrier dysfunction. We show that the application of 2ME leads to a dramatic increase in the level of ERM phosphorylation. This increase is attenuated in cells pretreated with the microtubule stabilizer taxol. In human pulmonary artery endothelial cells (HPAECs), the phosphorylation of ERM occurs in a p38-dependent and PKC-dependent manner. The activation of p38 appears to occur upstream from the activation of PKC, in response to 2ME. Phosphorylated ERM are localized at the cell periphery during the early phase of response to 2ME (15 minutes), and colocalize with F-actin branching points during the later phase of response (60 minutes). Using the short interfering RNA approach, we also showed that individual ERM depletion significantly attenuates 2ME-induced hyperpermeability. HPAEC monolayers, depleted of ERM proteins and monolayers, overexpressing phosphorylation-deficient ERM mutants, exhibit less attenuation of 2ME-induced barrier disruption in response to the PKC inhibitor Ro-31-7549. These results suggest a critical role of PKC activation in response to microtubule-disrupting agents, and implicate the phosphorylation of ERM in the barrier dysfunction induced by 2ME.

  14. [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.

  15. 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).

  16. Extendable retractable telescopic mast for deployable structures

    NASA Technical Reports Server (NTRS)

    Schmid, M.; Aguirre, M.

    1986-01-01

    The Extendable and Retractable Mast (ERM) which is presently developed by Dornier in the frame of an ESA-contract, will be used to deploy and retract large foldable structures. The design is based on a telescopic carbon-fiber structure with high stiffness, strength and pointing accuracy. To verify the chosen design, a breadboard model of an ERM was built and tested under thermal vacuum (TV)-conditions. It is planned as a follow-on development to manufacture and test an Engineering Model Mast. The Engineering Model will be used to establish the basis for an ERM-family covering a wide range of requirements.

  17. Positive esophageal proximal resection margin: an important prognostic factor for esophageal cancer that warrants adjuvant therapy

    PubMed Central

    Wang, Yun-Cang; Deng, Han-Yu; Wang, Wen-Ping; He, Du; Ni, Peng-Zhi; Hu, Wei-Peng; Wang, Zhi-Qiang

    2016-01-01

    Background Positive esophageal proximal resection margin (ERM+) following esophagectomy was considered as incomplete or R1 resection. The clinicopathological data and long-term prognosis of esophageal cancer (EC) patients with ERM+ after esophagectomy were still unknown. Therefore, the aim of this study was to assess the clinical significance of ERM+ and its therapeutic option. Methods From November 2008 to December 2014, 3,594 patients with histologically confirmed EC underwent radical resection in our department. Among them there were 37 patients (1.03%) who had ERM+. ERM+ was defined as carcinoma or atypical hyperplasia (severe or moderate) at the residual esophageal margin in our study. For comparison, another 74 patients with negative esophageal proximal resection margin (ERM−) were propensity-matched at a ratio of 1:2 as control group according to sex, age, tumor location and TNM staging. The relevant prognostic factors were investigated by univariate and multivariate regression analysis. Results In this large cohort of patients, the rate of ERM+ was 1.03%. The median survival time was 35.000 months in patients with ERM+, significantly worse than 68.000 months in those with ERM− (Chi-square =4.064, P=0.044). Survival in patients with esophageal residual atypical hyperplasia (severe or moderate) was similar to those with esophageal residual carcinoma. Survival rate in stage I–II was higher than that in stage III–IV (Chi-square =27.598, P=0.000) in ERM−; But there was no difference between the two subgroups of patients in ERM+. Furthermore, in those patients with ERM+, survival was better in those who having adjuvant therapy, compared to those without adjuvant therapy (Chi-square =5.480, P=0.019). And the average survival time which was improved to a well situation for ERM+ patients who have adjuvant therapy was 68.556 months which is comparable to average survival time (65.815 months) of ERM− for those patients who are at earlier stages

  18. Combined EMD-sLORETA Analysis of EEG Data Collected during a Contour Integration Task

    PubMed Central

    Al-Subari, Karema; Tomé, Ana Maria; Volberg, Gregor; Ludwig, Bernd; Lang, Elmar W.

    2016-01-01

    Lately, Ensemble Empirical Mode Decomposition (EEMD) techniques receive growing interest in biomedical data analysis. Event-Related Modes (ERMs) represent features extracted by an EEMD from electroencephalographic (EEG) recordings. We present a new approach for source localization of EEG data based on combining ERMs with inverse models. As the first step, 64 channel EEG recordings are pooled according to six brain areas and decomposed, by applying an EEMD, into their underlying ERMs. Then, based upon the problem at hand, the most closely related ERM, in terms of frequency and amplitude, is combined with inverse modeling techniques for source localization. More specifically, the standardized low resolution brain electromagnetic tomography (sLORETA) procedure is employed in this work. Accuracy and robustness of the results indicate that this approach deems highly promising in source localization techniques for EEG data. PMID:27936219

  19. Sheehan syndrome

    MedlinePlus

    ... GJ, Sibley CP, Jauniaux ERM. Placental anatomy and physiology. In: Gabbe SG, Niebyl JR, Simpson JL, et ... 2017:chap 1. Kaiser U, Ho KKY. Pitutary physiology and diagnostic evaluation. In: Melmed S, Polonsky KS, Larsen ...

  20. A Case of Secondary Epiretinal Membrane Spontaneous Release

    PubMed Central

    Andreev, Andrey N.; Bushuev, Alexey V.

    2016-01-01

    Purpose. To report a rare case of secondary epiretinal membrane (ERM) spontaneous separation with subsequent visual restoration. Case Summary. We are reporting a case with the history of branch retinal vein occlusion, peripheral retinal neovascularization, and retinal photocoagulation. Our examination revealed secondary ERM associated with relatively high visual acuity (0.6), and a watchful waiting strategy was chosen. During the follow-up, slight visual deterioration, progressive deformation of the retinal profile, and an increase in diffuse retinal edema were observed. No surgical or laser treatment was performed. On the next visit, the spontaneous ERM separation with residual parapapillary fixation, the increase in visual acuity (0.9), and the decrease in retinal thickness were revealed. Conclusion. Such cases present additional evidence to a deferral surgical strategy for the management of patients with ERM and relatively high visual acuity. PMID:27872778

  1. 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...

  2. Mean Sea Surface and Variability of the Gulf of Mexico Using Geosat Altimetry Data

    DTIC Science & Technology

    1990-03-15

    Geosat Exact Repeat Mission (ERM) altimetric measurements of the sea surface height in the Gulf of Mexico are used to determine the mean sea surface... Gulf of Mexico . Keywords: Altimetry; Mesoscale oceanography; Ocean forecasting; Reprints.

  3. Development of a risk-based environmental management tool for drilling discharges. Summary of a four-year project.

    PubMed

    Singsaas, Ivar; Rye, Henrik; Frost, Tone Karin; Smit, Mathijs G D; Garpestad, Eimund; Skare, Ingvild; Bakke, Knut; Veiga, Leticia Falcao; Buffagni, Melania; Follum, Odd-Arne; Johnsen, Ståle; Moltu, Ulf-Einar; Reed, Mark

    2008-04-01

    This paper briefly summarizes the ERMS project and presents the developed model by showing results from environmental fates and risk calculations of a discharge from offshore drilling operations. The developed model calculates environmental risks for the water column and sediments resulting from exposure to toxic stressors (e.g., chemicals) and nontoxic stressors (e.g., suspended particles, sediment burial). The approach is based on existing risk assessment techniques described in the European Union technical guidance document on risk assessment and species sensitivity distributions. The model calculates an environmental impact factor, which characterizes the overall potential impact on the marine environment in terms of potentially impacted water volume and sediment area. The ERMS project started in 2003 and was finalized in 2007. In total, 28 scientific reports and 9 scientific papers have been delivered from the ERMS project (http://www.sintef.no/erms).

  4. 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.

  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

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

    2012-03-30

    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.

  6. Examining Agencies' Satisfaction with Electronic Record Management Systems in e-Government: A Large-Scale Survey Study

    NASA Astrophysics Data System (ADS)

    Hsu, Fang-Ming; Hu, Paul Jen-Hwa; Chen, Hsinchun; Hu, Han-Fen

    While e-government is propelling and maturing steadily, advanced technological capabilities alone cannot guarantee agencies’ realizing the full benefits of the enabling computer-based systems. This study analyzes information systems in e-government settings by examining agencies’ satisfaction with an electronic record management system (ERMS). Specifically, we investigate key satisfaction determinants that include regulatory compliance, job relevance, and satisfaction with support services for using the ERMS. We test our model and the hypotheses in it, using a large-scale survey that involves a total of 1,652 government agencies in Taiwan. Our results show significant effects of regulatory compliance on job relevance and satisfaction with support services, which in turn determine government agencies’ satisfaction with an ERMS. Our data exhibit a reasonably good fit to our model, which can explain a significant portion of the variance in agencies’ satisfaction with an ERMS. Our findings have several important implications to research and practice, which are also discussed.

  7. Novel root-fungus symbiosis in Ericaceae: sheathed ericoid mycorrhiza formed by a hitherto undescribed basidiomycete with affinities to Trechisporales.

    PubMed

    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. 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

  9. Genotyping of erythromycin resistant group C & G streptococci isolated in Chennai, south India

    PubMed Central

    Prabu, D.; Menon, Thangam

    2013-01-01

    Background & objectives: Increasing resistance to erythromycin has been observed worldwide in group C and group G streptococci (GCS/GGS). The information available from India is scanty. The aim of the study was to identify erythromycin resistant GCS/GGS isolates in Chennai, south India, and to compare erythromycin resistant genotypes with emm types. Methods: One hundred and thirty one GCS/GGS isolates were tested for erythromycin resistance by disc diffusion and agar dilution methods. Erythromycin resistance genotypes [erm(A), erm(B) and mef(A)] were determined by a multiplex PCR. emm types of erythromycin resistant GCS/GGS isolates was also assessed using emm gene sequencing method. Results: Sixteen of the 131 isolates (12.21%) were resistant to erythromycin. Majority of the isolates were GGS (15/16). Eight of the 16 (50%) were S. dysgalactiae subsps. equisimilis. Twelve isolates (75%) were MLSB phenotype and four (25%) were M phenotype. Of the 12 isolates which exhibited MLSB resistance, seven showed cMLSB phenotype and were positive for erm(B) gene. The remaining five were iMLSB phenotype of which three were positive for erm(A) gene and two for erm(B) gene. erm(A) was common among carriers whereas erm(B) was common among clinical isolates. Interpretation & conclusions: MLSB was the predominant phenotype and erm(B) was the common genotype in the present study. The emm type stC1400.0 was frequently associated with erythromycin resistant GCS/GGS in our study. PMID:23481067

  10. Structure-Guided Insights into the Function of Merlin in Neurofibromatosis 2 (NF2)

    DTIC Science & Technology

    2011-08-01

    fragmented by collision -activated dissociation (CAD), and the MS/MS scans were acquired using the ion trap as the analyzer. Data were processed using a raw...spliced forms, merlin-1 and merlin-2, which differ at their C-termini. Merlin belongs to ERM (ezrin-radixin-moesin) family of proteins that localize to...domains. Finally the functions of ERM family proteins are regulated by alterations in their conformation triggered by binding to their partners

  11. Distribution of genes encoding erythromycin ribosomal methylases and an erythromycin efflux pump in epidemiologically distinct groups of staphylococci.

    PubMed

    Eady, E A; Ross, J I; Tipper, J L; Walters, C E; Cove, J H; Noble, W C

    1993-02-01

    Erythromycin-resistant staphylococci can be divided into two phenotypic classes based on their pattern of cross-resistance to other macrolides, lincosamides and type B streptogramins. Strains inducibly or constitutively resistant to all MLS antibiotics possess erythromycin ribosomal methylase (erm) genes, whereas strains inducibly resistant to only 14 and 15-membered ring macrolides and type B streptogramins harbour msrA, which encodes an ATP-dependent efflux pump. Dot-blot hybridization was used to study the distribution of ermA, ermB, ermC and msrA in five epidemiologically distinct groups of staphylococci. The most widely-distributed resistance determinant was ermC, which was detected in 112 (50.6%) of 221 isolates, alone in 106 isolates and in combination with a second erythromycin resistance determinant in six strains. MsrA was detected in 73 (33%) of isolates, alone in 65 and in combination with a methylase gene in eight strains. This determinant was responsible for erythromycin resistance in over one-third (36.4%) of clinical isolates of coagulase-negative staphylococci. ErmA and ermB were present in only a minority of isolates (5.9 and 7.2% of strains, respectively). The resistance determinants present in ten strains did not hybridize to any of the four probes although, in all cases, their resistance phenotype was consistent with the possession of a methylase gene. Interestingly, ermB was found exclusively in animal isolates of Staphylococcus intermedius, Staphylococcus xylosus and Staphylococcus hyicus, but not in coagulase-negative staphylococci of human origin. This determinant has previously only been found in a small number of epidemiologically related strains of Staphylococcus aureus.

  12. Toward Determination of Venous Thrombosis Ages by Using Fuzzy Logic and Supervised Bayes Classification

    DTIC Science & Technology

    2007-11-02

    Lim1,3, R. Debon1, B. Solaiman1, L. Bressollette2, B. Guias2, C. Roux1, H.T. Chuah3 1Département I.T.I, LaTIM ERM-0102, ENST Bretagne , BP 832, 29285...Departement I.T.I LaTIM ERM-0102 ENST Bretagne , BP 832 29285 Brest Cedex, France Performing Organization Report Number Sponsoring/Monitoring Agency Name(s

  13. Community Environmental Response Facilitation Act (CERFA) Report, Former Army Reserve Center, Gaithersburg, Maryland

    DTIC Science & Technology

    1994-04-01

    ORGANIZATION NAME(S) AND AOORESS(ES) L PERFORMING ORGANIZATION REPORT NUNSII ERM, Inc. 855 Springdale Drive N/ A Exton, PA 19341 9. SPONqSORING...Unlimited I& A &STRACT (Maamu, 200wow) . This report presents the results of the Community Environmental Response Facilitation Act (CERFA) investigation...conducted by Environmental Resources Management (ERM) at the former Army Reserve Center, Gaithersburg (ARC), a U.S. Government property selected for

  14. Heterogeneity of macrolide-lincosamide-streptogramin B resistance phenotypes in enterococci.

    PubMed

    Min, Yu-Hong; Jeong, Jae-Hee; Choi, Yun-Jeong; Yun, Hee-Jeong; Lee, Kyungwon; Shim, Mi-Ja; Kwak, Jin-Hwan; Choi, Eung-Chil

    2003-11-01

    We determined the macrolide resistance phenotypes of 241 clinical isolates of erythromycin-resistant enterococci (MICs, > or = 1 microg/ml), including 147 Enterococcus faecalis strains and 94 Enterococcus faecium strains, collected from a hospital in Seoul, Korea, between 1999 and 2000. By the erythromycin (40 micro g)-josamycin (100 microg) double-disk test, 93 strains were assigned to the constitutive macrolide, lincosamide, and streptogramin B (MLS(B)) resistance (cMLS(B)) phenotype, and the remaining 148 strains were assigned to the inducible MLS(B) resistance (iMLS(B)) phenotype. Of the strains with the iMLS(B) phenotype, 36 exhibited a reversibly inducible MLS(B) (riMLS(B)) phenotype, i.e., blunting of the erythromycin zone of inhibition, which indicates that the 16-membered-ring macrolide josamycin is a more effective inducer than the 14-membered-ring macrolide erythromycin. Sequence analysis of the regulatory regions of the erm(B) genes from all of the strains exhibiting the riMLS(B) phenotype revealed not only erm(Bv) [where v represents variant; previously erm(AMR)] (n = 13), as reported previously, but also three kinds of erm(B) variants, which were designated erm(Bv1) (n = 17), erm(Bv2) (n = 3), and erm(Bv3) (n = 3), respectively. In lacZ reporter gene assays of these variants, the 16-membered-ring macrolide tylosin had stronger inducibility than erythromycin at > or = 0.1 microg/ml. These findings highlight the versatility of erm(B) in induction specificity.

  15. Antimicrobial susceptibility of Bacillus strains isolated from primary starters for African traditional bread production and characterization of the bacitracin operon and bacitracin biosynthesis.

    PubMed

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

    2012-11-01

    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.

  16. 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.

  17. Transferable Antibiotic Resistances in Marketed Edible Grasshoppers (Locusta migratoria migratorioides).

    PubMed

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

    2017-03-24

    Grasshoppers are the most commonly eaten insects by humans worldwide, as they are rich in proteins and micronutrients. This study aimed to assess the occurrence of transferable antibiotic resistance genes in commercialized edible grasshoppers. To this end, the prevalence of 12 selected genes [aac(6')-Ie aph(2″)-Ia, blaZ, erm(A), erm(B), erm(C), mecA, tet(M), tet(O), tet(S), tet(K), vanA, vanB] coding for resistance to antibiotics conventionally used in clinical practice was determined. The majority of samples were positive for tet(M) (70.0%), tet(K) (83.3%) and blaZ (83.3%). A low percentage of samples were positive for erm(B) (16.7%), erm(C) (26.7%), and aac(6')-Ie aph(2″)-Ia (13.3%), whereas no samples were positive for erm(A), vanA, vanB, tet(O), and mecA. Cluster analysis identified 4 main clusters, allowing a separation of samples on the basis of their country of origin.

  18. Functional and Molecular Characterization of Ex Vivo Cultured Epiretinal Membrane Cells from Human Proliferative Diabetic Retinopathy

    PubMed Central

    Veréb, Zoltán; Lumi, Xhevat; Andjelic, Sofija; Globocnik-Petrovic, Mojca; Urbancic, Mojca; Hawlina, Marko; Facskó, Andrea; Petrovski, Goran

    2013-01-01

    Characterization of the cell surface marker phenotype of ex vivo cultured cells growing out of human fibrovascular epiretinal membranes (fvERMs) from proliferative diabetic retinopathy (PDR) can give insight into their function in immunity, angiogenesis, and retinal detachment. FvERMs from uneventful vitrectomies due to PDR were cultured adherently ex vivo. Surface marker analysis, release of immunity- and angiogenesis-pathway-related factors upon TNFα activation and measurement of the intracellular calcium dynamics upon mechano-stimulation using fluorescent dye Fura-2 were all performed. FvERMs formed proliferating cell monolayers when cultured ex vivo, which were negative for endothelial cell markers (CD31, VEGFR2), partially positive for hematopoietic- (CD34, CD47) and mesenchymal stem cell markers (CD73, CD90/Thy-1, and PDGFRβ), and negative for CD105. CD146/MCAM and CD166/ALCAM, previously unreported in cells from fvERMs, were also expressed. Secretion of 11 angiogenesis-related factors (DPPIV/CD26, EG-VEGF/PK1, ET-1, IGFBP-2 and 3, IL-8/CXCL8, MCP-1/CCL2, MMP-9, PTX3/TSG-14, Serpin E1/PAI-1, Serpin F1/PEDF, TIMP-1, and TSP-1) were detected upon TNFα activation of fvERM cells. Mechano-stimulation of these cells induced intracellular calcium propagation representing functional viability and role of these cells in tractional retinal detachment, thus serving as a model for studying tractional forces present in fvERMs in PDR ex vivo. PMID:24195074

  19. Functional and molecular characterization of ex vivo cultured epiretinal membrane cells from human proliferative diabetic retinopathy.

    PubMed

    Veréb, Zoltán; Lumi, Xhevat; Andjelic, Sofija; Globocnik-Petrovic, Mojca; Urbancic, Mojca; Hawlina, Marko; Facskó, Andrea; Petrovski, Goran

    2013-01-01

    Characterization of the cell surface marker phenotype of ex vivo cultured cells growing out of human fibrovascular epiretinal membranes (fvERMs) from proliferative diabetic retinopathy (PDR) can give insight into their function in immunity, angiogenesis, and retinal detachment. FvERMs from uneventful vitrectomies due to PDR were cultured adherently ex vivo. Surface marker analysis, release of immunity- and angiogenesis-pathway-related factors upon TNF α activation and measurement of the intracellular calcium dynamics upon mechano-stimulation using fluorescent dye Fura-2 were all performed. FvERMs formed proliferating cell monolayers when cultured ex vivo, which were negative for endothelial cell markers (CD31, VEGFR2), partially positive for hematopoietic- (CD34, CD47) and mesenchymal stem cell markers (CD73, CD90/Thy-1, and PDGFR β ), and negative for CD105. CD146/MCAM and CD166/ALCAM, previously unreported in cells from fvERMs, were also expressed. Secretion of 11 angiogenesis-related factors (DPPIV/CD26, EG-VEGF/PK1, ET-1, IGFBP-2 and 3, IL-8/CXCL8, MCP-1/CCL2, MMP-9, PTX3/TSG-14, Serpin E1/PAI-1, Serpin F1/PEDF, TIMP-1, and TSP-1) were detected upon TNF α activation of fvERM cells. Mechano-stimulation of these cells induced intracellular calcium propagation representing functional viability and role of these cells in tractional retinal detachment, thus serving as a model for studying tractional forces present in fvERMs in PDR ex vivo.

  20. The ER-mitochondria encounter structure contributes to hyphal growth, mitochondrial morphology and virulence of the pathogenic mold Aspergillus fumigatus.

    PubMed

    Geißel, Bernadette; Penka, Mirjam; Neubauer, Michael; Wagener, Johannes

    2017-01-01

    Aspergillus fumigatus is an opportunistic fungal pathogen and the primary causative species of invasive aspergillosis, a systemic disease associated with high mortality rates. Treatment of invasive fungal infection relies on a very limited number of antifungal drug classes. In order to extend the spectrum of antifungal drugs novel target structures have to be identified. The ER-mitochondria encounter structure (ERMES), a recently discovered tether that links mitochondria and endoplasmic reticulum, is a potential drug target based on its absence in Metazoa. Very recently, it was shown that ERMES is important for the fitness and immune evasion of the pathogenic yeast Candida albicans. We studied the role of the four ERMES core components Mdm10, Mdm12, Mdm34 and Mmm1 in the pathogenic mold A. fumigatus. By construction and characterizing conditional mutants of all four core components and deletion mutants of mdm10 and mdm12, we show that each component is of significant importance for growth of the fungal pathogen. While markedness of the individual mutant phenotypes differed slightly, all components are important for maintenance of the mitochondrial morphology and the intra-organellar distribution of nucleoids. Characterization of the Mmm1 ERMES mutant in a Galleria mellonella infection model indicates that ERMES contributes to virulence of A. fumigatus. Our results demonstrate that pharmacologic inhibition of ERMES could exert antifungal activity against this important pathogen.

  1. 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.

  2. 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

  3. 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

  4. The epidemiology of sports and exercise related injury in the United Kingdom.

    PubMed Central

    Nicholl, J P; Coleman, P; Williams, B T

    1995-01-01

    A national study of exercise related morbidity (ERM) in England and Wales was carried out using a postal questionnaire sent to 28,857 adults aged 16-45 years. The questionnaire asked about regular participation in sports or other recreational fitness activities involving physical exercise, and for details of any injuries occurring during a 28 d reference period. A return rate of 68% was achieved. Comparisons with other national data sources indicated that the information obtained was reliable. It is estimated that each year there are 29 million incidents resulting in new or recurrent injuries, however minor, of which 9.8 million (95% confidence interval 8.1 to 11.4 million) result in new 'substantive' injuries which are potentially serious, result in treatment, or in participants being unable to take part in their usual activities. Soccer accounted for more than 25% of all ERM, but the risk of a substantive injury in rugby was three times that in soccer. Over one third of ERM occurred in men aged 16-25 years. The most frequently reported injuries were sprains and strains of the lower limbs. Treatment was sought in approximately 25% of ERM incidents and 7% of all new ERM incidents involved attendance at a hospital accident and emergency department. The treatment provider most likely to be consulted was a general practitioner, but physiotherapists and complementary medicine practitioners were also consulted frequently. To maximize the health benefits of exercise, research strategies to reduce the volume and severity of ERM and to identify the most appropriate ways of managing ERM should be set. PMID:8808535

  5. NHERF-1 knockout mice have an attenuated hepatic inflammatory response and are protected from cholestatic liver injury

    PubMed Central

    Li, Man; Mennone, Albert; Soroka, Carol J.; Hagey, Lee R.; Ouyang, Xinshou; Weinman, Edward J.; Boyer, James L.

    2015-01-01

    The intercellular adhesion molecule-1 (ICAM-1) is induced in mouse liver after bile duct ligation (BDL) and plays a key role in neutrophil-mediated liver injury in BDL mice. ICAM-1 has been shown to interact with the cytoskeletal ezrin-radixin-moesin (ERM) proteins that also interact with the PDZ protein, Na+/H+ exchanger regulatory factor 1 (NHERF-1/EBP50). In NHERF-1−/− mice, ERM proteins are significantly reduced in brush border membranes from kidney and small intestine. ERM knockdown reduces ICAM-1 expression in response to TNF-α. Here we show that NHERF-1 assembles ERM proteins, ICAM-1 and F-actin into a macromolecule complex that is increased in mouse liver after BDL. Compared with wild-type (WT) mice, both sham-operated and BDL NHERF-1−/− mice have lower levels of activated ERM and ICAM-1 protein in the liver accompanied by significantly reduced hepatic neutrophil accumulation, serum ALT, and attenuated liver injury after BDL. However, total bile acid concentrations in the serum and liver of sham and BDL NHERF-1−/− mice were not significantly different from the WT controls, although hepatic tetrahydroxylated bile acids and Cyp3a11 mRNA levels were higher in NHERF-1−/− BDL mice. Conclusion NHERF-1 participates in the inflammatory response that is associated with BDL induced liver injury. Deletion of NHERF-1 in mice leads to disruption of the formation of ICAM-1-ERM-NHERF-1 complex and reduction of hepatic ERM proteins and ICAM-1, molecules that are up-regulated and are essential for neutrophil-mediated liver injury in cholestasis. Further study of the role of NHERF-1 in the inflammatory response in cholestasis and other forms of liver injury should lead to discovery of new therapeutic targets in hepatic inflammatory diseases. PMID:26108984

  6. 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

  7. 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

    2016-08-31

    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.

  8. Gene Expression Analysis of the Irrigation Solution Samples Collected during Vitrectomy for Idiopathic Epiretinal Membrane

    PubMed Central

    Myojin, Sayaka; Yoshida, Shigeo; Takeda, Atsunobu; Murakami, Yusuke; Kawano, Yoichi; Oshima, Yuji; Ishibashi, Tatsuro; Sonoda, Koh-Hei

    2016-01-01

    Purpose The analysis of gene expression in idiopathic epiretinal membranes (iERMs) may help elucidate ERM formation and its pathology. Here, we conducted a case-control study, in order to determine the expression levels of cytokines and other genes in eyes with macular hole (MH) or iERM. Methods Twenty eyes, obtained from seven male and 13 female patients, were included in the study. The average age of the study subjects was 69.1 ± 7.67 years, and 15 eyes had iERM, while five eyes had MH. Irrigation solution samples were collected during vitrectomy, centrifuged, and the levels of cytokine and other mRNAs in the sediment were assessed using real-time PCR. The expression level of 11 cytokine genes, four transcription factor genes, two cytoskeletal genes, and genes encoding two extracellular matrix proteins in eyes with MH or iERM were determined and compared. Results The expression levels of interleukin 6 (IL6), tumor growth factor B2 (TGFB2), vascular endothelial growth factor A (VEGFA), chemokine C-X-C motif ligand 1 (CXCL1), v-rel avian reticuloendotheliosis viral oncogene homolog A (RELA), glial fibrillary acidic protein (GFAP), and tenascin C (TNC) were significantly higher in eyes with iERM than in eyes with MH. The expression of these genes was not associated with the preoperative visual acuity of the investigated patients. Conclusions The obtained results indicate that real-time PCR analysis of irrigation solution samples collected during vitrectomy can help assess the expression levels of several genes, and that iERM is associated with the expression of pro-inflammatory genes and the genes expressed during angiogenesis and wound healing process (IL6, TGFB2, VEGFA, CXCL1, RELA, GFAP, and TNC). PMID:27736918

  9. Predictions of sediment toxicity using a database for Hyalella azteca and Chironomus riparius

    SciTech Connect

    Haverland, P.S.; Dwyer, F.J.; Henke, C.E.; Ingersoll, C.G.; Mount, D.R.; Field, J.; MacDonald, D.D.; Smith, S.L.

    1995-12-31

    A database was developed for calculating sediment effect concentrations (SECS) for various contaminants Associated with field-collected sediment using laboratory toxicity data for the amphipod H. azteca and the midge C. riparius. Three types of SECs were calculated: (1) Effect Range Low (ERL) and Effect Range Median (ERM), (2) Threshold Effect Level (TEL) and Probable Effect Level (PEL), and (3) No Effect Concentration (NEC). The predictive abilities of SECs were evaluated using independent toxicity data sets. For example, the predictive ability of ERMs was evaluated by first calculating ERMs using just the Great Lakes (GL) portion of the database. These GL ERMs were then used to predict responses in independent H. azteca 28-d tests and C. riparius 1 4-d tests with Clark Fork River sediments. About 70 to 90% of the samples were correctly classified at 1 to 2 exceedances of GL ERMS. At 1 to 2 exceedances of GL ERMS, Type 2 error (false negatives) was < 10% and Type 1 error (false positives) was 10 to 30%. Evaluations using GL PELs and GL NECs resulted in similar predictive ability compared to GL ERMS. When SECs are used to conduct a preliminary screening to predict the potential for toxicity in the absence of actual toxicity testing, a low number of SEC exceedances should be used to minimize the potential for false negatives; however, the risk of accepting higher false positives is increased. The authors are currently using SECs calculated from the entire database to predict the response of H. azteca and C. riparius in a variety of independent data sets generated by other laboratories.

  10. miRNAs in the vitreous humor of patients affected by idiopathic epiretinal membrane and macular hole

    PubMed Central

    Ragusa, Marco; Barbagallo, Cristina; Longo, Antonio; Avitabile, Teresio; Uva, Maurizio G.; Bonfiglio, Vincenza; Toro, Mario D.; Caltabiano, Rosario; Mariotti, Cesare; Boscia, Francesco; Romano, Mario; Di Pietro, Cinzia; Barbagallo, Davide; Purrello, Michele; Reibaldi, Michele

    2017-01-01

    Purpose The aim of the present study was to assess the expression of miRNAs in the Vitreous Humor (VH) of patients with Macular Hole (MH) and Epiretinal Membrane (ERM) compared to a control group. Methods In this prospective, comparative study, 2-ml of VH was extracted from the core of the vitreous chamber in consecutive patients who underwent standard vitrectomy for ERM and MH. RNA was extracted and TaqMan® Low Density Arrays (TLDAs) were used to profile the transcriptome of 754 miRNAs. Results were validated by single TaqMan® assays. Finally, we created a biological network of differentially expressed miRNA targets and their nearest neighbors. Results Overall 10 eyes with MH, 16 eyes with idiopathic ERM and 6 controls were enrolled in the study. Profiling data identified 5 miRNAs differentially expressed in patients affected by MH and ERM with respect to controls. Four were downregulated (miR-19b, miR-24, miR-155, miR-451) and 1 was downregulated (miR-29a); TaqMan® assays of the VH of patients affected by MH and ERM, with respect to controls, showed that the most differentially expressed were miR-19b (FC -9.13, p:<0.00004), mir-24 (FC -7.52, p:<0.004) and miR-142-3p (FC -5.32, p:<0.011). Our network data showed that deregulation of differentially expressed miRNAs induces an alteration of several pathways associated with genes involved in both MH and ERM. Conclusion The present study suggests that disregulation of miR-19b, miR-24 and miR-142-3p, might be related to the alterations that characterize patients affected by MH and ERM. PMID:28328945

  11. Technical note: Occurrence in fecal microbiota of genes conferring resistance to both macrolide-lincosamide-streptogramin B and tetracyclines concomitant with feeding of beef cattle with tylosin.

    PubMed

    Chen, J; Fluharty, F L; St-Pierre, N; Morrison, M; Yu, Z

    2008-09-01

    Development of antimicrobial resistance in food animals receiving antimicrobials has been well documented among bacterial isolates, especially pathogens, but information on development of antimicrobial resistance at the microbial community level during long-term feeding of antimicrobials is lacking. The objective of this study was to examine the association between inclusion of tylosin in feed and occurrence of resistance to macrolide-lincosamide-streptogramin B (MLS(B)) in the entire fecal microbial communities of beef cattle over a feeding study of 168 d. A completely randomized design included 6 pens housed together in 1 barn, with each pen housing 10 to 11 steers. The control and tylosin groups each had 3 pens, with the former receiving no antimicrobial whereas the latter received both tylosin and monensin (11 and 29.9 mg/ kg of feed, respectively, DM) in feed. The abundance of genes conferring resistance to MLS(B) (erm genes) and tetracyclines (tet genes) were quantified using class-specific, real-time PCR assays. The abundances of erm and tet genes were analyzed with pens as experimental units using the MIXED procedure of SAS. Correlations between abundance of different resistance genes were calculated using the CORR procedure of SAS. We identified 4 classes (B, F, T, and X) of erm genes in fresh fecal samples collected at wk 2, 17, and 21 of feeding. From wk 2 to 17, the abundance of erm(T) and erm(X) increased (P < 0.05), whereas that of erm(B) and erm(F) did not. The abundance of the erm genes did not further change from wk 17 to 21. The tet(A/C), tet(G), and tet gene variants encoding ribosomal protection proteins (including classes M, O, P, Q, S, T, and W) appeared to be co-selected by tylosin feeding. Such co-selection of multiresistance at community level by one antimicrobial drug used in animals has the important implication that future studies should examine resistance to not only the antimicrobials used in animals, but also other antimicrobials

  12. Epidermal growth factor-induced cellular invasion requires sphingosine-1-phosphate/sphingosine-1-phosphate 2 receptor-mediated ezrin activation

    PubMed Central

    Orr Gandy, K. Alexa; Adada, Mohamad; Canals, Daniel; Carroll, Brittany; Roddy, Patrick; Hannun, Yusuf A.; Obeid, Lina M.

    2013-01-01

    Ezrin, radixin, and moesin (ERM) proteins link cortical actin to the plasma membrane and coordinate cellular events that require cytoskeletal rearrangement, including cell division, migration, and invasion. While ERM proteins are involved in many important cellular events, the mechanisms regulating their function are not completely understood. Our laboratory previously identified reciprocal roles for the sphingolipids ceramide and sphingosine-1-phosphate (S1P) in the regulation of ERM proteins. We recently showed that ceramide-induced activation of PP1α leads to dephosphorylation and inactivation of ERM proteins, while S1P results in phosphorylation and activation of ERM proteins. Following these findings, we aimed to examine known inducers of the SK/S1P pathway and evaluate their ability to regulate ERM proteins. We examined EGF, a known inducer of the SK/S1P pathway, for its ability to regulate the ERM family of proteins. We found that EGF induces ERM c-terminal threonine phosphorylation via activation of the SK/S1P pathway, as this was prevented by siRNA knockdown or pharmacological inhibition of SK. Using pharmacological, as well as genetic, knockdown approaches, we determined that EGF induces ERM phosphorylation via activation of S1PR2. In addition, EGF led to cell polarization in the form of lamellipodia, and this occurred through a mechanism involving S1PR2-mediated phosphorylation of ezrin T567. EGF-induced cellular invasion was also found to be dependent on S1PR2-induced T567 ezrin phosphorylation, such that S1PR2 antagonist, JTE-013, and expression of a dominant-negative ezrin mutant prevented cellular invasion toward EGF. In this work, a novel mechanism of EGF-stimulated invasion is unveiled, whereby S1P-mediated activation of S1PR2 and phosphorylation of ezrin T567 is required.—Orr Gandy, K. A., Adada, M., Canals, D., Carroll, B., Roddy, P., Hannun, Y. A., Obeid, L. M. Epidermal growth factor-induced cellular invasion requires sphingosine-1-phosphate

  13. Impact of subtherapeutic administration of tylosin and chlortetracycline on antimicrobial resistance in farrow-to-finish swine.

    PubMed

    Holman, Devin B; Chénier, Martin R

    2013-07-01

    The use of antimicrobial agents in swine production at subtherapeutic concentrations for the purpose of growth promotion remains controversial due to the potential impact on public health. Beginning at weaning (3 weeks), pigs received either nonmedicated feed or feed supplemented with subtherapeutic levels of either tylosin (11-44 ppm) or chlortetracycline (5.5 ppm). After only 3 weeks, pigs given feed supplemented with tylosin had significantly higher levels of tylosin-resistant anaerobes (P < 0.0001) compared with the control group, increasing from 11.8% to 89.6%, a level which was stable for the duration of the study, even after a 2-week withdrawal prior to slaughter. Tylosin-fed pigs had a higher incidence of detection for erm(A), erm(F), and erm(G), as well as significantly (P < 0.001) higher concentrations of erm(B) in their feces. The continuous administration of chlortetracycline-supplemented feed, however, had no significant effect on the population of chlortetracycline-resistant anaerobes in comparison with nontreated pigs (P > 0.05). The resistance genes tet(O), tet(Q), and erm(B) were detected in all pigs at each sampling time, while tet(G), tet(L), and tet(M) were also frequently detected. Neither chlortetracycline nor tylosin increased the growth rate of pigs.

  14. An Hdac1/Rpd3-Poised Circuit Balances Continual Self-Renewal and Rapid Restriction of Developmental Potential during Asymmetric Stem Cell Division.

    PubMed

    Janssens, Derek H; Hamm, Danielle C; Anhezini, Lucas; Xiao, Qi; Siller, Karsten H; Siegrist, Sarah E; Harrison, Melissa M; Lee, Cheng-Yu

    2017-02-27

    How the developmental potential of differentiating stem cell progeny becomes rapidly and stably restricted following asymmetric stem cell division is unclear. In the fly larval brain, earmuff (erm) uniquely functions to restrict the developmental potential of intermediate neural progenitors (INPs) generated by asymmetrically dividing neural stem cells (neuroblasts). Here we demonstrate that the histone deacetylase Hdac1/Rpd3 functions together with self-renewal transcriptional repressors to maintain the erm immature INP enhancer in an inactive but poised state in neuroblasts. Within 2 hr of immature INP birth, downregulation of repressor activities alleviates Rpd3-mediated repression on the erm enhancer, enabling acetylation of multiple histone proteins and activating Erm expression. Erm restricts the developmental potential in immature INPs by repressing genes encoding neuroblast transcriptional activators. We propose that poising the fast-activating enhancers of master regulators of differentiation through continual histone deacetylation in stem cells enables self-renewal and rapid restriction of developmental potential following asymmetric division.

  15. Sphingolipid regulation of ezrin, radixin, and moesin proteins family: implications for cell dynamics.

    PubMed

    Adada, Mohamad; Canals, Daniel; Hannun, Yusuf A; Obeid, Lina M

    2014-05-01

    A key but poorly studied domain of sphingolipid functions encompasses endocytosis, exocytosis, cellular trafficking, and cell movement. Recently, the ezrin, radixin and moesin (ERM) family of proteins emerged as novel potent targets regulated by sphingolipids. ERMs are structural proteins linking the actin cytoskeleton to the plasma membrane, also forming a scaffold for signaling pathways that are used for cell proliferation, migration and invasion, and cell division. Opposing functions of the bioactive sphingolipid ceramide and sphingosine-1-phosphate (S1P), contribute to ERM regulation. S1P robustly activates whereas ceramide potently deactivates ERM via phosphorylation/dephosphorylation, respectively. This recent dimension of cytoskeletal regulation by sphingolipids opens up new avenues to target cell dynamics, and provides further understanding of some of the unexplained biological effects mediated by sphingolipids. In addition, these studies are providing novel inroads into defining basic mechanisms of regulation and action of bioactive sphingolipids. This review describes the current understanding of sphingolipid regulation of the cytoskeleton, it also describes the biologies in which ERM proteins have been involved, and finally how these two large fields have started to converge. This article is part of a Special Issue entitled New Frontiers in Sphingolipid Biology.

  16. Control of adipogenesis by ezrin, radixin and moesin-dependent biomechanics remodeling.

    PubMed

    Titushkin, Igor; Sun, Shan; Paul, Amit; Cho, Michael

    2013-02-01

    We have recently shown that altered stem cell biomechanics can regulate the lineage commitment through a family of the membrane-cytoskeleton linker proteins (ERM; ezrin, radixin, moesin). The ERM proteins not only modulate the cell stiffness and actin cytoskeleton organization, but also rearrange focal adhesions and therefore influence the biochemically-directed stem cell differentiation. Combining silencing RNA, atomic force microscopy, and fluorescence microscopy, the role of the ERM proteins involved in the regulation of stem cell biomechanics and adipogenic differentiation was quantitatively determined. Transient ERM knockdown by RNAi caused disassembly of actin stress fibers and focal adhesions and a decrease in the cell stiffness. The silencing RNA treatment not only induced mechanical changes in stem cells but impaired adipogenesis in a time-dependent manner. While siRNA ERM treatment at day 0 substantially interfered with adipogenesis, the same treatment at day 3 of adipogenic differentiation significantly facilitated adipogenesis, as assessed by the expression of adipocyte-specific markers. The intact biomechanics homeostasis appears to be critical for the adipogenic induction. These findings may lead to potential biomechanical intervention techniques and methodologies to control the fate and extent of adipogenesis that would likely be involved in stem cell-based therapeutics for soft tissue repair and regeneration.

  17. Moesin-deficient mice reveal a non-redundant role for moesin in lymphocyte homeostasis.

    PubMed

    Hirata, Takako; Nomachi, Akira; Tohya, Kazuo; Miyasaka, Masayuki; Tsukita, Sachiko; Watanabe, Takeshi; Narumiya, Shuh

    2012-11-01

    Moesin is a member of the ezrin-radixin-moesin (ERM) family of cytoskeletal proteins. These proteins organize membrane domains by interacting with plasma membrane proteins and the actin cytoskeleton. Because of their high sequence similarity, ERM proteins are usually thought to be functionally redundant. Lymphocytes express two ERM proteins, ezrin and moesin. Whether each ERM plays a specialized role in lymphocytes, particularly in vivo, remains unknown. Here, we show that moesin has a crucial, non-redundant role in lymphocyte homeostasis. Moesin-deficient mice exhibited decreases in both T and B cells in the peripheral blood and lymph nodes, but not in the spleen. This phenotype was recapitulated in bone marrow (BM) chimeras with a hematopoietic moesin deficiency. Although the T and B cells apparently developed without major defects in the moesin-deficient mice, T cell egress from the thymus and immature B cell egress from the BM were impaired. In the periphery, both T and B cells showed delayed egress from lymphoid organs. We showed that moesin is the primary phosphorylated ERM subject to dynamic regulation during cell shape changes and migration. Our findings identify a previously unknown, non-redundant function of moesin in lymphocyte homeostasis in regulating lymphocyte egress from lymphoid organs.

  18. 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.

  19. Macrolide resistance in Streptococcus pyogenes isolates from throat infections in the region of Aachen, Germany.

    PubMed

    Brandt, C M; Honscha, M; Truong, N D; Holland, R; Hövener, B; Bryskier, A; Lütticken, R; Reinert, R R

    2001-01-01

    Macrolide-resistance was assessed in 216 consecutive Streptococcus pyogenes isolates from throat infections in the region of Aachen, Germany. Seventeen isolates were resistant to erythromycin: 12 isolates revealed a macrolide (M) phenotype and harbored mefA, and five strains expressed an inducible macrolide-lincosamide-streptogramin B (MLSB) phenotype of which four strains harbored ermA(TR) and one strain contained ermB(AM). Telithromycin (HMR 3647) and quinupristin/dalfopristin remained active particularly against the ermA(TR)-containing S. pyogenes isolates studied. Random amplified polymorphic DNA analysis identified multiple clones among erythromycin-resistant strains, but did not discriminate beyond the emm-type. mefA was present in three isolates either with emm2, emm12, or emm75, and in nine isolates with emm4. All four strains with ermA(TR) contained emm77, and the single strain with ermB(AM) harbored emm1. Despite the relative low rate of macrolide-resistance, these data suggest that at least three different macrolide-resistance determinants are prevalent in Germany and that mefA has spread rapidly into multiple clones of S. pyogenes.

  20. CLIC proteins, ezrin, radixin, moesin and the coupling of membranes to the actin cytoskeleton: a smoking gun?

    PubMed

    Jiang, Lele; Phang, Juanita M; Yu, Jiang; Harrop, Stephen J; Sokolova, Anna V; Duff, Anthony P; Wilk, Krystyna E; Alkhamici, Heba; Breit, Samuel N; Valenzuela, Stella M; Brown, Louise J; Curmi, Paul M G

    2014-02-01

    The CLIC proteins are a highly conserved family of metazoan proteins with the unusual ability to adopt both soluble and integral membrane forms. The physiological functions of CLIC proteins may include enzymatic activity in the soluble form and anion channel activity in the integral membrane form. CLIC proteins are associated with the ERM proteins: ezrin, radixin and moesin. ERM proteins act as cross-linkers between membranes and the cortical actin cytoskeleton. Both CLIC and ERM proteins are controlled by Rho family small GTPases. CLIC proteins, ERM and Rho GTPases act in a concerted manner to control active membrane processes including the maintenance of microvillar structures, phagocytosis and vesicle trafficking. All of these processes involve the interaction of membranes with the underlying cortical actin cytoskeleton. The relationships between Rho GTPases, CLIC proteins, ERM proteins and the membrane:actin cytoskeleton interface are reviewed. Speculative models are proposed involving the formation of localised multi-protein complexes on the membrane surface that assemble via multiple weak interactions. This article is part of a Special Issue entitled: Reciprocal influences between cell cytoskeleton and membrane channels, receptors and transporters. Guest Editor: Jean Claude Hervé.

  1. Role of pars plana vitrectomy and membrane peel in vitreomacular traction associated with retinal vasoproliferative tumors

    PubMed Central

    Castro-Navarro, Verónica; Saktanasate, Jarin; Say, Emil Anthony T.; Chiang, Allen; Shields, Carol Lally

    2016-01-01

    To report a case of retinal vasoproliferative tumor (VPT) with secondary epiretinal membrane (ERM) formation and vitreo-macular traction managed by pars plana vitrectomy (PPV) and membrane peel. A 29-year-old male was referred for management of decreased vision in the right eye (OD) for 1 week. Presenting visual acuity was 20/50 Snellen feet (ft) OD, and fundus examination showed an ERM associated with a reddish-yellow mass in the inferotemporal quadrant with overlying exudation, hemorrhage, and subretinal fluid consistent with VPT, and cryotherapy was recommended. Two months later, there was complete tumor regression, but there was decreased vision from progressive vitreomacular traction to 20/400 ft. PPV with combined ERM and internal limiting membrane (ILM) peel were performed with resolution of vitreomacular traction and improvement of visual acuity to 20/50 ft at 6 months. PPV with combined ERM and ILM peel is effective for vision loss secondary to ERM and vitreomacular traction associated with retinal VPT. PMID:27843233

  2. Prevalence and mechanism of resistance against macrolides and lincosamides in Streptococcus suis isolates.

    PubMed

    Martel, A; Baele, M; Devriese, L A; Goossens, H; Wisselink, H J; Decostere, A; Haesebrouck, F

    2001-11-26

    Eighty-seven Streptococcus suis isolates recovered in 1999-2000 from diseased pigs, all from different farms, were screened for resistance against macrolide and lincosamide antibiotics by the disk diffusion and agar dilution test and a PCR assay, amplifying the ermB gene and the mefA/E gene. Seventy-one percent of the isolates showed constitutive resistance to macrolide and lincosamide antibiotics (MLS(B)-phenotype). All these isolates were positive for the ermB gene in the PCR, but negative for the mefA/E gene. For all strains minimum inhibitory concentrations (MIC) against five other antimicrobial agents were determined. All strains were susceptible to penicillin. Ninety-nine percent of the isolates were susceptible to enrofloxacin and tiamulin. Eighty-five percent of the strains were resistant to doxycycline. A 540bp fragment of the ermB genes of eight S. suis strains was sequenced and compared with ermB genes of five S. pneumoniae and five S. pyogenes strains of human origin. A 100% homology was found between these fragments in seven S. suis, one S. pneumoniae and three of the S. pyogenes isolates. This study demonstrates that resistance against macrolides, lincosamides and streptogramin B is widespread in S. suis and mediated by ribosome methylation, encoded by the ermB gene.

  3. The macrolide-lincosamide-streptogramin B resistance phenotypes characterized by using a specifically deleted, antibiotic-sensitive strain of Streptomyces lividans.

    PubMed Central

    Pernodet, J L; Fish, S; Blondelet-Rouault, M H; Cundliffe, E

    1996-01-01

    Genes conferring resistance to macrolide, lincosamide, and streptogramin B (MLS) antibiotics via ribosomal modification are widespread in bacteria, including clinical isolates and MLS-producing actinomycetes. Such erm-type genes encode enzymes that mono- or dimethylate residue A-2058 of 23S rRNA. The different phenotypes resulting from monomethylation (MLS-I phenotype, conferred by erm type I genes) or dimethylation (MLS-II phenotype due to erm type II genes) have been characterized by introducing tlrD or ermE, respectively, into an MLS-sensitive derivative of Streptomyces lividans TK21. This strain (designated OS456) was generated by specific replacement of the endogenous resistance genes lrm and mgt. The MLS-I phenotype is characterized by high-level resistance to lincomycin with only marginal resistance to macrolides such as chalcomycin or tylosin, whereas the MLS-II phenotype involves high-level resistance to all MLS drugs. Mono- and dimethylated ribosomes were introduced into a cell-free protein-synthesizing system prepared from S. lividans and compared with unmodified particles in their response to antibiotics. There was no simple correlation between the relative potencies of MLS drugs at the level of the target site (i.e., the ribosome) and their antibacterial activities expressed as MICs. PMID:8851574

  4. The macrolide-lincosamide-streptogramin B resistance phenotypes characterized by using a specifically deleted, antibiotic-sensitive strain of Streptomyces lividans.

    PubMed

    Pernodet, J L; Fish, S; Blondelet-Rouault, M H; Cundliffe, E

    1996-03-01

    Genes conferring resistance to macrolide, lincosamide, and streptogramin B (MLS) antibiotics via ribosomal modification are widespread in bacteria, including clinical isolates and MLS-producing actinomycetes. Such erm-type genes encode enzymes that mono- or dimethylate residue A-2058 of 23S rRNA. The different phenotypes resulting from monomethylation (MLS-I phenotype, conferred by erm type I genes) or dimethylation (MLS-II phenotype due to erm type II genes) have been characterized by introducing tlrD or ermE, respectively, into an MLS-sensitive derivative of Streptomyces lividans TK21. This strain (designated OS456) was generated by specific replacement of the endogenous resistance genes lrm and mgt. The MLS-I phenotype is characterized by high-level resistance to lincomycin with only marginal resistance to macrolides such as chalcomycin or tylosin, whereas the MLS-II phenotype involves high-level resistance to all MLS drugs. Mono- and dimethylated ribosomes were introduced into a cell-free protein-synthesizing system prepared from S. lividans and compared with unmodified particles in their response to antibiotics. There was no simple correlation between the relative potencies of MLS drugs at the level of the target site (i.e., the ribosome) and their antibacterial activities expressed as MICs.

  5. Enhanced proliferation, attachment and osteopontin expression by porcine periodontal cells exposed to Emdogain.

    PubMed

    Rincon, J C; Xiao, Y; Young, W G; Bartold, P M

    2005-12-01

    Emdogain (EMD) is an enamel matrix derivative extracted from developing porcine teeth with demonstrated periodontal regenerative potential. EMD has been shown to influence a number of properties of periodontal ligament cells including proliferation, cell attachment and matrix synthesis. To date, the effect of EMD on the epithelial cell rests of Malassez (ERM) is unknown. In this study, periodontal ligament fibroblasts, ERM, alveolar bone cells and gingival fibroblasts were obtained from porcine periodontal ligament, alveolar bone and gingiva. This study investigated, in vitro, the effect of EMD at three concentrations on proliferation, cell attachment and expression of mRNA for two mineralised tissue-related proteins (osteopontin and bone sialoprotein). As for other periodontal cells, the ERM proliferative response was enhanced by EMD. Attachment assays revealed a highly significant increase for ERM and gingival fibroblasts after EMD treatment at all concentrations. This study has also shown that EMD stimulated expression of osteopontin mRNA by ERM and alveolar bone cells. The results from this study provide evidence that EMD enhanced cellular events related with proliferation, attachment and osteopontin mRNA expression by porcine periodontal cells, in a manner consistent with its role in periodontal regenerative therapy.

  6. Aniseikonia associated with epiretinal membranes

    PubMed Central

    Ugarte, M; Williamson, T H

    2005-01-01

    Aims: To determine whether the computerised version of the new aniseikonia test (NAT) is a valid, reliable method to measure aniseikonia and establish whether aniseikonia occurs in patients with epiretinal membranes (ERM) with preserved good visual acuity. Methods: With a computerised version of the NAT, horizontal and vertical aniseikonia was measured in 16 individuals (mean 47 (SD 16.46) years) with no ocular history and 14 patients (mean 67.7 (14.36) years) with ERM. Test validity was evaluated by inducing aniseikonia with size lenses. Test reliability was assessed by the test-retest method. Results: In normal individuals, the mean percentage (SD) aniseikonia was −0.24% (0.71) horizontal and 0% (0.59) vertical. Validity studies revealed mean (SD) 0.990 (0.005) horizontal and 0.991 (0.004) vertical correlation coefficients, 0.985 (0.111) horizontal and 0.989 (0.102) vertical slope. Repeatability coefficients were 1.04 horizontal and 0.88 vertical. Aniseikonia in patients with ERM ranged from 4% to 14%. Eight patients showed 2% or more size difference between horizontal and vertical meridians. Conclusions: The aniseikonia test used in this study can be considered a simple, fast, valid and reliable method to measure the difference in image size perceived by each eye. Aniseikonia does occur in symptomatic patients with ERM. The effect of ERM on image size is heterogeneous across the retinal area affected. PMID:16299133

  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.

  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. 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.

  10. 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.

  11. 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

  12. Prototypic Enhanced Risk Monitor Framework and Evaluation - Advanced Reactor Technology Milestone: M3AT-15PN2301054

    SciTech Connect

    Ramuhalli, Pradeep; Hirt, Evelyn H.; Veeramany, Arun; Bonebrake, Christopher A.; Ivans, William J.; Coles, Garill A.; Coble, Jamie B.; Liu, X.; Wootan, David W.; Mitchell, Mark R.; Brass, Mary F.

    2015-09-24

    This research report summaries the development and evaluation of a prototypic enhanced risk monitor (ERM) methodology (framework) that includes alternative risk metrics and uncertainty analysis. This updated ERM methodology accounts for uncertainty in the equipment condition assessment (ECA), the prognostic result, and the probabilistic risk assessment (PRA) model. It is anticipated that the ability to characterize uncertainty in the estimated risk and update the risk estimates in real-time based on equipment condition assessment (ECA)will provide a mechanism for optimizing plant performance while staying within specified safety margins. These results (based on impacting active component O&M using real-time equipment condition information) are a step towards ERMs that, if integrated with AR supervisory plant control systems, can help control O&M costs and improve affordability of advanced reactors.

  13. 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

  14. Surgical Outcome of Idiopathic Epiretinal Membranes with Intraretinal Cystic Spaces

    PubMed Central

    Shiode, Yusuke; Morizane, Yuki; Toshima, Shinji; Kimura, Shuhei; Kumase, Fumiaki; Hosokawa, Mio; Hirano, Masayuki; Doi, Shinichiro; Takahashi, Kosuke; Hosogi, Mika; Fujiwara, Atsushi; Shiraga, Fumio

    2016-01-01

    Objective To investigate the occurrence ratio, localization, and surgical outcomes of intraretinal cystic spaces in idiopathic epiretinal membranes (ERMs). Methods We retrospectively reviewed the charts of 432 eyes of 398 consecutive patients with idiopathic ERM who underwent vitrectomy and ERM peeling from January 2012 to September 2015. We selected cases with intraretinal cystic space prior to surgery, detected by spectral-domain optical coherence tomography. We then evaluated the effects of ERM peeling on intraretinal cystic spaces, best corrected visual acuity, and central retinal thickness at 6 months after surgery. Results Twenty-four eyes (5.5%) showed intraretinal cystic spaces before surgery, present in the inner retinal layer (the inner group) in 9 eyes, in the outer retinal layer (the outer group) in 6 eyes, and in both the inner and the outer retinal layers (the combined group) in 9 eyes. Additionally, 30 eyes with ERM but without any presence of intraretinal cystic space were selected randomly and classified as the no cyst group. At 6 months after surgery, the disappearance rate of cystic spaces was significantly greater for the outer group than for the inner group (83.3% and 11.1%, respectively, P = 0.011). The mean best corrected visual acuity improved significantly after surgery in the inner group, the outer group, and the no cyst group (P < 0.05 for all three groups) but did not improve in the combined group (P = 0.58). The mean central retinal thickness decreased significantly after surgery in the inner group, the combined group, and the no cyst group (P < 0.05). Conclusions Intraretinal cystic spaces were observed in 5.5% of preoperative idiopathic ERM cases. Following surgery, the cystic spaces in the outer retinal layer disappeared at higher rates than those in the inner retinal layer, suggesting that the pathophysiologies of these cystic spaces are different. PMID:27992520

  15. [Phenotypes and genetic mechanisms of resistance to macrolides and lincosamides in viridans group streptococci].

    PubMed

    Artiles Campelo, F; Horcajada Herrera, I; Alamo Antúnez, I; Cãnas Pedrosa, A; Lafarga Capuz, B

    2007-09-01

    Viridans group streptococci (VGS) are part of the oropharyngeal, intestinal and genital flora, but they may cause endocarditis and bacteremia in susceptible patients. Penicillin- and macrolide-resistant strains are increasing every year. The aim of this study was to investigate genetic mechanisms of resistance to macrolides in clinically relevant isolates. We identified 85 isolates from January 2004 to June 2006. Susceptibility to penicillin, cefotaxime, erythromycin, clindamycin and gentamycin was determined. A resistance phenotype was assigned according to the disk approximation test (erythromycin-clindamycin). The mechanism of resistance was determined by PCR for the following genes: ermB, ermA, ermC, ermA (TR) and mefA/E. We identified 51 isolates belonging to Streptococcus anginosus species, most of which were obtained from abdominal abscesses, and 34 isolates belonging to other species, most of which were obtained from blood cultures. The macrolide resistance rate was 28.2% (24/85). The MLS(B) phenotype was observed in 66.7% of the isolates, primarily in the S. anginosus group. The M phenotype was predominant in S. mitis and S. oralis. Isolates that expressed the constitutive MLS(B) phenotype carried the ermB gene, and those that expressed the inducible MLSB phenotype carried the ermA gene. Isolates that expressed the M phenotype carried the mefA/E gene. There was coresistance with penicillin in 20.8% (5/24) of the isolates. Coresistance with penicillin was low. These results suggest that screening for macrolide resistance in VGS would be desirable because of the potential transmission of resistance genes to other pathogenic streptococci.

  16. Impacts of coexisting antibiotics, antibacterial residues, and heavy metals on the occurrence of erythromycin resistance genes in urban wastewater.

    PubMed

    Gao, Pin; He, Shi; Huang, Shenglin; Li, Kanzhu; Liu, Zhenhong; Xue, Gang; Sun, Weimin

    2015-05-01

    Antibiotic resistance is a global challenge and represents a growing threat on human health worldwide. Wastewater treatment plants (WWTPs) are generally considered as hotspots for control and/or dissemination of antibiotic resistance. The role of antibiotics, antibacterial residues, and heavy metals played on the evolution and spread of antibiotic resistance is still not well understood. Here, the occurrence of antibiotics (i.e., macrolides, tetracyclines, sulfonamides, and quinolones), antibacterial residues (i.e., triclosan), as well as heavy metals (i.e., cadmium, chromium, copper, zinc, lead, and nickel) in urban wastewater was investigated. Also, the abundances of erythromycin resistance genes (ERY-ARGs) including ere(A), ere(B), mef(A)/mef(E), erm(A), erm(B), erm(C), and msr(A)/msr(B) genes were screened. A relationship between certain antibiotics, antibacterial residues, and heavy metals and ERY-ARGs was demonstrated. ERY presented significant correlations (0.883 < r < 0.929, P < 0.05) with ere(A), ere(B), and mef(A)/mef(E) genes, while tetracycline exhibited a significant correlation (r = 0.829, P < 0.05) with erm(B) genes. It is noteworthy that triclosan correlated significantly (0.859 < r < 0.956, P < 0.05) with ere(A), ere(B), mef(A)/mef(E), and erm(B) genes. In addition, significantly positive correlations (0.823 < r < 0.871, P < 0.05) were observed between zinc and lead and certain ERY-ARGs (i.e., ere(B), mef(A)/mef(E), erm(B), etc.). Further investigations should be involved to elucidate the co-selection and/or cross-selection mechanisms due to co-existence of these selective factors in urban wastewater.

  17. 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.

  18. Earmuff restricts progenitor cell potential by attenuating the competence to respond to self-renewal factors

    PubMed Central

    Janssens, Derek H.; Komori, Hideyuki; Grbac, Daniel; Chen, Keng; Koe, Chwee Tat; Wang, Hongyan; Lee, Cheng-Yu

    2014-01-01

    Despite expressing stem cell self-renewal factors, intermediate progenitor cells possess restricted developmental potential, which allows them to give rise exclusively to differentiated progeny rather than stem cell progeny. Failure to restrict the developmental potential can allow intermediate progenitor cells to revert into aberrant stem cells that might contribute to tumorigenesis. Insight into stable restriction of the developmental potential in intermediate progenitor cells could improve our understanding of the development and growth of tumors, but the mechanisms involved remain largely unknown. Intermediate neural progenitors (INPs), generated by type II neural stem cells (neuroblasts) in fly larval brains, provide an in vivo model for investigating the mechanisms that stably restrict the developmental potential of intermediate progenitor cells. Here, we report that the transcriptional repressor protein Earmuff (Erm) functions temporally after Brain tumor (Brat) and Numb to restrict the developmental potential of uncommitted (immature) INPs. Consistently, endogenous Erm is detected in immature INPs but undetectable in INPs. Erm-dependent restriction of the developmental potential in immature INPs leads to attenuated competence to respond to all known neuroblast self-renewal factors in INPs. We also identified that the BAP chromatin-remodeling complex probably functions cooperatively with Erm to restrict the developmental potential of immature INPs. Together, these data led us to conclude that the Erm-BAP-dependent mechanism stably restricts the developmental potential of immature INPs by attenuating their genomic responses to stem cell self-renewal factors. We propose that restriction of developmental potential by the Erm-BAP-dependent mechanism functionally distinguishes intermediate progenitor cells from stem cells, ensuring the generation of differentiated cells and preventing the formation of progenitor cell-derived tumor-initiating stem cells. PMID

  19. 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

  20. Implementation of an enterprise risk-management program in a community teaching hospital.

    PubMed

    Behamdouni, Genefer; Millar, Kathy

    2010-01-01

    As the complexity of healthcare and expectations of comprehensive and transparent public accountability heighten, so too must a hospital's approach to assessing and managing risk. Over a period of two years, the area of patient safety and risk at our hospital has moved from a traditional focus on clinical risk management to an enterprise-wide risk management approach. One of the first community hospitals to embrace enterprise risk management (ERM), St. Joseph's Health Centre, in Toronto, Ontario, has seen early benefits in this transformational journey. This article discusses our approach to the development of an ERM program, tools used and lessons learned.

  1. Isolation of Genes Involved in Rac Induced Invasion and Metastasis of Breast Carcinoma Cells

    DTIC Science & Technology

    2001-08-01

    IQGAP Par-6 Dia - tLC LIM-I M- MLC- a-catenin kinase #phosphAr2/ z-catenin Par- Arp2/3 P-catenin aPKCs MLC-P cofilin adducin ERM NHE1 profilin E...a include MLC, the myosin-binding subunit of MLC sixth ROK substrate, namely NHE1 , is a ubiquitous phosphatase, LIM-kinase, adducin, the ERM (ezrin...Na÷/H’ exchange protein which potentiates stress fi- radixin/moesin) family of proteins, and a Na÷/IHI ex- ber formation [103]. change protein ( NHE1

  2. The Role of Drosophila Merlin in the Control of Mitosis Exit and Development

    DTIC Science & Technology

    2008-07-01

    gene / gene ?name=F42A10.2a;class=Transcript]. 52. The C . elegans Sequencing Consortium: Genome sequence of the nematode C . elegans : a...melanogaster merlin C . elegans nfm 1a C . briggsae merlin-like B. malayi merlin-like H. sapiens ezrin B. taurus ezrin G. gallus ezrin H. sapiens radixin G. gallus... elegans erm-like 1a C . briggsae erm-like D. melanogaster moesin T. saginata myosin-like E. multilocularis EM10 Actin-binding site

  3. Integrierte Segmentierung und Trajektorienberechnung mittels diffeomorpher Registrierung in räumlich-zeitlichen CT-Bildfolgen

    NASA Astrophysics Data System (ADS)

    Schmidt-Richberg, Alexander; Ehrhardt, Jan; Werner, René; Handels, Heinz

    Verfahren zur integrierten Segmentierung und Registrierung von 4D-Bilddaten ermöglichen die Berücksichtigung der gegenseitigen Abhängigkeiten beider Komponenten. Bestehende Ansätze konzentrieren sich dabei meist auf die Betrachtung zweier 3D-Bilder und lassen sich nicht direkt auf vollständige Bildfolgen übertragen. In dieser Arbeit wird ein Verfahren vorgestellt, in dem ein diffeomorpher Registrierungsansatz verwendet wird, um eine integrierte Berechnung von Segmentierungen und 3D-Voxeltrajektorien in 4D-Daten zu ermöglichen.

  4. Prevalence of resistance mechanisms against macrolides and lincosamides in methicillin-resistant coagulase-negative staphylococci in the Czech Republic and occurrence of an undefined mechanism of resistance to lincosamides.

    PubMed

    Novotna, Gabriela; Adamkova, Václava; Janata, Jiri; Melter, Oto; Spizek, Jaroslav

    2005-08-01

    High occurrence of the non-macrolide-lincosamide-streptogramin B resistance genes msrA (53%) and linA/linA' (30%) was found among 98 methicillin-resistant coagulase-negative staphylococci additionally resistant to macrolides and/or lincosamides. The gene msrA predominated in Staphylococcus haemolyticus (43 of 62 isolates). In Staphylococcus epidermidis, it was present in 7 of 27 isolates. A novel mechanism of resistance to lincosamides appears to be present in 10 genetically related isolates of S. haemolyticus in the absence of ermA, ermC, msrA, and linA/linA'.

  5. Phage ϕC2 Mediates Transduction of Tn6215, Encoding Erythromycin Resistance, between Clostridium difficile Strains

    PubMed Central

    Goh, Shan; Hussain, Haitham; Chang, Barbara J.; Emmett, Warren; Riley, Thomas V.; Mullany, Peter

    2013-01-01

    ABSTRACT In this work, we show that Clostridium difficile phage ϕC2 transduces erm(B), which confers erythromycin resistance, from a donor to a recipient strain at a frequency of 10−6 per PFU. The transductants were lysogenic for ϕC2 and contained the erm(B) gene in a novel transposon, Tn6215. This element is 13,008 bp in length and contains 17 putative open reading frames (ORFs). It could also be transferred at a lower frequency by filter mating. PMID:24255122

  6. 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

  7. Baroclinic internal wave energy distribution in the Baltic Sea derived from 45 years of circulation simulations

    NASA Astrophysics Data System (ADS)

    Rybin, Artem; Soomere, Tarmo; Kurkina, Oxana; Kurkin, Andrey; Rouvinskaya, Ekaterina; Markus Meier, H. E.

    2016-04-01

    Internal waves and internal tides are an essential component of the functioning of stratified shelf seas. They carry substantial amounts of energy through the water masses, drive key hydrophysical processes such as mixing and overturning and support the functioning of marine ecosystem in many ways. Their particular impact becomes evident near and at the bottom where they often create substantial loads to engineering structures and exert a wide range of impacts on the bottom sediments and evolution of the seabed. We analyse several properties of spatio-temporal distributions of energy of relatively long-period large-scale internal wave motions in the Baltic Sea. The analysis is based on numerically simulated pycnocline variations that are extracted from the hydrographic data calculated by the Rossby Centre Ocean circulation model (RCO) for the entire Baltic Sea for 1961-2005. This model has a horizontal resolution of 2 nautical miles and uses 41 vertical layers with a thickness between 3 m close to the surface and 12 m in 250 m depth. The model is forced with atmospheric data derived from the ERA-40 re-analysis using a regional atmosphere model with a horizontal resolution of 25 km. It also accounts for river inflow and water exchange through the Danish Straits. See (Meier, H.E.M., Höglund, A., 2013. Studying the Baltic Sea circulation with Eulerian tracers, in Soomere, T., Quak, E., eds., Preventive Methods for Coastal Protection, Springer, Cham, Heidelberg, 101-130) for a detailed description of the model and its forcing. The resolution of the model output used in this study (once in 6 hours) is sufficient for estimates of spectral amplitudes of the displacements of isopycnal surfaces with a typical period of 2-12 days. We provide the analysis of kinetic and potential energy of motions with these periods. The resulting maps of the maxima of energy and spatial distributions of near-bottom velocities have been evaluated for the entire simulation interval of 45

  8. Resistance to macrolides in Streptococcus pyogenes in France in pediatric patients.

    PubMed

    Bingen, E; Fitoussi, F; Doit, C; Cohen, R; Tanna, A; George, R; Loukil, C; Brahimi, N; Le Thomas, I; Deforche, D

    2000-06-01

    A total of 1,500 recent throat isolates of Streptococcus pyogenes collected between 1996 and 1999 from children throughout France were tested for their susceptibility to erythromycin, azithromycin, josamycin, clindamycin, and streptogramin B. The erythromycin-resistant isolates were further studied for their genetic mechanism of resistance, by means of PCR. The clonality of these strains was also investigated by means of serotyping and ribotyping. In all, 6.2% of the strains were erythromycin resistant, and 3.4 and 2.8% expressed the constitutive MLS(B) and M resistance phenotypes and harbored the ermB and mefA genes, respectively; ermTR was recovered from one isolate which also harbored the ermB gene. Ten serotypes and 8 ribotypes were identified, but we identified 17 strains by combining serotyping with ribotyping. Among the eight ribotypes, the mefA gene was recovered from six clusters, one being predominant, while the ermB gene was recovered from four clusters, of which two were predominant.

  9. Automated Information Systems Plan

    DTIC Science & Technology

    1989-10-01

    XNS-TCP/IP = Xerox Network Service-Transmission Control Protocol/ Internet Protocol 35 APPENDIX A SUMMARY OF USER SURVEY RESPONSES This appendix is...COVERED 14 DATE OF REPORT (Year, Month, Day) IS PAGE COUNT Fi FROM _____ _____ October 1989 40 16 53PPEMEN-AR’Y NOTATON 7 COSA -I CODES 13 SUBECT rERMS

  10. 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.

  11. Desert Storm’s Siren Song; Examining Revolution in Warfare

    DTIC Science & Technology

    1994-05-27

    develop a force structure which is unable to deal with the full gamut of twenty- first century threats. 14. SuluECT T1ERMS DeetRvlto nWraeIS. NUMBER Of...War a revolution could lead the U.S. military to develop a force structure which is unable to deal with the full gamut of twenty-first century

  12. 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.

  13. 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.

  14. 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,…

  15. Engineering of plant-specific phenylpropanoids biosynthesis in Streptomyces venezuelae.

    PubMed

    Park, Sung Ryeol; Yoon, Jin A; Paik, Ji Hye; Park, Je Won; Jung, Won Seok; Ban, Yeon-Hee; Kim, Eun Ji; Yoo, Young Ji; Han, Ah Reum; Yoon, Yeo Joon

    2009-05-20

    Phenylpropanoids, including flavonoids and stilbenes, are plant secondary metabolites with potential pharmacological and nutraceutical properties. To expand the applicability of Streptomyces venezuelae as a heterologous host to plant polyketide production, flavonoid and stilbene biosynthetic genes were expressed in an engineered strain of S. venezuelae DHS2001 bearing a deletion of native pikromycin polyketide synthase gene. A plasmid expressing the 4-coumarate/cinnamate:coenzyme A ligase from Streptomyces coelicolor (ScCCL) and the chalcone synthase from Arabidopsis thaliana (atCHS) under the control of a single ermE* promoter was constructed and introduced into S. venezuelae DHS2001. The resulting strain produced racemic naringenin and pinocembrin from 4-coumaric acid and cinnamic acid, respectively. Placement of an additional ermE* promoter upstream of the codon-optimized atCHS (atCHS(op)) gene significantly increased the yield of both flavanones. Expression of codon-optimized chalcone isomerase gene from Medicago sativa, together with ScCCL and atCHS(op) genes led to production of (2S)-flavanones, but the yield was reduced. On the other hand, a recombinant strain harboring the ScCCL and codon-optimized stilbene synthase gene from Arachis hypogaea generated stilbenes such as resveratrol and pinosylvin. This is the first report on the heterologous expression of plant phenylpropanoid biosynthetic pathways in Streptomyces genus.

  16. Macrolide-Resistant Streptococcus pyogenes in Norway: Population Structure and Resistance Determinants

    PubMed Central

    Littauer, P.; Caugant, D. A.; Sangvik, M.; Høiby, E. A.; Sundsfjord, A.; Simonsen, G. S.

    2006-01-01

    A 2.7% prevalence of macrolide resistance in 1,657 Norwegian clinical Streptococcus pyogenes isolates was primarily due to erm(TR) (59%) and mef(A) (20%). Four clonal complexes comprised 75% of the strains. Macrolide resistance in S. pyogenes in Norway is imported as resistant strains or locally selected in internationally disseminated susceptible clones. PMID:16641473

  17. 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…

  18. 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

  19. 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…

  20. 77 FR 76476 - Combined Notice of Filings #1

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-12-28

    .... Docket Numbers: ER13-357-001. Applicants: Cirrus Wind 1, LLC. Description: Substitute MBR Tariff to be...: CMS Energy Resource Management Company. Description: CMS ERM Company--MBR to be effective 12/19/2012... Station--MBR to be effective 12/19/2012. Filed Date: 12/19/12. Accession Number: 20121219-5000....

  1. Checks and Balances: Rpd3 Issues Executive Orders in Developmental Enhancer Regulation.

    PubMed

    Martire, Sara; Banaszynski, Laura

    2017-02-27

    Stem cells use poised enhancers of developmental regulators to maintain pluripotency and for subsequent activation in differentiating progeny. In this issue of Developmental Cell, Janssens et al. (2017) demonstrate that the erm enhancer is maintained in a poised state in neural stem cells by the histone deacetylase Hdac1/Rpd3.

  2. Targeting Thromboxane A2 Receptor for Anti-Metastasis Therapy of Breast Cancer

    DTIC Science & Technology

    2011-09-01

    phosphatase, LIM- kinase, adducin, ezrin/radixin/moesin (ERM) family of proteins, and Na+/H+ exchange protein ( NHE1 ). Among the six substrates, MLC...proteins and RhoA (Lamb, Roy et al. 2000). Finally, NHE1 is well known as a ubiquitous Na+/H+ exchange protein that enables stress fiber formation

  3. [The Educational Resources Management System: Application of PPBES Concepts to Education.

    ERIC Educational Resources Information Center

    Association of School Business Officials, Chicago, IL.

    This document reports the results of a survey of school business officials to determine the status of Educational Resource Management Systems (ERMS) or Planning-Programing-Budgeting-Evaluating Systems (PPBES) in school districts and colleges in the United States and Canada. Though the actual number of school districts developing management systems…

  4. 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.

  5. CERT Resilience Management Model: A Maturity Model Approach to Managing Operational Resilience

    DTIC Science & Technology

    2010-07-28

    for agile adoption 5 © 2010 Carnegie Mellon University Distinguishing features of CERT®-RMM CERT-RMM brings several innovative and advantageous...Specific Goals Specific Practices Sub- practices Codes of Practice: BS25999-1:2006 CMMI v1.2 CMMI for Services CobiT 4.1 COSO ERM DRII GAP FFIEC

  6. Ezrin and moesin expression within the developing human cerebrum and tuberous sclerosis-associated cortical tubers.

    PubMed

    Johnson, Michael W; Miyata, Hajime; Vinters, Harry V

    2002-08-01

    The ERM (ezrin, radixin, and moesin) proteins belong to the band-4.1 superfamily of membrane-cytoskeleton-linking proteins which bind to the actin cytoskeleton via their C-terminal sequences and bind ERM binding membrane proteins (ERMBMPs). We investigated the immunohistochemical expression of two of the ERM proteins (ezrin and moesin) in developing human cerebral cortex and in cortical tubers from patients with tuberous sclerosis (TSC), to assess possible consequences of TSC gene product malfunction or inactivation in the developing brain in relation to ERM protein expression. Ezrin is abundantly expressed within radial glia and migrating cells in the intermediate zone in the prenatal human cerebrum, while moesin is primarily expressed in vascular endothelial cells in developing and adult human brain and scattered microglia in adult brain. In addition, both ezrin and moesin are abundantly co-expressed with hamartin and tuberin within a population of abnormal cells in TSC-associated cortical tubers. The expression of these two proteins--primarily ezrin--suggests that they are developmentally regulated and abundantly expressed in germinal matrix and/or migrating cells during cerebral cortical development. In TSC-associated cortical tubers, both proteins appeared to be up-regulated and are co-localized within a population of abnormal neuroglial cells typical of those seen in tubers. Expression of these proteins and their co-localization with tuberin and hamartin in these cells may suggest a compensatory up-regulation in response to TSC gene mutation.

  7. Moesin is a glioma progression marker that induces proliferation and Wnt/β-catenin pathway activation via interaction with CD44.

    PubMed

    Zhu, Xiaoping; Morales, Fabiana C; Agarwal, Nitin Kumar; Dogruluk, Turgut; Gagea, Mihai; Georgescu, Maria-Magdalena

    2013-02-01

    Moesin is an ERM family protein that connects the actin cytoskeleton to transmembrane receptors. With the identification of the ERM family protein NF2 as a tumor suppressor in glioblastoma, we investigated roles for other ERM proteins in this malignancy. Here, we report that overexpression of moesin occurs generally in high-grade glioblastoma in a pattern correlated with the stem cell marker CD44. Unlike NF2, moesin acts as an oncogene by increasing cell proliferation and stem cell neurosphere formation, with its ectopic overexpression sufficient to shorten survival in an orthotopic mouse model of glioblastoma. Moesin was the major ERM member activated by phosphorylation in glioblastoma cells, where it interacted and colocalized with CD44 in membrane protrusions. Increasing the levels of moesin competitively displaced NF2 from CD44, increasing CD44 expression in a positive feedback loop driven by the Wnt/β-catenin signaling pathway. Therapeutic targeting of the moesin-CD44 interaction with the small-molecule inhibitor 7-cyanoquinocarcinol (DX-52-1) or with a CD44-mimetic peptide specifically reduced the proliferation of glioblastoma cells overexpressing moesin, where the Wnt/β-catenin pathway was activated. Our findings establish moesin and CD44 as progression markers and drugable targets in glioblastoma, relating their oncogenic effects to activation of the Wnt/β-catenin pathway.

  8. Expression of ezrin in subventricular zone neural stem cells and their progeny in adult and developing mice.

    PubMed

    Moon, Younghye; Kim, Joo Yeon; Choi, So Yoen; Cho, Hyo Min; Kim, Hyun; Sun, Woong

    2013-03-01

    Ezrin is a member of the ezrin-radixin-moesin (ERM) family of proteins, which link the cytoskeleton and cell membrane. ERM proteins are involved in pivotal cellular functions including cell-matrix recognition, cell-cell communication, and cell motility. Several recent studies have shown that ERM proteins are expressed in specific cell types of the adult rostral migratory stream (RMS). In this study, we found that ERM proteins are expressed highly in the early postnatal RMS and the ventricular zone of embryonic cerebral cortex, suggesting that these proteins may be expressed by neural progenitors. Furthermore, whereas ezrin previously was found to be expressed exclusively by astrocytes of the adult RMS, we found that ezrin-expressing cells also expressed the markers for indicating neuroblasts in vivo and in vitro, and that ezrin expression by neuroblasts decreases progressively as neuroblasts migrate. Using in vitro differentiation of adult neural stem cells, we found that ezrin is expressed by neural stem cells and their progeny (neuroblasts and astrocytes), but not by oligodendrocytic progeny. Collectively our findings demonstrate that adult neural stem cells and neuroblasts express ezrin and that ezrin may be involved in intracellular actin remodeling.

  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.

  10. Principles for Information Technology Investment in U.S. Federal Electronic Records Management.

    ERIC Educational Resources Information Center

    Van Wingen, Rachel Senner; Hathorn, Fred; Sprehe, J. Timothy

    1999-01-01

    The United States Environmental Protection Agency (EPA) underwent a business process reengineering (BPR) exercise with respect to future co-location of previously separate regulatory docket facilities. Their experience suggests that future mandatory electronic records management (ERM) requirements will cause federal agencies to take a more…

  11. 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 ...

  12. 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.

  13. Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Can Accurately Differentiate between Mycobacterium masilliense (M. abscessus subspecies bolletti) and M. abscessus (Sensu Stricto)

    PubMed Central

    Teng, Shih-Hua; Chen, Chung-Ming; Lee, Meng-Rui; Lee, Tai-Fen; Chien, Kun-Yi; Teng, Lee-Jene

    2013-01-01

    Among 36 Mycobacterium masilliense and 22 M. abscessus isolates identified by erm(41) PCR and sequencing analysis of rpoB and 23S rRNA genes, the rate of accurate differentiation between these two subspecies was 100% by cluster analysis of spectra generated by Bruker Biotyper matrix-assisted laser desorption ionization–time of flight mass spectrometry. PMID:23824775

  14. A Computer Simulation of an Adaptive Noise Canceler with a Single Input

    DTIC Science & Technology

    1991-06-01

    parameter values needed to cause the canceler to attenuate the strong signal to a user specified level and pass the weak signal. This study was...10 dBm. (continued) 71 rn -: - , erm utut at the kth interation X k - PTF tap amplptu2u vector at the kth iteration. When the input interfering

  15. Observations on continuously growing roots of the sloth and the K14-Eda transgenic mice indicate that epithelial stem cells can give rise to both the ameloblast and root epithelium cell lineage creating distinct tooth patterns.

    PubMed

    Tummers, Mark; Thesleff, Irma

    2008-01-01

    Root development is traditionally associated with the formation of Hertwig's epithelial root sheath (HERS), whose fragments give rise to the epithelial cell rests of Malassez (ERM). The HERS is formed by depletion of the core of stellate reticulum cells, the putative stem cells, in the cervical loop, leaving only a double layer of the basal epithelium with limited growth capacity. The continuously growing incisor of the rodent is subdivided into a crown analog half on the labial side, with a cervical loop containing a large core of stellate reticulum, and its progeny gives rise to enamel producing. The lingual side is known as the root analog and gives rise to ERM. We show that the lingual cervical loop contains a small core of stellate reticulum cells and suggest that it acts as a functional stem cell niche. Similarly we show that continuously growing roots represented by the sloth molar and K14-Eda transgenic incisor maintain a cervical loop with a small core of stellate reticulum cells around the entire circumference of the tooth and do not form a HERS, and still give rise to ERM. We propose that HERS is not a necessary structure to initiate root formation. Moreover, we conclude that crown vs. root formation, i.e. the production of enamel vs. cementum, and the differentiation of the epithelial cells into ameloblasts vs. ERM, can be regulated independently from the regulation of stem cell maintenance. This developmental flexibility may underlie the developmental and evolutionary diversity in tooth patterning.

  16. Negotiating with the North Vietnamese: A Military Perspective

    DTIC Science & Technology

    1975-06-06

    advancing toward national reunification and socialism . With regard to etruggle, we otand for Joint political end ermed etruggle, that Is to say, the...fighting. While nego- tiating, we will continue fighting the enemy more viqoroualy. It is poaaible that the North conducta nagatiatians while the South

  17. 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.

  18. 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 ...

  19. mef(A) is the predominant macrolide resistance determinant in Streptococcus pneumoniae and Streptococcus pyogenes in Germany.

    PubMed

    Bley, Christine; van der Linden, Mark; Reinert, Ralf René

    2011-05-01

    In this study, macrolide-resistant Streptococcus pneumoniae and Streptococcus pyogenes isolates from Germany were carefully characterised by susceptibility testing, phenotyping, polymerase chain reaction (PCR) and sequencing of macrolides resistance genes, and multilocus sequence typing (MLST). Of 2045 S. pneumoniae and 352 S. pyogenes isolates, 437 (21.4%) and 29 (8.2%), respectively, were found to be macrolide-resistant. Amongst the S. pneumoniae isolates, the most prevalent resistance marker was mef(A) (57.7%) followed by erm(B) (27.0%) and mef(E) (11.2%). Of note, the dual resistance mechanism mef(E)+erm(B) was found in a relatively high proportion (4.1%) of pneumococcal isolates. Amongst the S. pyogenes isolates, 31.0% carried mef(A), 34.5% erm(B) and 13.8% erm(A). Dissemination of a single clone [mef(A)-positive England(14)-9] has significantly contributed to the emergence of macrolide resistance amongst pneumococci in Germany.

  20. E-Resource Statistics: What to Do when You Have No Money

    ERIC Educational Resources Information Center

    Walker, Mary

    2009-01-01

    Libraries are moving toward electronic resource management systems (ERMSs) to track their usage statistics, but these can be expensive to purchase and maintain. For some libraries, an ERMS can be cost-prohibitive, but they still need to justify the renewal of databases and e-journals to their budget officers or determine which e-resources should…

  1. 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.

  2. Flagellar regulation in 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), a septicemia affecting primarily 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 a...

  3. 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...

  4. 77 FR 37038 - Clean Air Act Operating Permit Program; Petition for Objection to State Operating Permit for CF&I...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-20

    ... Permit Program; Petition for Objection to State Operating Permit for CF&I Steel, L.P. dba EVRAZ Rocky Mountain Steel AGENCY: Environmental Protection Agency (EPA). ACTION: Notice of final action. SUMMARY: This... December 28, 2010 Permit (Permit) issued to CF&I Steel, L.P. dba EVRAZ Rocky Mountain Steel (ERMS or...

  5. 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 ...

  6. 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.

  7. 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...

  8. 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

  9. Investigation of drug-drug interaction via mechanism-based inhibition of cytochrome P450 3A by macrolides in dexamethasone-treated female rats.

    PubMed

    Kanazu, Takushi; Sato, Norihito; Kadono, Kyoko; Touchi, Akira; Takeda, Yuri; Yamaguchi, Yoshitaka; Baba, Takahiko

    2012-05-01

    The in vitro and in vivo inhibition of cytochrome P450 (CYP) 3A with mechanism-based inhibition (MBI) by macrolides was investigated using dexamethasone-treated female rats (DEX-female rats). In the in vitro CYP inhibition studies using erythromycin (ERM) and clarithromycin (CAM), similar inhibition responses were observed between human and DEX-female rat liver microsomes, however, there were fewer effects in intact male rats. The ex vivo study showed that midazolam (MDZ) metabolism in liver microsomes of DEX-female rats was reduced by ERM administration and the inhibitory effect was increased with increasing ERM doses, indicating that metabolite intermediate complex formation caused irreversible inhibition of CYP3A activity in DEX-female rats as well as in humans. In the in vivo studies, ERM and CAM significantly increased the area under the plasma concentration-time curve of MDZ and decreased the total clearance in DEX-female rats. It was concluded that the DDIs via MBI of CYP3A following macrolide administration in humans could be reproduced in female rats, suggesting that DEX-female rats can serve as an in vivo model for assessing this DDI in humans.

  10. Addressing risk in the design of new healthcare services: a step-by-step methodology.

    PubMed

    Remus, Beth

    2008-01-01

    New service design can be done effectively through enterprise risk management (ERM) by identifying the clinical and administrative risks within the new service design. This article proposes a 10-step process describing the business team's tasks and integrating risk management, quality management and patient safety concepts.

  11. A screening model for depleted uranium testing using environmental radiation monitoring data

    SciTech Connect

    Dunfrund, F.L.; Ebinger, M.H.; Hansen, W.R.

    1996-06-01

    Information from an ecological risk assessment of depleted uranium test areas at Yuma Proving Ground (YPG) was used to update the required environmental radiation monitoring (ERM) plan. Data to be collected for the ERM can also be used to evaluate the potential for adverse radiological and toxicological effects to terrestrial reptiles and mammals in the affected areas. We developed a spreadsheet-based screening model that incorporates the ERM data and associated uncertainties. The purpose of the model is to provide a conservative estimate of radiological exposure of terrestrial, biota to DU using the ERM data. The uncertainty in the estimate is also predicted so that the variation in the radiological exposure can be used in assessing potential adverse effects from DU testing. Toxicological effects are evaluated as well as radiological effects in the same program using the same data. Our presentation shows an example data set, model calculations, and the report of expected radiation dose rates and probable kidney burdens of select mammals and reptiles. The model can also be used in an inverse mode to calculate the soil concentration required to give either a radiological dose that would produce a potential adverse effect such as fatal cancer or a toxicological dose that would result in nephrotoxic effects in mammals.

  12. The First Direct Measurements of Upper Oceanic Crustal Compressional Wave Attenuation

    DTIC Science & Technology

    1990-01-01

    glossesusingequation(10)and [ Ostle and Mensing, 1975]) indicate a significant difference in for xararnissicn . across the water-basemet interface, variances between...34:d scales of such heterogeneities 1973.0 Ostle , B., and R. W. Mensing. Statistics in Research, 596 pp., Iowa State cannot be de:erm,ned, but the s7

  13. Optimizing a Synthetic Signaling System, Using Mathematical Modeling to Direct Experimental Work

    DTIC Science & Technology

    2014-09-05

    Journal of Experimental Botany 53, 1871-1877, doi:10.1093/jxb/erf050 (2002). 52 Looger, L. L., Dwyer, M. A., Smith, J. J. & Hellinga, H. W...P. F., Christie, J. M. & Terry, M. J. Many hands make light work. Journal of Experimental Botany 58, 3071-3077, doi:10.1093/jxb/erm251 (2007).

  14. 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

  15. 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.

  16. 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…

  17. A comparison of sediment quality guidelines for toxicity assessment in the Sunderban wetlands (Bay of Bengal, India).

    PubMed

    Binelli, Andrea; Sarkar, Santosh Kumar; Chatterjee, Mousumi; Riva, Consuelo; Parolini, Marco; Bhattacharya, Bhaskar deb; Bhattacharya, Asok Kumar; Satpathy, Kamala Kanta

    2008-10-01

    The aim of this paper was to obtain the first screening ecotoxicological risk evaluation in the Sunderban wetlands, the largest prograding delta in the estuarine phase of the River Ganges. The characterization of exposure was conducted by means of an extensive survey of several persistent organic pollutants (PAHs, PCBs, DDTs, PBDEs, HCHs, HCB) measured in seven core sediments from the Sunderban wetlands, obtaining a dataset with more than 2200 analyses. The pollutant effects were assessed by the use of three different sediment quality guidelines (SQGs) previously developed in the literature to evaluate toxicity induced in sediment-dwelling organisms. The three different approaches chosen for risk assessment of the Sunderban were the consensus SQGs obtained by TEC (threshold effect concentration), PEC (probable effect concentration) and EEC (extreme effect concentration), the threshold/probable effect level (TEL/PEL) approach and, finally, the ERL-ERM guidelines, including the m-ERM-Q (mean ERM quotient). The evaluation of the toxicity induced by a mixture of the target pollutants indicated the importance of gamma-HCH contamination in the Sunderban sediments despite the very low concentrations measured in core sediments. A different sensitivity for toxicity assessment due to quality guidelines was obtained, as the consensus SQGs based on TEC were less conservative and protective than the TEL and ERL approaches, while the use of m-ERM-Q seems to be the most powerful tool to predict the toxicity related to a contaminant mixture.

  18. Prostate embryonal rhabdomyosarcoma in adults: Case report and review of literature

    PubMed Central

    Ciammella, Patrizia; Galeandro, Maria; D’Abbiero, Nunziata; Palmieri, Tamara; Donini, Elisa; Iotti, Cinzia

    2013-01-01

    Introduction Prostate embryonal rhabdomyosarcoma (ERMS) is a common tumour in infants and children, with a median occurrence age of 5 years, but it is rare in adults. It is characterized by a high degree of malignancy, both local rapid growth with formation of large pelvic masses, often leading to renal failure due to urethral obstruction, and systemic spread, commonly to the lungs, liver and bone. Several therapeutic approaches have been employed in the effort to treat prostate ERMS, but all of them have failed to gain a significant survival benefit in adult patients. Case report We report on a case of a stage IV prostate ERMS, approached with combined-modality treatment, with the administration of 5 courses of doxorubicin, ifosfamide and 2-mercaptoethane sulfonate sodium (mesna), and, subsequent radiotherapy to the prostatic bed (60 Gy/30 fxs). The patient remained free of progression of disease for about 1 year to finally experience a systemic relapse with multiple lung metastases and pleural effusion. The patient died for metastatic disease 27 months following the initial diagnosis. Conclusion While it remains questionable which therapeutic approach for prostate ERMS in adults is the most appropriate, our report demonstrates that a chemo-radiation combined treatment can control the prostate disease, reducing the symptoms and improving the quality of life of these patients, for the most part destined to die for systemic progression of disease. PMID:24416569

  19. 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…

  20. 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...

  1. 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…

  2. 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…

  3. Evolution and origin of merlin, the product of the Neurofibromatosis type 2 (NF2) tumor-suppressor gene

    PubMed Central

    Golovnina, Kseniya; Blinov, Alexander; Akhmametyeva, Elena M; Omelyanchuk, Leonid V; Chang, Long-Sheng

    2005-01-01

    Background Merlin, the product of the Neurofibromatosis type 2 (NF2) tumor suppressor gene, belongs to the ezrin-radixin-moesin (ERM) subgroup of the protein 4.1 superfamily, which links cell surface glycoproteins to the actin cytoskeleton. While merlin's functional activity has been examined in mammalian and Drosophila models, little is understood about its evolution, diversity, and overall distribution among different taxa. Results By combining bioinformatic and phylogenetic approaches, we demonstrate that merlin homologs are present across a wide range of metazoan lineages. While the phylogenetic tree shows a monophyletic origin of the ERM family, the origin of the merlin proteins is robustly separated from that of the ERM proteins. The derivation of merlin is thought to be in early metazoa. We have also observed the expansion of the ERM-like proteins within the vertebrate clade, which occurred after its separation from Urochordata (Ciona intestinalis). Amino acid sequence alignment reveals the absence of an actin-binding site in the C-terminal region of all merlin proteins from various species but the presence of a conserved internal binding site in the N-terminal domain of the merlin and ERM proteins. In addition, a more conserved pattern of amino acid residues is found in the region containing the so-called "Blue Box," although some amino acid substitutions in this region exist in the merlin sequences of worms, fish, and Ciona. Examination of sequence variability at functionally significant sites, including the serine-518 residue, the phosphorylation of which modulates merlin's intra-molecular association and function as a tumor suppressor, identifies several potentially important sites that are conserved among all merlin proteins but divergent in the ERM proteins. Secondary structure prediction reveals the presence of a conserved α-helical domain in the central to C-terminal region of the merlin proteins of various species. The conserved residues and

  4. 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.

  5. 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.

  6. In vitro activity of telithromycin against Spanish Streptococcus pneumoniae isolates with characterized macrolide resistance mechanisms.

    PubMed

    Morosini, M I; Cantón, R; Loza, E; Negri, M C; Galán, J C; Almaraz, F; Baquero, F

    2001-09-01

    The susceptibilities to telithromycin of 203 Streptococcus pneumoniae isolates prospectively collected during 1999 and 2000 from 14 different geographical areas in Spain were tested and compared with those to erythromycin A, clindamycin, quinupristin-dalfopristin, penicillin G, cefotaxime, and levofloxacin. Telithromycin was active against 98.9% of isolates (MICs, < or =0.5 microg/ml), with MICs at which 90% of isolates are inhibited being 0.06 microg/ml, irrespective of the resistance genotype. The corresponding values for erythromycin were 61.0% (MICs, < or =0.25 microg/ml) and >64 microg/ml. The erm(B) gene (macrolide-lincosamide-streptogramin B resistance phenotype) was detected in 36.4% (n = 74) of the isolates, which corresponded to 93.6% of erythromycin-intermediate and -resistant isolates, whereas the mef(A) gene (M phenotype [resistance to erythromycin and susceptibility to clindamycin and spiramycin without blunting]) was present in only 2.4% (n = 5) of the isolates. One of the latter isolates also carried erm(B). Interestingly, in one isolate for which the erythromycin MIC was 2 microg/ml, none of these resistance genes could be detected. Erythromycin MICs for S. pneumoniae erm(B)-positive isolates were higher (range, 0.5 to >64 microg/ml) than those for erm(B)- and mef(A)-negative isolates (range, 0.008 to 2 microg/ml). The corresponding values for telithromycin were lower for both groups, with ranges of 0.004 to 1 and 0.002 to 0.06 microg/ml, respectively. The erythromycin MIC was high for a large number of erm(B)-positive isolates, but the telithromycin MIC was low for these isolates. These results indicate the potential usefulness of telithromycin for the treatment of infections caused by erythromycin-susceptible and -resistant S. pneumoniae isolates when macrolides are indicated.

  7. 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

  8. Effects of bamboo charcoal on antibiotic resistance genes during chicken manure composting.

    PubMed

    Li, Haichao; Duan, Manli; Gu, Jie; Zhang, Yajun; Qian, Xun; Ma, Jun; Zhang, Ranran; Wang, Xiaojuan

    2017-06-01

    Composting is widely used for animal waste disposal, and bamboo charcoal (BC) can be used for nitrogen conservation during composting. However, the effects of BC on antibiotic resistance genes (ARGs) during chicken manure composting are still unclear. This study investigated the effects on ARGs of adding different proportions of BC (0%, 5%, 10%, and 20% w/w) to chicken manure compost. After 26 days, the relative abundances (RAs) of most ARGs (tetC, tetG, tetW, tetX, sul2, drfA1, drfA7, ermB, ermF, ermQ, and ermX) and intI1 declined by 21.6-99.5%, whereas sul1 increased by 7.5-17.7 times. The average RAs reductions with 0%, 5%, 10%, and 20% BC were 0.85, 1.05, 1.08, and 1.15 logs, respectively. The most important environmental factor for the ARG profiles was temperature according to redundancy analysis. Furthermore, BC significantly decreased the bio-Cu and bio-Zn levels, thereby reducing the co-selection pressure from heavy metals. Different proportions of BC had no significant effects on the removal of tetG, tetW, tetX, sul2, drfA1, and ermB. Supplementation with 10% BC was more effective at removing tetC and drfA7 compared with the other treatments. The results suggested that 10% BC supplementation is appropriate for reducing ARGs in chicken manure compost.

  9. Staphylococcus aureus nasal carriage, virulence traits, antibiotic resistance mechanisms, and genetic lineages in healthy humans in Spain, with detection of CC398 and CC97 strains.

    PubMed

    Lozano, Carmen; Gómez-Sanz, Elena; Benito, Daniel; Aspiroz, Carmen; Zarazaga, Myriam; Torres, Carmen

    2011-08-01

    S. aureus nasal carriage was investigated in 278 healthy humans, determining the antibiotic resistance mechanisms, virulence traits, and genetic lineages of recovered isolates. Nasal samples were cultured in specific media for S. aureus and methicillin-resistant S. aureus (MRSA) recovery. S. aureus was detected in 53 of 278 nasal samples (19.1%): MRSA was found in one sample (0.4%) and methicillin-susceptible S. aureus (MSSA) in the remaining 52 samples. The MRSA isolate was typed as ST1649-t701-agrI-SCCmec-IVc and only exhibited resistance to beta-lactams. A high diversity of spa types (n=37) was identified among the 52 MSSA, identifying 5 new spa-types. Multi-locus sequence typing (MLST) typing was performed in 30 selected MSSA, detecting 16 different sequence types, 2 of them being new. MSSA strains presented agr types I (30.2%), II (30.2%), III (34%), and IV (5.6%). Eleven strains showed erythromycin resistance and harbored different combinations of erm(A), erm(B), erm(C), erm(T), and msr(A) genes. Two strains exhibited ciprofloxacin resistance, and one of them presented amino acid changes in GyrA and GrlA proteins. The presence of 28 genes encoding staphylococcal toxins was investigated by PCR in all 53 S. aureus isolates. The toxic shock syndrome toxin 1 (TSST-1) gene was detected in 15 MSSA isolates (11 of them typed within the clonal complex CC30) and the gene of exfoliative toxin A in 2 strains. Different combinations of enterotoxin genes were identified among S. aureus strains. None of the S. aureus isolates harbored the Panton-Valentine leukocidin gene. Two MSSA presented the sequence-type ST398 [harboring erm(T) gene], and 2 additional isolates were typed as ST97. Interestingly, MSSA CC398 and CC97 isolates were detected. These clonal complexes are associated with food-producing animals.

  10. Effect of steam-flaked corn and soybeans on muscle and intramuscular fatty acid composition in Holstein calves.

    PubMed

    Zhang, Y Q; He, D C; Meng, Q X; Wang, D C

    2015-12-01

    This study aimed to evaluate the effects of steam-flaked corn grains and soybeans on muscle fatty acid composition. Thirty Holstein bull calves (21 ± 3 d) were divided into 3 groups according to birth date and BW and were randomly assigned to receive fresh milk and a commercial pelleted starter diet containing extruded corn and soybean (ECS), steam-flaked corn and soybean (SFCS), or ground corn and raw soybean (GCS). The calves were fed the designated diet from 3 to 13 wk of age, after which they were slaughtered. The supraspinatus (CTM), longissimus lumborum (RLM), and spinalis dorsi (ERM) were analyzed to determine the chemical and intramuscular fatty acid composition. The fatty acid composition of muscle and its deposition differed among calves fed different starter feeds. Medium-chain fatty acid levels of the RLM and CTM were greater in GCS-fed calves than in ECS- and SFCS-fed calves ( < 0.05). Extruded processing increased the content of linoleic, linolenic, and arachidonic acids of the RLM ( < 0.05). The palmitoleic and -vaccenic acid content of the ERM were greater in GCS-fed calves than in ECS- or SFCS-fed calves ( < 0.05). No significant differences were observed among the 3 diets with respect to the stearic, oleic, linoleic, -9 -11 CLA, or arachidonic acid content of the ERM ( > 0.05). The levels of -3 and -6 fatty acids were similar among the 3 groups; a lower -6:-3 PUFA ratio was observed in GCS-fed calves ( < 0.05). The cereal processing method of the calf starter feed had no significant effect on the chemical composition of the CTM, RLM, or ERM. Therefore, different methods of processing corn and soybean in calf starter feeds had no effect on the chemical composition of the RLM, CTM, or ERM but had a significant effect on the intramuscular fatty acid composition.

  11. Involvement of PtdIns(4,5)P2 in the regulatory mechanism of small intestinal P-glycoprotein expression.

    PubMed

    Kobori, Takuro; Harada, Shinichi; Nakamoto, Kazuo; Tokuyama, Shogo

    2014-02-01

    Previously, we reported that repeated oral administration of etoposide (ETP) activates the ezrin/radixin/moesin (ERM) scaffold proteins for P-glycoprotein (P-gp) via Ras homolog gene family member A (RhoA)/Rho-associated coiled-coil containing protein kinase (ROCK) signaling, leading to increased ileal P-gp expression. Recent studies indicate that phosphatidyl inositol 4,5-bisphosphate [PtdIns(4,5)P2] regulates the plasma-membrane localization of certain proteins, and its synthase, the type I phosphatidyl inositol 4-phosphate 5-kinase (PI4P5K), is largely controlled by RhoA/ROCK. Here, we examined whether PtdIns(4,5)P2 and PI4P5K are involved in the increased expression of ileal P-gp following the ERM activation by ETP treatment. Male ddY mice (4-week-old) were treated with ETP (10 mg/kg/day, per os, p.o.) for 5 days. Protein-expression levels were measured by either western blot or dot blot analysis and molecular interactions were assessed using immunoprecipitation assays. ETP treatment significantly increased PI4P5K, ERM, and P-gp expression in the ileal membrane. This effect was suppressed following the coadministration of ETP with rosuvastatin (a RhoA inhibitor) or fasudil (a ROCK inhibitor). Notably, the PtdIns(4,5)P2 expression in the ileal membrane, as well as both P-gp and ERM levels coimmunoprecipitated with anti-PtdIns(4,5)P2 antibody, were increased by ETP treatment. PtdIns(4,5)P2 and PI4P5K may contribute to the increase in ileal P-gp expression observed following the ETP treatment, possibly through ERM activation via the RhoA/ROCK pathway.

  12. 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

  13. Molecular basis of resistance to macrolides, lincosamides and streptogramins in Staphylococcus saprophyticus clinical isolates.

    PubMed

    Le Bouter, Anne; Leclercq, Roland; Cattoir, Vincent

    2011-02-01

    The aim of this study was to evaluate the prevalence of resistance to macrolide-lincosamide-streptogramin (MLS) antibiotics as well as to assess the molecular basis of this resistance amongst 72 Staphylococcus saprophyticus urinary isolates collected from 2005 to 2009 in University Hospital of Caen (France). Of the 72 strains studied, 33 (45.8%) were resistant to at least one MLS antibiotic, including 24 (72.7%) with an M phenotype, 5 (15.2%) with an inducible MLS(B) phenotype, 3 (9.1%) with a combined M+L phenotype and 1 (3.0%) with an L phenotype. All isolates were susceptible to the combination of streptogramins A and B. The resistance genes erm(A), erm(B), erm(C), msr(A) and lnu(A) were detected alone in 0, 0, 5 (15.2%), 24 (72.7%) and 1 (3.0%) of the 33 MLS-resistant isolates, respectively, whereas 2 strains (6.1%) were positive for both msr(A) and lnu(A). All msr(A)-positive isolates exhibited an M phenotype, whereas all five erm(C)-positive and all three lnu(A)-positive strains displayed, respectively, an inducible MLS(B) phenotype and an L phenotype with a positive Hodge test. Plasmid analysis indicated that erm(C) and lnu(A) genes were borne by small-size plasmids (ca. 2.5 kb), whereas larger plasmids (30-90 kb) harboured msr(A). In conclusion, these findings show a high prevalence of MLS resistance in S. saprophyticus, which was mainly associated with the presence of the msr(A) gene. Since S. saprophyticus colonises the gastrointestinal tract, it may constitute an unexpected reservoir for MLS resistance genes, in particular msr(A), amongst coagulase-negative staphylococci.

  14. RAGE signaling deficiency in rhabdomyosarcoma cells causes upregulation of PAX7 and uncontrolled proliferation.

    PubMed

    Riuzzi, Francesca; Sorci, Guglielmo; Sagheddu, Roberta; Sidoni, Angelo; Alaggio, Rita; Ninfo, Vito; Donato, Rosario

    2014-04-15

    Embryonal rhabdomyosarcomas (ERMSs) show elevated levels of PAX7, a transcription factor that marks quiescent adult muscle stem (satellite) cells and is important for proliferation and survival of activated satellite cells and whose timely repression is required for myogenic differentiation. However, the mechanism of PAX7 accumulation in ERMSs and whether high PAX7 causes uncontrolled proliferation in ERMS remains to be elucidated. The receptor for advanced glycation end-products (RAGE, encoded by AGER) transduces a myogenic and anti-proliferative signal in myoblasts, and stable transfection of the ERMS cell line TE671, which does not express RAGE, with AGER results in reduced proliferation and formation of tumor masses in vivo, and enhanced apoptosis and myogenic differentiation. Herein, we show that RAGE expression is low or absent in human ERMSs. We also show that in ERMS cells (1) PAX7 accumulates owing to absent or low RAGE signaling; (2) elevated PAX7 levels reduce RAGE expression and levels of MyoD and myogenin, muscle-specific transcription factors required for myoblast proliferation arrest and differentiation, respectively; (3) PAX7 supports myoblast proliferation by reducing the levels of MyoD, primarily by promoting its degradation; and (4), when ectopically expressed in ERMS cells, that RAGE upregulates myogenin which upregulates MyoD and downregulates PAX7, with consequent inhibition of proliferation and stimulation of differentiation. Thus, failure to express RAGE and, hence, MyoD and myogenin above a critical level in ERMS cells might result in deregulated PAX7 expression leading to uncontrolled proliferation and, potentially, to rhabdomyosarcomagenesis.

  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. 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.

  17. PubMed Central

    KHORAMIAN, B.; JABALAMELI, F.; BEIGVERDI, R.; ASADOLLAHI, K.; TAHERIKALANI, M.; LARI, A.R.

    2016-01-01

    Summary This study was designed to determine the molecular characteristics and antimicrobial resistance of enterococcal strains isolated from patients admitted to an Iranian Hospital. Enterococcal strains were isolated from the burn patients. All strains were screened for genes encoding resistance to aminoglycoside [aac(6')-Ie-aph(2'')-Ia, aph (3'), ant (4')], resistance to vancomycin (vanA, vanB), resistance to tetracycline (tetK, tetL, tetM, tetO), and resistance to erythromycin (ermA, ermB, ermC) by PCR and multiplex PCR-based methods. Genetic diversity was evaluated via Random Amplified Polymorphic DNA (RAPD)-PCR. All enterococcal isolates showed complete sensitivity to vancomycin with MIC ≤ 0.5μg/ml. Resistance to gentamicin, tetracycline, erythromycin, ciprofloxacin or quinopristin-dalfopristin was detected, whilst more than 96.2% of isolates were high-level gentamicinresistant (HLGR) and multiple drug resistant. The most prevalent aminoglycoside resistance gene was aac(6')-Ie-aph(2'')-Ia, that was found in 96.2% (26/27) of the isolates. The most prevalent tetracycline resistance genes were tetM, found in 85.1% (23/27) followed by tetL and tetO found in 7.4% (2/27) of the isolates. The ermA and ermB genes were detected in 33.3% (9/27) and 44.4% (12/27) of the isolates respectively. RAPD-PCR analysis yielded 17 distinct profiles among 27 investigated isolates. One cluster of isolates shared the same RAPD pattern, while 16 isolates had unique RAPD pattern. Our study showed that during the examination time period one RAPD genotype was the common type and was disseminated among patients in the burn unit. Interestingly, most of these strains had an identical or very similar antibiotic and gene resistance pattern. PMID:28167856

  18. Macrolide-resistance mechanisms in Streptococcus pneumoniae isolates from Chinese children in association with genes of tetM and integrase of conjugative transposons 1545.

    PubMed

    Shen, Xuzhuang; Yang, Hui; Yu, Shangjie; Yao, Kaihu; Wang, Yonghong; Yuan, Lin; Yang, Yonghong

    2008-06-01

    This study investigated macrolide-resistant Streptococcus pneumoniae carried by Beijing children presenting with respiratory tract infections. Nasopharyngeal S. pneumoniae strains were tested for sensitivity with 15 antibiotics and further analyzed for phenotypes of macrolide-resistant strains and by PCR for the macrolide-resistant genes ermB, mefA, tetM, and integrase of conjugative transposon (Tn1545) intTn. We found 185 strains of S. pneumoniae relatively highly resistant to erythromycin (78.9%), clindamycin (76.2%), tetracycline (86%), and SMZ-TMP (78.7%) but with relatively low resistance to amoxicillin (2.2%), cefaclor (15.5%), ceftriaxone (2.8%), and cefuroxime (14.1%). The 146 strains of erythromycin-resistant S. pneumoniae showed extensive cross-resistance to other macrolides like azithromycin (100%), clarithromycin (100%), acetylspiramycin (95.2%), and clindamycin (95.9%). Genes ermB and mefA were detected in all erythromycin-resistant strains, with ermB(+) 79.5%, ermB + mefA(+) 17.8%, and mefA(+) 2.7%. About 96.9% of tetracycline-resistant isolates were positive for tetM, compared to 26.9% of sensitive strains. Ninety percent of tetracycline-resistant strains were also erythromycin-resistant versus 11.5% of tetracycline-sensitive strains. The intTn gene was present in 87.6% of S. pneumoniae strains and correlated with erythromycin and tetracycline resistance. The close relationship between the conjugative transposon Tn1545 and the genes ermB and tetM is probably one of the important mechanisms explaining the multiple drug resistance of S. pneumoniae.

  19. 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.

  20. 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.

  1. 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

  2. Structural characterization suggests models for monomeric and dimeric forms of full-length ezrin.

    PubMed

    Phang, Juanita M; Harrop, Stephen J; Duff, Anthony P; Sokolova, Anna V; Crossett, Ben; Walsh, James C; Beckham, Simone A; Nguyen, Cuong D; Davies, Roberta B; Glöckner, Carina; Bromley, Elizabeth H C; Wilk, Krystyna E; Curmi, Paul M G

    2016-09-15

    Ezrin is a member of the ERM (ezrin-radixin-moesin) family of proteins that have been conserved through metazoan evolution. These proteins have dormant and active forms, where the latter links the actin cytoskeleton to membranes. ERM proteins have three domains: an N-terminal FERM [band Four-point-one (4.1) ERM] domain comprising three subdomains (F1, F2, and F3); a helical domain; and a C-terminal actin-binding domain. In the dormant form, FERM and C-terminal domains form a stable complex. We have determined crystal structures of the active FERM domain and the dormant FERM:C-terminal domain complex of human ezrin. We observe a bistable array of phenylalanine residues in the core of subdomain F3 that is mobile in the active form and locked in the dormant form. As subdomain F3 is pivotal in binding membrane proteins and phospholipids, these transitions may facilitate activation and signaling. Full-length ezrin forms stable monomers and dimers. We used small-angle X-ray scattering to determine the solution structures of these species. As expected, the monomer shows a globular domain with a protruding helical coiled coil. The dimer shows an elongated dumbbell structure that is twice as long as the monomer. By aligning ERM sequences spanning metazoan evolution, we show that the central helical region is conserved, preserving the heptad repeat. Using this, we have built a dimer model where each monomer forms half of an elongated antiparallel coiled coil with domain-swapped FERM:C-terminal domain complexes at each end. The model suggests that ERM dimers may bind to actin in a parallel fashion.

  3. Characterization of Methicillin-Resistant Staphylococcus aureus Isolated from Healthy Turkeys and Broilers Using DNA Microarrays.

    PubMed

    El-Adawy, Hosny; Ahmed, Marwa; Hotzel, Helmut; Monecke, Stefan; Schulz, Jochen; Hartung, Joerg; Ehricht, Ralf; Neubauer, Heinrich; Hafez, Hafez M

    2016-01-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is a major human health problem and recently, domestic animals are described as carriers and possible reservoirs. Twenty seven S. aureus isolates from five turkey farms (n = 18) and two broiler farms (n = 9) were obtained by culturing of choana and skin swabs from apparently healthy birds, identified by Taqman-based real-time duplex nuc-mecA-PCR and characterized by spa typing as well as by a DNA microarray based assay which covered, amongst others, a considerable number of antibiotic resistance genes, species controls, and virulence markers. The antimicrobial susceptibility profiles were tested by agar diffusion assays and genotypically confirmed by the microarray. Five different spa types (3 in turkeys and 2 in broilers) were detected. The majority of MRSA isolates (24/27) belonged to clonal complex 398-MRSA-V. The most frequently occurring spa types were accordingly t011, t034, and t899. A single CC5-MRSA-III isolated from turkey and CC398-MRSA with an unidentified/truncated SCCmec element in turkey and broiler were additionally detected. The phenotypic antimicrobial resistance profiles of S. aureus isolated from both turkeys and broilers against 14 different antimicrobials showed that all isolates were resistant to ampicillin, cefoxitin, oxacillin, doxycycline, and tetracycline. Moreover, all S. aureus isolated from broilers were resistant to erythromycin and azithromycin. All isolates were susceptible to gentamicin, chloramphenicol, sulphonamides, and fusidic acid. The resistance rate against ciprofloxacin was 55.6% in broiler isolates and 42.1% in turkey isolates. All tetracycline resistant isolates possessed genes tetK/M. All erythromycin-resistant broiler isolates carried ermA. Only one broiler isolate (11.1%) carried genes ermA, ermB, and ermC, while 55.6% of turkey isolates possessed ermA and ermB genes. Neither PVL genes (lukF/S-PV), animal-associated leukocidin (lukM and luk-P83) nor the gene encoding

  4. Characterization of Methicillin-Resistant Staphylococcus aureus Isolated from Healthy Turkeys and Broilers Using DNA Microarrays

    PubMed Central

    El-Adawy, Hosny; Ahmed, Marwa; Hotzel, Helmut; Monecke, Stefan; Schulz, Jochen; Hartung, Joerg; Ehricht, Ralf; Neubauer, Heinrich; Hafez, Hafez M.

    2016-01-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is a major human health problem and recently, domestic animals are described as carriers and possible reservoirs. Twenty seven S. aureus isolates from five turkey farms (n = 18) and two broiler farms (n = 9) were obtained by culturing of choana and skin swabs from apparently healthy birds, identified by Taqman-based real-time duplex nuc-mecA-PCR and characterized by spa typing as well as by a DNA microarray based assay which covered, amongst others, a considerable number of antibiotic resistance genes, species controls, and virulence markers. The antimicrobial susceptibility profiles were tested by agar diffusion assays and genotypically confirmed by the microarray. Five different spa types (3 in turkeys and 2 in broilers) were detected. The majority of MRSA isolates (24/27) belonged to clonal complex 398-MRSA-V. The most frequently occurring spa types were accordingly t011, t034, and t899. A single CC5-MRSA-III isolated from turkey and CC398-MRSA with an unidentified/truncated SCCmec element in turkey and broiler were additionally detected. The phenotypic antimicrobial resistance profiles of S. aureus isolated from both turkeys and broilers against 14 different antimicrobials showed that all isolates were resistant to ampicillin, cefoxitin, oxacillin, doxycycline, and tetracycline. Moreover, all S. aureus isolated from broilers were resistant to erythromycin and azithromycin. All isolates were susceptible to gentamicin, chloramphenicol, sulphonamides, and fusidic acid. The resistance rate against ciprofloxacin was 55.6% in broiler isolates and 42.1% in turkey isolates. All tetracycline resistant isolates possessed genes tetK/M. All erythromycin-resistant broiler isolates carried ermA. Only one broiler isolate (11.1%) carried genes ermA, ermB, and ermC, while 55.6% of turkey isolates possessed ermA and ermB genes. Neither PVL genes (lukF/S-PV), animal-associated leukocidin (lukM and luk-P83) nor the gene encoding

  5. 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

  6. 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.

  7. Environmentally relevant microorganisms.

    PubMed

    Watanabe, K; Baker, P W

    2000-01-01

    The development of molecular microbial ecology in the 1990s has allowed scientists to realize that microbial populations in the natural environment are much more diverse than microorganisms so far isolated in the laboratory. This finding has exerted a significant impact on environmental biotechnology, since knowledge in this field has been largely dependent on studies with pollutant-degrading bacteria isolated by conventional culture methods. Researchers have thus started to use molecular ecological methods to analyze microbial populations relevant to pollutant degradation in the environment (called environmentally relevant microorganisms, ERMs), although further effort is needed to gain practical benefits from these studies. This review highlights the utility and limitations of molecular ecological methods for understanding and advancing environmental biotechnology processes. The importance of the combined use of molecular ecological and physiological methods for identifying ERMs is stressed.

  8. Engineering a regulatory region of jadomycin gene cluster to improve jadomycin B production in Streptomyces venezuelae.

    PubMed

    Zheng, Jian-Ting; Wang, Sheng-Lan; Yang, Ke-Qian

    2007-09-01

    Streptomyces venezuelae ISP5230 produces a group of jadomycin congeners with cytotoxic activities. To improve jadomycin fermentation process, a genetic engineering strategy was designed to replace a 3.4-kb regulatory region of jad gene cluster that contains four regulatory genes (3' end 272 bp of jadW2, jadW3, jadR2, and jadR1) and the native promoter upstream of jadJ (P(J)) with the ermEp* promoter sequence so that ermEp* drives the expression of the jadomycin biosynthetic genes from jadJ in the engineered strain. As expected, the mutant strain produced jadomycin B without ethanol treatment, and the yield increased to about twofold that of the stressed wild-type. These results indicated that manipulation of the regulation of a biosynthetic gene cluster is an effective strategy to increase product yield.

  9. 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.

  10. 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.

  11. A simple method for establishing adherent ex vivo explant cultures from human eye pathologies for use in subsequent calcium imaging and inflammatory studies.

    PubMed

    Andjelic, Sofija; Lumi, Xhevat; Veréb, Zoltán; Josifovska, Natasha; Facskó, Andrea; Hawlina, Marko; Petrovski, Goran

    2014-01-01

    A novel, simple, and reproducible method for cultivating pathological tissues obtained from human eyes during surgery was developed using viscoelastic material as a tissue adherent to facilitate cell attachment and expansion and calcium imaging of cultured cells challenged by mechanical and acetylcholine (ACh) stimulation as well as inflammatory studies. Anterior lens capsule-lens epithelial cells (aLC-LECs) from cataract surgery and proliferative diabetic retinopathy (PDR) fibrovascular epiretinal membranes (fvERMs) from human eyes were used in the study. We hereby show calcium signaling in aLC-LECs by mechanical and acetylcholine (ACh) stimulation and indicate presence of ACh receptors in these cells. Furthermore, an ex vivo study model was established for measuring the inflammatory response in fvERMs and aLC-LECs upon TNFα treatment.

  12. A Simple Method for Establishing Adherent Ex Vivo Explant Cultures from Human Eye Pathologies for Use in Subsequent Calcium Imaging and Inflammatory Studies

    PubMed Central

    Veréb, Zoltán; Facskó, Andrea; Hawlina, Marko

    2014-01-01

    A novel, simple, and reproducible method for cultivating pathological tissues obtained from human eyes during surgery was developed using viscoelastic material as a tissue adherent to facilitate cell attachment and expansion and calcium imaging of cultured cells challenged by mechanical and acetylcholine (ACh) stimulation as well as inflammatory studies. Anterior lens capsule-lens epithelial cells (aLC-LECs) from cataract surgery and proliferative diabetic retinopathy (PDR) fibrovascular epiretinal membranes (fvERMs) from human eyes were used in the study. We hereby show calcium signaling in aLC-LECs by mechanical and acetylcholine (ACh) stimulation and indicate presence of ACh receptors in these cells. Furthermore, an ex vivo study model was established for measuring the inflammatory response in fvERMs and aLC-LECs upon TNFα treatment. PMID:25276840

  13. 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.

  14. Exploration of geosmin synthase from Streptomyces peucetius ATCC 27952 by deletion of doxorubicin biosynthetic gene cluster.

    PubMed

    Singh, Bijay; Oh, Tae-Jin; Sohng, Jae Kyung

    2009-10-01

    Thorough investigation of Streptomyces peucetius ATCC 27952 genome revealed a sesquiterpene synthase, named spterp13, which encodes a putative protein of 732 amino acids with significant similarity to S. avermitilis MA-4680 (SAV2163, GeoA) and S. coelicolor A3(2) (SCO6073). The proteins encoded by SAV2163 and SCO6073 produce geosmin in the respective strains. However, the spterp13 gene seemed to be silent in S. peucetius. Deletion of the doxorubicin gene cluster from S. peucetius resulted in increased cell growth rate along with detectable production of geosmin. When we over expressed the spterp13 gene in S. peucetius DM07 under the control of an ermE* promoter, 2.4 +/- 0.4-fold enhanced production of geosmin was observed.

  15. Dispensable ribosomal resistance to spiramycin conferred by srmA in the spiramycin producer Streptomyces ambofaciens.

    PubMed

    Pernodet, J L; Gourmelen, A; Blondelet-Rouault, M H; Cundliffe, E

    1999-09-01

    Streptomyces ambofaciens produces the macrolide antibiotic spiramycin, an inhibitor of protein synthesis, and possesses multiple resistance mechanisms to the produced antibiotic. Several resistance determinants have been isolated from S. ambofaciens and studies with one of them, srmA, which hybridized with ermE (the erythromycin-resistance gene from Saccharopolyspora erythraea), are detailed here. The nucleotide sequence of srmA was determined and the mechanism by which its product confers resistance was characterized. The SrmA protein is a methyltransferase which introduces a single methyl group into A-2058 (Escherichia coli numbering scheme) in the large rRNA, thereby conferring an MLS (macrolide-lincosamide-streptogramin type B) type I resistance phenotype. A mutant of S. ambofaciens in which srmA was inactivated was viable and still produced spiramycin, indicating that srmA is dispensable, at least in the presence of the other resistance determinants.

  16. An experimental comparison of proportional-integral, sliding mode, and robust adaptive control for piezo-actuated nanopositioning stages.

    PubMed

    Gu, Guo-Ying; Zhu, Li-Min

    2014-05-01

    This paper presents a comparative study of the proportional-integral (PI) control, sliding mode control (SMC), and robust adaptive control (RAC) for applications to piezo-actuated nanopositioning stages without the inverse hysteresis construction. For a fair comparison, the control parameters of the SMC and RAC are selected on the basis of the well-tuned parameters of the PI controller under same desired trajectories and sampling frequencies. The comparative results show that the RAC improves the tracking performance by 17 and 37 times than the PI controller in terms of the maximum tracking error e(m) and the root mean tracking error e(rms), respectively, while the RAC improves the tracking performance by 7 and 9 times than the SMC in terms of e(m) and e(rms), respectively.

  17. 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

  18. 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.

  19. Spatial and temporal distribution and pollution assessment of trace metals in marine sediments in Oyster Bay, NSW, Australia.

    PubMed

    Alyazichi, Yasir M; Jones, Brian G; McLean, Errol

    2015-01-01

    The disposal of untreated urban and industrial wastewater has a deleterious effect on both the water and sediment quality of Oyster Bay located in south Sydney, Australia. The present investigation was undertaken to evaluate the potential pollution of marine sediments in Oyster Bay. The results of metals were compared with adverse biological effect values effect range low (ERL) and effect range median (ERM). Spatial distribution of trace metals was estimated by applying geographic information system. The results indicated that the sediments were polluted with Cu, Zn, As and Pb, which exceeded ERL levels. However, these metals were still below ERM values, and other metals Cr and Ni were below ERL. Moreover, the highest concentrations of metals were around discharge points and in the inner bay. Further, trace metals could be attributed to human activities within the bay as they declined in concentrations with increasing sediment depth.

  20. 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

  1. 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

  2. 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.

  3. Activity of telithromycin against penicillin-resistant Streptococcus pneumoniae isolates recovered from French children with invasive and noninvasive infections.

    PubMed

    Bingen, Edouard; Doit, Catherine; Loukil, Chawki; Brahimi, Naima; Bidet, Philippe; Deforche, Dominique; Geslin, Pierre

    2003-07-01

    We compared the activities of telithromycin, erythromycin, azithromycin, josamycin, penicillin G, amoxicillin, cefpodoxime, and ceftriaxone against invasive and noninvasive non-penicillin-susceptible Streptococcus pneumoniae isolates recovered from children. Of the 186 isolates tested, 89% were positive for erm(B) by PCR. Telithromycin had the lowest MICs, with MICs at which 90% of the isolates tested are inhibited of 0.032 and 0.25 micro g/ml for erythromycin-sensitive and -resistant isolates, respectively.

  4. 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

  5. The Role of Drosophila Merlin in the Control of Mitosis Exit and Development

    DTIC Science & Technology

    2005-07-01

    like ancestors within the vertebrate clade that occurred after its separation 11 from Urochordata ( Ciona intestinalis ). Amino acid sequence alignment... Ciona intestinalis ). Amino-acid sequence alignment reveals the absence of an actin-binding site at the C- terminal domain of all merlin proteins...observed the expansion of the ERM-like ancestors within the vertebrate clade that occurred after its separation from Urochordata ( Ciona intestinalis

  6. Effects of Copper Addition on Copper Resistance, Antibiotic Resistance Genes, and intl1 during Swine Manure Composting

    PubMed Central

    Yin, Yanan; Gu, Jie; Wang, Xiaojuan; Song, Wen; Zhang, Kaiyu; Sun, Wei; Zhang, Xin; Zhang, Yajun; Li, Haichao

    2017-01-01

    Copper is one of the most abundant heavy metals present in swine manure. In this study, a laboratory-scale aerobic composting system was amended with Cu at three levels (0, 200, and 2000 mg kg-1, i.e., control, Cu200, and Cu2000 treatments, respectively) to determine its effect on the fate of copper resistance genes [copper resistance genes (CRGs): pcoA, cusA, copA, and tcrB], antibiotic resistance genes [antibiotic resistance genes (ARGs): erm(A) and erm(B)], and intl1. The results showed that the absolute abundances of pcoA, tcrB, erm(A), erm(B), and intl1 were reduced, whereas those of copA and cusA increased after swine manure composting. Redundancy analysis showed that temperature significantly affected the variations in CRGs, ARGs, and intl1. The decreases in CRGs, ARGs, and intI1 were positively correlated with the exchangeable Cu levels. The bacterial community could be grouped according to the composting time under different treatments, where the high concentration of copper had a more persistent effect on the bacterial community. Network analysis determined that the co-occurrence of CRGs, ARGs, and intI1, and the bacterial community were the main contributors to the changes in CRGs, ARG, and intl1. Thus, temperature, copper, and changes in the bacterial community composition had important effects on the variations in CRGs, ARGs, and intl1 during manure composting in the presence of added copper. PMID:28316595

  7. Differential Gene Expression in Rhododendron fortunei Roots Colonized by an Ericoid Mycorrhizal Fungus and Increased Nitrogen Absorption and Plant Growth

    PubMed Central

    Wei, Xiangying; Chen, Jianjun; Zhang, Chunying; Pan, Dongming

    2016-01-01

    Ericoid mycorrhizal (ERM) fungi are specifically symbiotic with plants in the family Ericaceae. Little is known thus far about their symbiotic establishment and subsequent nitrogen (N) uptake at the molecular level. The present study devised a system for establishing a symbiotic relationship between Rhododendron fortunei Lindl. and an ERM fungus (Oidiodendron maius var. maius strain Om19), quantified seedling growth and N uptake, and compared transcriptome profiling between colonized and uncolonized roots using RNA-Seq. The Om19 colonization induced 16,892 genes that were differentially expressed in plant roots, of which 14,364 were upregulated and 2,528 were downregulated. These genes included those homologous to ATP-binding cassette transporters, calcium/calmodulin-dependent kinases, and symbiosis receptor-like kinases. N metabolism was particularly active in Om19-colonized roots, and 51 genes were upregulated, such as nitrate transporters, nitrate reductase, nitrite reductase, ammonium transporters, glutamine synthetase, and glutamate synthase. Transcriptome analysis also identified a series of genes involving endocytosis, Fc-gamma R-mediated phagocytosis, glycerophospholipid metabolism, and Gonadotropin-releasing hormone (GnRH) signal pathway that have not been reported previously. Their roles in the symbiosis require further investigation. The Om19 colonization significantly increased N uptake and seedling growth. Total N content and dry weight of colonized seedlings were 36.6 and 46.6% greater than control seedlings. This is the first transcriptome analysis of a species from the family Ericaceae colonized by an ERM fungus. The findings from this study will shed light on the mechanisms underlying symbiotic relationships of ericaceous species with ERM fungi and the symbiosis-resultant N uptake and plant growth. PMID:27826312

  8. 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

  9. Phenotypes and genotypes of erythromycin-resistant Streptococcus pyogenes strains isolated from invasive and non-invasive infections from Mexico and the USA during 1999–2010

    PubMed Central

    Villaseñor-Sierra, Alberto; Katahira, Eva; Jaramillo-Valdivia, Abril N.; de los Angeles Barajas-García, María; Bryant, Amy; Morfín-Otero, Rayo; Márquez-Díaz, Francisco; Tinoco, Juan Carlos; Sánchez-Corona, José; Stevens, Dennis L.

    2012-01-01

    Summary Objective To compare the prevalence, phenotypes, and genes responsible for erythromycin resistance among Streptococcus pyogenes isolates from Mexico and the USA. Methods Eighty-nine invasive and 378 non-invasive isolates from Mexico, plus 148 invasive, 21 non-invasive, and five unclassified isolates from the USA were studied. Susceptibilities to penicillin, erythromycin, clindamycin, ceftriaxone, and vancomycin were evaluated according to Clinical and Laboratory Standards Institute (CLSI) standards. Phenotypes of erythromycin resistance were identified by triple disk test, and screening for mefA, ermTR, and ermB genes was carried out by PCR. Results All isolates were susceptible to penicillin, ceftriaxone, and vancomycin. Erythromycin resistance was found in 4.9% of Mexican strains and 5.2% of USA strains. Phenotypes in Mexican strains were 95% M and 5% cMLS; in strains from the USA, phenotypes were 33.3% iMLS, 33.3% iMLS-D, and 33.3% M. Erythromycin resistance genes in strains from Mexico were mefA (95%) and ermB (5%); USA strains harbored ermTR (56%), mefA (33%), and none (11%). In Mexico, all erythromycin-resistant strains were non-invasive, whereas 89% of strains from the USA were invasive. Conclusions Erythromycin resistance continues to exist at low levels in both Mexico and the USA, although the genetic mechanisms responsible differ between the two nations. These genetic differences may be related to the invasive character of the S. pyogenes isolated. PMID:22217469

  10. Oxidative transformation of micropollutants during municipal wastewater treatment: comparison of kinetic aspects of selective (chlorine, chlorine dioxide, ferrate VI, and ozone) and non-selective oxidants (hydroxyl radical).

    PubMed

    Lee, Yunho; von Gunten, Urs

    2010-01-01

    Chemical oxidation processes have been widely applied to water treatment and may serve as a tool to minimize the release of micropollutants (e.g. pharmaceuticals and endocrine disruptors) from municipal wastewater effluents into the aquatic environment. The potential of several oxidants for the transformation of selected micropollutants such as atenolol, carbamazepine, 17 alpha-ethinylestradiol (EE2), ibuprofen, and sulfamethoxazole was assessed and compared. The oxidants include chlorine, chlorine dioxide, ferrate(VI), and ozone as selective oxidants versus hydroxyl radicals as non-selective oxidant. Second-order rate constants (k) for the reaction of each oxidant show that the selective oxidants react only with some electron-rich organic moieties (ERMs), such as phenols, anilines, olefins, and deprotonated-amines. In contrast, hydroxyl radicals show a nearly diffusion-controlled reactivity with almost all organic moieties (k>or=10(9)M(-1) s(-1)). Due to a competition for oxidants between a target micropollutant and wastewater matrix (i.e. effluent organic matter, EfOM), a higher reaction rate with a target micropollutant does not necessarily translate into more efficient transformation. For example, transformation efficiencies of EE2, a phenolic micropollutant, in a selected wastewater effluent at pH 8 varied only within a factor of 7 among the selective oxidants, even though the corresponding k for the reaction of each selective oxidant with EE2 varied over four orders of magnitude. In addition, for the selective oxidants, the competition disappears rapidly after the ERMs present in EfOM are consumed. In contrast, for hydroxyl radicals, the competition remains practically the same during the entire oxidation. Therefore, for a given oxidant dose, the selective oxidants were more efficient than hydroxyl radicals for transforming ERMs-containing micropollutants, while hydroxyl radicals are capable of transforming micropollutants even without ERMs. Besides Ef

  11. 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

  12. Peroxisome proliferator-activated receptor alpha protects renal tubular cells from gentamicin-induced apoptosis via upregulating Na(+)/H(+) exchanger NHE1.

    PubMed

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

    2015-11-23

    Peroxisome proliferator-activated receptor alpha (PPARα) is a transcription factor which 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 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 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 pro-survival 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.

  13. Mississippi and Louisiana Estuarine Areas. Freshwater Diversion to Lake Pontchartrain Basin and Mississippi Sound. Feasibility Study. Volume 3. Technical Appendixes, E, F, G, H, I, J, K.

    DTIC Science & Technology

    1984-04-01

    8217).14. FccaI coliform data for the MR-CO at Bayou Dupre and Lake Bur,,une area imply uccasionaI contravent ions of the shell fish harvest ing standard...thhan 400 colonlies/mL.. Thle fcal echit erm datai for the Back Bay of El loxi at Ocean Springs, is ssipp i ugatno contravent tins of the primary

  14. The effect of radixin knockdown on the expression and efflux function of MRP2 in SGC-7901 cells.

    PubMed

    He, Xiao-Jie; Wang, Wei-Rong; Zhang, Yun; Yang, Qing

    2012-08-15

    Multidrug resistance-associated protein 2 (MRP2, ABCC2) is the second member of the MRP transporter family and functions physiologically as an organic anion transporter. Earlier studies have confirmed that radixin, which is a member of the ERM (ezrin/radixin/moesin) family, modulates MRP2 localization at the canalicular membrane in hepatocytes. The relationship between radixin and MRP2 - particularly, the effect of radixin on the expression and function of MRP2 in cells or tissues that co-express all three ERM proteins - has not been well studied. To examine the role of radixin in the expression and function of MRP2 and other MRPs, we chose human gastric carcinoma SGC-7901 cells that express all three ERM proteins rather than hepatocytes, which predominantly express radixin. Radixin stable knockdown SGC-7901 cells, which were constructed by RNAi, exhibited no compensatory up-regulation of ezrin or moesin. The mRNA expression profiles of MRPs in the radixin knockdown cells were primarily evaluated by RT-PCR. Real time quantitative RT-PCR and western blot analysis revealed that the radixin deficiency caused the mRNA and protein expression levels of MRP2 to be reduced by about 50%, respectively. Accordingly, efflux and MTT assays showed that the radixin knockdown cells exhibited lower efflux ability with respect to calcein but no significant change in cell viability. In conclusion, among the MRP1-6 family members, radixin selectively modulates the expression and function of MRP2 in a system co-expressing all three ERM proteins.

  15. Rational redesign of neutral endopeptidase binding to merlin and moesin proteins.

    PubMed

    Niv, Masha Y; Iida, Katsuyuki; Zheng, Rong; Horiguchi, Akio; Shen, Ruoqian; Nanus, David M

    2009-05-01

    Neutral endopeptidase (NEP) is a 90- to 110-kDa cell-surface peptidase that is normally expressed by numerous tissues but whose expression is lost or reduced in a variety of malignancies. The anti-tumorigenic function of NEP is mediated not only by its catalytic activity but also through direct protein-protein interactions of its cytosolic region with several binding partners, including Lyn kinase, PTEN, and ezrin/radixin/moesin (ERM) proteins. We have previously shown that mutation of the K(19)K(20)K(21) basic cluster in NEPs' cytosolic region to residues QNI disrupts binding to the ERM proteins. Here we show that the ERM-related protein merlin (NF2) does not bind NEP or its cytosolic region. Using experimental data, threading, and sequence analysis, we predicted the involvement of moesin residues E(159)Q(160) in binding to the NEP cytosolic domain. Mutation of these residues to NL (to mimic the corresponding N(159)L(160) residues in the nonbinder merlin) disrupted moesin binding to NEP. Mutation of residues N(159)L(160)Y(161)K(162)M(163) in merlin to the corresponding moesin residues resulted in NEP binding to merlin. This engineered NEP peptide-merlin interaction was diminished by the QNI mutation in NEP, supporting the role of the NEP basic cluster in binding. We thus identified the region of interaction between NEP and moesin, and engineered merlin into a NEP-binding protein. These data form the basis for further exploration of the details of NEP-ERM binding and function.

  16. 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).

  17. Survey of strain distribution and antibiotic resistance pattern of group B streptococci (Streptococcus agalactiae) isolated from clinical specimens

    PubMed Central

    Mousavi, Seyed Masoud; Nasaj, Mona; Hosseini, Seyed Mostafa; Arabestani, Mohammad Reza

    2016-01-01

    Aim: The aims of the present study were to determine the antibiotic susceptibility profils with particular emphasis on susceptible or resistant strains to macrolides and lincosamids antibiotics and to determine possible antibiotic resistance mechanisms occurring in group B streptococci (GBS) strains using PCR assay and disk diffusion method. Methods: A total of 62 clinical GBS strains were investigated. Antibacterial susceptibility testing was performed using the disk diffusion method and inducible resistance test for clindamycin by standard double disk diffusion or D-zone test for all isolates to differentiate macrolide resistance phenotype (M), constitutive macrolide-lincosamide-streptogramin B phenotype (cMLSB) and induced macrolide-lincosamide-streptogramin B phenotype (iMLSB). In addition, minimum inhibitory concentrations (MIC) of penicillin were determined for all isolates. Finally, possible existence of antibiotic resistance genes for erythromycin (ermTR, ermB and mefA/E) and for clindamycin (linB) were examined among isolates using PCR assay. Results: All 62 isolates were susceptible to penicillin, ampicillin, linezolid, cefazoline and vancomycin. However, 93.5% (n=58) of isolates showed an increased MIC to penicillin. The overall rate of erythromycin resistance was 35.5% (n=22). All erythromycin-resistant isolates displayed the M phenotype (100%, n=22). All three erythromycin resistance genes (i.e. ermTR, ermB and mefA/E) were found in erythromycin-resistant isolates. Conclusion: It was concluded that prescribing antibiotic without antibacterial susceptibility tests should be prevented because of the high prevalence of erythromycin-resistant GBS strains and the fact that erythromycin-resistant GBS strains has shown an increased MIC to penicillin, as the drug of choice for treating GBS infections. PMID:27648402

  18. 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.

  19. Trends in Childhood Rhabdomyosarcoma Incidence and Survival in the United States (1975–2005)

    PubMed Central

    Ognjanovic, Simona; Linabery, Amy M.; Charbonneau, Bridget; Ross, Julie A.

    2010-01-01

    Background Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in children and adolescents aged <20 years; its etiology remains largely unknown. Embryonal (ERMS) and alveolar rhabdomyosarcoma (ARMS), the most common subtypes, are thought to arise through distinct biological mechanisms. We evaluated incidence and survival trends by RMS demographic subgroups to inform future etiologic hypotheses. Methods Incidence and survival trends in RMS among children and adolescents aged <20 years were analyzed using data from the Surveillance, Epidemiology and End Results Program. Frequencies, age-adjusted incidence and survival rates, and jointpoint regression results, including annual percent change (APC) and 95% confidence intervals (CI), were calculated. Results Between 1975 and 2005, the incidence of ERMS was stable, while a significant increase in the incidence of ARMS was observed (APC=4.20%, 95%CI=2.60%–5.82%). This trend may be partially attributable to shifts in diagnosis, as a significant negative trend in RMS, not otherwise specified was observed concurrently. A bimodal age peak for ERMS was observed, with the second, smaller peak in adolescence noted for males only; ARMS incidence did not vary by age or sex. Five-year survival rates for RMS and ERMS increased from the period 1976–1980 (52.7% and 60.9%, respectively) to 1996–2000 (61.8% and 73.4%, respectively), while there was little improvement for ARMS (40.1% and 47.8%, respectively). Conclusions Observed differences in incidence and survival for two major RMS subtypes across gender and age subgroups further support unique underlying etiologies for these tumors. Exploration of these differences presents an opportunity to increase our knowledge of RMS. PMID:19536876

  20. Ezrin/radixin/moesin proteins are high affinity targets for ADP-ribosylation by Pseudomonas aeruginosa ExoS.

    PubMed

    Maresso, Anthony W; Baldwin, Michael R; Barbieri, Joseph T

    2004-09-10

    Pseudomonas aeruginosa ExoS is a bifunctional type III-secreted cytotoxin. The N terminus (amino acids 96-233) encodes a GTPase-activating protein activity, whereas the C terminus (amino acids 234-453) encodes a factor-activating ExoS-dependent ADP-ribosyltransferase activity. The GTPase-activating protein activity inactivates the Rho GTPases Rho, Rac, and Cdc42 in cultured cells and in vitro, whereas the ADP-ribosylation by ExoS is poly-substrate-specific and includes Ras as an early target for ADP-ribosylation. Infection of HeLa cells with P. aeruginosa producing a GTPase-activating protein-deficient form of ExoS rounded cells, indicating the ADP-ribosyltransferase domain alone is sufficient to elicit cytoskeletal changes. Examination of substrates modified by type III-delivered ExoS identified a 70-kDa protein as an early and predominant target for ADP-ribosylation. Matrix-assisted laser desorption ionization mass spectroscopy identified this protein as moesin, a member of the ezrin/radixin/moesin (ERM) family of proteins. ExoS ADP-ribosylated recombinant moesin at a linear velocity that was 5-fold faster and with a K(m) that was 2 orders of magnitude lower than Ras. Moesin homologs ezrin and radixin were also ADP-ribosylated, indicating the ERMs collectively represent high affinity targets of ExoS. Type III delivered ExoS ADP-ribosylated moesin and ezrin (and/or radixin) in cultured HeLa cells. The ERM proteins contribute to cytoskeleton dynamics, and the ability of ExoS to ADP-ribosylate the ERM proteins links ADP-ribosylation with the cytoskeletal changes associated with ExoS intoxication.

  1. Identification of Cytoplasmic Proteins Interacting with the Mammary Cell Transforming Domain of Ese-1

    DTIC Science & Technology

    2009-04-01

    and Nordheim, A. 1993. Gene regulation by Ets proteins. Biochim. Biophys. Acta 1155:346-356. 7. Chang, C , Scott, G, Kuo, W, Xiong, X, Suzdaltseva, Y...families, consisting of 27 ETS genes in humans, 26 in mice, 10 in Caenorhabditis elegans and 9 in Drosophila that can be structurally categorized...transcription factor genes in prostate cancer. Science 310, 644–648 40 Chen, C . et al. (2005) ERM is required for transcriptional control of the

  2. The Association between Smoking and Epiretinal Membrane

    PubMed Central

    Wang, Sheng-Zhan; Tong, Qi-Hu; Wang, Hui-Yun; Lu, Qin-Kang; Xu, Yu-feng

    2016-01-01

    We conducted a meta-analysis of analytic and observational studies to evaluate the association between smoking and epiretinal membrane (ERM). The pertinent studies were identified via a literature search using three databases (MEDLINE, Cochrane Library, Embase) and the reference lists of retrieved studies. Cohort, case-control and cross-sectional studies meeting the predefined criteria were included. We extracted the odds ratios (OR) and 95% confidence intervals (CI) from each study. Overall risk estimates were pooled using random-effects models. Subgroup analyses based on several stratified factors were also performed. Two cohort studies and six cross-sectional studies involving 46,837 subjects were included. The pooled effect of all eight studies showed an unexpected significant decreased association between smoking and the occurrence of ERM (OR, 0.72; 95% CI 0.61–0.84; p = 0.29, I2 = 17.9%). Subgroup analyses supported this finding, except for the age-unadjusted group (OR, 0.87; 95% CI 0.63–1.22), the ERM classification group (cellophane macular reflex (CMR) OR, 0.93; 95% CI 0.68–1.28; preretinal macular fibrosis (PMF) OR, 0.74; 95% CI 0.41–1.32), the Asian group (OR, 0.75; 95% CI 0.52–1.09) and the past smoker group (OR, 1.02; 95% CI 0.85–1.22). The pooled effects from the current literature suggested a declining association between smoking and ERM, which requires further studies to confirm. PMID:27897264

  3. Analysis for the Accuracy Definition of the Air Quality Assessment Model (AQAM) at Williams Air Force Base, Arizona. Volume II Appendices.

    DTIC Science & Technology

    1980-03-01

    VERIFICATION OF SHORT-’TtERM AIP QUALITY MODELS USING THE GAUSSIAN DISPERSION APPROACH by Karl ZcIler, EPA* (January 1977) 1. INTRODUCTION A need exists to... modeling of complex gi.o- metric situations, it is currently necessary to use empirical dispersion parameter values (standard deviations of plume spread as a...subjective modifications. For example, models used for airports or highways use the same dispersion parameters as models used for elevated area or point

  4. Fußball mit Wissenschaftlichem Maß: Bananenflanken, Flatterbälle, Kopfballkämpfe

    NASA Astrophysics Data System (ADS)

    Mathelitsch, Leopold; Thaller, Sigrid

    2006-05-01

    Die physikalische Betrachtung des Fußballs liefert spannende Resultate. Wegen der geringen Torzahl spielt der Zufall bei Sieg oder Niederlage stärker mit als in anderen Sportarten. Fußball ist daher mit gewissen statistischen Eigenschaften radioaktiver Quellen vergleichbar. Gleich mehrere physikalische Effekte beeinflussen die Flugbahn des Balls. Der Magnus-Effekt zum Beispiel ermöglicht erst Bananenflanken. Pfeift ein Schiedsrichter falsch, dann wird er nicht selten von seiner Sehperspektive getäuscht.

  5. Electronic Collection Management and Electronic Information Services

    DTIC Science & Technology

    2003-04-01

    library are relatively new concepts. The earliest literature dates to the 1970s and 1980s ( Harter and Kister 1981, Dowlin 1984). In 1984 Kenneth...they will have to. The Web may also serve as a viable substitute for expensive online databases and some traditional collections. Susan Lewis-Somers...pdf/erm0164.pdf [28 March 2002]. Harris, L.E. (2002) Licensing digital content. Chicago: American Library Association. Harter , S. and Kister, K

  6. 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.

  7. Problem Formulation in Knowledge Discovery via Data Analytics (KDDA) for Environmental Risk Management

    PubMed Central

    Li, Yan; Thomas, Manoj; Osei-Bryson, Kweku-Muata; Levy, Jason

    2016-01-01

    With the growing popularity of data analytics and data science in the field of environmental risk management, a formalized Knowledge Discovery via Data Analytics (KDDA) process that incorporates all applicable analytical techniques for a specific environmental risk management problem is essential. In this emerging field, there is limited research dealing with the use of decision support to elicit environmental risk management (ERM) objectives and identify analytical goals from ERM decision makers. In this paper, we address problem formulation in the ERM understanding phase of the KDDA process. We build a DM3 ontology to capture ERM objectives and to inference analytical goals and associated analytical techniques. A framework to assist decision making in the problem formulation process is developed. It is shown how the ontology-based knowledge system can provide structured guidance to retrieve relevant knowledge during problem formulation. The importance of not only operationalizing the KDDA approach in a real-world environment but also evaluating the effectiveness of the proposed procedure is emphasized. We demonstrate how ontology inferencing may be used to discover analytical goals and techniques by conceptualizing Hazardous Air Pollutants (HAPs) exposure shifts based on a multilevel analysis of the level of urbanization (and related economic activity) and the degree of Socio-Economic Deprivation (SED) at the local neighborhood level. The HAPs case highlights not only the role of complexity in problem formulation but also the need for integrating data from multiple sources and the importance of employing appropriate KDDA modeling techniques. Challenges and opportunities for KDDA are summarized with an emphasis on environmental risk management and HAPs. PMID:27983713

  8. Finite Memory Model for Haptic Recognition

    DTIC Science & Technology

    1991-12-01

    Slot 4 bu f fer s hort- term storel Slot N Long- ’erm store The model of memory proposed by Atkinson and Shiffrin . Primary memory here is as rehearsal...7 NAVAL POSTGRADUATE SCHOOL Monterey, Califormia AD-A245 342 THESIS Finite Memory Model for Haptic Recognition by Philip G. Beieri December 1991...ELEMEN1 No.) NO. No. ACCESSION NO. I1. TITLE (include Securitn Classification) FINITE MEMORY MODEL FOR HAPTIC RECOGNITION’ 12. PERSONALEAUTHOR(S) Philip

  9. 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

  10. Genetic Requirements for the Transformation of Human Cells

    DTIC Science & Technology

    2004-07-01

    apoptosis indicate that EIAACR2 can be functionally complemented cyclin E expression (A. Samuelson and S. Lowe, personal communication.) We therefore 4 SEGER...oncogenes such as c-myc or adenovirus El A sensi- moting factors, are not restricted by cell-cell contact, and are tizes primary cells to apoptosis (Debbas...which many cells undergo apoptosis or senes- cence. Cells that emerged from this crisis event IP-actin become telomerase-positive (ERM P.C.). The in

  11. In vivo evaluation of drug-drug interaction via mechanism-based inhibition by macrolide antibiotics in cynomolgus monkeys.

    PubMed

    Ogasawara, Akihito; Negishi, Isao; Kozakai, Kazumasa; Kume, Toshiyuki

    2009-11-01

    Irreversible inhibition, characterized as mechanism-based inhibition (MBI), of cytochrome P450 in drugs has to be avoided for their safe use. A comprehensive assessment of drug-drug interaction (DDI) potential is important during the drug discovery process. In the present study, we evaluated the effects of macrolide antibiotics, erythromycin (ERM), clarithromycin (CAM), and azithromycin (AZM), which are mechanism-based inhibitors of CYP3A, on biotransformation of midazolam (MDZ) in monkeys. These macrolides inhibited the formation of 1'-hydroxymidazolam in monkey microsomes as functions of incubation time and macrolide concentration. Furthermore, the inactivation potentials of macrolides (k(inact)/K(I): CAM congruent with ERM > AZM) were as effective as that observed in human samples. In in vivo studies, MDZ was administered orally (1 mg/kg) without or with multiple oral dosing of macrolides (15 mg/kg, twice a day on days 1-3). On day 3, the area under the plasma concentration-time curve (AUC) of MDZ increased 7.0-, 9.9-, and 2.0-fold with ERM, CAM, and AZM, respectively, compared with MDZ alone. Furthermore, the effects of ERM and CAM on the pharmacokinetics of MDZ were also observed on the day (day 4) after completion of macrolide treatments (AUC changes: 7.3- and 7.3-fold, respectively). Because the plasma concentrations of macrolides immediately before MDZ administration on day 4 were much lower than the IC(50) values for reversible CYP3A inhibition, the persistent effects may be predominantly caused by CYP3A inactivation. These results suggest that the monkey might be a suitable animal model to predict DDIs caused by MBI of CYP3A.

  12. Effects of Copper Addition on Copper Resistance, Antibiotic Resistance Genes, and intl1 during Swine Manure Composting.

    PubMed

    Yin, Yanan; Gu, Jie; Wang, Xiaojuan; Song, Wen; Zhang, Kaiyu; Sun, Wei; Zhang, Xin; Zhang, Yajun; Li, Haichao

    2017-01-01

    Copper is one of the most abundant heavy metals present in swine manure. In this study, a laboratory-scale aerobic composting system was amended with Cu at three levels (0, 200, and 2000 mg kg(-1), i.e., control, Cu200, and Cu2000 treatments, respectively) to determine its effect on the fate of copper resistance genes [copper resistance genes (CRGs): pcoA, cusA, copA, and tcrB], antibiotic resistance genes [antibiotic resistance genes (ARGs): erm(A) and erm(B)], and intl1. The results showed that the absolute abundances of pcoA, tcrB, erm(A), erm(B), and intl1 were reduced, whereas those of copA and cusA increased after swine manure composting. Redundancy analysis showed that temperature significantly affected the variations in CRGs, ARGs, and intl1. The decreases in CRGs, ARGs, and intI1 were positively correlated with the exchangeable Cu levels. The bacterial community could be grouped according to the composting time under different treatments, where the high concentration of copper had a more persistent effect on the bacterial community. Network analysis determined that the co-occurrence of CRGs, ARGs, and intI1, and the bacterial community were the main contributors to the changes in CRGs, ARG, and intl1. Thus, temperature, copper, and changes in the bacterial community composition had important effects on the variations in CRGs, ARGs, and intl1 during manure composting in the presence of added copper.

  13. Evaluation of disk approximation and single-well broth tests for detection of inducible clindamycin resistance in Streptococcus pneumoniae.

    PubMed

    Jorgensen, James H; McElmeel, M Leticia; Fulcher, Letitia C; McGee, Lesley; Glennen, Anita

    2011-09-01

    This study evaluated an agar disk diffusion D-zone test and an erythromycin-clindamycin (ERY + CLI) single-well broth test for inducible CLI resistance in Streptococcus pneumoniae. The standard CLSI disk approximation test and a single-well combination test incorporating 1 plus 0.5 μg/ml ERY + CLI detected >96% of isolates containing the ermB determinant.

  14. 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

  15. 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.

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

    PubMed

    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

    2014-05-27

    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.

  17. Role of the Porphyromonas gingivalis iron-binding protein PG1777 in oxidative stress resistance

    PubMed Central

    McKenzie, Rachelle M. E.; Henry, Leroy G.; Boutrin, Marie-Claire; Ximinies, Alexia

    2016-01-01

    Whole genome sequencing of the response of Porphyromonas gingivalis W83 to hydrogen peroxide revealed an upregulation of several uncharacterized, novel genes. Under conditions of prolonged oxidative stress in P. gingivalis, increased expression of a unique transcriptional unit carrying the grpE, dnaJ and three other hypothetical genes (PG1777, PG1778 and PG1779) was observed. The transcriptional start site of this operon appears to be located 91 bp upstream of the translational start, with a potential − 10 region at − 3 nt and a − 35 region at − 39 nt. Isogenic P. gingivalis mutants FLL273 (PG1777 : : ermF-ermAM) and FLL293 (PG1779 : : ermF-ermAM) showed increased sensitivity to and decreased survival after treatment with hydrogen peroxide. P. gingivalis FLL273 showed a fivefold increase in the formation of spontaneous mutants when compared with the parent strain after exposure to hydrogen peroxide. The recombinant PG1777 protein displayed iron-binding properties when incubated with FeSO4 and Fe(NH4)2(SO4).6H2O. The rPG1777 protein protected DNA from degradation when exposed to hydrogen peroxide in the presence of iron. Taken together, the data suggest that the grpE-dnaJ-PG1777-PG1778-PG1779 transcriptional unit may play an important role in oxidative stress resistance in P. gingivalis via its ability to protect against DNA damage. PMID:26581883

  18. 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.

  19. Problem Formulation in Knowledge Discovery via Data Analytics (KDDA) for Environmental Risk Management.

    PubMed

    Li, Yan; Thomas, Manoj; Osei-Bryson, Kweku-Muata; Levy, Jason

    2016-12-15

    With the growing popularity of data analytics and data science in the field of environmental risk management, a formalized Knowledge Discovery via Data Analytics (KDDA) process that incorporates all applicable analytical techniques for a specific environmental risk management problem is essential. In this emerging field, there is limited research dealing with the use of decision support to elicit environmental risk management (ERM) objectives and identify analytical goals from ERM decision makers. In this paper, we address problem formulation in the ERM understanding phase of the KDDA process. We build a DM³ ontology to capture ERM objectives and to inference analytical goals and associated analytical techniques. A framework to assist decision making in the problem formulation process is developed. It is shown how the ontology-based knowledge system can provide structured guidance to retrieve relevant knowledge during problem formulation. The importance of not only operationalizing the KDDA approach in a real-world environment but also evaluating the effectiveness of the proposed procedure is emphasized. We demonstrate how ontology inferencing may be used to discover analytical goals and techniques by conceptualizing Hazardous Air Pollutants (HAPs) exposure shifts based on a multilevel analysis of the level of urbanization (and related economic activity) and the degree of Socio-Economic Deprivation (SED) at the local neighborhood level. The HAPs case highlights not only the role of complexity in problem formulation but also the need for integrating data from multiple sources and the importance of employing appropriate KDDA modeling techniques. Challenges and opportunities for KDDA are summarized with an emphasis on environmental risk management and HAPs.

  20. 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.

  1. 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

  2. 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

  3. Pollution Prevention: National Trends, Forecasts and Options for the Army

    DTIC Science & Technology

    1992-06-01

    to ease the problems associated with municipal solid waste . In the long !erm. dastnbution of contaminants to vanous environmental media still results...potential impacts that hazardous waste generation may have on the Army. Municipal Solid Waste and Industrial Non-hazardous Waste The United States generates...landfilling (5,18). It is important to note that these tipping fees do not include costs to handle, transport and collect municipal solid waste . Using these

  4. Sorafenib blocks tumour growth, angiogenesis and metastatic potential in preclinical models of osteosarcoma through a mechanism potentially involving the inhibition of ERK1/2, MCL-1 and ezrin pathways

    PubMed Central

    2009-01-01

    Background Osteosarcoma (OS) is the most common primary bone tumour in children and young adults. Despite improved prognosis, metastatic or relapsed OS remains largely incurable and no significant improvement has been observed in the last 20 years. Therefore, the search for alternative agents in OS is mandatory. Results We investigated phospho-ERK 1/2, MCL-1, and phospho-Ezrin/Radixin/Moesin (P-ERM) as potential therapeutic targets in OS. Activation of these pathways was shown by immunohistochemistry in about 70% of cases and in all OS cell lines analyzed. Mutational analysis revealed no activating mutations in KRAS whereas BRAF gene was found to be mutated in 4/30 OS samples from patients. Based on these results we tested the multi-kinase inhibitor sorafenib (BAY 43-9006) in preclinical models of OS. Sorafenib inhibited OS cell line proliferation, induced apoptosis and downregulated P-ERK1/2, MCL-1, and P-ERM in a dose-dependent manner. The dephosphorylation of ERM was not due to ERK inhibition. The downregulation of MCL-1 led to an increase in apoptosis in OS cell lines. In chick embryo chorioallantoic membranes, OS supernatants induced angiogenesis, which was blocked by sorafenib and it was also shown that sorafenib reduced VEGF and MMP2 production. In addition, sorafenib treatment dramatically reduced tumour volume of OS xenografts and lung metastasis in SCID mice. Conclusion In conclusion, ERK1/2, MCL-1 and ERM pathways are shown to be active in OS. Sorafenib is able to inhibit their signal transduction, both in vitro and in vivo, displaying anti-tumoural activity, anti-angiogenic effects, and reducing metastatic colony formation in lungs. These data support the testing of sorafenib as a potential therapeutic option in metastatic or relapsed OS patients unresponsive to standard treatments. PMID:20003259

  5. Inoculation of cranberry (Vaccinium macrocarpon) with the ericoid mycorrhizal fungus Rhizoscyphus ericae increases nitrate influx.

    PubMed

    Kosola, Kevin R; Workmaster, Beth Ann A; Spada, Piero A

    2007-01-01

    Despite the ubiquitous presence of ericoid mycorrhizal (ERM) fungi in cranberry (Vaccinium macrocarpon), no prior studies have examined the effect of ERM colonization on NO(3)(-) influx kinetics. Here, (15)NO(3)(-) influx was measured in nonmycorrhizal and mycorrhizal cranberry in hydroponics. Mycorrhizal cranberry were inoculated with the ERM fungus Rhizoscyphus (syn. Hymenoscyphus) ericae. (15)NO(3)(-) influx by R. ericae in solution culture was also measured. Rhizoscyphus ericae NO(3)(-) influx kinetics were linear when mycelium was exposed for 24 h to 3.8 mm NH(4)(+), and saturable when pretreated with 3.8 mm NO(3)(-), 50 microm NO(3)(-), or 50 microm NH(4)(+). Both low-N pretreatments induced greater NO(3)(-) influx than either of the high-N pretreatments. Nonmycorrhizal cranberry exhibited linear NO(3)(-) influx kinetics. By contrast, mycorrhizal cranberry had saturable NO(3)(-) influx kinetics, with c. eightfold greater NO(3)(-) influx than nonmycorrhizal cranberry at NO(3)(-) concentrations from 20 microm to 2 mm. There was no influence of pretreatments on cranberry NO(3)(-) influx kinetics, regardless of mycorrhizal status. Inoculation with R. ericae increased the capacity of cranberry to utilize NO(3)(-)-N. This finding is significant both for understanding the potential nutrient niche breadth of cranberry and for management of cultivated cranberry when irrigation water sources contain nitrate.

  6. Comparison of VF-14 scores among different ophthalmic surgical interventions.

    PubMed

    Kishimoto, Fumiko; Ohtsuki, Hiroshi

    2012-01-01

    To clarify surgical outcomes for 5 ophthalmic diseases in terms of vision-related quality of life (QOL), we sent a self-administered Visual Function Questionnaire-14 (VF-14) to patients 3 months postoperatively, and the VF-14 scores for the surgical outcome of strabismus were compared with those of patients with diabetic macular edema (DME), cataract, glaucoma, and epiretinal membrane (ERM). Test-retest repeatability of VF-14 was evaluated with Bland-Altman analysis. Of the 625 eligible patients who were referred for enrollment, 48 with comitant strabismus, 50 with incomitant strabismus, 45 with DME, 38 with cataract, 129 with glaucoma, and 73 with ERM agreed to answer. Eighty percent of subjects showed 95% limits of agreement with the VF-14 evaluated by repeated measurements. The gain by surgery for incomitant strabismus was not different from that of cataract (p = 0.5551), but it was significantly better than those of DME (p = 0.0266), comitant strabismus (p = 0.0128), ERM (p = 0.0021), glaucoma with cataract (p < 0.0001), and glaucoma alone (p < 0.0001). The surgical outcome in terms of QOL for patients with incomitant strabismus was good and comparable to that of patients with cataract surgery.

  7. Precision orbit determination for the GEOSAT exact repeat mission

    NASA Astrophysics Data System (ADS)

    Smith, J. C.; Ries, J. C.; Shum, C. K.; Schutz, B. E.; Tapley, B. D.

    The Navy's Geodetic Satellite (GEOSAT) was launched on March 12, 1985, carrying a single-frequency microwave altimeter which measures the height of the satellite above the ocean surface to a precision of a few centimeters. The GEOSAT Exact Repeat Mission (ERM), which was initiated in November of 1986, placed the spacecraft in an exact 17 day repeat orbit. The Geophysical Data Records (GDR) for the ERM are available to the scientific community. GEOSAT is tracked by the Navy's OPNET and the Defense Mapping Agency's TRANET doppler tracking systems. The GDR orbits are computed using the OPNET tracking data and have an rms radial accuracy of one to two meters. The initial eighty days of the TRANET data during the ERM were made available for the assessment of the TRANET tracking system to perform precision orbit determination for the Topex/Poseidon Mission. This data was used to compute GEOSAT orbits using an improved gravity model which has been developed as part of the Topex gravity model improvement effort. Accuracy of the orbit was evaluated using altimeter crossover data. For a continuous 17 day GEOSAT orbit, the global crossover rms is at the 35 cm level, which suggests a radial orbit accuracy also on the order of 35 cm.

  8. Macrolide Resistance in Streptococcus pneumoniae

    PubMed Central

    Schroeder, Max R.; Stephens, David S.

    2016-01-01

    Streptococcus pneumoniae is a common commensal and an opportunistic pathogen. Suspected pneumococcal upper respiratory infections and pneumonia are often treated with macrolide antibiotics. Macrolides are bacteriostatic antibiotics and inhibit protein synthesis by binding to the 50S ribosomal subunit. The widespread use of macrolides is associated with increased macrolide resistance in S. pneumoniae, and the treatment of pneumococcal infections with macrolides may be associated with clinical failures. In S. pneumoniae, macrolide resistance is due to ribosomal dimethylation by an enzyme encoded by erm(B), efflux by a two-component efflux pump encoded by mef (E)/mel(msr(D)) and, less commonly, mutations of the ribosomal target site of macrolides. A wide array of genetic elements have emerged that facilitate macrolide resistance in S. pneumoniae; for example erm(B) is found on Tn917, while the mef (E)/mel operon is carried on the 5.4- or 5.5-kb Mega element. The macrolide resistance determinants, erm(B) and mef (E)/mel, are also found on large composite Tn916-like elements most notably Tn6002, Tn2009, and Tn2010. Introductions of 7-valent and 13-valent pneumococcal conjugate vaccines (PCV-7 and PCV-13) have decreased the incidence of macrolide-resistant invasive pneumococcal disease, but serotype replacement and emergence of macrolide resistance remain an important concern. PMID:27709102

  9. Epoxy resin monomers with reduced skin sensitizing potency.

    PubMed

    O'Boyle, Niamh M; Niklasson, Ida B; Tehrani-Bagha, Ali R; Delaine, Tamara; Holmberg, Krister; Luthman, Kristina; Karlberg, Ann-Therese

    2014-06-16

    Epoxy resin monomers (ERMs), especially diglycidyl ethers of bisphenol A and F (DGEBA and DGEBF), are extensively used as building blocks for thermosetting polymers. However, they are known to commonly cause skin allergy. This research describes a number of alternative ERMs, designed with the aim of reducing the skin sensitizing potency while maintaining the ability to form thermosetting polymers. The compounds were designed, synthesized, and assessed for sensitizing potency using the in vivo murine local lymph node assay (LLNA). All six epoxy resin monomers had decreased sensitizing potencies compared to those of DGEBA and DGEBF. With respect to the LLNA EC3 value, the best of the alternative monomers had a value approximately 2.5 times higher than those of DGEBA and DGEBF. The diepoxides were reacted with triethylenetetramine, and the polymers formed were tested for technical applicability using thermogravimetric analysis and differential scanning calorimetry. Four out of the six alternative ERMs gave polymers with a thermal stability comparable to that obtained with DGEBA and DGEBF. The use of improved epoxy resin monomers with less skin sensitizing effects is a direct way to tackle the problem of contact allergy to epoxy resin systems, particularly in occupational settings, resulting in a reduction in the incidence of allergic contact dermatitis.

  10. Yeast Vps13 promotes mitochondrial function and is localized at membrane contact sites

    PubMed Central

    Park, Jae-Sook; Thorsness, Mary K.; Policastro, Robert; McGoldrick, Luke L.; Hollingsworth, Nancy M.; Thorsness, Peter E.; Neiman, Aaron M.

    2016-01-01

    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

  11. Moesin-dependent cytoskeleton remodelling is associated with an anaplastic phenotype of pancreatic cancer.

    PubMed

    Abiatari, Ivane; Esposito, Irene; Oliveira, Tiago De; Felix, Klaus; Xin, Hong; Penzel, Roland; Giese, Thomas; Friess, Helmut; Kleeff, Jörg

    2010-05-01

    Cell motility is controlled by the dynamic cytoskeleton and its related proteins, such as members of the ezrin/radixin/moesin (ERM) family, which act as signalling molecules inducing cytoskeleton remodelling. Although ERM proteins have been identified as important factors in various malignancies, functional redundancy between these proteins has hindered the dissection of their individual contribution. The aim of the present study was to analyse the functional role of moesin in pancreatic malignancies. Cancer cells of different malignant lesions of human and transgenic mice pancreata were evaluated by immunohistochemistry. For functional analysis, cell growth, adhesion and invasion assays were carried out after transient and stable knock-down of moesin expression in pancreatic cancer cells. In vivo tumourigenicity was determined using orthotopic and metastatic mouse tumour models. We now show that moesin knock-down increases migration, invasion and metastasis and influences extracellular matrix organization of pancreatic cancer. Moesin-regulated migratory activities of pancreatic cancer cells were in part promoted through cellular translocation of beta-catenin, and re-distribution and organization of the cytoskeleton. Analysis of human and different transgenic mouse pancreatic cancers demonstrated that moesin is a phenotypic marker for anaplastic carcinoma, suggesting that this ERM protein plays a specific role in pancreatic carcinogenesis.

  12. Improvement of the Prediction Power of the CoMFA and CoMSIA Models on Histamine H3 Antagonists by Different Variable Selection Methods

    PubMed Central

    Ghasemi, Jahan B.; Tavakoli, Hossein

    2012-01-01

    The aim of this study is to enhance the predictivity power of CoMFA and CoMSIA models by means of different variable selection algorithms. The genetic algorithm (GA), successive projection algorithm (SPA), stepwise multiple linear regression (SW-MLR), and the enhanced replacement method (ERM) were used and tested as variable selection algorithms. Then, the selected variables were used to generate a simple and predictive model by the multilinear regression algorithm. A set of 74 histamine H3 antagonists were split into 40 compounds as a training set, and 17 compounds as a test set, by the Kennard-Stone algorithm. Before splitting the data, 17 compounds were randomly selected from the pool of the whole data set as an evaluation set without any supervision, pretreatment, or visual inspection. Among applied variable selection algorithms, ERM had noticeable improvement on the statistical parameters. The r2 values of training, test, and evaluation sets for the ERM-MLR model using CoMFA fields were 0.9560, 0.8630, and 0.8460 and using the CoMSIA fields were 0.9800, 0.8521, and 0.9080, respectively. In this study, the principles of organization for economic cooperation and development (OECD) for regulatory acceptability of QSARs are considered. PMID:23008805

  13. 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

  14. The composition of arbuscular mycorrhizal fungal communities differs among the roots, spores and extraradical mycelia associated with five Mediterranean plant species.

    PubMed

    Varela-Cervero, Sara; Vasar, Martti; Davison, John; Barea, José Miguel; Öpik, Maarja; Azcón-Aguilar, Concepción

    2015-08-01

    Arbuscular mycorrhizal fungi (AMF) are essential constituents of most terrestrial ecosystems. AMF species differ in terms of propagation strategies and the major propagules they form. This study compared the AMF community composition of different propagule fractions - colonized roots, spores and extraradical mycelium (ERM) - associated with five Mediterranean plant species in Sierra de Baza Natural Park (Granada, Spain). AMF were identified using 454 pyrosequencing of the SSU rRNA gene. A total of 96 AMF phylogroups [virtual taxa (VT)] were detected in the study site, including 31 novel VT. After per-sample sequencing depth standardization, 71 VT were recorded from plant roots, and 47 from each of the spore and ERM fractions. AMF communities differed significantly among the propagule fractions, and the root-colonizing fraction differed among host plant species. Indicator VT were detected for the root (13 Glomus VT), spore (Paraglomus VT281, VT336, Pacispora VT284) and ERM (Diversispora VT62) fractions. This study provides detailed evidence from a natural system that AMF taxa are differentially allocated among soil mycelium, soil spores and colonized root propagules. This has important implications for interpreting AMF diversity surveys and designing applications of AMF in vegetation restoration.

  15. The RhoE/ROCK/ARHGAP25 signaling pathway controls cell invasion by inhibition of Rac activity

    PubMed Central

    Thuault, Sylvie; Comunale, Franck; Hasna, Jessy; Fortier, Mathieu; Planchon, Damien; Elarouci, Nabila; De Reynies, Aurélien; Bodin, Stéphane; Blangy, Anne; Gauthier-Rouvière, Cécile

    2016-01-01

    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

  16. Dyes for Eyes™: hydrodynamics, biocompatibility and efficacy of 'heavy' (dual) dyes for chromovitrectomy.

    PubMed

    Mohr, Andreas; Bruinsma, Marieke; Oellerich, Silke; Frank, Hans; Gabel, Detlef; Melles, Gerrit R J

    2013-01-01

    As epiretinal membranes (ERMs), the internal limiting membrane (ILM) and the vitreous cortex are essentially transparent tissues, or translucent structures, nontraumatic removal may be challenging in various types of macular surgery. Vital dyes stain these thin tissues, thus allowing for better visualization of these structures during vitrectomy and selective 'membrane peeling' from the underlying retina. To avoid swirling of the dye within the fluid-filled vitreous cavity, and to better target the dye onto the macula, a fluid-air exchange is commonly performed. However, this may jeopardize visualization of the macula during peeling due to clouding of the posterior lens capsule, and may lead to postoperative visual field defects. Recently, a new dye solution for staining the ERM and ILM simultaneously has been developed that circumvents the need for fluid-air exchange, i.e. MembraneBlue-Dual™. This paper will focus on the hydrodynamics and biocompatibility of this 'heavy' dual dye and its efficacy for staining of the ILM and/or ERMs during posterior segment surgery in a multicenter clinical setting.

  17. Arp2/3 promotes junction formation and maintenance in the Caenorhabditis elegans intestine by regulating membrane association of apical proteins

    PubMed Central

    Bernadskaya, Yelena Y.; Patel, Falshruti B.; Hsu, Hsiao-Ting; Soto, Martha C.

    2011-01-01

    It has been proposed that Arp2/3, which promotes nucleation of branched actin, is needed for epithelial junction initiation but is less important as junctions mature. We focus here on how Arp2/3 contributes to the Caenorhabditis elegans intestinal epithelium and find important roles for Arp2/3 in the maturation and maintenance of junctions in embryos and adults. Electron microscope studies show that embryos depleted of Arp2/3 form apical actin-rich microvilli and electron-dense apical junctions. However, whereas apical/basal polarity initiates, apical maturation is defective, including decreased apical F-actin enrichment, aberrant lumen morphology, and reduced accumulation of some apical junctional proteins, including DLG-1. Depletion of Arp2/3 in adult animals leads to similar intestinal defects. The DLG-1/AJM-1 apical junction proteins, and the ezrin–radixin–moesin homologue ERM-1, a protein that connects F-actin to membranes, are required along with Arp2/3 for apical F-actin enrichment in embryos, whereas cadherin junction proteins are not. Arp2/3 affects the subcellular distribution of DLG-1 and ERM-1. Loss of Arp2/3 shifts both ERM-1 and DLG-1 from pellet fractions to supernatant fractions, suggesting a role for Arp2/3 in the distribution of membrane-associated proteins. Thus, Arp2/3 is required as junctions mature to maintain apical proteins associated with the correct membranes. PMID:21697505

  18. Antimicrobial resistance and virulence traits in Enterococcus strains isolated from dogs and cats.

    PubMed

    Iseppi, Ramona; Messi, Patrizia; Anacarso, Imacolata; Bondi, Moreno; Sabia, Carla; Condò, Carla; de Niederhausern, Simona

    2015-07-01

    We investigated presence and prevalence of antibiotic-resistances and other biological characters in enterococci isolated from faeces of healthy dogs and cats because these microorganisms represent important human and veterinary pathogens/opportunists, and a significant burden for healthcare systems. In all samples (n=115) we detected enterococci, with a predominance of Enterococcus faecium (42; 36.5%) and Enterococcus faecalis (36; 31.3%) species, endowed with virulence traits and multidrug-resistance. The two predominant resistance patterns (erythromycin, tetracycline) were examined by polymerase chain reaction for tet and erm genes. Only tetM for tetracycline, and ermA and ermB for erythromycin were detected. PCR for gelatinase gene (gelE) was positive in 62.6% of isolates, but only 26.1% produce gelatinase suggesting the existence of silent genes. efaAfs and efaAfm genes were found in E. faecalis and E. faecium respectively. 89.6% of isolates produced bacteriocin-like substances with a prevailing action against Listeria genus and, among these, 33.9% were positive for the bacteriocin structural genes entA, entL50 or entP. According to our study, pet animals can be considered a reservoir of potentially pathogenic enterococci and we cannot exclude that those microorganisms may be responsible for opportunistic infections in high-risk pet owners.

  19. Hedgehog Pathway Inhibition Hampers Sphere and Holoclone Formation in Rhabdomyosarcoma

    PubMed Central

    Almazán-Moga, Ana; Zarzosa, Patricia; Vidal, Isaac; Molist, Carla; Giralt, Irina; Navarro, Natalia; Soriano, Aroa; Alfranca, Arantza; Sánchez de Toledo, José

    2017-01-01

    Rhabdomyosarcoma (RMS) is the most common type of soft tissue sarcoma in children and can be divided into two main subtypes: embryonal (eRMS) and alveolar (aRMS). Among the cellular heterogeneity of tumors, the existence of a small fraction of cells called cancer stem cells (CSC), thought to be responsible for the onset and propagation of cancer, has been demonstrated in some neoplasia. Although the existence of CSC has been reported for eRMS, their existence in aRMS, the most malignant subtype, has not been demonstrated to date. Given the lack of suitable markers to identify this subpopulation in aRMS, we used cancer stem cell-enriched supracellular structures (spheres and holoclones) to study this subpopulation. This strategy allowed us to demonstrate the capacity of both aRMS and eRMS cells to form these structures and retain self-renewal capacity. Furthermore, cells contained in spheres and holoclones showed significant Hedgehog pathway induction, the inhibition of which (pharmacologic or genetic) impairs the formation of both holoclones and spheres. Our findings point to a crucial role of this pathway in the maintenance of these structures and suggest that Hedgehog pathway targeting in CSC may have great potential in preventing local relapses and metastases. PMID:28243259

  20. 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

  1. Effects of biochar on reducing the abundance of oxytetracycline, antibiotic resistance genes, and human pathogenic bacteria in soil and lettuce.

    PubMed

    Duan, Manli; Li, Haichao; Gu, Jie; Tuo, Xiaxia; Sun, Wei; Qian, Xun; Wang, Xiaojuan

    2017-05-01

    Antibiotics and antibiotic resistance genes (ARGs) in soil can affect human health via the food chain. Biochar is a soil amendment but its impacts on ARGs and the microbial communities associated with soil and vegetables are unclear. Therefore, we established three lettuce pot culture experiments, i.e., O300: 300 mg/kg oxytetracycline (OTC), BO300: 300 mg/kg OTC + 2% biochar, and a control without OTC or biochar. We found that under BO300, the relative abundances of ARGs were reduced by 51.8%, 43.4%, and 44.1% in lettuce leaves, roots, and soil, respectively, compared with O300. intI1 was highly abundant in soil and lettuce, and it co-occurred with some ARGs (tetW, ermF, and sul1). Redundancy analysis and network analysis indicated that the bacterial community succession was the main mechanism that affected the variations in ARGs and intI1. The reduction of Firmicutes due to the biochar treatment of soil and lettuce was the main factor responsible for the removal of tetracycline resistance genes in leaves. Biochar application led to the disappearance of human pathogenic bacteria (HPB), which was significantly correlated with the abundances of ermF and ermX. In summary, biochar is an effective farmland amendment for reducing the abundances of antibiotics, ARGs, and HPB in order to ensure the safety of vegetables and protect human health.

  2. QSAR studies on 4-anilino-3-quinolinecarbonitriles as Src kinase inhibitors using robust PCA and both linear and nonlinear models.

    PubMed

    Sun, Min; Zheng, Youguang; Wei, Hongtao; Chen, Junqing; Ji, Min

    2009-10-01

    Quantitative structure-activity relationship (QSAR) studies have been carried out on 4-anilino-3-quinolinecarbonitriles, a set of novel Src kinase inhibitors, with the aim of dissecting the structural requirements for Src inhibitory activities. After outlier identification using robust principal component analysis (robust PCA), linear models based on forward selection combined with multiple linear regression, (FS-MLR), enhanced replacement method followed by multiple linear regression (ERM) and a nonlinear model using support vector regression (SVR) were constructed and compared. All models were rigorously validated using leave-one-out cross-validation (LOOCV), 5-fold cross-validations (5-CV) and shuffling external validation (SEVs). ERM seems to outperform both FS-MLR and SVR evidenced by better prediction performance (n = 35, R(2)(training) = 0.918, R(2)(pred) = 0.928). Robustness and predictive ability of ERM model were also evaluated. The generated QASR model revealed that the Src inhibitory activity of 4-anilino-3-quinolinecarbonitriles could be associated with the size of substituents in the C7 position and the steric hindrance effect. The results of the present study may be of great help in designing novel 4-anilino-3-quinolinecarbonitriles with more potent Src kinase inhibitory activity.

  3. Energy restriction and potential energy restriction mimetics.

    PubMed

    Nikolai, Sibylle; Pallauf, Kathrin; Huebbe, Patricia; Rimbach, Gerald

    2015-12-01

    Energy restriction (ER; also known as caloric restriction) is the only nutritional intervention that has repeatedly been shown to increase lifespan in model organisms and may delay ageing in humans. In the present review we discuss current scientific literature on ER and its molecular, metabolic and hormonal effects. Moreover, criteria for the classification of substances that might induce positive ER-like changes without having to reduce energy intake are summarised. Additionally, the putative ER mimetics (ERM) 2-deoxy-d-glucose, metformin, rapamycin, resveratrol, spermidine and lipoic acid and their suggested molecular targets are discussed. While there are reports on these ERM candidates that describe lifespan extension in model organisms, data on longevity-inducing effects in higher organisms such as mice remain controversial or are missing. Furthermore, some of these candidates produce detrimental side effects such as immunosuppression or lactic acidosis, or have not been tested for safety in long-term studies. Up to now, there are no known ERM that could be recommended without limitations for use in humans.

  4. Extraradical mycelium of the arbuscular mycorrhizal fungus Glomus lamellosum can take up, accumulate and translocate radiocaesium under root-organ culture conditions.

    PubMed

    Declerck, Stéphane; Dupré de Boulois, Hervé; Bivort, Céline; Delvaux, Bruno

    2003-06-01

    Radiocaesium enters the food chain when plants absorb it from soil, in a process that is strongly dependent on soil properties and plant and microbial species. Among the microbial species, arbuscular mycorrhizal (AM) fungi are obligate symbionts that colonize the root cortex of many plants and develop an extraradical mycelial (ERM) network that ramifies in the soil. Despite the well-known involvement of this ERM network in mineral nutrition and uptake of some heavy metals, only limited data are available on its role in radiocaesium transport in plants. We used root-organ culture to demonstrate that the ERM of the AM fungus Glomus lamellosum can take up, possibly accumulate and unambiguously translocate radiocaesium from a 137Cs-labelled synthetic root-free compartment to a root compartment and within the roots. The accumulation of 137Cs by hyphae in the root-free compartment may be explained by sequestration in the hyphae or by a bottleneck effect resulting from a limited number of hyphae crossing the partition between the two compartments. Uptake and translocation resulted from the incorporation of 137Cs into the fungal hyphae, as no 137Cs was detected in mycorrhizal roots treated with formaldehyde. The importance of the translocation process was indicated by the correlation between 137Cs measured in the roots and the total hyphal length connecting the roots with the labelled compartment. 137Cs may be translocated via a tubular vacuolar system or by cytoplasmic streaming per se.

  5. 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.

  6. Multi-year elevation changes near the west margin of the Greenland ice sheet from satellite radar altimetry

    SciTech Connect

    Lingle, C.S.; Brenner, A.C.; Zwally, H.J.; DiMarzio, J.P.

    1992-03-01

    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 are also carried out using altimetry referenced to orbits not adjusted into a common ocean surface.

  7. 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.

  8. Macrolide resistance in Streptococcus pneumoniae isolated from Argentinian pediatric patients suffering from acute otitis media.

    PubMed

    Reijtman, Vanesa; Gagetti, Paula; Faccone, Diego; Fossati, Sofía; Sommerfleck, Patricia; Hernández, Claudia; Bernáldez, Patricia; Lopardo, Horacio; Corso, Alejandra

    2013-01-01

    Macrolide-resistant Streptococcus pneumoniae emerged in Argentina in 1995, representing 26% of invasive infection isolates in children under 5 years old. The objectives of this study were to describe the prevalence of ermB and mefA genes in macrolide-resistant S. pneumoniae isolates from acute otitis media (AOM) and to determine their genetic relatedness. Between May 2009 and August 2010, 126 S. pneumoniae isolates from 324 otherwise healthy children with a first episode of AOM were included. Twenty six of these isolates (20.6%) were resistant to erythromycin. Most frequent serotypes were: 14 (46.2%), 6A (23.1%), 19F (7.7%) and 9V (7.7%). Twenty (76.9%) carried the mefA gene, 5 (19.2%) have the ermB gene, and 1 (3.9%) both ermB + mefA. Ten clonal types were identified, mostly related to Sweden(15A)-25/ST782 (SLV63), CloneB(6A)/ST473 and England(14)-9/ ST9. This is the first study assessing the mechanisms of macrolide resistance in pneumococci isolates from pediatric AOM in Argentina and their genetic relatedness.

  9. 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

  10. 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.

  11. 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

  12. Importance of fumarate and nitrate reduction regulatory protein for intestinal proliferation of Vibrio vulnificus.

    PubMed

    Kado, Takehiro; Kashimoto, Takashige; Yamazaki, Kohei; Ueno, Shunji

    2017-01-01

    The sepsis caused by Vibrio vulnificus is characterized by an average incubation period of 26 h and a high mortality rate exceeding 50%. The fast growth and dissemination of V. vulnificus in vivo lead to poor clinical outcomes in patients. Therefore, elucidation of the proliferation mechanisms of this organism in vivo may lead to the development of an effective therapeutic strategy. In this study, we focused on the low oxygen concentration in the intestinal milieu because of its drastic difference from that in air. Fumarate and nitrate reduction regulatory protein (FNR) is known to be a global transcriptional regulator for adaptation to anaerobic conditions in various bacteria. We generated a strain of V. vulnificus in which the fnr gene was replaced with an erythromycin resistance gene (fnr::erm mutant). When the fnr::erm mutant was tested in a growth competition assay against the wild-type (WT) in vivo, the competitive index of fnr::erm mutant to WT in the intestinal loop and liver was 0.378 ± 0.192 (mean ± SD) and 0.243 ± 0.123, respectively. These data suggested that FNR is important for the proliferation of V. vulnificus in the intestine to achieve a critical mass to be able to invade the systemic circulation.

  13. 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

  14. Pathologic Roles of Receptor-Associated Prorenin System in Idiopathic Epiretinal Membrane

    PubMed Central

    Dong, Yoko; Kanda, Atsuhiro; Noda, Kousuke; Saito, Wataru; Ishida, Susumu

    2017-01-01

    Receptor-associated prorenin system (RAPS) refers to the pathogenic mechanism whereby prorenin binding to (pro)renin receptor [(P)RR] dually activates tissue renin-angiotensin system (RAS) and RAS-independent signaling via (P)RR. The aim of this study is to determine the association of RAPS with idiopathic epiretinal membrane (iERM). Reverse transcription-PCR indicated the expression of RAPS components, including (P)RR and Ang II type 1 receptor (AT1R), in iERM tissues and human Müller glial cell line. Double-labeling analyses demonstrated that (P)RR and AT1R were detected in cells positive for glial fibrillary acidic protein, a marker for glial cells, and co-localized with prorenin and angiotensinogen, respectively. Administration of prorenin to Müller glial cells enhanced mRNA expression of fibroblast growth factor 2, while Ang II application stimulated the expression of glial cell line-derived neurotrophic factor, nerve growth factor, and transforming growth factor-β1. These expression levels induced by prorenin or Ang II were reversed by (P)RR or AT1R blockade, respectively. Immunofluorescence revealed tissue co-localization of (P)RR and AT1R with the products of the upregulated genes in vitro. The present findings suggest the involvement of RAPS in the pathogenesis of iERM. PMID:28276504

  15. An evaluation of benthic community measures using laboratory-derived sediment effect concentrations

    SciTech Connect

    Dwyer, F.J.; Canfield, T.J.; Ingersoll, C.G.; Kemble, N.E.; Mount, D.R.

    1995-12-31

    Sediment effect concentrations (SECs) are contaminant sediment concentrations which are frequently associated with sediment toxicity. Recently, a number of different SECs have been calculated from laboratory toxicity tests with field collected sediments using Chironomus tentans, Chironomus riparius, and Hyalella azteca. Toxicity endpoints included (depending upon species) lethality, growth and sexual maturation. The authors selected the Effect Range Median (ERM) calculated for 28-d Hyalella azteca as an SEC for evaluating six different benthic community measures as indicators of contaminated sediment. The benthic measures included: taxa richness, chironomid genera richness, percent chironomid deformity, chironomid biotic index, ratio of chironomids/oligochaetes, and oligochaete biotic index. Benthic measures were obtained for 31 stations from the Great Lakes and 13 stations from Milltown Reservoir and Clark Fork River, MT. Each benthic measure was ranked from 1 to 100 and individual ranks and various combinations of ranks were plotted against the ratio of chemical concentration at the site/ERM calculated for that chemical (similar to a toxic unit approach) and the sum of the ERM ratios (sum of toxic units). Preliminary analysis indicates that, in general, benthic measures varied widely in relatively uncontaminated stations, confounding any underlying relationship that may have existed. The absence of chironomids, in areas with suitable habitat, seems to be indicative of grossly contaminated stations, but not an endpoint useful for discriminating stations with contaminant concentrations closer to the SEC. The usefulness of benthic measures as diagnostic tools for contaminated sediments and potential ways to improve these measures will be discussed.

  16. Contamination profiles of antibiotic resistance genes in the sediments at a catchment scale.

    PubMed

    Su, Hao-Chang; Pan, Chang-Gui; Ying, Guang-Guo; Zhao, Jian-Liang; Zhou, Li-Jun; Liu, You-Sheng; Tao, Ran; Zhang, Rui-Quan; He, Liang-Ying

    2014-08-15

    The aim of this study was to investigate the contamination profiles of tetracycline, sulfonamide, and macrolide resistance genes, as well as integrons in sediments of Dongjiang River basin of South China by real time quantitative polymerase chain reaction. sul2 was the most abundant resistance gene, with the average concentration of 6.97×10(8) copies/g and 1.00×10(8) copies/g in the dry and wet seasons, respectively, followed by ermF, sul3, sul1, intI1, tetA, ermB, tetX, tetM, tetQ, tetO, tetW, tetS, ermC, and tetB. The abundance of intI2 gene was the lowest in the sediment samples. Significant correlations existed between the ARGs and sediment properties as well as metals (Cu and Zn) and corresponding antibiotic classes, suggesting that the contamination of ARGs is related to chemical pollution of the sediments in the river basin. Principal component analysis showed distinct groupings of the sampling sites, reflecting that human activities are the key player in the dissemination of ARGs in the catchment environment.

  17. 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

  18. Mean sea surface and variability of the Gulf of Mexico using Geosat altimetry data

    NASA Technical Reports Server (NTRS)

    Leben, Robert R.; Born, George H.; Fox, Chad A.; Thompson, Dana J.

    1990-01-01

    Geosat Exact Repeat Mission (ERM) altimetric measurements of the sea surface height in the Gulf of Mexico are used to determine the mean sea surface height with respect to the ellipsoid and mesoscale variability along Geosat ground tracks in the gulf for the time period from November 8, 1986 to November 25, 1988. A mean surface generated using the Geosat ERM along-track mean is calculated and contrasted with a previously derived mean surface determined using GEOS 3 and Seasat crossover differences. This provides a first look at the variability in the mean between the time periods of 1987-1988 and 1975-1978. In addition, the along-track mesoscale variability time series has been produced from the Geosat ERM data set by using a robust orbit-error removal algorithm to determine the variability of the sea-surface height with respect to the along-track mean. Good qualitative and quantitative agreement with previous in situ observations in the region is found. This study demonstrates the potential of satellite altimetry for oceanographic studies of the Gulf of Mexico.

  19. Noise Reduction Effect of Multiple-Sampling-Based Signal-Readout Circuits for Ultra-Low Noise CMOS Image Sensors.

    PubMed

    Kawahito, Shoji; Seo, Min-Woong

    2016-11-06

    This paper discusses the noise reduction effect of multiple-sampling-based signal readout circuits for implementing ultra-low-noise image sensors. The correlated multiple sampling (CMS) technique has recently become an important technology for high-gain column readout circuits in low-noise CMOS image sensors (CISs). This paper reveals how the column CMS circuits, together with a pixel having a high-conversion-gain charge detector and low-noise transistor, realizes deep sub-electron read noise levels based on the analysis of noise components in the signal readout chain from a pixel to the column analog-to-digital converter (ADC). The noise measurement results of experimental CISs are compared with the noise analysis and the effect of noise reduction to the sampling number is discussed at the deep sub-electron level. Images taken with three CMS gains of two, 16, and 128 show distinct advantage of image contrast for the gain of 128 (noise(median): 0.29 e(-)rms) when compared with the CMS gain of two (2.4 e(-)rms), or 16 (1.1 e(-)rms).

  20. Normal and Malignant Muscle Cell Transplantation into Immune Compromised Adult Zebrafish

    PubMed Central

    Moore, John C.; Langenau, David M.

    2014-01-01

    Zebrafish have become a powerful tool for assessing development, regeneration, and cancer. More recently, allograft cell transplantation protocols have been developed that permit engraftment of normal and malignant cells into irradiated, syngeneic, and immune compromised adult zebrafish. These models when coupled with optimized cell transplantation protocols allow for the rapid assessment of stem cell function, regeneration following injury, and cancer. Here, we present a method for cell transplantation of zebrafish adult skeletal muscle and embryonal rhabdomyosarcoma (ERMS), a pediatric sarcoma that shares features with embryonic muscle, into immune compromised adult rag2E450fs homozygous mutant zebrafish. Importantly, these animals lack T cells and have reduced B cell function, facilitating engraftment of a wide range of tissues from unrelated donor animals. Our optimized protocols show that fluorescently labeled muscle cell preparations from α-actin-RFP transgenic zebrafish engraft robustly when implanted into the dorsal musculature of rag2 homozygous mutant fish. We also demonstrate engraftment of fluorescent-transgenic ERMS where fluorescence is confined to cells based on differentiation status. Specifically, ERMS were created in AB-strain myf5-GFP; mylpfa-mCherry double transgenic animals and tumors injected into the peritoneum of adult immune compromised fish. The utility of these protocols extends to engraftment of a wide range of normal and malignant donor cells that can be implanted into dorsal musculature or peritoneum of adult zebrafish. PMID:25591079

  1. 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

  2. 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.

  3. Quantitative proteomics of aqueous and vitreous fluid from patients with idiopathic epiretinal membranes.

    PubMed

    Pollreisz, Andreas; Funk, Marion; Breitwieser, Florian P; Parapatics, Katja; Sacu, Stefan; Georgopoulos, Michael; Dunavoelgyi, Roman; Zlabinger, Gerhard J; Colinge, Jacques; Bennett, Keiryn L; Schmidt-Erfurth, Ursula

    2013-03-01

    The purpose of the study was to perform a detailed, quantitative protein analysis of the aqueous (AF) and vitreous fluids (VF) from human eyes with idiopathic epiretinal membranes (iERM). The complementary approaches of quantitative liquid chromatography mass spectrometry (LCMS) and multiplex protein assays were utilised to reveal the protein composition in ocular fluids of this retinal disorder. In a prospective clinical trial, AF and VF was collected during surgery from twenty four eyes corresponding to twenty four patients with iERM. VF and AF from eight patients were labelled with the 4-plex iTRAQ reagent and analysed by LCMS. Each iTRAQ 4-plex experiment consisted of the AH and VH from two patients. A total of 323 proteins were identified in the AF and VF from eyes with iERM. Grouping the proteins according to involvement in biological processes, showed that the majority were involved in the classical and alternative pathway of complement activation (n = 27), proteolysis (n = 26) and cell adhesion (n = 28). iTRAQ relative quantitation revealed minimal variation in the protein content between both ocular compartments with only 3.96% of the identified proteins significantly, differentially-expressed. Eight proteins were expressed at a higher level in the VF compared to the AF; and 4 proteins were expressed at a lower level in the VF compared to the AF. For the multiplex bead assays, 29 growth factors and cytokines were assessed in the AF and VF from 16 additional patients with iERM. The protein profile was shown to be similar between VF and AF for the majority of factors except PDGF-AA. This factor was expressed at a higher level in the VF compared to the AF. The data presented in this study revealed that the majority of the proteins identified by LCMS and multiplex bead assays were present in both ocular compartments in similar quantities. This study is a first step, 'discovery phase' towards revealing and quantitating the protein content in the aqueous

  4. [Serotype distribution and antibiotic susceptibilities of Streptococcus pneumoniae causing acute exacerbations and pneumonia in children with chronic respiratory diseases].

    PubMed

    Altınkanat Gelmez, Gülşen; Soysal, Ahmet; Kuzdan, Canan; Karadağ, Bülent; Hasdemir, Ufuk; Bakır, Mustafa; Söyletir, Güner

    2013-10-01

    This study aimed to investigate serotype distribution and antimicrobial resistance of Streptococcus pneumoniae isolates obtained from children with chronic respiratory diseases admitted to hospital with a diagnosis of acute exacerbations between 2008-2010 at Marmara University Hospital, Istanbul, Turkey. Sixty one S.pneumoniae strains isolated from the respiratory samples of patients were studied for erythromycin, clindamycin, tetracyline, trimethoprim-sulphametoxazole (TMP-SMX), vancomycin, levofloxacin susceptibilities by disk diffusion method; MIC values of penicillin and ceftriaxone were determined by E-test (AB Biodisk, Sweden). Results were evaluated according to the CLSI standards. The erythromycin-clindamycin double disc method was applied for the detection of macrolide resistance phenotypes. The presence of macrolide resistance genes, ermB, mef(A)/(E), ermTR were determined by PCR using specific primers for each gene. The serotypes were determined by multiplex PCR using specific primers for 40 different serotypes. According to CLSI criteria, penicillin resistance in S.pneumoniae isolates were found to be 8.2% (5/61) and intermediate resistance rate was 54% (33/61) for oral penicillin. Penicillin resistance were found to be only 1.6% (1/61) for parenteral penicillin. Resistance rates of erythromycin, clindamycin, tetracyline, TMP-SMX were detected as 55.8%, 46%, 47.5% and 67.2%; respectively. No resistance was detected to vancomycin and levofloxacin. Constitutive macrolide-lincosamide-streptogramin B (cMLSB) phenotype and M phenotype were observed in 82.4% (n= 28) and 17.6% (n= 6) of the macrolide resistant isolates, respectively. Inducible macrolide-lincosamide-streptogramin B (iMLSB) phenotype was not detected. The macrolid resistance genotypes, ermB, mef(A)/(E), were positive 50% and 14.7%; respectively. Both ermB and mef(A)/(E) genes were detected 35.3% of the macrolid resistant isolates. None of the isolates were positive for ermTR gene. The most

  5. A Low Prevalence of Inducible Macrolide, Lincosamide, and Streptogramin B Resistance Phenotype among Methicillin-Susceptible Staphylococcus aureus Isolated from Malaysian Patients and Healthy Individuals

    PubMed Central

    Mesbah Elkammoshi, Asma; Ghasemzadeh-Moghaddam, Hamed; Amin Nordin, Syafinaz; Mohd Taib, Niazlin; Kumar Subbiah, Suresh; Neela, Vasanthakumari; Awang Hamat, Rukman

    2016-01-01

    Background Antibiotic resistance among Staphylococcus aureus is of great concern worldwide. This resistance is further complicated by the ability of S. aureus to confer cross-resistance to other antibiotics due to the presence of resistance genes, such as erythromycin resistance methylase (erm) genes, which render the bacterium resistant to macrolide-lincosamide-streptogramin B (MLSB) antibiotics. Resistance to these antibiotics can lead to therapeutic failure, resulting in significant morbidity and mortality in patients with S. aureus infections. Objectives This study was performed to examine the distribution of MLSB-resistant strains of methicillin-susceptible S. aureus (MSSA), which were obtained from hospitalized patients and normal healthy individuals (carriers) using phenotypic methods, such as the double-disk diffusion (D-test) and the genotypic method by polymerase chain reaction (PCR). Methods A total of 183 nonduplicative MSSA isolates obtained from hospitalized patients (133) and carriers (50) in our previous studies were randomly selected for the D-test. The guidelines of the Clinical and Laboratory Standards Institute (CLSI) were used for the interpretation of the results of this test. The detection of ermA, ermB, ermC and msrA genes by PCR was performed for isolates that had positive D-test results and that were resistant to erythromycin. Results Of the 183 MSSA isolates, 97.2% and 98.4% were highly susceptible to erythromycin and clindamycin, respectively. MSLB resistance was detected in four isolates (2.2%). Of the 133 MSSA isolated from hospitalized patients, only 3.0% (4/133) and 2.3% (3/133) exhibited resistance to erythromycin and clindamycin, respectively. With regard to the MLSB resistance phenotypes, only 1.6% and 0.6% exhibited inducible MLSB (iMLSB) and MS phenotypes, respectively. The ermC gene was detected in all three iMLSB phenotypes, and the msrA gene was detected in the MS phenotype. Surprisingly, all MSSA isolates (100%) from

  6. Prevalence and characteristics of hyporeflective preretinal tissue in vitreomacular interface disorders

    PubMed Central

    Itoh, Yuji; Levison, Ashleigh L.; Kaiser, Peter K.; Srivastava, Sunil K.; Singh, Rishi P.; Ehlers, Justis P.

    2016-01-01

    Aims To assess prevalence and characteristics of hyporeflective preretinal tissue on spectral domain optical coherence tomography (SDOCT) in eyes with vitreomacular interface disorders. Methods 4037 eyes (3195 patients) with diagnosis of lamellar macular hole (LMH), full-thickness macular hole (FTMH), epiretinal membrane (ERM) and vitreomacular traction (VMT) were included. Quantitative analysis was performed including volume and area of the epiretinal proliferation, as well as the brightness of the hyporeflective band. Clinical characteristics were also collected and analyzed. Results A hyporeflective preretinal tissue layer was identified in 204 of 4037 eyes (5.1%); 162 eyes in LMH (79.4 %), 23 eyes in FTMH (11.3%) and 19 eyes in ERM (9.3 %). In LMH, the visual acuity was significantly different between the cases with and without epiretinal proliferation at the initial and the final visit, (P = 0.012, 0.046, respectively). The maximum thickness, area, volume of hyporeflective preretinal tissue became significantly larger during observation period (P < 0.001). Brightness of the preretinal tissue (109.3 ± 21.1 a.u.) was close to retinal ganglion cell layer (112.0 ± 19.5) and retinal outer plexiform layer (117.7 ± 19.5). Conclusions Hyporeflective preretinal tissue was found with significant frequency of eyes with LMH, FTMH and ERM, with a particularly high incidence in LMH. The increased presence of this tissue in cases of LMH may signify a particular subtype of LMH. More research is needed to better understand the implications of the presence of this tissue for visual and surgical outcomes. PMID:26206790

  7. 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

  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.

  9. 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.

  10. 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

  11. DNMT3B in vitro knocking-down is able to reverse embryonal rhabdomyosarcoma cell phenotype through inhibition of proliferation and induction of myogenic differentiation.

    PubMed

    Megiorni, Francesca; Camero, Simona; Ceccarelli, Simona; McDowell, Heather P; Mannarino, Olga; Marampon, Francesco; Pizer, Barry; Shukla, Rajeev; Pizzuti, Antonio; Marchese, Cinzia; Clerico, Anna; Dominici, Carlo

    2016-11-29

    Aberrant DNA methylation has been frequently observed in many human cancers, including rhabdomyosarcoma (RMS), the most common soft tissue sarcoma in children. To date, the expression and function of the de novo DNA methyltransferase (DNMT) 3B in RMS have not yet been investigated. Our study show for the first time a significant up-regulation of DNMT3B levels in 14 RMS tumour samples and 4 RMS cell lines in comparison to normal skeletal muscle. Transfection of RD and TE671 cells, two in vitro models of embryonal RMS (ERMS), with a synthetic DNMT3B siRNA decreased cell proliferation by arresting cell cycle at G1 phase, as demonstrated by the reduced expression of Cyclin B1, Cyclin D1 and Cyclin E2, and by the concomitant up-regulation of the checkpoint regulators p21 and p27. DNMT3B depletion also impaired RB phosphorylation status and decreased migratory capacity and clonogenic potential. Interestingly, DNMT3B knock-down was able to commit ERMS cells towards myogenic terminal differentiation, as confirmed by the acquisition of a myogenic-like phenotype and by the increased expression of the myogenic markers MYOD1, Myogenin and MyHC. Finally, inhibition of MEK/ERK signalling by U0126 resulted in a reduction of DNMT3B protein, giving evidence that DNMT3B is a down-stream molecule of this oncogenic pathway.Taken together, our data indicate that altered expression of DNMT3B plays a key role in ERMS development since its silencing is able to reverse cell cancer phenotype by rescuing myogenic program. Epigenetic therapy, by targeting the DNA methylation machinery, may represent a novel therapeutic strategy against RMS.

  12. 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.

  13. Phosphorylation of myosin-binding subunit (MBS) of myosin phosphatase by Rho-kinase in vivo.

    PubMed

    Kawano, Y; Fukata, Y; Oshiro, N; Amano, M; Nakamura, T; Ito, M; Matsumura, F; Inagaki, M; Kaibuchi, K

    1999-11-29

    Rho-associated kinase (Rho-kinase), which is activated by the small GTPase Rho, phosphorylates myosin-binding subunit (MBS) of myosin phosphatase and thereby inactivates the phosphatase activity in vitro. Rho-kinase is thought to regulate the phosphorylation state of the substrates including myosin light chain (MLC), ERM (ezrin/radixin/moesin) family proteins and adducin by their direct phosphorylation and by the inactivation of myosin phosphatase. Here we identified the sites of phosphorylation of MBS by Rho-kinase as Thr-697, Ser-854 and several residues, and prepared antibody that specifically recognized MBS phosphorylated at Ser-854. We found by use of this antibody that the stimulation of MDCK epithelial cells with tetradecanoylphorbol-13-acetate (TPA) or hepatocyte growth factor (HGF) induced the phosphorylation of MBS at Ser-854 under the conditions in which membrane ruffling and cell migration were induced. Pretreatment of the cells with Botulinum C3 ADP-ribosyltransferase (C3), which is thought to interfere with Rho functions, or Rho-kinase inhibitors inhibited the TPA- or HGF-induced MBS phosphorylation. The TPA stimulation enhanced the immunoreactivity of phosphorylated MBS in the cytoplasm and membrane ruffling area of MDCK cells. In migrating MDCK cells, phosphorylated MBS as well as phosphorylated MLC at Ser-19 were localized in the leading edge and posterior region. Phosphorylated MBS was localized on actin stress fibers in REF52 fibroblasts. The microinjection of C3 or dominant negative Rho-kinase disrupted stress fibers and weakened the accumulation of phosphorylated MBS in REF52 cells. During cytokinesis, phosphorylated MBS, MLC and ERM family proteins accumulated at the cleavage furrow, and the phosphorylation level of MBS at Ser-854 was increased. Taken together, these results indicate that MBS is phosphorylated by Rho-kinase downstream of Rho in vivo, and suggest that myosin phosphatase and Rho-kinase spatiotemporally regulate the

  14. Coagulase-negative staphylococci: pathogenesis, occurrence of antibiotic resistance genes and in vitro effects of antimicrobial agents on biofilm-growing bacteria.

    PubMed

    Szczuka, Ewa; Jabłońska, Lucyna; Kaznowski, Adam

    2016-12-01

    Coagulase-negative staphylococci (CoNS) are opportunistic pathogens that particularly cause infections in patients with implanted medical devices. The present research was performed to study the virulence potential of 53 clinical isolates of Staphylococcus capitis, Staphylococcus auricularis, Staphylococcus lugdunensis, Staphylococcus simulans, Staphylococcus cohnii and Staphylococcus caprae. All clinical strains were clonally unrelated. Isolates carried genes encoding resistance to β-lactam (mecA) (15 %), aminoglycoside [aac(6')/aph(2″)(11 %), aph (3')-IIIa (15 %), ant(4')-Ia (19 %)] and macrolide, lincosamide and streptogramin B (MLSB) [erm(A) (4 %), erm(B) (13 %), erm(C) (41 %), msr(A) (11 %)] antibiotics. CoNS isolates (64 %) were able to form biofilms. Confocal laser scanning microscopy revealed that these biofilms formed a three-dimensional structure composed mainly of living cells. All biofilm-positive strains carried the ica operon. In vitro studies demonstrated that a combination treatment with tigecycline and rifampicin was more effective against biofilms than one with ciprofloxacin and rifampicin. The minimum biofilm eradication concentration values were 0.062-0.5 µg ml-1 for tigecycline/rifampicin and 0.250-2 µg ml-1 for ciprofloxacin/rifampicin. All CoNS strains adhered to the human epithelial cell line HeLa, and more than half of the isolates were able to invade the HeLa cells, although most invaded relatively poorly. The virulence of CoNS is also attributed to their cytotoxic effects on HeLa cells. Incubation of HeLa cells with culture supernatant of the CoNS isolates resulted in cell death. The results indicate that the pathogenicity of S. capitis, S. auricularis, S. lugdunensis, S. cohnii and S. caprae is multi-factorial, involving the ability of these bacteria to adhere to human epithelial cells, form biofilms and invade and destroy human cells.

  15. Detection of Yersinia ruckeri in rainbow trout blood by use of the polymerase chain reaction.

    PubMed

    Altinok, I; Grizzle, J M; Liu, Z

    2001-01-26

    We evaluated a polymerase chain reaction (PCR) method for detecting Yersinia ruckeri, the bacterial pathogen causing enteric redmouth disease (ERM), in blood of rainbow trout Oncorhynchus mykiss. Identification of the PCR product was confirmed by Southern blot hybridization with a 32P-labeled oligonucleotide probe matching a sequence within the small subunit ribosomal RNA gene of Y. ruckeri. Following a 1 h immersion of rainbow trout in water with 4.5 x 10(6) colony-forming units of Y. ruckeri l(-1), the PCR was positive for all blood samples from 1 h (first sample) to 5 d and was negative from 9 to 30 d (last sample). Fish in this experiment did not show signs of disease, probably because they had been vaccinated against Y. ruckeri. To test this method with naturally infected fish, 42 rainbow trout from hatcheries were examined. Four of these fish had clinical signs of ERM and were infected with Y. ruckeri based on bacteriological culture. The PCR method detected Y. ruckeri in blood, intestine, liver, and trunk kidney from the 4 fish with ERM and from 5 additional rainbow trout that were bacteriologically negative for Y. ruckeri. Three of 5 rainbow trout from streams receiving effluent from hatcheries were positive for Y. ruckeri when tested with PCR, although there was no growth of Y. ruckeri on culture plates inoculated with the same samples. Samples were successfully stored for 1 wk in lysis buffer at 25 degrees C. This study demonstrated that a non-lethal blood sample can be used with PCR to detect Y. ruckeri.

  16. 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.

  17. Molecular resistance mechanisms of macrolide-resistant invasive Streptococcus pneumoniae isolates from Alaska, 1986 to 2010.

    PubMed

    Rudolph, Karen; Bulkow, Lisa; Bruce, Michael; Zulz, Tammy; Reasonover, Alisa; Harker-Jones, Marcella; Hurlburt, Debby; Hennessy, Thomas

    2013-11-01

    The rapid emergence of antibiotic-resistant pneumococcal strains has reduced treatment options. The aim of this study was to determine antimicrobial susceptibilities, serotype distributions, and molecular resistance mechanisms among macrolide-resistant invasive pneumococcal isolates in Alaska from 1986 to 2010. We identified cases of invasive pneumococcal disease in Alaska from 1986 to 2010 through statewide population-based laboratory surveillance. All invasive pneumococcal isolates submitted to the Arctic Investigations Program laboratory were confirmed by standard microbiological methods and serotyped by slide agglutination and the Quellung reaction. MICs were determined by the broth microdilution method, and macrolide-resistant genotypes were determined by multiplex PCR. Among 2,923 invasive pneumococcal isolates recovered from 1986 to 2010, 270 (9.2%) were nonsusceptible to erythromycin; 177 (66%) erythromycin-nonsusceptible isolates demonstrated coresistance to penicillin, and 167 (62%) were multidrug resistant. The most frequent serotypes among the macrolide-resistant isolates were serotypes 6B (23.3%), 14 (20.7%), 19A (16.7%), 9V (8.9%), 19F (6.3%), 6A (5.6%), and 23F (4.8%). mef and erm(B) genes were detected in 207 (77%) and 32 (12%) of the isolates, respectively. Nineteen (7%) of the erythromycin-nonsusceptible isolates contained both mef and erm(B) genotypes; 15 were of serotype 19A. There was significant year-to-year variation in the proportion of isolates that were nonsusceptible to erythromycin (P < 0.001). Macrolide resistance among pneumococcal isolates from Alaska is mediated predominantly by mef genes, and this has not changed significantly over time. However, there was a statistically significant increase in the proportion of isolates that possess both erm(B) and mef, primarily due to serotype 19A isolates.

  18. Unraveling Antimicrobial Resistance Genes and Phenotype Patterns among Enterococcus faecalis Isolated from Retail Chicken Products in Japan

    PubMed Central

    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

  19. 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.

  20. Expression of ezrin radixin moesin proteins in the adult subventricular zone and the rostral migratory stream.

    PubMed

    Persson, A; Lindwall, C; Curtis, M A; Kuhn, H G

    2010-05-05

    Continuous proliferation occurs in the adult subventricular zone (SVZ) of the lateral ventricles throughout life. In the SVZ, progenitor cells differentiate into neuroblasts, which migrate tangentially along the rostral migratory stream (RMS) to reach their final destination in the olfactory bulb. These progenitor cells mature and integrate into the existing neural network of the olfactory bulb. Long distance migration of neuroblasts in the RMS requires a highly dynamic cytoskeleton with the ability to respond to surrounding stimuli. Radixin is a member of the ERM (Ezrin, Radixin, Moesin) family, which connect the actin cytoskeleton to the extracellular matrix through transmembrane proteins. The membrane-cytoskeleton linker proteins of the ERM family may regulate cellular events with a high demand on cytoskeleton plasticity, such as cell motility. Recently, specific expression of the ERM protein ezrin was shown in the RMS. Radixin however has not been characterized in this region. Here we used immunohistochemistry and confocal microscopy to examine the expression of radixin in the different cell types of the adult subventricular zone niche and in the RMS. Our findings indicate that radixin is strongly expressed in neuroblasts of the adult RMS and subventricular zone, and also in Olig2-positive cells. We also demonstrate the presence of radixin in the cerebral cortex, striatum, cerebellum, thalamus, hippocampus as well as the granular and periglomerular layers of the olfactory bulb. Our studies also reveal the localization of radixin in neurosphere culture studies and we reveal the specificity of our labeling using Western blotting. The expression pattern demonstrated here suggests a role for radixin in neuronal migration and differentiation in the adult RMS. Understanding how adult neuronal migration is regulated is of importance for the development of new therapeutic interventions using endogenous repair for neurodegenerative diseases.

  1. 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.

  2. Small interfering RNA library screen of human kinases and phosphatases identifies polo-like kinase 1 as a promising new target for the treatment of pediatric rhabdomyosarcomas.

    PubMed

    Hu, Kaiji; Lee, Cathy; Qiu, Dexin; Fotovati, Abbas; Davies, Alastair; Abu-Ali, Samah; Wai, Daniel; Lawlor, Elizabeth R; Triche, Timothy J; Pallen, Catherine J; Dunn, Sandra E

    2009-11-01

    Rhabdomyosarcoma, consisting of alveolar (aRMS) and embryonal (eRMS) subtypes, is the most common type of sarcoma in children. Currently, there are no targeted drug therapies available for rhabdomyosarcoma. In searching for new molecular therapeutic targets, we carried out genome-wide small interfering RNA (siRNA) library screens targeting human phosphatases (n = 206) and kinases (n = 691) initially against an aRMS cell line, RH30. Sixteen phosphatases and 50 kinases were identified based on growth inhibition after 72 hours. Inhibiting polo-like kinase 1 (PLK1) had the most remarkable impact on growth inhibition (approximately 80%) and apoptosis on all three rhabdomyosarcoma cell lines tested, namely, RH30, CW9019 (aRMS), and RD (eRMS), whereas there was no effect on normal muscle cells. The loss of PLK1 expression and subsequent growth inhibition correlated with decreased p-CDC25C and Cyclin B1. Increased expression of WEE 1 was also noted. The induction of apoptosis after PLK1 silencing was confirmed by increased p-H2AX, propidium iodide uptake, and chromatin condensation, as well as caspase-3 and poly(ADP-ribose) polymerase cleavage. Pediatric Ewing's sarcoma (TC-32), neuroblastoma (IMR32 and KCNR), and glioblastoma (SF188) models were also highly sensitive to PLK1 inhibition. Finally, based on cDNA microarray analyses, PLK1 mRNA was overexpressed (>1.5 fold) in 10 of 10 rhabdomyosarcoma cell lines and in 47% and 51% of primary aRMS (17 of 36 samples) and eRMS (21 of 41 samples) tumors, respectively, compared with normal muscles. Similarly, pediatric Ewing's sarcoma, neuroblastoma, and osteosarcoma tumors expressed high PLK1. We conclude that PLK1 could be a promising therapeutic target for the treatment of a wide range of pediatric solid tumors including rhabdomyosarcoma.

  3. Vibrio cholerae O1 with Reduced Susceptibility to Ciprofloxacin and Azithromycin Isolated from a Rural Coastal Area of Bangladesh

    PubMed Central

    Rashed, Shah M.; Hasan, Nur A.; Alam, Munirul; Sadique, Abdus; Sultana, Marzia; Hoq, Md. Mozammel; Sack, R. Bradley; Colwell, Rita R.; Huq, Anwar

    2017-01-01

    Cholera outbreaks occur each year in the remote coastal areas of Bangladesh and epidemiological surveillance and routine monitoring of cholera in these areas is challenging. In this study, a total of 97 Vibrio cholerae O1 isolates from Mathbaria, Bangladesh, collected during 2010 and 2014 were analyzed for phenotypic and genotypic traits, including antimicrobial susceptibility. Of the 97 isolates, 95 possessed CTX-phage mediated genes, ctxA, ace, and zot, and two lacked the cholera toxin gene, ctxA. Also both CTX+ and CTX− V. cholerae O1 isolated in this study carried rtxC, tcpAET, and hlyA. The classical cholera toxin gene, ctxB1, was detected in 87 isolates, while eight had ctxB7. Of 95 CTX+ V. cholerae O1, 90 contained rstRET and 5 had rstRCL. All isolates, except two, contained SXT related integrase intSXT. Resistance to penicillin, streptomycin, nalidixic acid, sulfamethoxazole-trimethoprim, erythromycin, and tetracycline varied between the years of study period. Most importantly, 93% of the V. cholerae O1 were multidrug resistant. Six different resistance profiles were observed, with resistance to streptomycin, nalidixic acid, tetracycline, and sulfamethoxazole-trimethoprim predominant every year. Ciprofloxacin and azithromycin MIC were 0.003–0.75 and 0.19–2.00 μg/ml, respectively, indicating reduced susceptibility to these antibiotics. Sixteen of the V. cholerae O1 isolates showed higher MIC for azithromycin (≥0.5 μg/ml) and were further examined for 10 macrolide resistance genes, erm(A), erm(B), erm(C), ere(A), ere(B), mph(A), mph(B), mph(D), mef(A), and msr(A) with none testing positive for the macrolide resistance genes. PMID:28270803

  4. Presence of drug resistance in intestinal lactobacilli of dairy and human origin in Turkey.

    PubMed

    Cataloluk, Osman; Gogebakan, Bulent

    2004-07-01

    The prevalence of different resistance genes was investigated in lactobacilli of human and dairy origin by PCR. The presence of erm, van, tet, and cat-TC genes were determined in 16 raw milk, 15 cream, 10 yogurt, 50 hand-made cheese, and 20 industrially produced white-cheese samples of dairy origin and 16 mouth, 32 fecal, and 36 vaginal samples from different subjects of human origin. Lactobacilli of dairy and human origin were found to carry only erm(B) and tet(M) genes. The majority of the isolates, Lactobacillus crispatus (61), Lactobacillus gasseri (49), Lactobacillus plantarum (80) studied were found to harbor either erm(B) or tet(M) gene or both. No resistant lactobacilli was found in raw-milk and cream samples. All the human fecal samples and the majority of vaginal (29 of 36) and mouth (10 of 14) samples were found to carry the resistance genes. While a third of the hand-made cheeses carried resistant lactobacilli only one industrially produced cheese was found to carry resistant lactobacilli. Furthermore, the genes were found in the non-starter species, Lactobacillus acidophilus and Lb. plantarum, indicating that industrially produced cheeses in this respect could be considered more favorable. These results indicate that drug resistance seems to be very common in Turkey. Even though the number of dairy samples harboring the resistance genes (17 of 111) is smaller in regards to human samples, 10% of them were still found to carry the resistance genes as well. The presence of the resistance genes in majority of the samples of human origin and in minority of the samples of dairy origin indicates that drug resistance may be acquired in the intestinal tract during passage and spread to dairy products by the hands of workers during production.

  5. 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.

  6. The role of flagella in Clostridium difficile pathogenesis: comparison between a non-epidemic and an epidemic strain.

    PubMed

    Baban, Soza T; Kuehne, Sarah A; Barketi-Klai, Amira; Cartman, Stephen T; Kelly, Michelle L; Hardie, Kim R; Kansau, Imad; Collignon, Anne; Minton, Nigel P

    2013-01-01

    Clostridium difficile is a major cause of healthcare-associated infection and inflicts a considerable financial burden on healthcare systems worldwide. Disease symptoms range from self-limiting diarrhoea to fatal pseudomembranous colitis. Whilst C. difficile has two major virulence factors, toxin A and B, it is generally accepted that other virulence components of the bacterium contribute to disease. C. difficile colonises the gut of humans and animals and hence the processes of adherence and colonisation are essential for disease onset. Previously it has been suggested that flagella might be implicated in colonisation. Here we tested this hypothesis by comparing flagellated parental strains to strains in which flagella genes were inactivated using ClosTron technology. Our focus was on a UK-outbreak, PCR-ribotype 027 (B1/NAP1) strain, R20291. We compared the flagellated wild-type to a mutant with a paralyzed flagellum and also to mutants (fliC, fliD and flgE) that no longer produce flagella in vitro and in vivo. Our results with R20291 provide the first strong evidence that by disabling the motor of the flagellum, the structural components of the flagellum rather than active motility, is needed for adherence and colonisation of the intestinal epithelium during infection. Comparison to published data on 630Δerm and our own data on that strain revealed major differences between the strains: the R20291 flagellar mutants adhered less than the parental strain in vitro, whereas we saw the opposite in 630Δerm. We also showed that flagella and motility are not needed for successful colonisation in vivo using strain 630Δerm. Finally we demonstrated that in strain R20291, flagella do play a role in colonisation and adherence and that there are striking differences between C. difficile strains. The latter emphasises the overriding need to characterize more than just one strain before drawing general conclusions concerning specific mechanisms of pathogenesis.

  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. Species diversity and antibiotic resistance properties of Staphylococcus of farm animal origin in Nkonkobe Municipality, South Africa.

    PubMed

    Adegoke, Anthony A; Okoh, Anthony I

    2014-03-01

    The occurrence and antibiotic susceptibility profile of Staphylococcus isolates of healthy farm animal origin in Nkonkobe Municipality as well as the prevalence of putative antibiotic resistance genes were investigated using phenotypic and molecular methods. A total of 120 Staphylococcus isolates were isolated from 150 animal samples and consisted of Staphylococcus haemolyticus (30 %) and Staphylococcus aureus (23.3 %) from pig, Staphylococcus capitis (15 %) from goat, S. haemolyticus (5 %) and Staphylococcus xylosus (15 %) from cattle, and other staphylococci (11.7 %) from dead chicken and pigs. Besides this, the presence of these isolates was observed from the animal dung, showing that the organisms are shed to the environment. About 23.3 % of these isolates were coagulase-positive and 76.7 % were coagulase-negative Staphylococcus. Between 75 and 100 % of the isolates were resistant to penicillin G, tetracycline, sulfamethoxazole, and nalidixic acid; about 38 % were methicillin-resistant staphylococci, including 12.6 % methicillin-resistant S. aureus from pigs. In total, 12 % of all isolates were vancomycin resistant. Also, 12 % of the isolates were erythromycin resistant, while 40.2 % were resistant to ceftazidime. Only the genes mecA and mphC could be confirmed, whereas the genes vanA, vanB, ermA, ermB, and ermC could not be detected. The high phenotypic antibiotic resistance and the presence of some associated resistance genes is a potential threat to public health and suggest the animals to be important reservoirs of antibiotic resistance determinants in the environment.

  9. Synergistic effects of antimicrobial peptide DP7 combined with antibiotics against multidrug-resistant bacteria

    PubMed Central

    Wu, Xiaozhe; Li, Zhan; Li, Xiaolu; Tian, Yaomei; Fan, Yingzi; Yu, Chaoheng; Zhou, Bailing; Liu, Yi; Xiang, Rong; Yang, Li

    2017-01-01

    Antibiotic-resistant bacteria present a great threat to public health. In this study, the synergistic effects of antimicrobial peptides (AMPs) and antibiotics on several multidrug-resistant bacterial strains were studied, and their synergistic effects on azithromycin (AZT)-resistance genes were analyzed to determine the relationships between antimicrobial resistance and these synergistic effects. A checkerboard method was used to evaluate the synergistic effects of AMPs (DP7 and CLS001) and several antibiotics (gentamicin, vancomycin [VAN], AZT, and amoxicillin) on clinical bacterial strains (Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter baumannii, and Escherichia coli). The AZT-resistance genes (ermA, ermB, ermC, mefA, and msrA) were identified in the resistant strains using quantitative polymerase chain reaction. For all the clinical isolates tested that were resistant to different antibiotics, DP7 had high antimicrobial activity (≤32 mg/L). When DP7 was combined with VAN or AZT, the effect was most frequently synergistic. When we studied the resistance genes of the AZT-resistant isolates, the synergistic effect of DP7–AZT occurred most frequently in highly resistant strains or strains carrying more than two AZT-resistance genes. A transmission electron microscopic analysis of the S. aureus strain synergistically affected by DP7–AZT showed no noteworthy morphological changes, suggesting that a molecular-level mechanism plays an important role in the synergistic action of DP7–AZT. AMP DP7 plus the antibiotic AZT or VAN is more effective, especially against highly antibiotic-resistant strains. PMID:28356719

  10. DNMT3B in vitro knocking-down is able to reverse embryonal rhabdomyosarcoma cell phenotype through inhibition of proliferation and induction of myogenic differentiation

    PubMed Central

    Megiorni, Francesca; Camero, Simona; Ceccarelli, Simona; McDowell, Heather P.; Mannarino, Olga; Marampon, Francesco; Pizer, Barry; Shukla, Rajeev; Pizzuti, Antonio; Marchese, Cinzia; Clerico, Anna; Dominici, Carlo

    2016-01-01

    Aberrant DNA methylation has been frequently observed in many human cancers, including rhabdomyosarcoma (RMS), the most common soft tissue sarcoma in children. To date, the expression and function of the de novo DNA methyltransferase (DNMT) 3B in RMS have not yet been investigated. Our study show for the first time a significant up-regulation of DNMT3B levels in 14 RMS tumour samples and 4 RMS cell lines in comparison to normal skeletal muscle. Transfection of RD and TE671 cells, two in vitro models of embryonal RMS (ERMS), with a synthetic DNMT3B siRNA decreased cell proliferation by arresting cell cycle at G1 phase, as demonstrated by the reduced expression of Cyclin B1, Cyclin D1 and Cyclin E2, and by the concomitant up-regulation of the checkpoint regulators p21 and p27. DNMT3B depletion also impaired RB phosphorylation status and decreased migratory capacity and clonogenic potential. Interestingly, DNMT3B knock-down was able to commit ERMS cells towards myogenic terminal differentiation, as confirmed by the acquisition of a myogenic-like phenotype and by the increased expression of the myogenic markers MYOD1, Myogenin and MyHC. Finally, inhibition of MEK/ERK signalling by U0126 resulted in a reduction of DNMT3B protein, giving evidence that DNMT3B is a down-stream molecule of this oncogenic pathway. Taken together, our data indicate that altered expression of DNMT3B plays a key role in ERMS development since its silencing is able to reverse cell cancer phenotype by rescuing myogenic program. Epigenetic therapy, by targeting the DNA methylation machinery, may represent a novel therapeutic strategy against RMS. PMID:27764816

  11. Ezrin: a regulator of actin microfilaments in cell junctions of the rat testis

    PubMed Central

    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.

  13. Antibiotic Resistance Genes in Freshwater Biofilms May Reflect Influences from High-Intensity Agriculture.

    PubMed

    Winkworth-Lawrence, Cynthia; Lange, Katharina

    2016-11-01

    Antibiotic resistance is a major public health concern with growing evidence of environmental gene reservoirs, especially in freshwater. However, the presence of antibiotic resistance genes in freshwater, in addition to the wide spectrum of land use contaminants like nitrogen and phosphate, that waterways are subjected to is inconclusive. Using molecular analyses, freshwater benthic rock biofilms were screened for genes conferring resistance to antibiotics used in both humans and farmed animals (aacA-aphD to aminoglycosides; mecA to ß-lactams; ermA and ermB to macrolides; tetA, tetB, tetK, and tetM to tetracyclines; vanA and vanB to glycopeptides). We detected widespread low levels of antibiotic resistance genes from 20 waterways across southern New Zealand throughout the year (1.3 % overall detection rate; 480 samples from three rocks per site, 20 sites, eight occasions; July 2010-May 2011). Three of the ten genes, ermB, tetK, and tetM, were detected in 62 of the 4800 individual screens; representatives confirmed using Sanger sequencing. No distinction could be made between human and agricultural land use contamination sources based on gene presence distribution alone. However, land use pressures are suggested by moderate correlations between antibiotic resistance genes and high-intensity farming in winter. The detection of antibiotic resistance genes at several sites not subject to known agricultural pressures suggests human sources of resistance, like waterway contamination resulting from unsatisfactory toilet facilities at recreational sites.

  14. 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

  15. 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

  16. 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.

  17. 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.

  18. 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.

  19. Impact of temperature on the arbuscular mycorrhizal (AM) symbiosis: growth responses of the host plant and its AM fungal partner.

    PubMed

    Heinemeyer, A; Fitter, A H

    2004-02-01

    The growth response of the hyphae of mycorrhizal fungi has been determined, both when plant and fungus together and when only the fungus was exposed to a temperature change. Two host plant species, Plantago lanceolata and Holcus lanatus, were grown separately in pots inoculated with the mycorrhizal fungus Glomus mosseae at 20/18 degrees C (day/night); half of the pots were then transferred to 12/10 degrees C. Plant and fungal growth were determined at six sequential destructive harvests. A second experiment investigated the direct effect of temperature on the length of the extra-radical mycelium (ERM) of three mycorrhizal fungal species. Growth boxes were divided in two equal compartments by a 20 micro m mesh, allowing only the ERM and not roots to grow into a fungal compartment, which was either heated (+8 degrees C) or kept at ambient temperature. ERM length (LERM) was determined on five sampling dates. Growth of H. lanatus was little affected by temperature, whereas growth of P. lanceolata increased with temperature, and both specific leaf area (SLA) and specific root length (SRL) increased independently of plant size. Percentage of colonized root (LRC) and LERM were positively correlated with temperature when in symbiosis with P. lanceolata, but differences in LRC were a function of plant biomass. Colonization was very low in H. lanatus roots and there was no significant temperature effect. In the fungal compartment LERM increased over time and was greatest for Glomus mosseae. Heating the fungal compartment significantly increased LERM in two of the three species but did not affect LRC. However, it significantly increased SRL of roots in the plant compartment, suggesting that the fungus plays a regulatory role in the growth dynamics of the symbiosis. These temperature responses have implications for modelling carbon dynamics under global climate change.

  20. Minimum current requirement for confirming the localization of an epiradicular catheter placement

    PubMed Central

    Jeong, Ji Seon; Shim, Jae Hang; Kim, Dong Won; Kang, Min Serk

    2012-01-01

    Background Based on the necessity to confirm the epiradicular catheter misplacement, epiradicular threshold current for the confirmation of catheter tip localization is required. Methods Thirty-four adult patients with low extremity radiating pain were to receive epiradicular catheterization at the lumbosacral level. The epidural space was accessed percutaneously in cranial to caudal direction. A metal coil-reinforced epidural catheter was inserted and advanced caudolaterally toward the target neural foramen until the catheter tip was located below the bisection of pedicle. The electrical stimulation was performed after catheter placement in epidural and epiradicular space. Using the constant current nerve stimulator, the stimulating current was increased from 0 to 5 mA (pulse width of 0.3 ms; frequency of 2 Hz) until adequate motor contraction was evident. The threshold current for motor response with epidural space (EDmA) and epiradicular space (ERmA) placement were recorded upon electrical stimulation. In addition, the threshold charge for motor response with epidural (EDnC) and epiradicular (ERnC) placement were recorded. Results Of 34 catheters intentionally placed in the epiradicular space, ERmA was 0.53 ± 0.48 mA. The ERnC was significantly lower than EDnC (P < 0.05). The EDmA and ERmA were below 1 mA in 3 patients and above 1 mA in 4 patients, respectively. Conclusions We conclude that, threshold current for motor response seems to be lower for epiradicular compared with epidural placement, although we were not able to directly investigate the epidural threshold current. The threshold current of epiradicular space overlap that in the epidural space. PMID:23060981

  1. Morphision: A method for subjective evaluation of metamorphopsia in patients with unilateral macular pathology (i.e., full thickness macular hole and epiretinal membrane)

    PubMed Central

    Ugarte, Marta; Shunmugam, Manoharan; Laidlaw, D Alistair H; Williamson, Tom H

    2013-01-01

    Background: Lack of clinical tests to quantify spatial components of distortion in patients with full thickness macular holes (FTMH) and epiretinal membranes (ERM). Aim: To develop a test for subjective evaluation of visual distortion in the central visual field around fixation in patients with unilateral FTMH or ERM. Settings and Design: Prospective case-control study carried out at tertiary referral center. Materials and Methods: Twenty-five patients with unilateral macular disease (13 macular epiretinal membranes, 12 full-thickness macular holes), and nine controls (without ocular pathology) underwent ophthalmological examination with logMAR ETDRS visual acuity, near vision and contrast sensitivity assessed. Macular optical coherence tomography and metamorphopsia assessment using Morphision test was also carried out. This test consists of a set of modified Amsler charts for detection, identification, and subjective quantification of visual distortion in the central visual field around fixation. Morphision test content and construct validity, and reliability (test-retest method) were evaluated. Sixteen patients completed an unstructured survey on test performance and preference. Results: Every patient with unilateral FTMH or ERM identified a particular chart using Morphision test (content validity). None of the normal subjects without symptoms of metamorphopsia identified any distortion (construct validity). Test-retest showed a 100% consistency for frequency and 67% for amplitude. The mean amplitude difference between measurements was 0.02 degrees (SD = 0.038). The coefficient of repeatability was 0.075. There was a correlation between Morphision amplitude score and visual acuity and contrast sensitivity, individually. Conclusions: Morphision test allowed detection and subjective quantification of metamorphopsia in the clinical setting in our patients with unilateral macular epiretinal membranes and full thickness macular holes. PMID:24008785

  2. Ecological and health risk-based characterization of agricultural soils contaminated with polycyclic aromatic hydrocarbons in the vicinity of a chemical plant in China.

    PubMed

    Liu, Geng; Niu, Junjie; Guo, Wenjiong; An, Xiangsheng; Zhao, Long

    2016-11-01

    Polycyclic aromatic hydrocarbons (PAHs) from chemical plants can cause serious pollution of surrounding agricultural soils. A comprehensive study of agricultural soils was conducted in the vicinity of a chemical plant in China to characterize the soil PAH concentration, as well as their composition and sources. Human health and a screening-level ecological risk assessment were conducted for PAH contamination in agricultural soils. The results showed that the total concentrations of 16 priority PAHs ranged from 250.49 to 9387.26 ng g(-1), with an average of 2780.42 ng g(-1). High molecular weight PAHs (four to six rings) were the dominant component, accounting for more than 60% of all PAHs. Principal component analysis (PCA) and positive matrix factorization model (PMF) suggested that diesel emissions, coal combustion, coke ovens, and fuel combustion and gasoline emissions were the main sources of PAHs in agricultural soils. The ecological risk assessment results based on the effects range-low (ERL), the effects range-median (ERM), and the ecological screening levels (ESL) indicated that the exposure to ∑PAH16 was >ERL, >ERM, and ≥ERL and <ERM at 21.9, 0, and 21.9% of the soil sampling stations, the exposure to ∑PAH16 was >ESL at 78.1% of the soil sampling stations, and could induce biological effects in mammals. The Bapeq concentrations posed a potential carcinogenic risk to humans. Further risk management and control of soil PAHs in these agricultural soils is required to ensure the safety of the biocoenosis and human health.

  3. First record of Rhizoscyphus ericae in Southern Hemisphere's Ericaceae.

    PubMed

    Bruzone, M Clara; Fehrer, Judith; Fontenla, Sonia B; Vohník, Martin

    2017-02-01

    Ericoid mycorrhiza is arguably the least investigated mycorrhizal type, particularly when related to the number of potential hosts and the ecosystems they inhabit. Little is known about the global distribution of ericoid mycorrhizal (ErM) fungi, and this holds true even for the prominent ErM mycobiont Rhizoscyphus ericae. Earlier studies suggested R. ericae might be low in abundance or absent in the roots of Southern Hemisphere's Ericaceae, and our previous investigations in two Argentine Patagonian forests supported this view. Here, we revisited the formerly investigated area, albeit at a higher altitude, and screened fungi inhabiting hair roots of Gaultheria caespitosa and Gaultheria pumila at a treeless alpine site using the same methods as previously. We obtained 234 isolates, most of them belonging to Ascomycota. In contrast to previous findings, however, among 37 detected operational taxonomic units (OTUs), OTU 1 (=R. ericae s. str.) comprised the highest number of isolates (87, ∼37 %). Most of the OTUs and isolates belonged to the Helotiales, and 82.5 % of isolates belonged to OTUs shared between both Gaultheria species. At the alpine site, ericoid mycorrhizal fungi dominated, followed by dark septate endophytes and aquatic hyphomycetes probably acting as root endophytes. Our results suggest that the distribution of R. ericae is influenced, among others, by factors related to altitude such as soil type and presence/absence and type of the neighboring vegetation. Our study is the first report on R. ericae colonizing Ericaceae roots in the Southern Hemisphere and extends the known range of this prominent ErM species to NW Patagonia.

  4. Defects Along Blood Vessels in Glaucoma Suspects and Patients

    PubMed Central

    Hood, Donald C.; De Cuir, Nicole; Mavrommatis, Maria A.; Xin, Daiyan; Muhammad, Hassan; Reynaud, Juan; Ritch, Robert; Fortune, Brad

    2016-01-01

    Purpose To examine the relationship between small hypodense regions (“holes”) in the retinal nerve fiber layer (RNFL) seen on circumpapillary optical coherence tomography (OCT) images of glaucoma patients and suspects and the paravascular inner retinal defects (PIRDs) seen with OCT line scans near blood vessels in individuals without optic nerve disease but with high myopia and/or epiretinal membranes (ERMs). Methods Based upon the availability of wide-field, swept-source OCT scans, 19 eyes from 15 glaucoma patients or suspects were selected from a larger group of eyes with holes on circumpapillary frequency-domain OCT scans. Paravascular defects (PDs) were identified using en face slab images generated (ATL 3D-Suite) from 52-μm slabs just below the vitreal border of the inner limiting membrane by averaging reflective intensity. Paravascular defects were confirmed with B-scans from these images. Results For 13 of the 19 eyes, the appearance of the PDs fit the previously described PIRDs and extended well beyond the circumpapillary region. In the other 6 eyes, the PDs were restricted to a small region and did not fit the previously described appearance of PIRDs. In these eyes, the holes were associated with an arcuate defect of the RNFL. Of the 13 with PIRDs, 9 had ERMs and/or high myopia previously associated with PIRDs in otherwise healthy eyes. Conclusions Holes seen on circumpapillary OCT scans of glaucoma patients and suspects are associated with local glaucomatous damage, as well as with PIRDs associated with high myopia and ERMs. PMID:27054521

  5. Diesel exhaust particles induce aberrant alveolar epithelial directed cell movement by disruption of polarity mechanisms.

    PubMed

    LaGier, Adriana J; Manzo, Nicholas D; Dye, Janice A

    2013-01-01

    Disruption of the respiratory epithelium contributes to the progression of a variety of respiratory diseases that are aggravated by exposure to air pollutants, specifically traffic-based pollutants such as diesel exhaust particles (DEP). Recognizing that lung repair following injury requires efficient and directed alveolar epithelial cell migration, this study's goal was to understand the mechanisms underlying alveolar epithelial cells response to DEP, particularly when exposure is accompanied with comorbid lung injury. Separate mechanistic steps of directed migration were investigated in confluent murine LA-4 cells exposed to noncytotoxic concentrations (0-100 μg/cm(2)) of either automobile-emitted diesel exhaust particles (DEP(A)) or carbon black (CB) particles. A scratch wound model ascertained how DEP(A) exposure affected directional cell migration and BCECF ratio fluorimetry-monitored intracellular pH (pHi). Cells were immunostained with giantin to assess cell polarity, and with paxillin to assess focal cell adhesions. Cells were immunoblotted for ezrin/radixin/moesin (ERM) to assess cytoskeletal anchoring. Data demonstrate herein that exposure of LA-4 cells to DEP(A) (but not CB) resulted in delayed directional cell migration, impaired de-adhesion of the trailing edge cell processes, disrupted regulation of pHi, and altered Golgi polarity of leading edge cells, along with modified focal adhesions and reduced ERM levels, indicative of decreased cytoskeletal anchoring. The ability of DEP(A) to disrupt directed cell migration at multiple levels suggests that signaling pathways such as ERM/Rho are critical for transduction of ion transport signals into cytoskeletal arrangement responses. These results provide insights into the mechanisms by which chronic exposure to traffic-based emissions may result in decrements in lung capacity.

  6. The First Report of a Fully Sequenced Resistance Plasmid from Shigella boydii

    PubMed Central

    Wang, Li; Liu, Lei; Liu, Dong; Yin, Zhe; Feng, Jiao; Zhang, Defu; Fang, Haihong; Qiu, Yefeng; Chen, Weijun; Yang, Ruisheng; Wang, Jinglin; Fa, Yunzhi; Zhou, Dongsheng

    2016-01-01

    The purpose of this study was to characterize mechanisms of plasmid-mediated antimicrobial resistance in Shigella boydii. S. boydii strain 2246 with resistance to ciprofloxacin, ceftriaxone and azithromycin was isolated from a human case of watery diarrhea in a Chinese public hospital. Resistance in strain 2246 to ceftriaxone and azithromycin was attributable to the presence of blaCTX-M-14, and erm(B) and mph(A), respectively, which were co-located on a multidrug-resistant (MDR) plasmid p2246-CTXM. p2246-CTXM represented a novel IncFII-type MDR plasmid with a very complex chimera structure. Its master backbone was genetically closely related to the R100 plasmid, but p2246-CTXM had evolved to integrate additional R100-unrelated backbone regions as well as massive exogenous mobile elements that carried multiple resistance determinants. In p2246-CTXM, erm(B) together with its leading peptide gene erm(C), mph(A) together with its regulatory genes mrx and mphR(A), and blaCTX-M-14 were captured by three different mobile elements Tn6295, the IS26-mph(A)-mrx-mphR(A)-IS6100 unit, and a truncated ISEcp1-blaCTX-M-14-IS903D-iroN transposition unit, respectively, all of which were harbored in a large Tn3-family transposon Tn6285. p2246-CTXM still carried additional resistance determinants mer (mercury resistance), aacA4 (aminoglycoside resistance), cmlA1 (chloramphenicol resistance), and qacED1 (quaternary ammonium compound resistance). This is the first report of identifying a clinical S. boydii strain simultaneously resistant to ciprofloxacin, ceftriaxone, and azithromycin, and determining the complete sequence of a resistance plasmid from S. boydii. PMID:27766094

  7. Resistance to macrolides in Group A streptococci from the European section of Turkey: genetic and phenotypic characterization.

    PubMed

    Akata, Filiz; Oztürk, Devrim; Tansel, Ozlem; Tatman-Otkun, Müserref; Otkun, Metin; Fitoussi, Frederic; Bingen, Edouard; Tugrul, Murat

    2002-12-01

    The aim of this study was to determine the susceptibilities to macrolides of Group A streptococcal isolates from the European section of Turkey. In the case of resistant isolates, the patterns and genetic mechanisms of erythromycin resistance were studied. Seven (2.7%) of the 260 isolates were resistant to erythromycin. Four of them showed the M phenotype and harboured mefA genes whereas three isolates showed the inducible macrolide, lincosamide and streptogramin B resistance phenotype and harboured ermTR genes. In the European section of Turkey, the current resistance rate of Group A streptococci to macrolides remains low.

  8. [Emergence of macrolide resistant Streptococcus pyogenes strains in pediatric patients in France].

    PubMed

    Mariani-Kurkdjian, P; Doit, C; Deforche, D; Brahimi, N; Francois, M; Van den Abbeele, T; Bingen, E

    2004-10-01

    A total of 206 recent throat isolates of Streptococcus pyogenes collected between 2002 and 2004 from children were tested for their susceptibility to penicillin, amoxycillin, erythromycin, clarythromycin and clindamycin. The erythromycin resistant isolates were further studied for their genetic mechanism of resistance by means of PCR. In all, 14.5% of the strains were erythromycin resistant and 13.5 and 1% expressed the constitutive MLS(B) and M resistance phenotypes and harbored the ermB and mef A genes respectively.

  9. An ecological response model for the Cache la Poudre River through Fort Collins

    USGS Publications Warehouse

    Shanahan, Jennifer; Baker, Daniel; Bledsoe, Brian P.; Poff, LeRoy; Merritt, David M.; Bestgen, Kevin R.; Auble, Gregor T.; Kondratieff, Boris C.; Stokes, John; Lorie, Mark; Sanderson, John

    2014-01-01

    The ERM was designed to represent the multi-dimensional ecological character of the contemporary urban Poudre River. It provides a scientific foundation that can serve as a decision support tool and foster a more informed community discussion about the future of the river as it provides a better understanding of the likely response of the Poudre River ecosystem to environmental flow management and other stewardship activities. In particular, model results can assist managers in developing specific management actions to achieve desirable goals for key indicators of river health.

  10. Innovative Training Concepts for Use in Distributed Interactive Simulation Environments

    DTIC Science & Technology

    1993-03-22

    unmanned vehicles and aircraft, both friendly (BLUFOR) and enemy (OPFOR); a Plan View Display (PVD) for providing a "birds eye view" of the battlefield which...expressed in :erms of table top mn-basked exeises designed to be conducted in a classroom setting. The Info-rmationI Management Exercise (IMEX) training...cx-i~e, the following equipment is needed: I • a Managemet , Command and Control System (MCC); 1 SPARCS Worksd-ion with. CCD software; • a SEND station

  11. Detailed Requirements Analysis for a Management Information System for the Department of Family Practice and Community Medicine at Silas B. Hays Army Community Hospital, Fort Ord, California

    DTIC Science & Technology

    1989-03-01

    U.S. Govcrnnct. i -- I - Cosati Codes IS Subct [ erms rco,,thv on rrti erse f neessarv and ide,t’v bl. Ho..k zeuto , r Hospital Administratio.h i...lume riaw, Jr. Approved By: Q/W 7L’),? fk ’L _ "Thomas P. Moore, Thesis Advisor T e7g Reader David R . Whipple, Jr., Chairman, Department of...Lnsh //ew C.n onliution.upen o Opiion SolutriReues Trainingfo I Proposals NCWDsg Proposals System SpecficationsL 1 r 8 -7Vendors Deliver Computer

  12. Novel Plasmid-Borne Multidrug Resistance Gene Cluster Including lsa(E) from a Linezolid-Resistant Enterococcus faecium Isolate of Swine Origin

    PubMed Central

    Si, Hongbin; Zhang, Wan-Jiang; Chu, Shengbo; Wang, Xiu-Mei; Dai, Lei; Hua, Xin; Dong, Zhimin

    2015-01-01

    A novel nonconjugative plasmid of 28,489 bp from a porcine linezolid-resistant Enterococcus faecium isolate was completely sequenced. This plasmid harbored a novel type of multiresistance gene cluster that comprised the resistance genes lnu(B), lsa(E), spw, aadE, aphA3, and two copies of erm(B), which account for resistance to macrolides, lincosamides, streptogramins, pleuromutilins, streptomycin, spectinomycin, and kanamycin/neomycin. Structural comparisons suggested that this plasmid might have developed from other enterococcal plasmids by insertion element (IS)-mediated interplasmid recombination processes. PMID:26324271

  13. Methodology for Risk-informed O&M decision making in power plants

    SciTech Connect

    2016-10-06

    •Incorporation of real-time component information using equipment condition assessment (ECA) through the developmentof enhanced risk monitors (ERM) for active components in advanced reactor (AR) and advanced small modular reactor (SMR) designs •We incorporate time-dependent failure probabilities from prognostic health management (PHM) systems to dynamically update the risk metric of interest •This information is used to augment data used for supervisory control and plant-wide coordination of multiple modules by providing the incremental risk incurred due to aging and demands placed on components that support mission requirements.

  14. Dynamische Satzgenerierung und Sprachausgabe für einen mobilen Serviceroboter

    NASA Astrophysics Data System (ADS)

    Parlitz, Christopher; Amann, Bernd; Hägele, Martin

    Die vorliegende Arbeit befasst sich mit der Entwicklung einer Sprachausgabe für ein mobiles Robotersystem. Dazu wurde zum einen ein auf die Anwendung zugeschittenes Text-To-Speech Verfahren und zum anderen eine Grammatik, die eine dynamische Satzgenerierung ermöglicht, implementiert. Die Verfahren arbeiten unabhängig voneinander und können daher auch gegen andere Komponenten ausgetauscht werden. Die Programme wurden als Client-Server Applikationen programmiert, um möglichst flexibel die Sprachausgabe einsetzen zu können.

  15. Medical Image Indexing and Compression Based on Vector Quantization: Image Retrieval Efficiency Evaluation

    DTIC Science & Technology

    2007-11-02

    Puentes, B. Solaiman, J. M. Cauvin, C. Roux LATIM ERM 0102 - INSERM, Bat 2 bis, CHU Morvan, Brest, France Département I.T.I., ENST de Bretagne , B.P. 832...s) and Address(es) Departement I.T.I., ENST de Bretagne B.P. 832, 29285 Brest-Cedex, France Performing Organization Report Number Sponsoring...Latin America – NR: 6.0130.9), and in part by the ENST de Bretagne – Groupe des Écoles de Télécommunications (GET). REFERENCES [1] F. Idris, S

  16. 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.

  17. Tumor-Wachstumsmodellierung als parametrisches Bildregistrierproblem

    NASA Astrophysics Data System (ADS)

    Becker, Stefan; Jungmann, Jan Ole; Mang, Andreas; Buzug, Thorsten M.

    In der vorliegenden Arbeit stellen wir ein neues Modell zur Kopplung des Tumormasseeffektes an die, der Wachstumsmodellierung unterliegende, anisotrope Reaktionsdiffusionsgleichung vor. Die Gleichung wird auf einem hochaufgelösten Voxelgitter diskretisiert. Eine Modellierung der Anisotropie des Diffusionsprozesses ermöglicht die Integration von Diffusions-Tensor-Bildgebungsdaten. Die raumfordernde Wirkung des Tumors wird als parametrisches Bildregistrierproblem aufgefasst. Hierbei wird die resultierende Verteilung der Tumorzellkonzentration in die zu optimierende Zielfunktion integriert. Erste qualitative Ergebnisse zeigen, dass eine Minimierung der aufgestellten Zielfunktion zu einer plausiblen Modellierung des Masseeffektes führt.

  18. Schichtübergreifende Früherkennung von Verbindungsausfällen in drahtlosen Mesh-Netzwerken

    NASA Astrophysics Data System (ADS)

    Lindhorst, Timo

    Drahtlose Mesh-Netzwerke (WMNs) bilden aufgrund ihrer Selbstorganisation ein flexibles Kommunikationssystem. Bei der Erkennung einzelner Verbindungsausfälle, die bei der Integration mobiler Teilnehmer unvermeidbar sind, ist bei aktuellen Routingprotokollen die Kommunikation jedoch für mehrere Sekunden unterbrochen. In dieser Arbeit wird gezeigt, wie durch Nutzung von Informationen aus der MAC-Schicht ein Ausfall frühzeitig erkannt und dadurch die Anzahl der Paketverluste von mehreren Hundert auf maximal vier reduziert werden kann. Hierdurch wird die Zuverlässigkeit und Verfügbarkeit von WMNs erhöht und ihr Einsatz für Echtzeit-Anwendungen ermöglicht.

  19. Indikatorenbasierte Bewertung von Entwurfsentscheidungen auf Systemebene

    NASA Astrophysics Data System (ADS)

    Freier, M.; Wenzler, A.; Mayer, T.; Gerlach, J.; Rosenstiel, W.

    2013-07-01

    Bei dem Entwurf von anwendungsspezifischen integrierten Schaltungen muss ein Entwickler zusätzliche Funktionen integrieren und die zunehmende Komplexität bewältigen. Für die Reduzierung der Kosten bleibt weiterhin die Verkürzung der benötigten Entwicklungszeit ein Ziel. Ein entscheidender Faktor dabei ist die zuverlässige Absicherung von Entwurfsentscheidungen in einer frühen Entwurfsphase. In der vorliegenden Arbeit wird eine Methode vorgestellt, die eine Bewertung von Entwurfsentscheidungen bei Systemmodellen mittels automatisch ermittelter Indikatoren ermöglichen soll. Die Systemmodelle können mit der Entwurfsumgebung MATLAB/Simulink oder in SystemC beschrieben sein.

  20. Probabilistisches Belegtheitsfilter zur Schätzung dynamischer Umgebungen unter Verwendung multipler Bewegungsmodelle

    NASA Astrophysics Data System (ADS)

    Brechtel, Sebastian; Gindele, Tobias; Vogelgesang, Jan; Dillmann, Rüdiger

    In dieser Arbeit wird eine Erweiterung des zeilbasierten Belegtheitsfilters BOFUM1 um Objektgruppen zum BOFUG (Bayesian Occupancy Filtering using Groups) vorgenommen. Diese ermöglicht die Einteilung und Klassifikation der Gruppenzugehörigkeit von Belegtheit, allein auf Basis von statischen Belegtheitsmessungen. Exemplarisch wird für Fußgänger und Fahrzeuge gezeigt, dass die Definition unterschiedlicher Dynamikmodelle ausreicht, um auf Objektinformationen zu schließen und das Filterergebnis nachhaltig zu verbessern. Die implizite Gruppeninferenz stellt einen ersten Schritt zur Vereinigung von Objekt- und Zellebene dar.

  1. Molecular epidemiology, antimicrobial susceptibilities and resistance mechanisms of Streptococcus pyogenes isolates resistant to erythromycin and tetracycline in Spain (1994–2006)

    PubMed Central

    2012-01-01

    Background Group A Streptococcus (GAS) causes human diseases ranging in severity from uncomplicated pharyngitis to life-threatening necrotizing fasciitis and shows high rates of macrolide resistance in several countries. Our goal is to identify antimicrobial resistance in Spanish GAS isolates collected between 1994 and 2006 and to determine the molecular epidemiology (emm/T typing and PFGE) and resistance mechanisms of those resistant to erythromycin and tetracycline. Results Two hundred ninety-five out of 898 isolates (32.8%) were erythromycin resistant, with the predominance of emm4T4, emm75T25, and emm28T28, accounting the 67.1% of the 21 emm/T types. Spread of emm4T4, emm75T25 and emm28T28 resistant clones caused high rates of macrolide resistance. The distribution of the phenotypes was M (76.9%), cMLSB (20.3%), iMLSB (2.7%) with the involvement of the erythromycin resistance genes mef(A) (89.5%), msr(D) (81.7%), erm(B) (37.3%) and erm(A) (35.9%). Sixty-one isolates were tetracycline resistant, with the main representation of the emm77T28 among 20 emm/T types. To note, the combination of tet(M) and tet(O) tetracycline resistance genes were similar to tet(M) alone reaching values close to 40%. Resistance to both antibiotics was detected in 19 isolates of 7 emm/T types, being emm11T11 and the cMLSB phenotype the most frequent ones. erm(B) and tet(M) were present in almost all the strains, while erm(A), mef(A), msr(D) and tet(O) appeared in less than half of them. Conclusions Spanish GAS were highly resistant to macrolides meanwhile showed minor resistance rate to tetracycline. A remarkable correlation between antimicrobial resistance and emm/T type was noticed. Clonal spread of emm4T4, emm75T25 and emm28T28 was the main responsable for macrolide resistance where as that emm77T28 clones were it to tetraclycline resistance. A wide variety of macrolide resistance genes were responsible for three macrolide resistance phenotypes. PMID:22998619

  2. Definition of the Cellular Mechanisms which Distinguish Between Estrogen Receptor Agonists and Antagonists

    DTIC Science & Technology

    2001-07-01

    performed a yeast two-hybrid screen using the ER-3x ligand binding domain as bait and identified several ER-interacting factors that interacted with ER...M) Figure 4. Identification of known ER-interacting proteins, SRC-1A and RIP140, validates the strategy of using ER-HBD3x as bait in the yeast two...hybrid screen. The ER-HBD3x was used as bait in yeast two-hybrid system to screen for ER-interacting proteins from human brain cDNA library. The C

  3. First report of swine-associated methicillin-resistant Staphylococcus aureus ST398 in Lithuania.

    PubMed

    Ruzauskas, M; Couto, N; Belas, A; Klimiene, I; Siugzdiniene, R; Pomba, C

    2013-01-01

    During 2011, 160 nasal samples were taken from pigs on 8 different farms in Lithuania. Four methicillin-resistant Staphylococcus aureus (MRSA) isolates were obtained. The isolates were ST398, spa type t011 and SCCmec V and none carried the lukF/lukS genes. Strains were resistant to tetracycline, attributed to tetK and tetM genes, and to erythromycin owing to the ermB gene. One MRSA strain was resistant to trimethoprim/sulfamethoxazole and carried the dfrK gene. This is the first report on the presence and characteristics of livestock-associated MRSA isolated from pigs in Lithuania.

  4. 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

  5. Identification of livestock-associated methicillin-resistant Staphylococcus aureus isolates in Korea and molecular comparison between isolates from animal carcasses and slaughterhouse workers.

    PubMed

    Moon, Dong Chan; Tamang, Migma Dorji; Nam, Hyang-Mi; Jeong, Jin-Ha; Jang, Geum-Chan; Jung, Suk-Chan; Park, Yong-Ho; Lim, Suk-Kyung

    2015-04-01

    This study was undertaken to screen methicillin-resistant Staphylococcus aureus (MRSA) in animal carcasses and slaughterhouse workers and characterize MRSA isolates identified during 2010-2012 in Korea. A total of 830 (16.4%) S. aureus and 65 (1.3%) MRSA were isolated from 9669 carcass samples. MRSA was more frequently detected in chicken carcasses (1.2%) than in cattle (0.3%) and pig carcasses (0.6%). The prevalence of MRSA in workers was 6.9% (4/58) in chicken slaughterhouse workers, but no MRSA was detected in pig and cattle slaughterhouse workers (0/41). Two different lineages of MRSA were identified (i.e., human-associated type [ST5, ST59, and ST72] and livestock-associated [LA] type [ST398, ST541, and ST692]); only LA MRSA was observed in chicken carcasses, whereas both types were found in cattle and pig carcasses and workers. All human-associated MRSA isolates carried enterotoxin and/or leukotoxin genes, whereas LA MRSA types did not carry these genes, except ST692 type. However, all LA MRSA isolates were multiresistant, whereas human-associated types were susceptible or resistant to fewer than two antimicrobials except ST5. Furthermore, one or more resistance genes were attributed for resistance to aminoglycosides (aac(6')-Ie-aph(2″), ant(4')-Ia, and aph(3')-IIIa), tetracycline [tet(K), tet(L), tet(M), and tet(S)], macrolide-lincosamide-streptogramin B (ermA, ermB, ermC, and ermT), lincosamide [lnu(B)], phenicol-lincosamide-oxazolidinone-pleuromutilin-streptogramin A (cfr), chloramphenicol (fexA), and fusidic acid [fus(C)]. To our knowledge, this is the first report of tet(S) gene in MRSA isolates and first detection of a unique (ST692) type of MRSA in occupational workers. Detection of new types of human-associated and LA MRSA with multiple resistance and virulence genes in food animal products constitutes a potential threat to public health.

  6. 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

  7. Chemiluminescence of Organic Peroxides. Thermal Generation of an o-Xylylene Peroxide.

    DTIC Science & Technology

    1981-04-07

    AD-A098 078 ILLINOIS LIIIV AT URBANA DEPT OF CHEMISTRY F/G 7/4 CHEMILLUMINESCENSE OF ORGANIC PEROXIDES - THERMAL GENERATION OF A--ETC (U) APR 81 J P...11’r- 4. 1 L E (ad Subtitle) or REPORT or R1 0 CRED emilum’i-nescence of Organic Peroxides . Thrai ehia Generation of an o-Xylylene Peroxide ,,, erm...Iq It. KEY WORDS (Countinue oni tow.e*ole At neesar did tffoiltl by *lack Mmber) ?T. chemil uminesceflce AR~~ 1 * peroxides A CIEEL therniol1ys is b

  8. Calculation and evaluation of sediment effect concentrations for the amphipod Hyalella azteca and the midge Chironomus riparius

    USGS Publications Warehouse

    Ingersoll, Christopher G.; Haverland, Pamela S.; Brunson, Eric L.; Canfield, Timothy J.; Dwyer, F. James; Henke, Chris; Kemble, Nile E.; Mount, David R.; Fox, Richard G.

    1996-01-01

    Procedures are described for calculating and evaluating sediment effect concentrations (SECs) using laboratory data on the toxicity of contaminants associated with field-collected sediment to the amphipod Hyalella azteca and the midge Chironomus riparius. SECs are defined as the concentrations of individual contaminants in sediment below which toxicity is rarely observed and above which toxicity is frequently observed. The objective of the present study was to develop SECs to classify toxicity data for Great Lake sediment samples tested with Hyalella azteca and Chironomus riparius. This SEC database included samples from additional sites across the United States in order to make the database as robust as possible. Three types of SECs were calculated from these data: (1) Effect Range Low (ERL) and Effect Range Median (ERM), (2) Threshold Effect Level (TEL) and Probable Effect Level (PEL), and (3) No Effect Concentration (NEC). We were able to calculate SECs primarily for total metals, simultaneously extracted metals, polychlorinated biphenyls (PCBs), and polycyclic aromatic hydrocarbons (PAHs). The ranges of concentrations in sediment were too narrow in our database to adequately evaluate SECs for butyltins, methyl mercury, polychlorinated dioxins and furans, or chlorinated pesticides. About 60 to 80% of the sediment samples in the database are correctly classified as toxic or not toxic depending on type of SEC evaluated. ERMs and ERLs are generally as reliable as paired PELs and TELs at classifying both toxic and non-toxic samples in our database. Reliability of the SECs in terms of correctly classifying sediment samples is similar between ERMs and NECs; however, ERMs minimize Type I error (false positives) relative to ERLs and minimize Type II error (false negatives) relative to NECs. Correct classification of samples can be improved by using only the most reliable individual SECs for chemicals (i.e., those with a higher percentage of correct classification). SECs

  9. Methodology for Long-Term Permeation Test Periods for HD in High-Density Polyethylene: Universal Munitions Storage Container for the Non-Stockpile Chemical Materiel Program

    DTIC Science & Technology

    2016-05-01

    Regression lines intersect at about 1330–1340 min. 0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 0 120 240 360 480 600 720 840 960 1080 1200 1320 1440 1560 M as s P er...m ea te d , n g Time, min y = 5E-05x + 0.7842 R² = 0.2619 y = 0.0076x - 9.3979 R² = 0.9817 0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 600 720 840 960 1080 1200

  10. National Dam Safety Program. Mach Lake Dam (MO 30133), Mississippi - Kaskaskia - St. Louis River Basin, Perry County, Missouri. Phase I Inspection Report.

    DTIC Science & Technology

    1981-04-01

    embankment was in fair condition. Deficiencies vilcuallv Ubse (rvrd by the inspection team were: (1) brUSh and briar ’rowth on upf [rem and~ rdownst-r-tm face...The slopes of the embankment were briar and )rush covered. Several ]is.. (12 to 15 inches diameter) tree stumps were noted on the upstream fare. No...perfriri ILV ai(n ~er~Me 0 in the construction of dams. (2) The slough area at midhei’Lht on rhe do)wustfre--ir fiS o Sa 1 (3) The briar and brush

  11. 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.

  12. Rapid Determination of Macrolide and Lincosamide Resistance in Group B Streptococcus Isolated from Vaginal-Rectal Swabs

    PubMed Central

    Dela Cruz, Wilfred P.; Richardson, Joann Y.; Broestler, Judith M.; Thornton, Jennifer A.; Danaher, Patrick J.

    2007-01-01

    Objective. Our objective was to assess the ability of real-time PCR to predict in vitro resistance in isolates of group B streptococcus (GBS). Methods. The first real-time PCR assays for the genes known to confer resistance to erythromycin and clindamycin in GBS were developed. Three hundred and forty clinical GBS isolates were assessed with these assays and compared with conventional disk diffusion. Results. The presence of an erythromycin ribosome methylation gene (ermB or ermTR variant A) predicted in vitro constitutive or inducible resistance to clindamycin with a sensitivity of 93% (95% CI 86%–97%), specificity of 90% (95% CI 85%–93%), positive predictive value of 76% (95% CI 67%–84%), and negative predictive value of 97% (95% CI 94%–99%). Conclusion. This rapid and simple assay can predict in vitro susceptibility to clindamycin within two hours of isolation as opposed to 18–24 hours via disk diffusion. The assay might also be used to screen large numbers of batched isolates to establish the prevalence of resistance in a given area. PMID:17710240

  13. 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

  14. Radionuclides (210Po and 210Pb) and Some Heavy Metals in Fish and Sediments in Lake Bafa, Turkey, and the Contribution of 210Po to the Radiation Dose

    PubMed Central

    Manav, Ramazan; Uğur Görgün, Aysun; Filizok, Işık

    2016-01-01

    The pollution level of Lake Bafa was investigated by collecting fish samples {Dicentrarchus labrax (sea bass), Liza ramada (mullet) and Anguilla anguilla (eel)}, surface sediment, and core samples. In all these samples, 210Po and 210Pb concentrations were estimated, and total annual dose rates were obtained for each species. Some heavy metal (Cr, Ni, Pb, Cd, Mn, Fe, and Zn) concentration levels were obtained for the fish and a core sample. The sediment mass accumulation rate was found to be 3.27 g·m−2·day−1 (0.119 g·cm−2·y−1) from a core sample. The heavy metal concentrations in the vertical profile of samples from the core were also observed. The measured concentration of Zn, Pb, Cd, and Cr were between the ERL (effects range low) and ERM (effects range median) limits, while Ni concentrations were higher than the ERM limit. The observed concentrations of Cd, Pb, and Zn in fish samples did not exceed the limits in accordance with Turkish Food Regulations. Further, the maximum effective dose equivalent of 210Po in the area was found to be 1.169 µSv·y−1. PMID:27834886

  15. Antimicrobial Resistance Profiles of Listeria monocytogenes and Listeria innocua Isolated from Ready-to-Eat Products of Animal Origin in Spain.

    PubMed

    Escolar, Cristina; Gómez, Diego; Rota García, María Del Carmen; Conchello, Pilar; Herrera, Antonio

    2017-03-29

    The objective of this work was to investigate the antimicrobial resistance in Listeria spp. isolated from food of animal origin. A total of 50 Listeria strains isolated from meat and dairy products, consisting of 7 Listeria monocytogenes and 43 Listeria innocua strains, were characterized for antimicrobial susceptibility against nine antimicrobials. The strains were screened by real-time PCR for the presence of antimicrobial resistance genes: tet M, tet L, mef A, msr A, erm A, erm B, lnu A, and lnu B. Multidrug resistance was identified in 27 Listeria strains, 4 belonging to L. monocytogenes. Resistance to clindamycin was the most common resistance phenotype and was identified in 45 Listeria strains; the mechanisms of resistance are still unknown. A medium prevalence of resistance to tetracycline (15 and 9 resistant and intermediate strains) and ciprofloxacin (13 resistant strains) was also found. Tet M was detected in Listeria strains with reduced susceptibility to tetracycline, providing evidence that both L. innocua and L. monocytogenes displayed acquired resistance. The presence of antimicrobial resistance genes in L. innocua and L. monocytogenes indicates that these genes may be transferred to commensal and pathogenic bacteria via the food chain; besides this, antibiotic resistance in L. monocytogenes could compromise the effective treatment of listeriosis in humans.

  16. [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.

  17. Determination of fluorine, chlorine and bromine in household products by means of oxygen bomb combustion and ion chromatography.

    PubMed

    Zhang, Shuai; Zhao, Tianbo; Wang, Jia; Qu, Xiaoling; Chen, Wei; Han, Yin

    2013-01-01

    A method for routine determination of fluorine, chlorine and bromine in household products was developed and validated. In this work, halogen analyses were made based on oxygen bomb combustion followed by ion chromatography (IC). The chromatographic analysis was performed by an IonPac AS19 hydroxide-selective anion-exchange column, a reagent free ion chromatograph eluent generator and an anion self-regenerating suppressor in 10 min. The response was linear (r ≥ 0.9995) in the entire investigated domain. The limit of detection for the halogens was in the range of 2 to 9 × 10(-3) mg/L and the limit of quantification was lower than 8 mg/Kg with 20 µL of injection volume. The certified reference material of ERM-EC 681k was pretreated using an oxygen bomb combustion procedure to demonstrate the precision of the proposed method. The quantitative analysis results obtained by IC for the target elements were 797 ± 9 mg/Kg chlorine and 786 ± 25 mg/Kg bromine, which were in good agreement with the certified values of 800 ± 4 mg/Kg chlorine, 770 ± 5 mg/Kg bromine for ERM-EC 681k, respectively. This validated method was successfully applied for the analysis of fluorine, chlorine and bromine in household product samples, and the variation of halogen contained among the tested samples was remarkable.

  18. Differentially Private Empirical Risk Minimization.

    PubMed

    Chaudhuri, Kamalika; Monteleoni, Claire; Sarwate, Anand D

    2011-03-01

    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.

  19. 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.

  20. Bacteroides mobilizable and conjugative genetic elements: antibiotic resistance among clinical isolates.

    PubMed

    Quesada-Gómez, Carlos

    2011-12-01

    The conjugation is one of the most important mechanisms of horizontal gene transfer in prokaryotes, leading to genetic variation within a species and the acquisition of new traits, such as antibiotic resistance. Bacteroides is an obligate anaerobe of the colon and a significant opportunistic pathogen. Antibiotic resistance among Bacteroides spp. is rapidly increasing, largely due to the dissemination of DNA transfer factors (plasmids and transposons) harbored by members of this genus. Transfer factors can be divided into two classes, conjugative and mobilizable. Species of the intestinal Bacteroides have yielded different resistance plasmids, all of which have been intensely studied, the plasmids encode high-level MLS resistance conferred by a conserved erm gene. It has been reported an interesting observation associated with the transfer of several of these types of elements, all of which conferred Tcr and displayed greatly increased transfer efficiency following exposure to tetracycline. Many of the conjugative transposons (CTns) in Bacteroides are related to various genetic elements (such as CTnDOT, CTnERL, NBU and others). CTnDOT carries a tetracycline resistance gene, tetQ, and an erythromycin resistance gene, ermF. Resistance to drugs used to treat Bacteroides infections, such as clindamycin, has also been increasing. These conjugal elements have been found in Bacteroides clinical isolates. Thus, horizontal gene transfer could conceivably have played a role in the rising incidence of resistance in this bacterial group.

  1. Enhancement of doxorubicin production by expression of structural sugar biosynthesis and glycosyltransferase genes in Streptomyces peucetius.

    PubMed

    Malla, Sailesh; Niraula, Narayan Prasad; Liou, Kwangkyoung; Sohng, Jae Kyung

    2009-08-01

    To enhance doxorubicin (DXR) production, the structural sugar biosynthesis genes desIII and desIV from Streptomyces venezuelae ATCC 15439 and the glycosyltransferase pair dnrS/dnrQ from Streptomyces peucetius ATCC 27952 were cloned into the expression vector pIBR25, which contains a strong ermE promoter. The recombinant plasmids pDnrS25 and pDnrQS25 were constructed for overexpression of dnrS and the dnrS/dnrQ pair, whereas pDesSD25 and pDesQS25 were constructed to express desIII/desIV and dnrS/dnrQ-desIII/desIV, respectively. All of these recombinant plasmids were introduced into S. peucetius ATCC 27952. The recombinant strains produced more DXR than the S. peucetius parental strain: a 1.2-fold increase with pDnrS25, a 2.8-fold increase with pDnrQS25, a 2.6-fold increase with pDesSD25, and a 5.6-fold increase with pDesQS25. This study showed that DXR production was significantly enhanced by overexpression of potential biosynthetic sugar genes and glycosyltransferase.

  2. Phenotypic and genotypic antimicrobial resistance traits of foodborne Staphylococcus aureus isolates from Shanghai.

    PubMed

    Xu, Jie; Shi, Chunlei; Song, Minghui; Xu, Xuebin; Yang, Puyu; Paoli, George; Shi, Xianming

    2014-04-01

    Staphylococcus aureus is a recognized pathogen in humans, which causes nosocomial infections and food poisoning. The transmission of antibiotic resistant S. aureus (ARSA), especially methicillin-resistant S. aureus, between food products and humans has become a serious problem. Hence, it is necessary to monitor S. aureus through the food supply chain. In this study, the disk diffusion method and epsilometer test were performed to determine the prevalence of ARSA in 78 foodborne isolates using 18 antibiotics. The highest resistance frequency was found for penicillin G (74.4%), followed by erythromycin (59.0%) and clindamycin (44.9%), whereas no vancomycin-resistant isolates were found. The 78 isolates could be subtyped into 31 resistance profiles and 11 clusters based on their antimicrobial susceptibility. Furthermore, Polymerase chain reaction (PCR) screening for the presence of 13 genes conferring antibiotic resistance was conducted. The presence of resistance genes was relatively high: blaTEM (80.8%), ermB (41.0%), grlA (38.5%), ermC (35.9%), and aac6'/aph2" (35.9%). The incidence of antibiotic resistance was significantly correlated to food types (p = 0.018), with isolates from meat and raw milk more resistant to antibiotics than those from frozen food and vegetables.

  3. 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.

  4. Recurrence Interval and Event Age Data for Type A Faults

    USGS Publications Warehouse

    Dawson, Timothy E.; Weldon, Ray J.; Biasi, Glenn P.

    2008-01-01

    This appendix summarizes available recurrence interval, event age, and timing of most recent event data for Type A faults considered in the Earthquake Rate Model 2 (ERM 2) and used in the ERM 2 Appendix C analysis as well as Appendix N (time-dependent probabilities). These data have been compiled into an Excel workbook named Appendix B A-fault event ages_recurrence_V5.0 (herein referred to as the Appendix B workbook). For convenience, the Appendix B workbook is attached to the end of this document as a series of tables. The tables within the Appendix B workbook include site locations, event ages, and recurrence data, and in some cases, the interval of time between earthquakes is also reported. The Appendix B workbook is organized as individual worksheets, with each worksheet named by fault and paleoseismic site. Each worksheet contains the site location in latitude and longitude, as well as information on event ages, and a summary of recurrence data. Because the data has been compiled from different sources with different presentation styles, descriptions of the contents of each worksheet within the Appendix B spreadsheet are summarized.

  5. Challenges in the size analysis of a silica nanoparticle mixture as candidate certified reference material

    NASA Astrophysics Data System (ADS)

    Kestens, Vikram; Roebben, Gert; Herrmann, Jan; Jämting, Åsa; Coleman, Victoria; Minelli, Caterina; Clifford, Charles; De Temmerman, Pieter-Jan; Mast, Jan; Junjie, Liu; Babick, Frank; Cölfen, Helmut; Emons, Hendrik

    2016-06-01

    A new certified reference material for quality control of nanoparticle size analysis methods has been developed and produced by the Institute for Reference Materials and Measurements of the European Commission's Joint Research Centre. The material, ERM-FD102, consists of an aqueous suspension of a mixture of silica nanoparticle populations of distinct particle size and origin. The characterisation relied on an interlaboratory comparison study in which 30 laboratories of demonstrated competence participated with a variety of techniques for particle size analysis. After scrutinising the received datasets, certified and indicative values for different method-defined equivalent diameters that are specific for dynamic light scattering (DLS), centrifugal liquid sedimentation (CLS), scanning and transmission electron microscopy (SEM and TEM), atomic force microscopy (AFM), particle tracking analysis (PTA) and asymmetrical-flow field-flow fractionation (AF4) were assigned. The value assignment was a particular challenge because metrological concepts were not always interpreted uniformly across all participating laboratories. This paper presents the main elements and results of the ERM-FD102 characterisation study and discusses in particular the key issues of measurand definition and the estimation of measurement uncertainty.

  6. Specific requirements of nonbilayer phospholipids in mitochondrial respiratory chain function and formation

    PubMed Central

    Baker, Charli D.; Basu Ball, Writoban; Pryce, Erin N.; Gohil, Vishal M.

    2016-01-01

    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

  7. 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

  8. 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.

  9. 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

  10. 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

  11. 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.

  12. Antimicrobial Chemicals Are Associated with Elevated Antibiotic Resistance Genes in the Indoor Dust Microbiome

    PubMed Central

    2016-01-01

    Antibiotic resistance is increasingly widespread, largely due to human influence. Here, we explore the relationship between antibiotic resistance genes and the antimicrobial chemicals triclosan, triclocarban, and methyl-, ethyl-, propyl-, and butylparaben in the dust microbiome. Dust samples from a mixed-use athletic and educational facility were subjected to microbial and chemical analyses using a combination of 16S rRNA amplicon sequencing, shotgun metagenome sequencing, and liquid chromatography tandem mass spectrometry. The dust resistome was characterized by identifying antibiotic resistance genes annotated in the Comprehensive Antibiotic Resistance Database (CARD) from the metagenomes of each sample using the Short, Better Representative Extract Data set (ShortBRED). The three most highly abundant antibiotic resistance genes were tet(W), blaSRT-1, and erm(B). The complete dust resistome was then compared against the measured concentrations of antimicrobial chemicals, which for triclosan ranged from 0.5 to 1970 ng/g dust. We observed six significant positive associations between the concentration of an antimicrobial chemical and the relative abundance of an antibiotic resistance gene, including one between the ubiquitous antimicrobial triclosan and erm(X), a 23S rRNA methyltransferase implicated in resistance to several antibiotics. This study is the first to look for an association between antibiotic resistance genes and antimicrobial chemicals in dust. PMID:27599587

  13. 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.

  14. 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.

  15. 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.

  16. Heterologous Expression of the Thiopeptide Antibiotic GE2270 from Planobispora rosea ATCC 53733 in Streptomyces coelicolor Requires Deletion of Ribosomal Genes from the Expression Construct

    PubMed Central

    Flinspach, Katrin; Kapitzke, Claudia; Tocchetti, Arianna; Sosio, Margherita; Apel, Alexander K.

    2014-01-01

    GE2270 is a thiopeptide antibiotic generated by extensive posttranslational modifications of a ribosomally generated precursor peptide. Thiopeptides are especially active against Gram-positive bacteria, including methicillin resistant Staphylococcus aureus (MRSA). In this study the GE2270 biosynthetic gene cluster (pbt) from Planobispora rosea ATCC 53733 was successfully expressed in the heterologous host strain Streptomyces coelicolor M1146. Notably, exconjugants containing the pbt gene cluster could only be obtained after deletion of the major part of the ribosomal genes flanking the gene cluster. This is a striking example that genes belonging to primary metabolism can prevent the successful conjugative transfer of DNA from phylogenetic distant species and thus complicate heterologous expression of secondary metabolite gene clusters. GE2270 production in the heterologous producer strain increased after introduction of the constitutive ermE* promoter upstream of the GE2270 resistance gene tuf from P. rosea. Insertion of the inducible tcp830 promoter resulted in inducible GE2270 production. When the regulatory gene pbtR was deleted, the resulting strain ceased to produce GE2270, suggesting an essential role of PbtR as a putative transcriptional activator of GE2270 expression. PMID:24598591

  17. An engineered strong promoter for streptomycetes.

    PubMed

    Wang, Weishan; Li, Xiao; Wang, Juan; Xiang, Sihai; Feng, Xiaozhou; Yang, Keqian

    2013-07-01

    Well-characterized promoters are essential tools for metabolic engineering and synthetic biology. In Streptomyces coelicolor, the native kasOp is a temporally expressed promoter strictly controlled by two regulators, ScbR and ScbR2. In this work, first, kasOp was engineered to remove a common binding site of ScbR and ScbR2 upstream of its core region, thus generating a stronger promoter, kasOp3. Second, another ScbR binding site internal to the kasOp3 core promoter region was abolished by random mutation and screening of the mutant library to obtain the strongest promoter, kasOp* (where the asterisk is used to distinguish the engineered promoter from the native promoter). The activities of kasOp* were compared with those of two known strong promoters, ermEp* and SF14p, in three Streptomyces species. kasOp* showed the highest activity at the transcription and protein levels in all three hosts. Furthermore, relative to ermEp* and SF14p, kasOp* was shown to confer the highest actinorhodin production level when used to drive the expression of actII-ORF4 in S. coelicolor. Therefore, kasOp* is a simple and well-defined strong promoter useful for gene overexpression in streptomycetes.

  18. 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

  19. CPI-17 drives oncogenic Ras signaling in human melanomas via Ezrin-Radixin-Moesin family proteins

    PubMed Central

    Riecken, Lars Björn; Zoch, Ansgar; Wiehl, Ulrike; Reichert, Sabine; Scholl, Ingmar; Cui, Yan; Ziemer, Mirjana; Anderegg, Ulf; Hagel, Christian; Morrison, Helen

    2016-01-01

    Hyperactive Ras signaling has strong oncogenic effects causing several different forms of cancer. Hyperactivity is frequently induced by mutations within Ras itself, which account for up to 30% of all human cancers. In addition, hyperactive Ras signaling can also be triggered independent of Ras by either mutation or by misexpression of various upstream regulators and immediate downstream effectors. We have previously reported that C-kinase potentiated protein phosphatase-1 inhibitor of 17 kDa (CPI-17) can drive Ras activity and promote tumorigenic transformation by inhibition of the tumor suppressor Merlin. We now describe an additional element of this oncogenic mechanism in the form of the ezrin-radixin-moesin (ERM) protein family, which exhibits opposing roles in Ras activity control. Thus, CPI-17 drives Ras activity and tumorigenesis in a two-fold way; inactivation of the tumor suppressor merlin and activation of the growth promoting ERM family. The in vivo significance of this oncogenic switch is highlighted by demonstrating CPI-17's involvement in human melanoma pathogenesis. PMID:27793041

  20. Determination of nitrogen in boron carbide by instrumental photon activation analysis.

    PubMed

    Merchel, Silke; Berger, Achim

    2007-05-01

    Boron carbide is widely used as industrial material, because of its extreme hardness, and as a neutron absorber. As part of a round-robin exercise leading to certification of a new reference material (ERM-ED102) which was demanded by the industry we analysed nitrogen in boron carbide by inert gas fusion analysis (GFA) and instrumental photon activation analysis (IPAA) using the 14N(gamma,n)13N nuclear reaction. The latter approach is the only non-destructive method among all the methods applied. By using photons with energy below the threshold of the 12C(gamma,n)11C reaction, we hindered activation of matrix and other impurities. A recently installed beam with a very low lateral activating flux gradient enabled us to homogeneously activate sample masses of approximately 1 g. Taking extra precautions, i.e. self-absorption correction and deconvolution of the complex decay curves, we calculated a nitrogen concentration of 2260+/-100 microg g-1, which is in good agreement with our GFA value of 2303+/-64 microg g-1. The values are the second and third highest of a rather atypical (non-S-shape) distribution of data of 14 round-robin participants. It is of utmost importance for the certification process that our IPAA value is the only one not produced by inert gas fusion analysis and, therefore, the only one which is not affected by a possible incomplete release of nitrogen from high-melting boron carbide.

  1. Involvement of moesin in the development of morphine analgesic tolerance through P-glycoprotein at the blood-brain barrier.

    PubMed

    Kobori, Takuro; Fujiwara, Shuhei; Miyagi, Kei; Harada, Shinichi; Nakamoto, Kazuo; Nakagawa, Takayuki; Takahashi, Hideo; Narita, Minoru; Tokuyama, Shogo

    2014-01-01

    Altered expression of P-glycoprotein (P-gp), a drug efflux transporter expressed by brain capillary endothelial cells (BCECs), may contribute to the development of opioid analgesic tolerance, as demonstrated by cumulative evidence from research. However, the detailed mechanism by which chronic morphine treatment increases P-gp expression remains unexplained. Ezrin/radixin/moesin (ERM) are scaffold proteins that are known to regulate the plasma membrane localization of some drug transporters such as P-gp in peripheral tissues, although a few reports suggest its role in the central nervous system as well. In this study, we investigated the involvement of ERM in the development of morphine analgesic tolerance through altered P-gp expression in BCECs. Repeated treatment with morphine (10 mg/kg/day, s.c. for 5 days) decreased its analgesic effect in the tail-flick test and increased P-gp protein expression in BCECs, as determined by Western blotting. Furthermore, moesin protein expression increased in the same fraction whereas that of ezrin decreased; no change was observed in the radixin expression. Furthermore, immunoprecipitation and immunofluorescence assays revealed interaction between moesin and P-gp molecules, along with co-localization, in BCECs. In conclusion, an increase in moesin expression may contribute to the increased expression of P-gp in BCECs, leading to the development of morphine analgesic tolerance.

  2. Ezrin and moesin expression in canine and feline osteosarcoma.

    PubMed

    Hlavaty, Juraj; Wolfesberger, Birgitt; Hauck, Marlene; Obermayer-Pietsch, Barbara; Fuchs-Baumgartinger, Andrea; Miller, Ingrid; Walter, Ingrid

    2016-11-30

    Biological features of canine osteosarcomas (OS) differ markedly from those found in feline and resemble more human osteosarcomas, in particular for their high rate of metastasis and poor prognosis. Ezrin, radixin and moesin are members of the ERM protein family and link the actin cytoskeleton with the cell membrane. Ezrin and moesin have been shown to be of prognostic significance in tumor progression due to their role in the metastatic process. The objective of this study was to analyze ezrin and moesin protein expression in a series of dog (n = 16) and cat (n = 8) osteosarcoma samples using immunohistochemistry and western blot techniques. We found that cat OS have a higher moesin expression compared to dog OS, however, the active phosphorylated forms of moesin and ezrin Tyr353 were more abundant in the dog samples. A statistically significant difference was found for the low and high immunohistochemical scores of ezrin and pan-phospho-ERM proteins between cat and dog. Although phospho-ezrin Thr567 was higher in feline OS, the membranous localization in dog OS samples indicates the presence of the biologically active form. Therefore, the observed differences in phosphorylated forms of ezrin and moesin status should be further studied to demonstrate if they are relevant for different biological behavior between dog and cat OS.

  3. PDZD8 is a novel moesin-interacting cytoskeletal regulatory protein that suppresses infection by herpes simplex virus type 1.

    PubMed

    Henning, Matthew S; Stiedl, Patricia; Barry, Denis S; McMahon, Robert; Morham, Scott G; Walsh, Derek; Naghavi, Mojgan H

    2011-07-05

    The host cytoskeleton plays a central role in the life cycle of many viruses yet our knowledge of cytoskeletal regulators and their role in viral infection remains limited. Recently, moesin and ezrin, two members of the ERM (Ezrin/Radixin/Moesin) family of proteins that regulate actin and plasma membrane cross-linking and microtubule (MT) stability, have been shown to inhibit retroviral infection. To further understand how ERM proteins function and whether they also influence infection by other viruses, we identified PDZD8 as a novel moesin-interacting protein. PDZD8 is a poorly understood protein whose function is unknown. Exogenous expression of either moesin or PDZD8 reduced the levels of stable MTs, suggesting that these proteins functioned as part of a cytoskeletal regulatory complex. Additionally, exogenous expression or siRNA-mediated knockdown of either factor affected Herpes Simplex Virus type 1 (HSV-1) infection, identifying a cellular function for PDZD8 and novel antiviral properties for these two cytoskeletal regulatory proteins.

  4. Effect of knockdown of ezrin, radixin, and moesin on P-glycoprotein function in HepG2 cells.

    PubMed

    Kano, Takashi; Wada, Sho; Morimoto, Kaori; Kato, Yukio; Ogihara, Takuo

    2011-12-01

    Ezrin, radixin, and moesin (ERM) proteins regulate functional expression of certain transporters, but little is known about their effect on P-glycoprotein (P-gp). Here, we investigated the influence of ERM proteins on the expression and activity of P-gp at the transcriptional, translational, and posttranslational levels, using HepG2 as a model cell line. Knockdown of ezrin with RNA interference decreased the level of P-gp messenger RNA. On the contrary, knockdown of radixin caused a decrease of the P-gp gene product at the cell surface, but not in whole cell lysate. Furthermore, a significant increase in accumulation of rhodamine123, a typical P-gp substrate, was observed in radixin knockdown cells, compared with control cells. Knockdown of moesin did not influence the expression or function of P-gp. These results indicate that ezrin influences the expression of P-gp at the translational level, whereas radixin is involved in membrane localization of P-gp in HepG2 cells.

  5. Dysregulation of lung injury and repair in moesin-deficient mice treated with intratracheal bleomycin.

    PubMed

    Hashimoto, Soshi; Amaya, Fumimasa; Matsuyama, Hiroki; Ueno, Hiroshi; Kikuchi, Shojiro; Tanaka, Masaki; Watanabe, Yoshihisa; Ebina, Masahito; Ishizaka, Akitoshi; Tsukita, Sachiko; Hashimoto, Satoru

    2008-10-01

    Moesin belongs to the ezrin/radixin/moesin (ERM) protein family and participates in cellular functions, such as morphogenesis and motility, by cross-linking between the actin cytoskeleton and plasma membranes. Although moesin seems necessary for tissue construction and repair, its function at the whole body level remains elusive, perhaps because of redundancy among ERM proteins. To determine the role played by moesin in the modulation of pulmonary alveolar structure associated with lung injury and repair, we examined the morphological changes in the lung and the effect of bleomycin-induced lung injury and fibrosis in moesin-deficient (Msn(-/Y)) and control wild-type mice (Msn(+/Y)). Immunohistochemical analysis revealed that moesin was specifically localized in the distal lung epithelium, where ezrin and radixin were faintly detectable in Msn(+/Y) mice. Compared with Msn(+/Y) mice, Msn(-/Y) mice displayed abnormalities of alveolar architecture and, when treated with bleomycin, developed more prominent lung injury and fibrosis and lower body weight and survival rate. Furthermore, Msn(-/Y) mice had abnormal cytokine and chemokine gene expression as shown by real-time PCR. This is the first report of a functional involvement of moesin in the regulation of lung inflammation and repair. Our observations show that moesin critically regulates the preservation of alveolar structure and lung homeostasis.

  6. Viscum album agglutinin-I (VAA-I) increases cell surface expression of cytoskeletal proteins in apoptotic human neutrophils: moesin and ezrin are two novel targets of VAA-I.

    PubMed

    Simon, M M; Simard, J C; Girard, D

    2013-10-01

    Viscum album agglutinin-I (VAA-I) is a plant lectin, which possesses anti-inflammatory properties, including the ability to induce neutrophil apoptosis by a mechanism that is not completely understood. Among the three actin-binding membrane-anchoring proteins ezrin/radixin/moesin (ERM), neutrophils are known to express ezrin and moesin. The behavior of these proteins in apoptotic neutrophils is not well established. In the present study, the expression and localization of ezrin and moesin by Western blot and immunofluorescence revealed a clear degradation and relocalization of both the proteins during VAA-I-induced apoptosis. Also, flow cytometry analysis revealed that VAA-I markedly and significantly induced the cell surface expression of ezrin and moesin and this was reversed when cells were pretreated with the Syk inhibitor piceatannol. The expression of ezrin and moesin on the cell surface of apoptotic neutrophils may represent a mechanism responsible for the appearance of autoantibodies directed against ERM proteins, which have been found in the serum of patients suffering from autoimmune diseases. Therefore, the ability of VAA-I to increase cell surface expression of cytoskeletal proteins in apoptotic neutrophils provides important insight into a possible toxic mechanism of this plant lectin and this has to be considered for its potential utilization for in vivo treatment.

  7. Extraradical mycelium of arbuscular mycorrhizal fungi radiating from large plants depresses the growth of nearby seedlings in a nutrient deficient substrate.

    PubMed

    Janoušková, Martina; Rydlová, Jana; Püschel, David; Száková, Jiřina; Vosátka, Miroslav

    2011-10-01

    The effect of arbuscular mycorrhiza (AM) on the interaction of large plants and seedlings in an early succession situation was investigated in a greenhouse experiment using compartmented rhizoboxes. Tripleurospermum inodorum, a highly mycorrhiza-responsive early coloniser of spoil banks, was cultivated either non-mycorrhizal or inoculated with AM fungi in the central compartment of the rhizoboxes. After two months, seedlings of T. inodorum or Sisymbrium loeselii, a non-host species colonising spoil banks simultaneously with T. inodorum, were planted in lateral compartments, which were colonised by the extraradical mycelium (ERM) of the pre-cultivated T. inodorum in the inoculated treatments. The experiment comprised the comparison of two AM fungal isolates and two substrates: spoil bank soil and a mixture of this soil with sand. As expected based on the low nutrient levels in the substrates, the pre-cultivated T. inodorum plants responded positively to mycorrhiza, the response being more pronounced in phosphorus uptake than in nitrogen uptake and growth. In contrast, the growth of the seedlings, both the host and the non-host species, was inhibited in the mycorrhizal treatments. Based on the phosphorus and nitrogen concentrations in the biomass of the experimental plants, this growth inhibition was attributed to nitrogen depletion in the lateral compartments by the ERM radiating from the central compartment. The results point to an important aspect of mycorrhizal effects on the coexistence of large plants and seedlings in nutrient deficient substrates.

  8. Challenges in the size analysis of a silica nanoparticle mixture as candidate certified reference material.

    PubMed

    Kestens, Vikram; Roebben, Gert; Herrmann, Jan; Jämting, Åsa; Coleman, Victoria; Minelli, Caterina; Clifford, Charles; De Temmerman, Pieter-Jan; Mast, Jan; Junjie, Liu; Babick, Frank; Cölfen, Helmut; Emons, Hendrik

    A new certified reference material for quality control of nanoparticle size analysis methods has been developed and produced by the Institute for Reference Materials and Measurements of the European Commission's Joint Research Centre. The material, ERM-FD102, consists of an aqueous suspension of a mixture of silica nanoparticle populations of distinct particle size and origin. The characterisation relied on an interlaboratory comparison study in which 30 laboratories of demonstrated competence participated with a variety of techniques for particle size analysis. After scrutinising the received datasets, certified and indicative values for different method-defined equivalent diameters that are specific for dynamic light scattering (DLS), centrifugal liquid sedimentation (CLS), scanning and transmission electron microscopy (SEM and TEM), atomic force microscopy (AFM), particle tracking analysis (PTA) and asymmetrical-flow field-flow fractionation (AF4) were assigned. The value assignment was a particular challenge because metrological concepts were not always interpreted uniformly across all participating laboratories. This paper presents the main elements and results of the ERM-FD102 characterisation study and discusses in particular the key issues of measurand definition and the estimation of measurement uncertainty.

  9. Prevalence, diversity and characterization of enterococci from three coraciiform birds.

    PubMed

    Splichalova, Petra; Svec, Pavel; Ghosh, Anuradha; Zurek, Ludek; Oravcova, Veronika; Radimersky, Tomas; Bohus, Mirko; Literak, Ivan

    2015-05-01

    Coraciiform birds hoopoe (Upupa epops), common kingfisher (Alcedo atthis) and European roller (Coracius garrulus) were examined for enterococci in their cloacae and uropygial glands. The enterococcal isolates were identified at the species level using several genomic and proteomic methods, screened for antibiotic susceptibility and genotyped by pulsed-field gel electrophoresis (PFGE). Clonality of isolates from the common kingfisher was also assessed by multi-locus sequence typing (MLST). Using selective media, putative enterococcal isolates (n = 117) were recovered from 74% (32 out of a total of 43) of the bird samples and 114 isolates were confirmed as enterococci. Overall, among the total of 6 different species detected, Enterococcus faecalis was dominant (59%) in all three bird species. The second most frequently isolated species was Enterococcus casseliflavus (32%). PFGE revealed great diversity of strains from different bird species and anatomic location. Closely related strains were found only from nestlings from the same nest. No genes conferring resistance to vancomycin (vanA, vanB, vanC1 and van C2/C3) or erythromycin (erm A, ermB and mefA/E) were detected. MLST analysis and eBURST clustering revealed that sequence types of E. faecalis from the common kingfisher were identical to those of isolates found previously in water, chickens, and humans.

  10. Involvement of phosphatidylinositol 3-kinase in stromal cell-derived factor-1 alpha-induced lymphocyte polarization and chemotaxis.

    PubMed

    Vicente-Manzanares, M; Rey, M; Jones, D R; Sancho, D; Mellado, M; Rodriguez-Frade, J M; del Pozo, M A; Yáñez-Mó, M; de Ana, A M; Martínez-A, C; Mérida, I; Sánchez-Madrid, F

    1999-10-01

    The role of phosphatidylinositol 3-kinase (PI3-kinase), an important enzyme involved in signal transduction events, has been studied in the polarization and chemotaxis of lymphocytes induced by the chemokine stromal cell-derived factor-1 alpha (SDF-1 alpha). This chemokine was able to directly activate p85/p110 PI3-kinase in whole human PBL and to induce the association of PI3-kinase to the SDF-1 alpha receptor, CXCR4, in a pertussis toxin-sensitive manner. Two unrelated chemical inhibitors of PI3-kinase, wortmannin and Ly294002, prevented ICAM-3 and ERM protein moesin polarization as well as the chemotaxis of PBL in response to SDF-1 alpha. However, they did not interfere with the reorganization of either tubulin or the actin cytoskeleton. Moreover, the transient expression of a dominant negative form of the PI3-kinase 85-kDa regulatory subunit in the constitutively polarized Peer T cell line inhibited ICAM-3 polarization and markedly reduced SDF-1 alpha-induced chemotaxis. Conversely, overexpression of a constitutively activated mutant of the PI3-kinase 110-kDa catalytic subunit in the round-shaped PM-1 T cell line induced ICAM-3 polarization. These results underline the role of PI3-kinase in the regulation of lymphocyte polarization and motility and indicate that PI3-kinase plays a selective role in the regulation of adhesion and ERM proteins redistribution in the plasma membrane of lymphocytes.

  11. Staphylococcus saprophyticus surface-associated protein (Ssp) is associated with lifespan reduction in Caenorhabditis elegans.

    PubMed

    Szabados, Florian; Mohner, Amelie; Kleine, Britta; Gatermann, Sören G

    2013-10-01

    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.

  12. 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.

  13. Fluorescence Resonance Energy Transfer Analysis of Merlin Conformational Changes ▿

    PubMed Central

    Hennigan, Robert F.; Foster, Lauren A.; Chaiken, Mary F.; Mani, Timmy; Gomes, Michelle M.; Herr, Andrew B.; Ip, Wallace

    2010-01-01

    Neurofibromatosis type 2 is an inherited autosomal disorder caused by biallelic inactivation of the NF2 tumor suppressor gene. The NF2 gene encodes a 70-kDa protein, merlin, which is a member of the ezrin-radixin-moesin (ERM) family. ERM proteins are believed to be regulated by a transition between a closed conformation, formed by binding of their N-terminal FERM domain and C-terminal tail domain (CTD), and an open conformation, in which the two domains do not interact. Previous work suggests that the tumor suppressor function of merlin is similarly regulated and that only the closed form is active. Therefore, understanding the mechanisms that control its conformation is crucial. We have developed a series of probes that measures merlin conformation by fluorescence resonance energy transfer, both as purified protein and in live cells. Using these tools, we find that merlin exists predominately as a monomer in a stable, closed conformation that is mediated by the central α-helical domain. The contribution from the FERM-CTD interaction to the closed conformation appears to be less important. Upon phosphorylation or interaction with an effector protein, merlin undergoes a subtle conformational change, suggesting a novel mechanism that modulates the interaction between the FERM domain and the CTD. PMID:19884346

  14. Knockdown of ezrin suppresses the migration and angiogenesis of human umbilical vein endothelial cells in vitro.

    PubMed

    Zhao, Liang-ping; Huang, Lei; Tian, Xun; Liang, Feng-qi; Wei, Jun-cheng; Zhang, Xian; Li, Sha; Zhang, Qing-hua

    2016-04-01

    Progressive tumor growth is dependent on angiogenesis. The mechanisms by which endothelial cells (ECs) are incorporated to develop new blood vessels are not well understood. Recent studies reveal that the ezrin radixin moesin (ERM) family members are key regulators of cellular activities such as adhesion, morphogenetic change, and migration. We hypothesized that ezrin, one of the ERM family members, may play important roles in ECs organization during angiogenesis, and new vessels formation in preexisting tissues. To test this hypothesis, in this study, we investigated the effects of ezrin gene silencing on the migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) in vitro. HUVECs were transfected with plasmids with ezrin-targeting short hairpin RNA by using the lipofectamine-2000 system. Wound assay in vitro and three-dimensional culture were used to detect the migration and angiogenesis capacity of HUVECs. The morphological changes of transfected cells were observed by confocal and phase contrast microscopy. Our results demonstrated that the decreased expression of ezrin in HUVECs significantly induced the morphogenetic changes and cytoskeletal reorganization of the transfected cells, and also reduced cell migration and angiogenesis capacity in vitro, suggesting that ezrin play an important role in the process of HUVECs migration and angiogenesis.

  15. 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.

  16. Macrolones Are a Novel Class of Macrolide Antibiotics Active against Key Resistant Respiratory Pathogens In Vitro and In Vivo

    PubMed Central

    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

    2016-01-01

    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

  17. Virulence factors, antimicrobial susceptibility and molecular characterization of Streptococcus agalactiae isolated from pregnant women.

    PubMed

    Beigverdi, Reza; Jabalameli, Fereshteh; Mirsalehian, Akbar; Hantoushzadeh, Sedigheh; Boroumandi, Shahram; Taherikalani, Morovat; Emaneini, Mohammad

    2014-12-01

    Forty-one Streptococcus agalactiae isolates collected from pregnant women at 35-37 weeks of gestation were analysed for their capsular types, antimicrobial resistance determinants, distribution of virulence factors and genetic relatedness using PCR and multiplex PCR. Capsular type III was predominant (65.8%), followed by capsular type II (14.6%), Ib (7.3%), and V(4.9%). All isolates were susceptible to penicillin, vancomycin, linezolid and quinupristin-dalfopristin. Resistance to tetracycline, erythromycin and clindamycin were found in 97.6%, 24.4%, and 14.6% of isolates, respectively. The most common antimicrobial resistance gene was tetM found in 97.6% of the isolates followed by ermTR and ermB found in 12% and 7.3% of isolates, respectively. The most common virulence gene was hly (100%), followed by scpB (97.6%), bca (97.6%), rib (53.65%) and bac (4.9%). The insertion sequence IS1548 was found in 63.4% of isolates. By multi locus variable number of tandem repeat analysis (MLVA) typing, 30 different allelic profiles or MLVA types (MTs) were identified. The most frequent was the MT1 (5/41, 12.2%) and followed by MT2 (4/41, 9.75%). Our data revealed that population structure of these isolates is highly diverse and indicates different MLVA types.

  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 t