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Sample records for selectively isolate phosphopeptides

  1. The use of titanium dioxide micro-columns to selectively isolate phosphopeptides from proteolytic digests

    DEFF Research Database (Denmark)

    Thingholm, Tine E; Larsen, Martin R

    2009-01-01

    Titanium dioxide has very high affinity for phosphopeptides and it has become an efficient alternative to already existing methods for phosphopeptide enrichment from complex samples. Peptide loading in a highly acidic environment in the presence of 2,5-dihydroxybenzoic acid (DHB), phthalic acid......, or glycolic acid has been shown to improve selectivity significantly by reducing unspecific binding from nonphosphorylated peptides. The enriched phosphopeptides bound to the titanium dioxide are subsequently eluted from the micro-column using an alkaline buffer. Titanium dioxide chromatography is extremely...... tolerant towards most buffers used in biological experiments. It is highly robust and as such it has become one of the methods of choice in large-scale phospho-proteomics. Here we describe the protocol for phosphopeptide enrichment using titanium dioxide chromatography followed by desalting...

  2. A novel strategy for phosphopeptide enrichment using lanthanide phosphate co-precipitation.

    Science.gov (United States)

    Mirza, Munazza Raza; Rainer, Matthias; Güzel, Yüksel; Choudhary, Iqbal M; Bonn, Günther K

    2012-08-01

    Reversible phosphorylation of proteins is a common theme in the regulation of important cellular functions such as growth, metabolism, and differentiation. The comprehensive understanding of biological processes requires the characterization of protein phosphorylation at the molecular level. Although, the number of cellular phosphoproteins is relatively high, the phosphorylated residues themselves are generally of low abundance due to the sub-stoichiometric nature. However, low abundance of phosphopeptides and low degree of phosphorylation typically necessitates isolation and concentration of phosphopeptides prior to mass spectrometric analysis. In this study, we used trivalent lanthanide ions (LaCl(3), CeCl(3), EuCl(3), TbCl(3), HoCl(3), ErCl(3), and TmCl(3)) for phosphopeptide enrichment and cleaning-up. Due to their low solubility product, lanthanide ions form stable complexes with the phosphate groups of phosphopeptides and precipitate out of solution. In a further step, non-phosphorylated compounds can easily be removed by simple centrifugation and washing before mass spectrometric analysis using Matrix-assisted laser desorption/ionisation-time of flight. The precipitation method was applied for the isolation of phosphopeptides from standard proteins such as ovalbumin, α-casein, and β-casein. High enrichment of phosphopeptides could also be achieved for real samples such as fresh milk and egg white. The technology presented here represents an excellent and highly selective tool for phosphopeptide recovery; it is easily applicable and shows several advantages as compared with standard approaches such as TiO(2) or IMAC.

  3. Proteolytic Digestion and TiO2 Phosphopeptide Enrichment Microreactor for Fast MS Identification of Proteins

    Science.gov (United States)

    Deng, Jingren; Lazar, Iulia M.

    2016-04-01

    The characterization of phosphorylation state(s) of a protein is best accomplished by using isolated or enriched phosphoprotein samples or their corresponding phosphopeptides. The process is typically time-consuming as, often, a combination of analytical approaches must be used. To facilitate throughput in the study of phosphoproteins, a microreactor that enables a novel strategy for performing fast proteolytic digestion and selective phosphopeptide enrichment was developed. The microreactor was fabricated using 100 μm i.d. fused-silica capillaries packed with 1-2 mm beds of C18 and/or TiO2 particles. Proteolytic digestion-only, phosphopeptide enrichment-only, and sequential proteolytic digestion/phosphopeptide enrichment microreactors were developed and tested with standard protein mixtures. The protein samples were adsorbed on the C18 particles, quickly digested with a proteolytic enzyme infused over the adsorbed proteins, and further eluted onto the TiO2 microreactor for enrichment in phosphopeptides. A number of parameters were optimized to speed up the digestion and enrichments processes, including microreactor dimensions, sample concentrations, digestion time, flow rates, buffer compositions, and pH. The effective time for the steps of proteolytic digestion and enrichment was less than 5 min. For simple samples, such as standard protein mixtures, this approach provided equivalent or better results than conventional bench-top methods, in terms of both enzymatic digestion and selectivity. Analysis times and reagent costs were reduced ~10- to 15-fold. Preliminary analysis of cell extracts and recombinant proteins indicated the feasibility of integration of these microreactors in more advanced workflows amenable for handling real-world biological samples.

  4. Molecularly Imprinted Porous Monolithic Materials from Melamine-Formaldehyde for Selective Trapping of Phosphopeptides

    DEFF Research Database (Denmark)

    Liu, Mingquan; Tran, Tri Minh; Abbas Elhaj, Ahmed Awad

    2017-01-01

    monoliths, chosen based on the combination of meso- and macropores providing optimal percolative flow and accessible surface area, was synthesized in the presence of N-Fmoc and O-Et protected phosphoserine and phosphotyrosine to prepare molecularly imprinted monoliths with surface layers selective...... for phosphopeptides. These imprinted monoliths were characterized alongside nonimprinted monoliths by a variety of techniques and finally evaluated by liquid chromatography-mass spectrometry in the capillary format to assess their abilities to trap and release phosphorylated amino acids and peptides from partly...

  5. Comparing Multi-Step IMAC and Multi-Step TiO2 Methods for Phosphopeptide Enrichment

    Science.gov (United States)

    Yue, Xiaoshan; Schunter, Alissa; Hummon, Amanda B.

    2016-01-01

    Phosphopeptide enrichment from complicated peptide mixtures is an essential step for mass spectrometry-based phosphoproteomic studies to reduce sample complexity and ionization suppression effects. Typical methods for enriching phosphopeptides include immobilized metal affinity chromatography (IMAC) or titanium dioxide (TiO2) beads, which have selective affinity and can interact with phosphopeptides. In this study, the IMAC enrichment method was compared with the TiO2 enrichment method, using a multi-step enrichment strategy from whole cell lysate, to evaluate their abilities to enrich for different types of phosphopeptides. The peptide-to-beads ratios were optimized for both IMAC and TiO2 beads. Both IMAC and TiO2 enrichments were performed for three rounds to enable the maximum extraction of phosphopeptides from the whole cell lysates. The phosphopeptides that are unique to IMAC enrichment, unique to TiO2 enrichment, and identified with both IMAC and TiO2 enrichment were analyzed for their characteristics. Both IMAC and TiO2 enriched similar amounts of phosphopeptides with comparable enrichment efficiency. However, phosphopeptides that are unique to IMAC enrichment showed a higher percentage of multi-phosphopeptides, as well as a higher percentage of longer, basic, and hydrophilic phosphopeptides. Also, the IMAC and TiO2 procedures clearly enriched phosphopeptides with different motifs. Finally, further enriching with two rounds of TiO2 from the supernatant after IMAC enrichment, or further enriching with two rounds of IMAC from the supernatant TiO2 enrichment does not fully recover the phosphopeptides that are not identified with the corresponding multi-step enrichment. PMID:26237447

  6. Highly selective manganese-doped zinc sulfide quantum dots based label free phosphorescent sensor for phosphopeptides in presence of zirconium (IV).

    Science.gov (United States)

    Gong, Yan; Fan, Zhefeng

    2015-04-15

    We report a room-temperature phosphorescence (RTP) sensor for phosphopeptides based on zirconium (IV)-modulated mercaptopropionic acid (MPA)-capped Mn-doped ZnS quantum dots (QDs). This sensor incorporates the advantages of the well-known Zr(4+)-phosphopeptide affinity pair and the RTP properties of doped QDs. The RTP of Mn-doped ZnS QDs capped with MPA can be effectively quenched by Zr(4+). The high affinity of phosphopeptides to Zr(4+) enables the dissociation of the ion from the surface of MPA-capped ZnS QDs, thereby forming a stable complex with phosphopeptides in the solution, and recovering the RTP of the QDs. The Zr(4+)-induced RTP quenching and subsequent phosphopeptide-induced RTP recovery for MPA-capped ZnS QDs provide a solid basis for the present RTP sensor based on QDs for the detection of phosphopeptides. The detection limit for phosphopeptides is 0.9ngmL(-1), the relative standard deviations is 2.5%, and the recovery of urine and serum samples with phosphopeptides addition rangs from 96% to 105% at optimal conditions. The proposed method was successfully applied to biological fluids and obtained satisfactory results. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Enhanced MALDI-TOF MS Analysis of Phosphopeptides Using an Optimized DHAP/DAHC Matrix

    Science.gov (United States)

    Hou, Junjie; Xie, Zhensheng; Xue, Peng; Cui, Ziyou; Chen, Xiulan; Li, Jing; Cai, Tanxi; Wu, Peng; Yang, Fuquan

    2010-01-01

    Selecting an appropriate matrix solution is one of the most effective means of increasing the ionization efficiency of phosphopeptides in matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). In this study, we systematically assessed matrix combinations of 2, 6-dihydroxyacetophenone (DHAP) and diammonium hydrogen citrate (DAHC), and demonstrated that the low ratio DHAP/DAHC matrix was more effective in enhancing the ionization of phosphopeptides. Low femtomole level of phosphopeptides from the tryptic digests of α-casein and β-casein was readily detected by MALDI-TOF-MS in both positive and negative ion mode without desalination or phosphopeptide enrichment. Compared with the DHB/PA matrix, the optimized DHAP/DAHC matrix yielded superior sample homogeneity and higher phosphopeptide measurement sensitivity, particularly when multiple phosphorylated peptides were assessed. Finally, the DHAP/DAHC matrix was applied to identify phosphorylation sites from α-casein and β-casein and to characterize two phosphorylation sites from the human histone H1 treated with Cyclin-Dependent Kinase-1 (CDK1) by MALDI-TOF/TOF MS. PMID:20339515

  8. Enhanced MALDI-TOF MS Analysis of Phosphopeptides Using an Optimized DHAP/DAHC Matrix

    Directory of Open Access Journals (Sweden)

    Junjie Hou

    2010-01-01

    Full Text Available Selecting an appropriate matrix solution is one of the most effective means of increasing the ionization efficiency of phosphopeptides in matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS. In this study, we systematically assessed matrix combinations of 2, 6-dihydroxyacetophenone (DHAP and diammonium hydrogen citrate (DAHC, and demonstrated that the low ratio DHAP/DAHC matrix was more effective in enhancing the ionization of phosphopeptides. Low femtomole level of phosphopeptides from the tryptic digests of α-casein and β-casein was readily detected by MALDI-TOF-MS in both positive and negative ion mode without desalination or phosphopeptide enrichment. Compared with the DHB/PA matrix, the optimized DHAP/DAHC matrix yielded superior sample homogeneity and higher phosphopeptide measurement sensitivity, particularly when multiple phosphorylated peptides were assessed. Finally, the DHAP/DAHC matrix was applied to identify phosphorylation sites from α-casein and β-casein and to characterize two phosphorylation sites from the human histone H1 treated with Cyclin-Dependent Kinase-1 (CDK1 by MALDI-TOF/TOF MS.

  9. Hydrophilic Nb{sup 5+}-immobilized magnetic core–shell microsphere – A novel immobilized metal ion affinity chromatography material for highly selective enrichment of phosphopeptides

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Xueni; Liu, Xiaodan; Feng, Jianan [Pharmaceutical Analysis Department, School of Pharmacy, Fudan University, Shanghai 201203 (China); Li, Yan, E-mail: yanli@fudan.edu.cn [Pharmaceutical Analysis Department, School of Pharmacy, Fudan University, Shanghai 201203 (China); Deng, Chunhui [Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433 (China); Duan, Gengli [Pharmaceutical Analysis Department, School of Pharmacy, Fudan University, Shanghai 201203 (China)

    2015-06-23

    Highlights: • A new IMAC material (Fe{sub 3}O{sub 4}@PD-Nb{sup 5+}) was synthesized. • The strong magnetic behaviors of the microspheres ensure fast and easy separation. • The enrichment ability was tested by human serum and nonfat milk. • The results were compared with other IMAC materials including the commercial kits. • All results proved the good enrichment ability, especially for multiphosphopeptides. - Abstract: Rapid and selective enrichment of phosphopeptides from complex biological samples is essential and challenging in phosphorylated proteomics. In this work, for the first time, niobium ions were directly immobilized on the surface of polydopamine-coated magnetic microspheres through a facile and effective synthetic route. The Fe{sub 3}O{sub 4}@polydopamine-Nb{sup 5+} (denoted as Fe{sub 3}O{sub 4}@PD-Nb{sup 5+}) microspheres possess merits of high hydrophilicity and good biological compatibility, and demonstrated low limit of detection (2 fmol). The selectivity was also basically satisfactory (β-casein:BSA = 1:500) to capture phosphopeptides. They were also successfully applied for enrichment of phosphopeptides from real biological samples such as human serum and nonfat milk. Compared with Fe{sub 3}O{sub 4}@PD-Ti{sup 4+} microspheres, the Fe{sub 3}O{sub 4}@PD-Nb{sup 5+} microspheres exhibit superior selectivity to multi-phosphorylated peptides, and thus may be complementary to the conventional IMAC materials.

  10. Phosphopeptide Enrichment by Covalent Chromatography after Derivatization of Protein Digests Immobilized on Reversed-Phase Supports

    Science.gov (United States)

    Nika, Heinz; Nieves, Edward; Hawke, David H.; Angeletti, Ruth Hogue

    2013-01-01

    A rugged sample-preparation method for comprehensive affinity enrichment of phosphopeptides from protein digests has been developed. The method uses a series of chemical reactions to incorporate efficiently and specifically a thiol-functionalized affinity tag into the analyte by barium hydroxide catalyzed β-elimination with Michael addition using 2-aminoethanethiol as nucleophile and subsequent thiolation of the resulting amino group with sulfosuccinimidyl-2-(biotinamido) ethyl-1,3-dithiopropionate. Gentle oxidation of cysteine residues, followed by acetylation of α- and ε-amino groups before these reactions, ensured selectivity of reversible capture of the modified phosphopeptides by covalent chromatography on activated thiol sepharose. The use of C18 reversed-phase supports as a miniaturized reaction bed facilitated optimization of the individual modification steps for throughput and completeness of derivatization. Reagents were exchanged directly on the supports, eliminating sample transfer between the reaction steps and thus, allowing the immobilized analyte to be carried through the multistep reaction scheme with minimal sample loss. The use of this sample-preparation method for phosphopeptide enrichment was demonstrated with low-level amounts of in-gel-digested protein. As applied to tryptic digests of α-S1- and β-casein, the method enabled the enrichment and detection of the phosphorylated peptides contained in the mixture, including the tetraphosphorylated species of β-casein, which has escaped chemical procedures reported previously. The isolates proved highly suitable for mapping the sites of phosphorylation by collisionally induced dissociation. β-Elimination, with consecutive Michael addition, expanded the use of the solid-phase-based enrichment strategy to phosphothreonyl peptides and to phosphoseryl/phosphothreonyl peptides derived from proline-directed kinase substrates and to their O-sulfono- and O-linked β-N-acetylglucosamine (O

  11. Phosphopeptide enrichment by immobilized metal affinity chromatography

    DEFF Research Database (Denmark)

    Thingholm, Tine E.; Larsen, Martin R.

    2016-01-01

    Immobilized metal affinity chromatography (IMAC) has been the method of choice for phosphopeptide enrichment prior to mass spectrometric analysis for many years and it is still used extensively in many laboratories. Using the affinity of negatively charged phosphate groups towards positively...... charged metal ions such as Fe3+, Ga3+, Al3+, Zr4+, and Ti4+ has made it possible to enrich phosphorylated peptides from peptide samples. However, the selectivity of most of the metal ions is limited, when working with highly complex samples, e.g., whole-cell extracts, resulting in contamination from...

  12. A novel tantalum-based sol-gel packed microextraction syringe for highly specific enrichment of phosphopeptides in MALDI-MS applications.

    Science.gov (United States)

    Çelikbıçak, Ömür; Atakay, Mehmet; Güler, Ülkü; Salih, Bekir

    2013-08-07

    A new tantalum-based sol-gel material was synthesized using a unique sol-gel synthesis pathway by PEG incorporation into the sol-gel structure without performing a calcination step. This improved its chemical and physical properties for the high capacity and selective enrichment of phosphopeptides from protein digests in complex biological media. The specificity of the tantalum-based sol-gel material for phosphopeptides was evaluated and compared with tantalum(V) oxide (Ta2O5) in different phosphopeptide enrichment applications. The tantalum-based sol-gel and tantalum(V) oxide were characterized in detail using FT-IR spectroscopy, X-ray diffraction (XRD) and scanning electron microscopy (SEM), and also using a surface area and pore size analyzer. In the characterization studies, the surface morphology, pore volume, crystallinity of the materials and PEG incorporation into the sol-gel structure to produce a more hydrophilic material were successfully demonstrated. The X-ray diffractograms of the two different materials were compared and it was noted that the broad signals of the tantalum-based sol-gel clearly represented the amorphous structure of the sol-gel material, which was more likely to create enough surface area and to provide more accessible tantalum atoms for phosphopeptides to be easily adsorbed when compared with the neat and more crystalline structure of Ta2O5. Therefore, the phosphopeptide enrichment performance of the tantalum-based sol-gels was found to be remarkably higher than the more crystalline Ta2O5 in our studies. Phosphopeptides at femtomole levels could be selectively enriched using the tantalum-based sol-gel and detected with a higher signal-to-noise ratio by matrix-assisted laser desorption/ionization-mass spectrometer (MALDI-MS). Moreover, phosphopeptides in a tryptic digest of non-fat bovine milk as a complex real-world biological sample were retained with higher yield using a tantalum-based sol-gel. Additionally, the sol-gel material

  13. Functionalized diamond nanopowder for phosphopeptides enrichment from complex biological fluids

    Energy Technology Data Exchange (ETDEWEB)

    Hussain, Dilshad [Division of Analytical Chemistry, Institute of Chemical Sciences, Bahauddin Zakariya University, Multan 60800 (Pakistan); Najam-ul-Haq, Muhammad, E-mail: najamulhaq@bzu.edu.pk [Division of Analytical Chemistry, Institute of Chemical Sciences, Bahauddin Zakariya University, Multan 60800 (Pakistan); Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University, Innrain 80-82, A-6020 Innsbruck (Austria); Jabeen, Fahmida; Ashiq, Muhammad N.; Athar, Muhammad [Division of Analytical Chemistry, Institute of Chemical Sciences, Bahauddin Zakariya University, Multan 60800 (Pakistan); Rainer, Matthias; Huck, Christian W.; Bonn, Guenther K. [Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University, Innrain 80-82, A-6020 Innsbruck (Austria)

    2013-05-02

    Graphical abstract: -- Highlights: •Derivatization of diamond nanopowder as IMAC and RP. •Characterization with SEM, EDX and FT-IR. •Phosphopeptide enrichment from standard as well as real samples. •Desalting and human serum profiling with reproducible results. •MALDI-MS analysis with database identification. -- Abstract: Diamond is known for its high affinity and biocompatibility towards biomolecules and is used exclusively in separation sciences and life science research. In present study, diamond nanopowder is derivatized as Immobilized Metal Ion Affinity Chromatographic (IMAC) material for the phosphopeptides enrichment and as Reversed Phase (C-18) media for the desalting of complex mixtures and human serum profiling through MALDI-TOF-MS. Functionalized diamond nanopowder is characterized by Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM) and energy dispersive X-ray (EDX) spectroscopy. Diamond-IMAC is applied to the standard protein (β-casein), spiked human serum, egg yolk and non-fat milk for the phosphopeptides enrichment. Results show the selectivity of synthesized IMAC-diamond immobilized with Fe{sup 3+} and La{sup 3+} ions. To comprehend the elaborated use, diamond-IMAC is also applied to the serum samples from gall bladder carcinoma for the potential biomarkers. Database search is carried out by the Mascot program ( (www.matrixscience.com)) for the assignment of phosphorylation sites. Diamond nanopowder is thus a separation media with multifunctional use and can be applied to cancer protein profiling for the diagnosis and biomarker identification.

  14. On-bead chemical synthesis and display of phosphopeptides for affinity pull-down proteomics

    DEFF Research Database (Denmark)

    Malene, Brandt; Madsen, Jens C.; Bunkenborg, Jakob

    2006-01-01

    We describe a new method for phosphopeptide proteomics based on the solid-phase synthesis of phosphopeptides on beads suitable for affinity pull-down experiments. Peptide sequences containing the Bad Ser112 and Ser136 phosphorylation motifs were used as bait in affinity pull-down experiments...... (aldehyde) at the C terminus for potential activity-based proteomics. The synthetic support-bound Bad phosphopeptides were able to pull down 14-3-3zeta. Furthermore, Bad phosphopeptides bound endogenous 14-3-3 proteins, and all seven members of the 14-3-3 family were identified by mass spectrometry....... In control experiments, none of the unphosphorylated Bad peptides bound transfected 14-3-3zeta or endogenous 14-3-3. We conclude that the combined synthesis and display of phosphopeptides on-bead is a fast and efficient method for affinity pull-down proteomics....

  15. Phosphopeptide derivatization signatures to identify serine and threonine phosphorylated peptides by mass spectrometry.

    Science.gov (United States)

    Molloy, M P; Andrews, P C

    2001-11-15

    The development of rapid, global methods for monitoring states of protein phosphorylation would provide greater insight for understanding many fundamental biological processes. Current best practices use mass spectrometry (MS) to profile digests of purified proteins for evidence of phosphorylation. However, this approach is beset by inherent difficulties in both identifying phosphopeptides from within a complex mixture containing many other unmodified peptides and ionizing phosphopeptides in positive-ion MS. We have modified an approach that uses barium hydroxide to rapidly eliminate the phosphoryl group of serine and threonine modified amino acids, creating dehydroamino acids that are susceptible to nucleophilic derivatization. By derivatizing a protein digest with a mixture of two different alkanethiols, phosphopeptide-specific derivatives were readily distinguished by MS due to their characteristic ion-pair signature. The resulting tagged ion pairs accommodate simple and rapid screening for phosphopeptides in a protein digest, obviating the use of isotopically labeled samples for qualitative phosphopeptide detection. MALDI-MS is used in a first pass manner to detect derivatized phosphopeptides, while the remaining sample is available for tandem MS to reveal the site of derivatization and, thus, phosphorylation. We demonstrated the technique by identifying phosphopeptides from beta-casein and ovalbumin. The approach was further used to examine in vitro phosphorylation of recombinant human HSP22 by protein kinase C, revealing phosphorylation of Thr-63.

  16. Specific phosphopeptide binding regulates a conformational change in the PI 3-kinase SH2 domain associated with enzyme activation.

    Science.gov (United States)

    Shoelson, S E; Sivaraja, M; Williams, K P; Hu, P; Schlessinger, J; Weiss, M A

    1993-01-01

    SH2 (src-homology 2) domains define a newly recognized binding motif that mediates the physical association of target phosphotyrosyl proteins with downstream effector enzymes. An example of such phosphoprotein-effector coupling is provided by the association of phosphatidylinositol 3-kinase (PI 3-kinase) with specific phosphorylation sites within the PDGF receptor, the c-Src/polyoma virus middle T antigen complex and the insulin receptor substrate IRS-1. Notably, phosphoprotein association with the SH2 domains of p85 also stimulates an increase in catalytic activity of the PI 3-kinase p110 subunit, which can be mimicked by phosphopeptides corresponding to targeted phosphoprotein phosphorylation sites. To investigate how phosphoprotein binding to the p85 SH2 domain stimulates p110 catalytic activation, we have examined the differential effects of phosphotyrosine and PDGF receptor-, IRS-1- and c-Src-derived phosphopeptides on the conformation of an isolated SH2 domain of PI 3-kinase. Although phosphotyrosine and both activating and non-activating phosphopeptides bind to the SH2 domain, activating phosphopeptides bind with higher affinity and induce a qualitatively distinct conformational change as monitored by CD and NMR spectroscopy. Amide proton exchange and protease protection assays further show that high affinity, specific phosphopeptide binding induces non-local dynamic SH2 domain stabilization. Based on these findings we propose that specific phosphoprotein binding to the p85 subunit induces a change in SH2 domain structure which is transmitted to the p110 subunit and regulates enzymatic activity by an allosteric mechanism. Images PMID:8382612

  17. Hierarchically templated beads with tailored pore structure for phosphopeptide capture and phosphoproteomics

    DEFF Research Database (Denmark)

    Wierzbicka, Celina; Torsetnes, Silje B.; Jensen, Ole N.

    2017-01-01

    Two templating approaches to produce imprinted phosphotyrosine capture beads with a controllable pore structure are reported and compared with respect to their ability to enrich phosphopeptides from a tryptic peptide mixture. The beads were prepared by the polymerization of urea-based host monomers...... and crosslinkers inside the pores of macroporous silica beads with both free and immobilized template. In the final step the silica was removed by fluoride etching resulting in mesoporous polymer replicas with narrow pore size distributions, pore diameters ≈ 10 nm and surface area > 260 m2 g-1. The beads displayed...... pronounced phosphotyrosine affinity and selectivity in binding tests using model peptides in acetonitrile rich solutions with a performance surpassing solution polymerized bulk imprinted materials. Tests of the beads for the enrichment of phosphopeptides from tryptic digests of twelve proteins revealed both...

  18. An Internal Standard for Assessing Phosphopeptide Recovery from Metal Ion/Oxide Enrichment Strategies

    Science.gov (United States)

    Paulo, Joao A.; Navarrete-Perea, Jose; Erickson, Alison R.; Knott, Jeffrey; Gygi, Steven P.

    2018-04-01

    Phosphorylation-mediated signaling pathways have major implications in cellular regulation and disease. However, proteins with roles in these pathways are frequently less abundant and phosphorylation is often sub-stoichiometric. As such, the efficient enrichment, and subsequent recovery of phosphorylated peptides, is vital. Mass spectrometry-based proteomics is a well-established approach for quantifying thousands of phosphorylation events in a single experiment. We designed a peptide internal standard-based assay directed toward sample preparation strategies for mass spectrometry analysis to understand better phosphopeptide recovery from enrichment strategies. We coupled mass-differential tandem mass tag (mTMT) reagents (specifically, TMTzero and TMTsuper-heavy), nine mass spectrometry-amenable phosphopeptides (phos9), and peak area measurements from extracted ion chromatograms to determine phosphopeptide recovery. We showcase this mTMT/phos9 recovery assay by evaluating three phosphopeptide enrichment workflows. Our assay provides data on the recovery of phosphopeptides, which complement other metrics, namely the number of identified phosphopeptides and enrichment specificity. Our mTMT/phos9 assay is applicable to any enrichment protocol in a typical experimental workflow irrespective of sample origin or labeling strategy. [Figure not available: see fulltext.

  19. Highly efficient enrichment of phosphopeptides from HeLa cells using hollow magnetic macro/mesoporous TiO2 nanoparticles.

    Science.gov (United States)

    Hong, Yayun; Zhan, Qiliang; Pu, Chenlu; Sheng, Qianying; Zhao, Hongli; Lan, Minbo

    2018-09-01

    In this work, hollow magnetic macro/mesoporous TiO 2 nanoparticles (denoted as Fe 3 O 4 @H-fTiO 2 ) were synthesized by a facile "hydrothermal etching assisted crystallization" route to improve the phosphopeptide enrichment efficiency. The porous nanostructure of TiO 2 shell and large hollow space endowed the Fe 3 O 4 @H-fTiO 2 with a high surface area (144.71 m 2 g -1 ) and a large pore volume (0.52 cm 3 g -1 ), which could provide more affinity sites for phosphopeptide enrichment. Besides, the large pore size of TiO 2 nanosheets and large hollow space could effectively prevent the "shadow effect", thereby facilitating the diffusion and release of phosphopeptides. Compared with the hollow magnetic mesoporous TiO 2 with small and deep pores (denoted as Fe 3 O 4 @H-mTiO 2 ) and solid magnetic macro/mesoporous TiO 2 , the Fe 3 O 4 @H-fTiO 2 nanoparticles showed a better selectivity (molar ratio of α-casein/BSA up to 1:10000) and a higher sensitivity (0.2 fmol/μL α-casein) for phosphopeptide enrichment. Furthermore, 1485 unique phosphopeptides derived from 660 phosphoproteins were identified from HeLa cell extracts after enrichment with Fe 3 O 4 @H-fTiO 2 nanoparticles, further demonstrating that the Fe 3 O 4 @H-fTiO 2 nanoparticles had a high-efficiency performance for phosphopeptide enrichment. Taken together, the Fe 3 O 4 @H-fTiO 2 nanoparticles will have unique advantages in phosphoproteomics analysis. Copyright © 2018 Elsevier B.V. All rights reserved.

  20. Phosphopeptide occupancy and photoaffinity cross-linking of the v-Src SH2 domain attenuates tyrosine kinase activity.

    Science.gov (United States)

    Garcia, P; Shoelson, S E; Drew, J S; Miller, W T

    1994-12-02

    Phosphorylation of c-Src at carboxyl-terminal Tyr-527 suppresses tyrosine kinase activity and transforming potential, presumably by facilitating the intramolecular interaction of the C terminus of Src with its SH2 domain. In addition, it has been shown previously that occupancy of the c-Src SH2 domain with a phosphopeptide stimulates c-Src kinase catalytic activity. We have performed analogous studies with v-Src, the transforming protein from Rous sarcoma virus, which has extensive homology with c-Src. v-Src lacks an autoregulatory phosphorylation site, and its kinase domain is constitutively active. Phosphopeptides corresponding to the sequences surrounding c-Src Tyr-527 and a Tyr-Glu-Glu-Ile motif from the hamster polyoma virus middle T antigen inhibit tyrosine kinase activity of baculovirus-expressed v-Src 2- and 4-fold, respectively. To determine the mechanism of this regulation, the Tyr-527 phosphopeptide was substituted with the photoactive amino acid p-benzoylphenylalanine at the adjacent positions (N- and C-terminal) to phosphotyrosine. These peptides photoinactivate the v-Src tyrosine kinase 5-fold in a time- and concentration-dependent manner. Furthermore, the peptides cross-link an isolated Src SH2 domain with similar rates and specificity. These data indicate that occupancy of the v-Src SH2 domain induces a conformational change that is transmitted to the kinase domain and attenuates tyrosine kinase activity.

  1. Phosphoric acid as a matrix additive for MALDI MS analysis of phosphopeptides and phosphoproteins

    DEFF Research Database (Denmark)

    Kjellström, Sven; Jensen, Ole Nørregaard

    2004-01-01

    Phosphopeptides are often detected with low efficiency by MALDI MS analysis of peptide mixtures. In an effort to improve the phosphopeptide ion response in MALDI MS, we investigated the effects of adding low concentrations of organic and inorganic acids during peptide sample preparation in 2,5-di...... acid to 2,5-DHB were also observed in LC-MALDI-MS analysis of tryptic phosphopeptides of B. subtilis PrkC phosphoprotein. Finally, the mass resolution of MALDI mass spectra of intact proteins was significantly improved by using phosphoric acid in 2,5-DHB matrix....

  2. Development of an enrichment method for endogenous phosphopeptide characterization in human serum.

    Science.gov (United States)

    La Barbera, Giorgia; Capriotti, Anna Laura; Cavaliere, Chiara; Ferraris, Francesca; Laus, Michele; Piovesana, Susy; Sparnacci, Katia; Laganà, Aldo

    2018-01-01

    The work describes the development of an enrichment method for the analysis of endogenous phosphopeptides in serum. Endogenous peptides can play significant biological roles, and some of them could be exploited as future biomarkers. In this context, blood is one of the most useful biofluids for screening, but a systematic investigation of the endogenous peptides, especially phosphorylated ones, is still lacking, mainly due to the lack of suitable analytical methods. Thus, in this paper, different phosphopeptide enrichment strategies were pursued, based either on metal oxide affinity chromatography (MOAC, in the form of commercial TiO 2 spin columns or magnetic graphitized carbon black-TiO 2 composite), or on immobilized metal ion affinity chromatography (IMAC, in the form of Ti 4+ -IMAC magnetic material or commercial Fe 3+ -IMAC spin columns). While MOAC strategies proved completely unsuccessful, probably due to interfering phospholipids displacing phosphopeptides, the IMAC materials performed very well. Different sample preparation strategies were tested, comprising direct dilution with the loading buffer, organic solvent precipitation, and lipid removal from the matrix, as well as the addition of phosphatase inhibitors during sample handling for maximized endogenous phosphopeptide enrichment. All data were acquired by a shotgun peptidomics approach, in which peptide samples were separated by reversed-phase nanoHPLC hyphenated with high-resolution tandem mass spectrometry. The devised method allowed the identification of 176 endogenous phosphopeptides in fresh serum added with inhibitors by the direct dilution protocol and the Ti 4+ -IMAC magnetic material enrichment, but good results could also be obtained from the commercial Fe 3+ -IMAC spin column adapted to the batch enrichment protocol.

  3. Assessing the iron chelation capacity of goat casein digest isolates.

    Science.gov (United States)

    Smialowska, A; Matia-Merino, L; Carr, A J

    2017-04-01

    We isolated goat phosphopeptides via calcium and ethanol precipitation from a caseinate digest and investigated their feasibility as an iron-fortification ingredient in nutritional foods. Goat tryptic-digested phosphopeptides could bind 54.37 ± 0.50 mg of Fe/g of protein compared with goat milk, which could bind 3.83 ± 0.01 mg of Fe/g of protein, indicating that isolation did increase iron binding. However, the >13-fold increase in iron binding was only partly explained by the increased concentration of phosphoserine-rich residues in the isolated fraction: we observed a 77% increase in serine residue content and a 5.9-fold increase in phosphorus in the goat peptide isolate compared with the starting caseinate material. We investigated the effect of potential industrial processing conditions (including heating, cooling, holding time, and processing order) on iron binding by the tryptic-digested phosphopeptides. In addition, we tested the effect of ionic strength and the addition of peptides to a milk system to understand how food formulations could affect iron binding. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  4. Highly selective enrichment of phosphorylated peptides using titanium dioxide

    DEFF Research Database (Denmark)

    Thingholm, Tine; Jørgensen, Thomas J D; Jensen, Ole N

    2006-01-01

    -column. Although phosphopeptide enrichment can be achieved by using TFA and acetonitrile alone, the selectivity is dramatically enhanced by adding DHB or phthalic acid since these compounds, in conjunction with the low pH caused by TFA, prevent binding of nonphosphorylated peptides to TiO2. Using an alkaline...... a protocol for selective phosphopeptide enrichment using titanium dioxide (TiO2) chromatography. The selectivity toward phosphopeptides is obtained by loading the sample in a 2,5-dihydroxybenzoic acid (DHB) or phthalic acid solution containing acetonitrile and trifluoroacetic acid (TFA) onto a TiO2 micro...... solution (pH > or = 10.5) both monophosphorylated and multiphosphorylated peptides are eluted from the TiO2 beads. This highly efficient method for purification of phosphopeptides is well suited for the characterization of phosphoproteins from both in vitro and in vivo studies in combination with mass...

  5. Facile Synthesis of Mesocrystalline SnO2 Nanorods on Reduced Graphene Oxide Sheets: An Appealing Multifunctional Affinity Probe for Sequential Enrichment of Endogenous Peptides and Phosphopeptides.

    Science.gov (United States)

    Ma, Wen; Zhang, Feng; Li, Liping; Chen, Shuai; Qi, Limin; Liu, Huwei; Bai, Yu

    2016-12-28

    A novel multifunctional composite comprising mesocrystalline SnO 2 nanorods (NRs) vertically aligned on reduced graphene oxide (rGO) sheets was synthesized and developed for sequential capture of endogenous peptides and phosphopeptides. With the hydrophobicity of rGO and high affinity of SnO 2 nanorods, sequential enrichment of endogenous peptides and phosphopeptides could be easily achieved through a modulation of elution buffer. With this multifunctional nanomaterial, 36 peptides were observed from diluted bovine serum albumin (BSA) tryptic digest and 4 phosphopeptides could be selectively captured from β-casein digest. The detection limit of tryptic digest of β-casein was low to 4 × 10 -10 M, and the selectivity was up to 1:500 (molar ratio of β-casein and BSA digest). The effectiveness and robustness of rGO-SnO 2 NRs in a complex biological system was also confirmed by using human serum as a real sample. Our work is promising for small peptide enrichment and identification especially in complicated biological sample preparation, which also opens a new perspective in the design of multifunctional affinity probes for proteome or peptidome.

  6. High-Throughput Quantification of SH2 Domain-Phosphopeptide Interactions with Cellulose-Peptide Conjugate Microarrays.

    Science.gov (United States)

    Engelmann, Brett W

    2017-01-01

    The Src Homology 2 (SH2) domain family primarily recognizes phosphorylated tyrosine (pY) containing peptide motifs. The relative affinity preferences among competing SH2 domains for phosphopeptide ligands define "specificity space," and underpins many functional pY mediated interactions within signaling networks. The degree of promiscuity exhibited and the dynamic range of affinities supported by individual domains or phosphopeptides is best resolved by a carefully executed and controlled quantitative high-throughput experiment. Here, I describe the fabrication and application of a cellulose-peptide conjugate microarray (CPCMA) platform to the quantitative analysis of SH2 domain specificity space. Included herein are instructions for optimal experimental design with special attention paid to common sources of systematic error, phosphopeptide SPOT synthesis, microarray fabrication, analyte titrations, data capture, and analysis.

  7. Neighbor-directed histidine N(τ) alkylation. A route to imidazolium-containing phosphopeptide macrocycles

    Energy Technology Data Exchange (ETDEWEB)

    Qian, Wen-Jian [National Cancer Inst., Frederick, MD (United States); Park, Jung-Eun [National Cancer Inst., Bethesda, MD (United States); Grant, Robert [Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Lai, Christopher C. [National Cancer Inst., Frederick, MD (United States); Kelley, James A. [National Cancer Inst., Frederick, MD (United States); Yaffe, Michael B. [Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Lee, Kyung S. [National Cancer Inst., Bethesda, MD (United States); Burke, Terrence R. [National Cancer Inst., Frederick, MD (United States)

    2015-07-07

    Our recently discovered, selective, on-resin route to N(τ)-alkylated imidazolium-containing histidine residues affords new strategies for peptide mimetic design. In this, we demonstrate the use of this chemistry to prepare a series of macrocyclic phosphopeptides, in which imidazolium groups serve as ring-forming junctions. These cationic moieties subsequently serve to charge-mask the phosphoamino acid group that directed their formation. Furthermore, neighbor-directed histidine N(τ)-alkylation opens the door to new families of phosphopeptidomimetics for use in a range of chemical biology contexts.

  8. In vivo Phosphoproteome of Human Skeletal Muscle Revealed by Phosphopeptide Enrichment and HPLC-ESI-MS/MS

    DEFF Research Database (Denmark)

    Højlund, Kurt; Bowen, Benjamin P; Hwang, Hyonson

    2009-01-01

    volunteers. Trypsin digestion of 3-5 mg human skeletal muscle protein was followed by phosphopeptide enrichment using SCX and TiO2. The resulting phosphopeptides were analyzed by HPLC-ESI-MS/MS. Using this unbiased approach, we identified 306 distinct in vivo phosphorylation sites in 127 proteins, including...

  9. Optimized IMAC-IMAC protocol for phosphopeptide recovery from complex biological samples

    DEFF Research Database (Denmark)

    Ye, Juanying; Zhang, Xumin; Young, Clifford

    2010-01-01

    using Fe(III)-NTA IMAC resin and it proved to be highly selective in the phosphopeptide enrichment of a highly diluted standard sample (1:1000) prior to MALDI MS analysis. We also observed that a higher iron purity led to an increased IMAC enrichment efficiency. The optimized method was then adapted...... to phosphoproteome analyses of cell lysates of high protein complexity. From either 20 microg of mouse sample or 50 microg of Drosophila melanogaster sample, more than 1000 phosphorylation sites were identified in each study using IMAC-IMAC and LC-MS/MS. We demonstrate efficient separation of multiply phosphorylated...... characterization of phosphoproteins in functional phosphoproteomics research projects....

  10. Search for phosphopeptides in the feces of axenic rats fed radioactive ovine casein

    International Nuclear Information System (INIS)

    Pelissier, J.P.; Dubos, F.; Daburon, F.

    1981-01-01

    Radioactive ovine casein was obtained by injecting 100 μCi of 14 C-Ser into the jugular vein of an ewe. The milk collected 17 and 24 h after this injection contained 12% of the radioactivity injected in protein form. The seryl residues were specificially labelled. This casein was used as the only protein source fed to axenic rats; 0.30% of the tracer ingested was found in the feces of those rats. Since phosphoserine represented 25% of the total casein seryl residues, the phosphopeptides may not be selectively unabsorbable [fr

  11. Parallel reaction monitoring on a Q Exactive mass spectrometer increases reproducibility of phosphopeptide detection in bacterial phosphoproteomics measurements.

    Science.gov (United States)

    Taumer, Christoph; Griesbaum, Lena; Kovacevic, Alen; Soufi, Boumediene; Nalpas, Nicolas C; Macek, Boris

    2018-03-29

    Increasing number of studies report the relevance of protein Ser/Thr/Tyr phosphorylation in bacterial physiology, yet the analysis of this type of modification in bacteria still presents a considerable challenge. Unlike in eukaryotes, where tens of thousands of phosphorylation events likely occupy more than two thirds of the proteome, the abundance of protein phosphorylation is much lower in bacteria. Even the state-of-the-art phosphopeptide enrichment protocols fail to remove the high background of abundant unmodified peptides, leading to low signal intensity and undersampling of phosphopeptide precursor ions in consecutive data-dependent MS runs. Consequently, large-scale bacterial phosphoproteomic datasets often suffer from poor reproducibility and a high number of missing values. Here we explore the application of parallel reaction monitoring (PRM) on a Q Exactive mass spectrometer in bacterial phosphoproteome analysis, focusing especially on run-to-run sampling reproducibility. In multiple measurements of identical phosphopeptide-enriched samples, we show that PRM outperforms data-dependent acquisition (DDA) in terms of detection frequency, reaching almost complete sampling efficiency, compared to 20% in DDA. We observe a similar trend over multiple heterogeneous phosphopeptide-enriched samples and conclude that PRM shows a great promise in bacterial phosphoproteomics analyses where reproducible detection and quantification of a relatively small set of phosphopeptides is desired. Bacterial phosphorylated peptides occur in low abundance compared to their unmodified counterparts, and are therefore rarely reproducibly detected in shotgun (DDA) proteomics measurements. Here we show that parallel reaction monitoring complements DDA analyses and makes detection of known, targeted phosphopeptides more reproducible. This will be of significance in replicated MS measurements that have a goal to reproducibly detect and quantify phosphopeptides of interest. Copyright

  12. Simultaneous quantification of protein phosphorylation sites using liquid chromatography-tandem mass spectrometry-based targeted proteomics: a linear algebra approach for isobaric phosphopeptides.

    Science.gov (United States)

    Xu, Feifei; Yang, Ting; Sheng, Yuan; Zhong, Ting; Yang, Mi; Chen, Yun

    2014-12-05

    As one of the most studied post-translational modifications (PTM), protein phosphorylation plays an essential role in almost all cellular processes. Current methods are able to predict and determine thousands of phosphorylation sites, whereas stoichiometric quantification of these sites is still challenging. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS)-based targeted proteomics is emerging as a promising technique for site-specific quantification of protein phosphorylation using proteolytic peptides as surrogates of proteins. However, several issues may limit its application, one of which relates to the phosphopeptides with different phosphorylation sites and the same mass (i.e., isobaric phosphopeptides). While employment of site-specific product ions allows for these isobaric phosphopeptides to be distinguished and quantified, site-specific product ions are often absent or weak in tandem mass spectra. In this study, linear algebra algorithms were employed as an add-on to targeted proteomics to retrieve information on individual phosphopeptides from their common spectra. To achieve this simultaneous quantification, a LC-MS/MS-based targeted proteomics assay was first developed and validated for each phosphopeptide. Given the slope and intercept of calibration curves of phosphopeptides in each transition, linear algebraic equations were developed. Using a series of mock mixtures prepared with varying concentrations of each phosphopeptide, the reliability of the approach to quantify isobaric phosphopeptides containing multiple phosphorylation sites (≥ 2) was discussed. Finally, we applied this approach to determine the phosphorylation stoichiometry of heat shock protein 27 (HSP27) at Ser78 and Ser82 in breast cancer cells and tissue samples.

  13. Treatment of post-orthodontic white spot lesions with casein phosphopeptide-stabilised amorphous calcium phosphate

    DEFF Research Database (Denmark)

    Bröchner, Ann; Christensen, Carsten; Kristensen, Bjarne

    2010-01-01

    This study aims to investigate the effect of topical applications of 10% casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) on white spot lesions (WSL) detected after treatment with fixed orthodontic appliances. Sixty healthy adolescents with >/=1 clinically visible WSL at debonding were...... findings were largely reflected by the clinical scores. No side effects were reported. Topical treatment of white spot lesions after debonding of orthodontic appliances with a casein phosphopeptide-stabilised amorphous calcium phosphate agent resulted in significantly reduced fluorescence and a reduced...

  14. Phosphopeptide enrichment with inorganic nanofibers prepared by forcespinning technology

    Czech Academy of Sciences Publication Activity Database

    Křenková, Jana; Morávková, J.; Buk, J.; Foret, František

    2016-01-01

    Roč. 1427, JAN (2016), s. 8-15 ISSN 0021-9673 R&D Projects: GA ČR(CZ) GA14-06319S; GA ČR(CZ) GBP206/12/G014 Institutional support: RVO:68081715 Keywords : nanofibers * enrichment * phosphopeptides Subject RIV: CB - Analytical Chemistry , Separation Impact factor: 3.981, year: 2016

  15. Phosphopeptide enrichment with inorganic nanofibers prepared by forcespinning technology

    Czech Academy of Sciences Publication Activity Database

    Křenková, Jana; Morávková, J.; Buk, J.; Foret, František

    2016-01-01

    Roč. 1427, JAN (2016), s. 8-15 ISSN 0021-9673 R&D Projects: GA ČR(CZ) GA14-06319S; GA ČR(CZ) GBP206/12/G014 Institutional support: RVO:68081715 Keywords : nanofibers * enrichment * phosphopeptides Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 3.981, year: 2016

  16. Cyclic phosphopeptides for interference with Grb2 SH2 domain signal transduction prepared by ring-closing metathesis and phosphorylation

    NARCIS (Netherlands)

    Dekker, Frank J; de Mol, Nico J; Fischer, Marcel J E; Kemmink, Johan; Liskamp, Rob M J; Dekker, Frank

    2003-01-01

    Cyclic phosphopeptides were prepared using ring-closing metathesis followed by phosphorylation. These cyclic phosphopeptides were designed to interact with the SH2 domain of Grb2, which is a signal transduction protein of importance as a target for antiproliferative drug development. Binding of

  17. Neighbor-Directed Histidine N (s)–Alkylation: A Route to Imidazolium-Containing Phosphopeptide Macrocycles-Biopolymers | Center for Cancer Research

    Science.gov (United States)

    Our recently discovered, selective, on-resin route to N(s)-alkylated imidazolium-containing histidine residues affords new strategies for peptide mimetic design. In this, we demonstrate the use of this chemistry to prepare a series of macrocyclic phosphopeptides, in which imidazolium groups serve as ring-forming junctions. Interestingly, these cationic moieties subsequently serve to charge-mask the phosphoamino acid group that directed their formation.

  18. Casein phosphopeptides drastically increase the secretion of extracellular proteins in Aspergillus awamori. Proteomics studies reveal changes in the secretory pathway.

    Science.gov (United States)

    Kosalková, Katarina; García-Estrada, Carlos; Barreiro, Carlos; Flórez, Martha G; Jami, Mohammad S; Paniagua, Miguel A; Martín, Juan F

    2012-01-10

    The secretion of heterologous animal proteins in filamentous fungi is usually limited by bottlenecks in the vesicle-mediated secretory pathway. Using the secretion of bovine chymosin in Aspergillus awamori as a model, we found a drastic increase (40 to 80-fold) in cells grown with casein or casein phosphopeptides (CPPs). CPPs are rich in phosphoserine, but phosphoserine itself did not increase the secretion of chymosin. The stimulatory effect is reduced about 50% using partially dephosphorylated casein and is not exerted by casamino acids. The phosphopeptides effect was not exerted at transcriptional level, but instead, it was clearly observed on the secretion of chymosin by immunodetection analysis. Proteomics studies revealed very interesting metabolic changes in response to phosphopeptides supplementation. The oxidative metabolism was reduced, since enzymes involved in fermentative processes were overrepresented. An oxygen-binding hemoglobin-like protein was overrepresented in the proteome following phosphopeptides addition. Most interestingly, the intracellular pre-protein enzymes, including pre-prochymosin, were depleted (most of them are underrepresented in the intracellular proteome after the addition of CPPs), whereas the extracellular mature form of several of these secretable proteins and cell-wall biosynthetic enzymes was greatly overrepresented in the secretome of phosphopeptides-supplemented cells. Another important 'moonlighting' protein (glyceraldehyde-3-phosphate dehydrogenase), which has been described to have vesicle fusogenic and cytoskeleton formation modulating activities, was clearly overrepresented in phosphopeptides-supplemented cells. In summary, CPPs cause the reprogramming of cellular metabolism, which leads to massive secretion of extracellular proteins.

  19. Synthesis of Thermally Switchable Chromatographic Materials with Immobilized Ti4+ for Enrichment of Phosphopeptides by Reversible Addition Fragmentation Chain Transfer Polymerization

    Science.gov (United States)

    Wang, Di; Cao, Zhihan; Pang, Xinzhu; Deng, Yulin; Li, Bo; Dai, Rongji

    2018-01-01

    Reversible phosphorylation of proteins is one of the most crucial types of post-translational modifications (PTMs). And it shows significant work in diversified biological processes. However, the separation technology of phosphorylated peptides is still an analytical challenge in phosphoproteomics, because phosphopeptides are alway in low stoichiometry. Thus, enrichment of phosphopeptides before detection is indispensable. In this study, a novel temperature regulated separation protocol was developed. Silica@p (NIPAAm-co-IPPA)-Ti4+, a new Ti(IV)-IMAC (Immobilized Metal Affinity chromatography) materials was synthesized by reversible addition fragmentation chain transfer polymerization (RAFT). By the unique thermally responsive properties of poly(N-isopropylacrylamide) (PNIPAAm), the captured phosphorylated peptides could be released by changing temperature only without applying any other eluant which could damage the phosphopeptides. We employed isopropanol phosphonic acid (IPPA) as an IMAC ligand for the immobilization of Ti(IV) which could increase the specific adsorption of phosphopeptides. The enrichment and release properties were examined by treatment with pyridoxal 5’-phosphate (PLP) and casein phosphopeptides (CPP). Two phosphorylated compounds above have temperature-stimulated binding to Ti4+. Finally, silica@p (NIPAAm-co-IPPA)-Ti4+ was successfully employed in pretreatment of phosphopeptides in a tryptic digest of a-casein and human serum albumin (HSA). The results indicated a great potential of this new temperature-responsive material in phosphoproteomics study.

  20. Casein phosphopeptides drastically increase the secretion of extracellular proteins in Aspergillus awamori. Proteomics studies reveal changes in the secretory pathway

    Directory of Open Access Journals (Sweden)

    Kosalková Katarina

    2012-01-01

    Full Text Available Abstract Background The secretion of heterologous animal proteins in filamentous fungi is usually limited by bottlenecks in the vesicle-mediated secretory pathway. Results Using the secretion of bovine chymosin in Aspergillus awamori as a model, we found a drastic increase (40 to 80-fold in cells grown with casein or casein phosphopeptides (CPPs. CPPs are rich in phosphoserine, but phosphoserine itself did not increase the secretion of chymosin. The stimulatory effect is reduced about 50% using partially dephosphorylated casein and is not exerted by casamino acids. The phosphopeptides effect was not exerted at transcriptional level, but instead, it was clearly observed on the secretion of chymosin by immunodetection analysis. Proteomics studies revealed very interesting metabolic changes in response to phosphopeptides supplementation. The oxidative metabolism was reduced, since enzymes involved in fermentative processes were overrepresented. An oxygen-binding hemoglobin-like protein was overrepresented in the proteome following phosphopeptides addition. Most interestingly, the intracellular pre-protein enzymes, including pre-prochymosin, were depleted (most of them are underrepresented in the intracellular proteome after the addition of CPPs, whereas the extracellular mature form of several of these secretable proteins and cell-wall biosynthetic enzymes was greatly overrepresented in the secretome of phosphopeptides-supplemented cells. Another important 'moonlighting' protein (glyceraldehyde-3-phosphate dehydrogenase, which has been described to have vesicle fusogenic and cytoskeleton formation modulating activities, was clearly overrepresented in phosphopeptides-supplemented cells. Conclusions In summary, CPPs cause the reprogramming of cellular metabolism, which leads to massive secretion of extracellular proteins.

  1. Improved detection of hydrophilic phosphopeptides using graphite powder microcolumns and mass spectrometry: evidence for in vivo doubly phosphorylated dynamin I and dynamin III

    DEFF Research Database (Denmark)

    Larsen, Martin Røssel; Graham, Mark E; Robinson, Phillip J

    2004-01-01

    A common strategy in proteomics to improve the number and quality of peptides detected by mass spectrometry (MS) is to desalt and concentrate proteolytic digests using reversed phase (RP) chromatography prior to analysis. However, this does not allow for detection of small or hydrophilic peptides...... a large improvement in the detection of small amounts of phosphopeptides by MS and the approach has major implications for both small- and large-scale projects in phosphoproteomics.......A common strategy in proteomics to improve the number and quality of peptides detected by mass spectrometry (MS) is to desalt and concentrate proteolytic digests using reversed phase (RP) chromatography prior to analysis. However, this does not allow for detection of small or hydrophilic peptides......, or peptides altered in hydrophilicity such as phosphopeptides. We used microcolumns to compare the ability of RP resin or graphite powder to retain phosphopeptides. A number of standard phosphopeptides and a biologically relevant phosphoprotein, dynamin I, were analyzed. MS revealed that some phosphopeptides...

  2. Incorporation of casein phosphopeptide-amorphous calcium phosphate into a glass-ionomer cement.

    Science.gov (United States)

    Mazzaoui, S A; Burrow, M F; Tyas, M J; Dashper, S G; Eakins, D; Reynolds, E C

    2003-11-01

    Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) nanocomplexes have been shown to prevent demineralization and promote remineralization of enamel subsurface lesions in animal and in situ caries models. The aim of this study was to determine the effect of incorporating CPP-ACP into a self-cured glass-ionomer cement (GIC). Incorporation of 1.56% w/w CPP-ACP into the GIC significantly increased microtensile bond strength (33%) and compressive strength (23%) and significantly enhanced the release of calcium, phosphate, and fluoride ions at neutral and acidic pH. MALDI mass spectrometry also showed casein phosphopeptides from the CPP-ACP nanocomplexes to be released. The release of CPP-ACP and fluoride from the CPP-ACP-containing GIC was associated with enhanced protection of the adjacent dentin during acid challenge in vitro.

  3. The decreased of Streptococcus Mutans growth after topical application of phosphopeptide amorphous calcium phosphate paste

    Directory of Open Access Journals (Sweden)

    Tika Faradina Araf

    2011-07-01

    Full Text Available Casein Phosphopeptide-Amorphous Calcium Phosphate (CPP-ACP paste is a topical application substance that consisted of a series of milk derivative peptide as a result of phosphorylation and has an antibacterial activity. The objective of this research was to find out the difference of Streptococcus mutans growth before and after CPP-ACP paste given topically to child's teeth. The method of the research was a quasi-experiment. Research samples were 10 students of MI Al Falah Islamic Boarding School, Jatinangor, West Jawa Indonesia and collected with purposive sampling technique. This research used dental plaque from child's teeth before and after applicated by CPP-ACP paste. The plaque was cultivated in selective media Tryptone Yeast Cysteine Sucrose Bacitracin (TYCSB with repeated twice. Streptococcus mutans colony in TYCSB were counted by Stuart colony counter and statistically analyzed based on paired t-test. The results showed the average of Streptococcus mutans growth before applicated CPP-ACP paste was 57.05, whereas after applicated CPP-ACP paste for 1 days was 9.4; for 3 days was 2.85, and for 14 days was 1.7 colony. The research concluded that there was a decrease of Streptococcus mutans growth in isolate plaque after CPP-ACP paste topically given to child's teeth.

  4. Evaluation of phosphopeptide enrichment strategies for quantitative TMT analysis of complex network dynamics in cancer-associated cell signalling

    Directory of Open Access Journals (Sweden)

    Benedetta Lombardi

    2015-03-01

    Full Text Available Defining alterations in signalling pathways in normal and malignant cells is becoming a major field in proteomics. A number of different approaches have been established to isolate, identify and quantify phosphorylated proteins and peptides. In the current report, a comparison between SCX prefractionation versus an antibody based approach, both coupled to TiO2 enrichment and applied to TMT labelled cellular lysates, is described. The antibody strategy was more complete for enriching phosphopeptides and allowed the identification of a large set of proteins known to be phosphorylated (715 protein groups with a minimum number of not previously known phosphorylated proteins (2.

  5. Enzymatic Dissolution of Biocomposite Solids Consisting of Phosphopeptides to Form Supramolecular Hydrogels

    KAUST Repository

    Shi, Junfeng; Yuan, Dan; Haburcak, Richard; Zhang, Qiang; Zhao, Chao; Zhang, Xixiang; Xu, Bing

    2015-01-01

    Enzyme-catalyzed dephosphorylation is essential for biomineralization and bone metabolism. Here we report the exploration of using enzymatic reaction to transform biocomposites of phosphopeptides and calcium (or strontium) ions to supramolecular hydrogels as a mimic of enzymatic dissolution of biominerals. 31P NMR shows that strong affinity between the phosphopeptides and alkaline metal ions (e.g., Ca2+ or Sr2+) induces the formation of biocomposites as precipitates. Electron microscopy reveals that the enzymatic reaction regulates the morphological transition from particles to nanofibers. Rheology confirms the formation of a rigid hydrogel. As the first example of enzyme-instructed dissolution of a solid to form supramolecular nanofibers/hydrogels, this work provides an approach to generate soft materials with desired properties, expands the application of supramolecular hydrogelators, and offers insights to control the demineralization of calcified soft tissues.

  6. Enzymatic Dissolution of Biocomposite Solids Consisting of Phosphopeptides to Form Supramolecular Hydrogels

    KAUST Repository

    Shi, Junfeng

    2015-10-14

    Enzyme-catalyzed dephosphorylation is essential for biomineralization and bone metabolism. Here we report the exploration of using enzymatic reaction to transform biocomposites of phosphopeptides and calcium (or strontium) ions to supramolecular hydrogels as a mimic of enzymatic dissolution of biominerals. 31P NMR shows that strong affinity between the phosphopeptides and alkaline metal ions (e.g., Ca2+ or Sr2+) induces the formation of biocomposites as precipitates. Electron microscopy reveals that the enzymatic reaction regulates the morphological transition from particles to nanofibers. Rheology confirms the formation of a rigid hydrogel. As the first example of enzyme-instructed dissolution of a solid to form supramolecular nanofibers/hydrogels, this work provides an approach to generate soft materials with desired properties, expands the application of supramolecular hydrogelators, and offers insights to control the demineralization of calcified soft tissues.

  7. The use of titanium dioxide for selective enrichment of phosphorylated peptides

    DEFF Research Database (Denmark)

    Thingholm, Tine E.; Larsen, Martin R.

    2016-01-01

    acid (DHB), phthalic acid, lactic acid, or glycolic acid has been shown to improve selectivity significantly by reducing unspecific binding of non-phosphorylated peptides. The phosphopeptides bound to the TiO2 are subsequently eluted from the chromatographic material using an alkaline buffer. TiO2......Titanium dioxide (TiO2) has very high affinity for phosphopeptides and in recent years it has become one of the most popular methods for phosphopeptide enrichment from complex biological samples. Peptide loading onto TiO2 resin in a highly acidic environment in the presence of 2,5-dihydroxybenzoic...... chromatography is extremely tolerant towards most buffers used in biological experiments, highly robust and as such it has become the method of choice in large-scale phosphoproteomics. Here we describe a batch mode protocol for phosphopeptide enrichment using TiO2 chromatographic material followed by desalting...

  8. Comparative evaluation of Nano-hydroxyapatite and casein Phosphopeptide-amorphous calcium phosphate on the remineralization potential of early enamel lesions: An in vitro study

    Directory of Open Access Journals (Sweden)

    Anshul Sharma

    2017-01-01

    Full Text Available Background: Benefits of remineralizing agents in a wide variety of formulations have been proved beneficial in caries management. Casein phosphopeptide-amorphous calcium phosphate (CPP–ACP nanocomplex has been recommended and used as remineralizing agent. Nano-hydroxyapatite (n-HAp is one of the most biocompatible and bioactive material having wide range of application in dentistry, but does it excel better compared to CPP-ACP. Aims: To evaluate and compare the remineralizing efficiency of the paste containing hydroxyapatite and casein phosphopeptide-amorphous calcium phosphate. Settings and Design: The study was an in vitro single blinded study with lottery method of randomization approved by the Institutional Ethics Committee. Materials and methods: 30 non carious premolar teeth. The teeth were demineralized and divided into 2 groups and subjected to remineralization. The samples were analysed for surface hardness and mineral content. Statistical Analysis: Student t’ test and repeated measures of ANOVA was applied. Results: Average hardness in Nano-hydroxyapatite group increased to 340 ± 31.70 SD and 426 ± 50.62 SD for 15 and 30 days respectively and that of (CPP–ACP, 355.83 ± 38.55 SD and 372.67 ± 53.63 SD. The change in the hardness values was not statistically significant with P value of 0.39 (P > 0.05. Calcium and Phosphorous levels increased in both the groups but was not significant. Conclusion: Both the agents used are effective in causing remineralization of enamel. Nano-hydroxyapatite is more effective as compared to Casein phosphopeptide-amorphous calcium phosphate, in increasing the Calcium and Phosphorus content of enamel, and this effect is more evident over a longer treatment period. Key Message: Remineralizing agents are a boon for caries management. With the advent of many formulations it is difficult to clinically select the agent. This study compares the remineralizing potential of Casein

  9. A Multidimensional System for Phosphopeptide Analysis Using TiO{sub 2} Enrichment and Ion-exchange Chromatography with Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Cho, Kun; Yoo, Jisun; Kim, Eunmin; Kim, Jin Young; Kim, Young Hwan; Yoo, Jong Shin [Korea Basic Science Institute, Ochang (Korea, Republic of); Oh, Han Bin [Sogang Univ., Seoul (Korea, Republic of)

    2012-10-15

    Although offline enrichment of phosphorylated peptides is widely used, enrichment for phosphopeptides using TiO{sub 2} is often performed manually, which is labor-intensive and can lead to irreproducible results. To address the problems associated with offline enrichment and to improve the effectiveness of phosphopeptide detection, we developed an automated online enrichment system for phosphopeptide analysis. A standard protein mixture comprising BSA, fetuin, crystalline, α-casein and β-casein, and ovalbumin was assessed using our new system. Our multidimensional system has four main parts: a sample pump, a 20-mm TiO{sub 2}-based column, a weak anion-exchange, and a strong cation-exchange (2:1 WAX:SCX) separation column with LC/MS. Phosphorylated peptides were successfully detected using the TiO{sub 2}-based online system with little interference from nonphosphorylated peptides. Our results confirmed that our online enrichment system is a simple and efficient method for detecting phosphorylated peptides.

  10. Structural insights into the recognition of phosphopeptide by the FHA domain of kanadaptin

    DEFF Research Database (Denmark)

    Xu, Qingping; Deller, Marc C; Nielsen, Tine K

    2014-01-01

    with a phosphopeptide mimic derived from a peptide segment from the N-terminus of a symmetry-related molecule as well as a sulfate bound to the structurally conserved phosphothreonine recognition cleft. This structure provides insights into the molecular recognition features utilized by this family of proteins...

  11. Comparison of Zirconium Phosphonate-Modified Surfaces for Immobilizing Phosphopeptides and Phosphate-Tagged Proteins.

    Science.gov (United States)

    Forato, Florian; Liu, Hao; Benoit, Roland; Fayon, Franck; Charlier, Cathy; Fateh, Amina; Defontaine, Alain; Tellier, Charles; Talham, Daniel R; Queffélec, Clémence; Bujoli, Bruno

    2016-06-07

    Different routes for preparing zirconium phosphonate-modified surfaces for immobilizing biomolecular probes are compared. Two chemical-modification approaches were explored to form self-assembled monolayers on commercially available primary amine-functionalized slides, and the resulting surfaces were compared to well-characterized zirconium phosphonate monolayer-modified supports prepared using Langmuir-Blodgett methods. When using POCl3 as the amine phosphorylating agent followed by treatment with zirconyl chloride, the result was not a zirconium-phosphonate monolayer, as commonly assumed in the literature, but rather the process gives adsorbed zirconium oxide/hydroxide species and to a lower extent adsorbed zirconium phosphate and/or phosphonate. Reactions giving rise to these products were modeled in homogeneous-phase studies. Nevertheless, each of the three modified surfaces effectively immobilized phosphopeptides and phosphopeptide tags fused to an affinity protein. Unexpectedly, the zirconium oxide/hydroxide modified surface, formed by treating the amine-coated slides with POCl3/Zr(4+), afforded better immobilization of the peptides and proteins and efficient capture of their targets.

  12. Enrichment and characterization of phosphopeptides by immobilized metal affinity chromatography (IMAC) and mass spectrometry

    DEFF Research Database (Denmark)

    Thingholm, Tine E; Jensen, Ole N

    2009-01-01

    The combination of immobilized metal affinity chromatography (IMAC) and mass spectrometry is a widely used technique for enrichment and sequencing of phosphopeptides. In the IMAC method, negatively charged phosphate groups interact with positively charged metal ions (Fe3+, Ga3+, and Al3...

  13. An SH2 domain model of STAT5 in complex with phospho-peptides define "STAT5 Binding Signatures".

    Science.gov (United States)

    Gianti, Eleonora; Zauhar, Randy J

    2015-05-01

    The signal transducer and activator of transcription 5 (STAT5) is a member of the STAT family of proteins, implicated in cell growth and differentiation. STAT activation is regulated by phosphorylation of protein monomers at conserved tyrosine residues, followed by binding to phospho-peptide pockets and subsequent dimerization. STAT5 is implicated in the development of severe pathological conditions, including many cancer forms. However, nowadays a few STAT5 inhibitors are known, and only one crystal structure of the inactive STAT5 dimer is publicly available. With a view to enabling structure-based drug design, we have: (1) analyzed phospho-peptide binding pockets on SH2 domains of STAT5, STAT1 and STAT3; (2) generated a model of STAT5 bound to phospho-peptides; (3) assessed our model by docking against a class of known STAT5 inhibitors (Müller et al. in ChemBioChem 9:723-727, 2008); (4) used molecular dynamics simulations to optimize the molecular determinants responsible for binding and (5) proposed unique "Binding Signatures" of STAT5. Our results put in place the foundations to address STAT5 as a target for rational drug design, from sequence, structural and functional perspectives.

  14. An SH2 domain model of STAT5 in complex with phospho-peptides define ``STAT5 Binding Signatures''

    Science.gov (United States)

    Gianti, Eleonora; Zauhar, Randy J.

    2015-05-01

    The signal transducer and activator of transcription 5 (STAT5) is a member of the STAT family of proteins, implicated in cell growth and differentiation. STAT activation is regulated by phosphorylation of protein monomers at conserved tyrosine residues, followed by binding to phospho-peptide pockets and subsequent dimerization. STAT5 is implicated in the development of severe pathological conditions, including many cancer forms. However, nowadays a few STAT5 inhibitors are known, and only one crystal structure of the inactive STAT5 dimer is publicly available. With a view to enabling structure-based drug design, we have: (1) analyzed phospho-peptide binding pockets on SH2 domains of STAT5, STAT1 and STAT3; (2) generated a model of STAT5 bound to phospho-peptides; (3) assessed our model by docking against a class of known STAT5 inhibitors (Müller et al. in ChemBioChem 9:723-727, 2008); (4) used molecular dynamics simulations to optimize the molecular determinants responsible for binding and (5) proposed unique "Binding Signatures" of STAT5. Our results put in place the foundations to address STAT5 as a target for rational drug design, from sequence, structural and functional perspectives.

  15. 3-Aminoquinoline/p-coumaric acid as a MALDI matrix for glycopeptides, carbohydrates, and phosphopeptides.

    Science.gov (United States)

    Fukuyama, Yuko; Funakoshi, Natsumi; Takeyama, Kohei; Hioki, Yusaku; Nishikaze, Takashi; Kaneshiro, Kaoru; Kawabata, Shin-Ichirou; Iwamoto, Shinichi; Tanaka, Koichi

    2014-02-18

    Glycosylation and phosphorylation are important post-translational modifications in biological processes and biomarker research. The difficulty in analyzing these modifications is mainly their low abundance and dissociation of labile regions such as sialic acids or phosphate groups. One solution in matrix-assisted laser desorption/ionization (MALDI) mass spectrometry is to improve matrices for glycopeptides, carbohydrates, and phosphopeptides by increasing the sensitivity and suppressing dissociation of the labile regions. Recently, a liquid matrix 3-aminoquinoline (3-AQ)/α-cyano-4-hydroxycinnamic acid (CHCA) (3-AQ/CHCA), introduced by Kolli et al. in 1996, has been reported to increase sensitivity for carbohydrates or phosphopeptides, but it has not been systematically evaluated for glycopeptides. In addition, 3-AQ/CHCA enhances the dissociation of labile regions. In contrast, a liquid matrix 1,1,3,3-tetramethylguanidium (TMG, G) salt of p-coumaric acid (CA) (G3CA) was reported to suppress dissociation of sulfate groups or sialic acids of carbohydrates. Here we introduce a liquid matrix 3-AQ/CA for glycopeptides, carbohydrates, and phosphopeptides. All of the analytes were detected as [M + H](+) or [M - H](-) with higher or comparable sensitivity using 3-AQ/CA compared with 3-AQ/CHCA or 2,5-dihydroxybenzoic acid (2,5-DHB). The sensitivity was increased 1- to 1000-fold using 3-AQ/CA. The dissociation of labile regions such as sialic acids or phosphate groups and the fragmentation of neutral carbohydrates were suppressed more using 3-AQ/CA than using 3-AQ/CHCA or 2,5-DHB. 3-AQ/CA was thus determined to be an effective MALDI matrix for high sensitivity and the suppression of dissociation of labile regions in glycosylation and phosphorylation analyses.

  16. Three-dimensional ordered titanium dioxide-zirconium dioxide film-based microfluidic device for efficient on-chip phosphopeptide enrichment.

    Science.gov (United States)

    Zhao, De; He, Zhongyuan; Wang, Gang; Wang, Hongzhi; Zhang, Qinghong; Li, Yaogang

    2016-09-15

    Microfluidic technology plays a significant role in separating biomolecules, because of its miniaturization, integration, and automation. Introducing micro/nanostructured functional materials can improve the properties of microfluidic devices, and extend their application. Inverse opal has a three-dimensional ordered net-like structure. It possesses a large surface area and exhibits good mass transport, making it a good candidate for bio-separation. This study exploits inverse opal titanium dioxide-zirconium dioxide films for on-chip phosphopeptide enrichment. Titanium dioxide-zirconium dioxide inverse opal film-based microfluidic devices were constructed from templates of 270-, 340-, and 370-nm-diameter poly(methylmethacrylate) spheres. The phosphopeptide enrichments of these devices were determined by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. The device constructed from the 270-nm-diameter sphere template exhibited good comprehensive phosphopeptide enrichment, and was the best among these three devices. Because the size of opal template used in construction was the smallest, the inverse opal film therefore had the smallest pore sizes and the largest surface area. Enrichment by this device was also better than those of similar devices based on nanoparticle films and single component films. The titanium dioxide-zirconium dioxide inverse opal film-based device provides a promising approach for the efficient separation of various biomolecules. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. Iminodiacetic acid-modified magnetic poly(2-hydroxyethyl methacrylate)-based microspheres for phosphopeptide enrichment

    Czech Academy of Sciences Publication Activity Database

    Novotná, L.; Emmerová, T.; Horák, Daniel; Kučerová, Z.; Tichá, M.

    2010-01-01

    Roč. 1217, č. 51 (2010), s. 8032-8040 ISSN 0021-9673 R&D Projects: GA AV ČR(CZ) KAN401220801; GA ČR GA203/09/0857; GA ČR GAP503/10/0664 Institutional research plan: CEZ:AV0Z40500505 Keywords : IMAC phosphopeptide separation * IDA-modified magnetic microspheres * Porcine pepsin A Subject RIV: EE - Microbiology, Virology Impact factor: 4.194, year: 2010

  18. Direct effects of casein phosphopeptides on growth and differentiation of in vitro cultured osteoblastic cells (MC3T3-E1).

    Science.gov (United States)

    Tulipano, Giovanni; Bulgari, Omar; Chessa, Stefania; Nardone, Alessandro; Cocchi, Daniela; Caroli, Anna

    2010-02-25

    Casein phosphopeptides (CPPs) obtained by enzymatic hydrolysis in vitro of caseins, have been shown to enhance calcium solubility and to increase the calcification of embryonic rat bones in their diaphyseal area. Little is known about the direct effects of CPPs on cultured osteoblastic cells. Calcium in the microenvironment surrounding bone cells is not only important for the mineralization of the extracellular matrix, but it is believed to provide preosteblasts with a signal that modulates their proliferation and differentiation. The aim of the present study was to investigate the direct effects of four selected casein phosphopeptides on osteoblastic cell (MC3T3-E1 cells) viability and differentiation. The selected peptides have been obtained by chemical synthesis and differed in the number of phosphorylated sites and in the amino acid spacing out two phosphorylated sites, in order to further characterize the relationship between structure and function. The results obtained in this work demonstrated that CPPs may directly affect osteoblast-like cell growth, calcium uptake and ultimately calcium deposition in the extracellular matrix. The effects exerted by distinct CPPs on osteogenesis in vitro can be either stimulatory or inhibitory. Differential short amino acid sequences in their molecules, like the -SpEE- and the -SpTSpEE-motifs, are likely the molecular determinants for their biological activities on osteoblastic cells. Moreover, two genetic variants of CPPs showing one amino acid change in their sequence may profoundly differ in their biological activities. Finally, our data may also suggest important clues about the role of intrinsic phosphorylated peptides derived from endogenous phosphorylated proteins in bone metabolism, apart from extrinsic CPPs. Copyright 2009 Elsevier B.V. All rights reserved.

  19. Selection and characterisation of monepantel resistance in Teladorsagia circumcincta isolates

    Directory of Open Access Journals (Sweden)

    D.J. Bartley

    2015-08-01

    To address these questions three MPTL-resistant Teladorsagia circumcincta isolates (MTci2-11, MTci5-13 and MTci7-12 have been artificially selected in vivo from phenotypically characterised parent isolates (MTci2, MTci5, MTci7 respectively. The selection process involved collecting and culturing eggs from surviving worms from sheep administered sub-optimal dosages of MPTL (Zolvix® to provide infective larvae to infect further sheep until resistant isolates were generated (between 9 and 13 rounds of selection. A controlled efficacy test was conducted using the original parental isolates and the newly generated MPTL resistant isolates (n = 5 per group. Selected isolates were assessed both under anthelmintic stress (Zolvix®, 2.5 mg/kg bodyweight; MTci-MPTL and at rest (untreated, MTci-CON. A number of life-history traits were assessed, namely, worm establishment rates, time to patency, faecal egg output, body length of adults and eggs in utero. The estimated resistance status of the selected isolates was confirmed with 48%, 28% and 9% reductions in worm burden at 7-days post Zolvix® administration for MTci2-11-MPTL, MTci5-13-MPTL and MTci7-12-MPTL, respectively, compared with untreated controls. One of the selected isolates MTci7-12-CON showed significantly greater total worm burden (p = 0.025, greater establishment rate (p = 0.033, decreased time to patency (p = 0.048, higher cumulative egg outputs (p = 0.002 compared with its parental derivative MTci7. The trial results suggest that anthelmintic selection in T. circumcincta, albeit under experimental conditions, can select for more prolific/fecund and quicker maturing populations. These data provide an insight into how parasites evolve in response to anthelmintic pressure.

  20. Selection and characterisation of monepantel resistance in Teladorsagia circumcincta isolates.

    Science.gov (United States)

    Bartley, D J; Devin, L; Nath, M; Morrison, A A

    2015-08-01

    Monepantel (MPTL) is one of two new anthelmintic compounds introduced onto the sheep market to control gastro-intestinal nematodes. Resistance to this compound is rare but has been reported. In order to preserve the efficacy of this and other anthelmintics, it is essential to understand both (a) the mechanisms involved in the selection of resistance and (b) how the parasites evolve to deal with these compounds. To address these questions three MPTL-resistant Teladorsagia circumcincta isolates (MTci2-11, MTci5-13 and MTci7-12) have been artificially selected in vivo from phenotypically characterised parent isolates (MTci2, MTci5, MTci7 respectively). The selection process involved collecting and culturing eggs from surviving worms from sheep administered sub-optimal dosages of MPTL (Zolvix®) to provide infective larvae to infect further sheep until resistant isolates were generated (between 9 and 13 rounds of selection). A controlled efficacy test was conducted using the original parental isolates and the newly generated MPTL resistant isolates (n = 5 per group). Selected isolates were assessed both under anthelmintic stress (Zolvix®, 2.5 mg/kg bodyweight; MTci-MPTL) and at rest (untreated, MTci-CON). A number of life-history traits were assessed, namely, worm establishment rates, time to patency, faecal egg output, body length of adults and eggs in utero. The estimated resistance status of the selected isolates was confirmed with 48%, 28% and 9% reductions in worm burden at 7-days post Zolvix® administration for MTci2-11-MPTL, MTci5-13-MPTL and MTci7-12-MPTL, respectively, compared with untreated controls. One of the selected isolates MTci7-12-CON showed significantly greater total worm burden (p = 0.025), greater establishment rate (p = 0.033), decreased time to patency (p = 0.048), higher cumulative egg outputs (p = 0.002) compared with its parental derivative MTci7. The trial results suggest that anthelmintic selection in T

  1. Selective medium for the isolation of Bacteroides gingivalis.

    Science.gov (United States)

    Hunt, D E; Jones, J V; Dowell, V R

    1986-03-01

    Bacteroides gingivalis has been implicated in various forms of periodontal disease and may be responsible for other diseases in humans. The role of B. gingivalis in disease has been difficult to assess, because it is inhibited by most selective media commonly used by clinical laboratories to aid in isolating gram-negative, nonsporeforming anaerobes. We have developed a new medium, Bacteroides gingivalis agar, which contains bacitracin, colistin, and nalidixic acid as selective agents. This medium allowed B. gingivalis to be isolated from oral specimens with little difficulty and also allowed B. gingivalis to be isolated from phenotypically similar Bacteroides species, such as B. asaccharolyticus and B. endodontalis, with which it can easily be confused.

  2. Effect of Casein Phosphopeptide-Amorphous Calcium Phosphate and Three Calcium Phosphate on Enamel Microhardness.

    Science.gov (United States)

    Haghgou, En Hr; Haghgoo, Roza; Roholahi, Mohamad R; Ghorbani, Zahra

    2017-07-01

    This study aims to investigate the effect of casein phos-phopeptide-amorphous calcium phosphate and three calcium phosphate (CPP-ACP and TCP) on increasing the microhardness of human enamel after induction of erosion. A total of 26 healthy human-impacted third molar teeth were chosen, and their hardness measured using a microhardness testing machine. The samples were immersed in Coca Cola (pH = 4.7) for 8 minutes. Then, micro-hardness was measured again, and these samples were randomly divided into four groups (two control groups and two experimental groups). (1) Negative control group: Artificial saliva was used for 10 minutes, (2) positive control group: Fluoride gel was used for 10 minutes, (3) β-TCP group: TCP was used for 10 minutes, (4) CCP-ACP group: CCP-ACP was used for 10 minutes. The final microhardness of those samples was measured, and the changes in microhardness of teeth within group and between groups were analyzed using the paired and analysis of variance tests respectively. Results were considered statistically significant at a level of p < 0.05. No significant difference was observed in microhard-ness between CPP-ACP group and TCP group (p = 0.368) during the time microhardness significantly dropped after soaking in soda. Casein phosphopeptide-amorphous calcium phosphate and TCP increased the microhardness of teeth. The increase in hardness in the TCP group was higher than in the CPP-ACP group, but this difference was not significant (p = 0.36). Casein phosphopeptide-amorphous calcium phosphate and TCP can affect the remineralization of erosive lesions.

  3. Structure of a 14-3-3σ–YAP phosphopeptide complex at 1.15 Å resolution

    International Nuclear Information System (INIS)

    Schumacher, Benjamin; Skwarczynska, Malgorzata; Rose, Rolf; Ottmann, Christian

    2010-01-01

    The first structure of a 14-3-3 protein–phosphopeptide complex is reported at 1.15 Å resolution. The YAP 14-3-3-binding motif is revealed for the first time using crystallographic tools. The 14-3-3 proteins are a class of eukaryotic acidic adapter proteins, with seven isoforms in humans. 14-3-3 proteins mediate their biological function by binding to target proteins and influencing their activity. They are involved in pivotal pathways in the cell such as signal transduction, gene expression, enzyme activation, cell division and apoptosis. The Yes-associated protein (YAP) is a WW-domain protein that exists in two transcript variants of 48 and 54 kDa in humans. By transducing signals from the cytoplasm to the nucleus, YAP is important for transcriptional regulation. In both variants, interaction with 14-3-3 proteins after phosphorylation of Ser127 is important for nucleocytoplasmic trafficking, via which the localization of YAP is controlled. In this study, 14-3-3σ has been cloned, purified and crystallized in complex with a phosphopeptide from the YAP 14-3-3-binding domain, which led to a crystal that diffracted to 1.15 Å resolution. The crystals belonged to space group C222 1 , with unit-cell parameters a = 82.3, b = 112.1, c = 62.9 Å

  4. An improved culture method for selective isolation of Campylobacter jejuni from wastewater

    Directory of Open Access Journals (Sweden)

    Jinyong Kim

    2016-08-01

    Full Text Available Campylobacter jejuni is one of the leading foodborne pathogens worldwide. C. jejuni is isolated from a wide range of foods, domestic animals, wildlife, and environmental sources. The currently-available culture-based isolation methods are not highly effective for wastewater samples due to the low number of C. jejuni in the midst of competing bacteria. To detect and isolate C. jejuni from wastewater samples, in this study, we evaluated a few different enrichment conditions using five different antibiotics (i.e., cefoperazone, vancomycin, trimethoprim, polymyxin B, and rifampicin, to which C. jejuni is intrinsically resistant. The selectivity of each enrichment condition was measured with Ct value using quantitative real-time PCR (qRT-PCR, and multiplex PCR to determine Campylobacter species. In addition, the efficacy of Campylobacter isolation on different culture media after selective enrichment was examined by growing on Bolton and Preston agar plates. The addition of polymyxin B, rifampicin, or both to the Bolton selective supplements enhanced the selective isolation of C. jejuni. In particular, rifampicin supplementation and an increased culture temperature (i.e., 42°C had a decisive effect on the selective enrichment of C. jejuni from wastewater. The results of 16S rDNA sequencing also revealed that Enterococcus spp. and Pseudomonas aeruginosa are major competing bacteria in the enrichment conditions. Although it is known to be difficult to isolate Campylobacter from samples with heavy contamination, this study well exhibited that the manipulation of antibiotic selective pressure improves the isolation efficiency of fastidious Campylobacter from wastewater.

  5. Evaluation of novel supports for selective and efficient enrichment of phosphorylated peptides

    CSIR Research Space (South Africa)

    Bensadek, D

    2011-06-01

    Full Text Available A series of novel emulsion based supports MagReSynTM TiO 2 (A-D) were evaluated using 32P labelled phosphopeptides and nano LC-MS/MS. The performance of MagReSynTM TiO 2 (D) was found to be significantly better in terms of selectivity and specificity...

  6. Production of Microbial Protease from Selected Soil Fungal Isolates ...

    African Journals Online (AJOL)

    Production of Microbial Protease from Selected Soil Fungal Isolates. ... Nigerian Journal of Biotechnology ... and 500C. The optimal pH on the enzyme production was observed to be between pH 3.5 and 5.5 for the organisms. Keywords: Soil microorganism, fungal isolate, incubation period, microbial enzyme. Nig J. Biotech.

  7. Global proteomic profiling of phosphopeptides using electron transfer dissociation tandem mass spectrometry

    DEFF Research Database (Denmark)

    Molina, Henrik; Horn, David M; Tang, Ning

    2007-01-01

    Electron transfer dissociation (ETD) is a recently introduced mass spectrometric technique that provides a more comprehensive coverage of peptide sequences and posttranslational modifications. Here, we evaluated the use of ETD for a global phosphoproteome analysis. In all, we identified a total...... of 1,435 phosphorylation sites from human embryonic kidney 293T cells, of which 1,141 ( approximately 80%) were not previously described. A detailed comparison of ETD and collision-induced dissociation (CID) modes showed that ETD identified 60% more phosphopeptides than CID, with an average of 40% more...... fragment ions that facilitated localization of phosphorylation sites. Although our data indicate that ETD is superior to CID for phosphorylation analysis, the two methods can be effectively combined in alternating ETD and CID modes for a more comprehensive analysis. Combining ETD and CID, from this single...

  8. Fully automated synthesis of (phosphopeptide arrays in microtiter plate wells provides efficient access to protein tyrosine kinase characterization

    Directory of Open Access Journals (Sweden)

    Goldstein David J

    2005-01-01

    Full Text Available Abstract Background Synthetic peptides have played a useful role in studies of protein kinase substrates and interaction domains. Synthetic peptide arrays and libraries, in particular, have accelerated the process. Several factors have hindered or limited the applicability of various techniques, such as the need for deconvolution of combinatorial libraries, the inability or impracticality of achieving full automation using two-dimensional or pin solid phases, the lack of convenient interfacing with standard analytical platforms, or the difficulty of compartmentalization of a planar surface when contact between assay components needs to be avoided. This paper describes a process for synthesis of peptides and phosphopeptides on microtiter plate wells that overcomes previous limitations and demonstrates utility in determination of the epitope of an autophosphorylation site phospho-motif antibody and utility in substrate utilization assays of the protein tyrosine kinase, p60c-src. Results The overall reproducibility of phospho-peptide synthesis and multiplexed EGF receptor (EGFR autophosphorylation site (pY1173 antibody ELISA (9H2 was within 5.5 to 8.0%. Mass spectrometric analyses of the released (phosphopeptides showed homogeneous peaks of the expected molecular weights. An overlapping peptide array of the complete EGFR cytoplasmic sequence revealed a high redundancy of 9H2 reactive sites. The eight reactive phospopeptides were structurally related and interestingly, the most conserved antibody reactive peptide motif coincided with a subset of other known EGFR autophosphorylation and SH2 binding motifs and an EGFR optimal substrate motif. Finally, peptides based on known substrate specificities of c-src and related enzymes were synthesized in microtiter plate array format and were phosphorylated by c-Src with the predicted specificities. The level of phosphorylation was proportional to c-Src concentration with sensitivities below 0.1 Units of

  9. Characterization of casein phosphopeptides from fermented milk products.

    Science.gov (United States)

    Kawahara, Takeshi; Aruga, Kaori; Otani, Hajime

    2005-10-01

    This study dealt with the potential of fermented milk products as a source of functional casein phosphopeptides (CPPs) using plain yogurts and Camembert cheeses. The CPPs were prepared by tryptic digestion from four commercially available plain yogurts (P1-P4), five Camembert cheeses (C1-C5), and raw milk. From portions with a 1-g protein content of the plain yogurts, the Camembert cheeses, and the raw milk, 171 mg, 139 mg, and 146 mg of CPPs were obtained, respectively. The Camembert cheeses retained high amounts of organic phosphorus (32 microg) per 1 mg CPPs compared to the raw milk (15 microg) and plain yogurts (16 microg). Reverse-phase high-performance liquid chromatographic analysis showed that the elution patterns and retention times of the three major peaks of CPPs from P1 and C1 were similar to those from raw milk. Moreover, CPPs from P1 and C1 showed a mitogenic effect, while CPPs from C1 showed an IgA-enhancing effect in mouse spleen cell cultures. These results suggest that fermented milk products such as plain yogurts and Camembert cheeses generate functional CPPs in the body and exert beneficial effects on the immune system.

  10. Microalgae Isolation and Selection for Prospective Biodiesel Production

    Directory of Open Access Journals (Sweden)

    Peer M. Schenk

    2012-06-01

    Full Text Available Biodiesel production from microalgae is being widely developed at different scales as a potential source of renewable energy with both economic and environmental benefits. Although many microalgae species have been identified and isolated for lipid production, there is currently no consensus as to which species provide the highest productivity. Different species are expected to function best at different aquatic, geographical and climatic conditions. In addition, other value-added products are now being considered for commercial production which necessitates the selection of the most capable algae strains suitable for multiple-product algae biorefineries. Here we present and review practical issues of several simple and robust methods for microalgae isolation and selection for traits that maybe most relevant for commercial biodiesel production. A combination of conventional and modern techniques is likely to be the most efficient route from isolation to large-scale cultivation.

  11. Isolation and characterization of Acinetobacter baumannii recovered from Campylobacter selective medium.

    Directory of Open Access Journals (Sweden)

    Dinesh M Fernando

    2016-11-01

    Full Text Available Acinetobacter baumannii, a Gram-negative opportunistic pathogen, is known to cause multidrug resistant infections. This organism has primarily been isolated from clinical environments and its environmental reservoirs remain largely unknown. In the present study, we recovered seven isolates of A. baumannii growing under conditions selective for Campylobacter spp. (microaerophilic at 42 oC and in the presence of antibiotics from dairy cattle manure storage tank or surface water impacted by livestock effluents. Antibiotic susceptibility tests revealed that all of these isolates were less susceptible to at least two different clinically relevant antibiotics, compared to the type strain A. baumannii ATCC17978. Expression of resistance-nodulation-division efflux pumps, an important mechanism of intrinsic resistance in these organisms, was analyzed and adeB was found to be overexpressed in one and adeJ was overexpressed in three isolates. Comparison of these isolates using genomic DNA Macro-Restriction Fragment Pattern Analysis (MRFPA revealed relatively low relatedness among themselves or with some of the clinical isolates from previous studies. This study suggests that A. baumannii isolates are capable of growing under selective conditions for Campylobacter spp. and that this organism can be present in manure and water.

  12. Selective medium for the isolation of Bacteroides gingivalis.

    OpenAIRE

    Hunt, D E; Jones, J V; Dowell, V R

    1986-01-01

    Bacteroides gingivalis has been implicated in various forms of periodontal disease and may be responsible for other diseases in humans. The role of B. gingivalis in disease has been difficult to assess, because it is inhibited by most selective media commonly used by clinical laboratories to aid in isolating gram-negative, nonsporeforming anaerobes. We have developed a new medium, Bacteroides gingivalis agar, which contains bacitracin, colistin, and nalidixic acid as selective agents. This me...

  13. Phosphotyrosine-based-phosphoproteomics scaled-down to biopsy level for analysis of individual tumor biology and treatment selection.

    Science.gov (United States)

    Labots, Mariette; van der Mijn, Johannes C; Beekhof, Robin; Piersma, Sander R; de Goeij-de Haas, Richard R; Pham, Thang V; Knol, Jaco C; Dekker, Henk; van Grieken, Nicole C T; Verheul, Henk M W; Jiménez, Connie R

    2017-06-06

    Mass spectrometry-based phosphoproteomics of cancer cell and tissue lysates provides insight in aberrantly activated signaling pathways and potential drug targets. For improved understanding of individual patient's tumor biology and to allow selection of tyrosine kinase inhibitors in individual patients, phosphoproteomics of small clinical samples should be feasible and reproducible. We aimed to scale down a pTyr-phosphopeptide enrichment protocol to biopsy-level protein input and assess reproducibility and applicability to tumor needle biopsies. To this end, phosphopeptide immunoprecipitation using anti-phosphotyrosine beads was performed using 10, 5 and 1mg protein input from lysates of colorectal cancer (CRC) cell line HCT116. Multiple needle biopsies from 7 human CRC resection specimens were analyzed at the 1mg-level. The total number of phosphopeptides captured and detected by LC-MS/MS ranged from 681 at 10mg input to 471 at 1mg HCT116 protein. ID-reproducibility ranged from 60.5% at 10mg to 43.9% at 1mg. Per 1mg-level biopsy sample, >200 phosphopeptides were identified with 57% ID-reproducibility between paired tumor biopsies. Unsupervised analysis clustered biopsies from individual patients together and revealed known and potential therapeutic targets. This study demonstrates the feasibility of label-free pTyr-phosphoproteomics at the tumor biopsy level based on reproducible analyses using 1mg of protein input. The considerable number of identified phosphopeptides at this level is attributed to an effective down-scaled immuno-affinity protocol as well as to the application of ID propagation in the data processing and analysis steps. Unsupervised cluster analysis reveals patient-specific profiles. Together, these findings pave the way for clinical trials in which pTyr-phosphoproteomics will be performed on pre- and on-treatment biopsies. Such studies will improve our understanding of individual tumor biology and may enable future p

  14. Genomic signatures of geographic isolation and natural selection in coral reef fishes.

    Science.gov (United States)

    Gaither, Michelle R; Bernal, Moisés A; Coleman, Richard R; Bowen, Brian W; Jones, Shelley A; Simison, W Brian; Rocha, Luiz A

    2015-04-01

    The drivers of speciation remain among the most controversial topics in evolutionary biology. Initially, Darwin emphasized natural selection as a primary mechanism of speciation, but the architects of the modern synthesis largely abandoned that view in favour of divergence by geographic isolation. The balance between selection and isolation is still at the forefront of the evolutionary debate, especially for the world's tropical oceans where biodiversity is high, but isolating barriers are few. Here, we identify the drivers of speciation in Pacific reef fishes of the genus Acanthurus by comparative genome scans of two peripheral populations that split from a large Central-West Pacific lineage at roughly the same time. Mitochondrial sequences indicate that populations in the Hawaiian Archipelago and the Marquesas Islands became isolated approximately 0.5 Ma. The Hawaiian lineage is morphologically indistinguishable from the widespread Pacific form, but the Marquesan form is recognized as a distinct species that occupies an unusual tropical ecosystem characterized by upwelling, turbidity, temperature fluctuations, algal blooms and little coral cover. An analysis of 3737 SNPs reveals a strong signal of selection at the Marquesas, with 59 loci under disruptive selection including an opsin Rh2 locus. While both the Hawaiian and Marquesan populations indicate signals of drift, the former shows a weak signal of selection that is comparable with populations in the Central-West Pacific. This contrast between closely related lineages reveals one population diverging due primarily to geographic isolation and genetic drift, and the other achieving taxonomic species status under the influence of selection. © 2015 The Authors. Molecular Ecology Published by John Wiley & Sons Ltd.

  15. Selective isolation and characterization of agriculturally beneficial endopytic bacteria from wild hemp using canola

    International Nuclear Information System (INIS)

    Afzal, I.; Iqrar, I.

    2015-01-01

    Endophytic bacteria can provide a useful alternative to synthetic fertilizers to improve plant growth. Wild plants are little investigated as a source of growth promoting endophytic bacteria for commercial application to crops. In present study, endophytic bacteria were isolated from Cannabis sativa L. (hemp) using two different methods to examine their ability to promote canola growth. Besides direct isolation from the roots, endophytic bacteria were also selectively isolated from the rhizosphere of C. sativa using canola. Under gnotobiotic conditions, six bacteria from the selective isolation significantly improved canola root growth, as compared to the two bacteria isolated from direct method. Overall, three isolates performed distinctly well, namely, Pantoea vagans MOSEL-t13, Pseudomonas geniculata MOSEL-tnc1, and Serratia marcescens MOSEL-w2. These bacteria tolerated high salt concentrations and promoted canola growth under salt stress. Further, the isolated bacteria possessed plant growth promoting traits like IAA production, phosphate solubilization, and siderophore production. Most isolates produced plant cell-wall degrading enzymes, cellulase and pectinase. Some isolates were also effective in hindering the growth of two phytopathogenic fungi in dual culture assay, and displayed chitinase and protease activity. Paenibacillus sp. MOSEL-w13 displayed the greatest antifungal activity among all the isolates. Present findings conclude that wild plants can be a good source for isolating beneficial microbes, and validates the employed selective isolation for improved isolation of plant-beneficial endophytic bacteria. (author)

  16. Glycine-containing selective medium for isolation of Legionellaceae from environmental specimens.

    Science.gov (United States)

    Wadowsky, R M; Yee, R B

    1981-11-01

    Glycine, at a final concentration of 0.3%, has been shown to be an excellent selective agent for the isolation of Legionellaceae. Stock cultures of Legionella pneumophila were not inhibited on buffered charcoal-yeast extract agar containing the amino acid. Among the other Legionellaceae tested, only one of two strains of L. dumoffii and two of six strains of L. micdadei were appreciably inhibited. This medium permitted the isolation of L. pneumophila from environmental specimens with marked inhibition of many non-Legionellaceae bacteria. The selectivity of the medium was subsequently improved by the incorporation of vancomycin (5 microgram/ml) and polymyxin B (100 U/ml). This selective medium, glycine-vancomycin-polymyxin B agar, should facilitate the recovery of Legionellaceae from environmental sources.

  17. Comparison of a novel TiO₂/diatomite composite and pure TiO₂ for the purification of phosvitin phosphopeptides.

    Science.gov (United States)

    Zhang, Yang; Li, Junhua; Niu, Fuge; Sun, Jun; Dou, Yuan; Liu, Yuntao; Su, Yujie; Zhou, Bei; Xu, Qinqin; Yang, Yanjun

    2014-06-01

    A novel TiO2/diatomite composite (TD) was prepared and then characterized by scanning electron microscope (SEM) and Fourier Transform Infrared (FTIR). The results of SEM showed that after modification, the porous surface of diatomite was covered with TiO2. Both diatomite and TD had clear disc-shaped structures with average grain diameters of around 25 μm. Then TD and pure TiO2 were applied in the purification of phosvitin phosphopeptides (PPPs) from the digest of egg yolk protein, and a comparative study of adsorption properties of PPPs on TD and TiO2 was performed. In the study of adsorption kinetics, the adsorption equilibrium of PPPs on TD and TiO2 fitted well with the Langmuir model, and the time needed to reach adsorption equilibrium were both around 10 min. The maximum dynamic adsorption capacity of TD (8.15 mg/g) was higher than that of TiO2 (4.96 mg/g). The results of repeated use showed that TD and TiO2 were very stable after being subjected to ten repeated adsorption-elution cycles, and TD could easily be separated from aqueous solution by filtration. On the other hand, the present synthetic technology of TD was very simple, cost-effective, organic solvent-free and available for large-scale preparation. Thus, this separation method not only brings great advantages in the purification of PPPs from egg yolk protein but also provides a promising purification material for the enrichment of phosphopeptides in proteomic researches. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. Preparation of Affinity Column Based on Zr4+ Ion for Phosphoproteins Isolation

    International Nuclear Information System (INIS)

    Lee, Seon Mi; Bae, In Ae; Park, Jung Hyen; Kim, Tae Dong; Choi, Seong Ho

    2009-01-01

    This paper has described about preparation of Zr 4+ affinity column based on the poly(styreneco- glycidyl methacrylate) prepared by emulsion polymerization of styrene and glycidyl methacrylate in order to isolate phosphopeptide. The Zr 4+ ions were introduced after the phophonation of an epoxy group on polymeric microspheres. The successful preparation of Zr 4+ -immobilized polymeric microsphere stationary phase was confirmed through Fourier transform infrared spectra, optical microscopy, scanning electron microscopy, X-ray photoelectron spectra and inductively coupled plasma-atomic emission spectrometer. The separation efficiency for Zr 4+ affinity column prepared by slurry packing was tested to phosphonated casein and dephosphonated casein. The resolution time (min) of the phosphonated casein was higher than that of dephosphated casein for Zr 4+ affinity polymeric microsphere by liquid chromatography. This Zr 4+ affinity column can be used for isolation of phosphonated casein from casein using liquid chromatography

  19. [Isolation of Actinomycetales from the soil of Kazakhstan on selective media with antibiotics].

    Science.gov (United States)

    Vetlugina, L A; Adiiatova, Zh F; Khozhamuratova, S Sh; Rymzhanova, Z A; Trenozhnikova, L P; Kopytina, M N

    1990-02-01

    About 3000 actinomycetes were isolated from various soil samples collected in 11 regions of Kazakhstan. 62.7 per cent of them proved to be antagonists. For isolation of the strains, selective media supplemented with antibiotics were used. Kanamycin promoted growth of Actinomadura and Nocardia. Rubomycin promoted growth of Actinomadura. Tavromycetin and roseofungin were used as selective agents for the first time. Tavromycetin favoured isolation of Actinomadura and Nocardia. Roseofungin favoured isolation of Actinomadura. Light chestnut and serozemic soils were the most rich in antagonists (67.1 and 61.3 per cent, respectively) while saline and chestnut soils were the poorest in antagonists (32.2 and 30.6 per cent, respectively). Actinomadura were more frequent in light-chestnut light-loamy and serozemic soils. Half of the antibiotics isolated in the form of concentrates were identified with the known antibiotics or classified as belonging to various groups. A culture producing a novel antibiotic was isolated.

  20. Characterization of phosphorylation sites in the cytoplasmic domain of the 300 kDa mannose-6-phosphate receptor

    DEFF Research Database (Denmark)

    Rosorius, O; Mieskes, G; Issinger, O G

    1993-01-01

    The human 300 kDa mannose-6-phosphate receptor (MPR 300) is phosphorylated in vivo at serine residues of its cytoplasmic domain. Two-dimensional separation can resolve tryptic phosphopeptides into four major species. To identify the kinases involved in MPR 300 phosphorylation and the phosphorylat......The human 300 kDa mannose-6-phosphate receptor (MPR 300) is phosphorylated in vivo at serine residues of its cytoplasmic domain. Two-dimensional separation can resolve tryptic phosphopeptides into four major species. To identify the kinases involved in MPR 300 phosphorylation...... and the phosphorylation sites the entire coding sequence of the cytoplasmic tail was expressed in Escherichia coli. The isolated cytoplasmic domain was used as a substrate for four purified serine/threonine kinases [casein kinase II (CK II), protein kinase A (PKA), protein kinase C and Ca2+/calmodulin kinase]. All...... kinases phosphorylate the cytoplasmic tail exclusively on serine residues. Inhibition studies using synthetic peptides, partial sequencing of isolated tryptic phosphopeptides and co-migration with tryptic phosphopeptides from MPR 300 labelled in vivo showed that (i) PKA phosphorylates the cytoplasmic MPR...

  1. Evidence for estrogen receptor beta-selective activity of Vitex agnus-castus and isolated flavones.

    Science.gov (United States)

    Jarry, Hubertus; Spengler, Barbara; Porzel, Andrea; Schmidt, Juergen; Wuttke, Wolfgang; Christoffel, Volker

    2003-10-01

    Recent cell culture experiments indicated that extracts of Vitex agnus-castus (VAC) may contain yet unidentified phytoestrogens. Estrogenic actions are mediated via estrogen receptors (ER). To investigate whether VAC compounds bind to the currently known isoforms ERalpha or ERss, ligand binding assays (LBA) were performed. Subtype specific ER-LBA revealed a binding of VAC to ERss only. To isolate the ERss-selective compounds, the extract was fractionated by bio-guidance. The flavonoid apigenin was isolated and identified as the most active ERss-selective phytoestrogen in VAC. Other isolated compounds were vitexin and penduletin. These data demonstrate that the phytoestrogens in VAC are ERss-selective.

  2. Selection, isolation, and identification of fungi for bioherbicide production

    Directory of Open Access Journals (Sweden)

    Angélica Rossana Castro de Souza

    Full Text Available Abstract Production of a bioherbicide for biological control of weeds requires a series of steps, from selection of a suitable microbial strain to final formulation. Thus, this study aimed to select fungi for production of secondary metabolites with herbicidal activity using biological resources of the Brazilian Pampa biome. Phytopathogenic fungi were isolated from infected tissues of weeds in the Pampa biome. A liquid synthetic culture medium was used for production of metabolites. The phytotoxicity of fungal metabolites was assessed via biological tests using the plant Cucumis sativus L., and the most promising strain was identified by molecular analysis. Thirty-nine fungi were isolated, and 28 presented some phytotoxic symptoms against the target plant. Fungus VP51 belonging to the genus Diaporthe showed the most pronounced herbicidal activity. The Brazilian Pampa biome is a potential resource for the development of new and sustainable chemical compounds for modern agriculture.

  3. Selective Osmotic Shock (SOS)-Based Islet Isolation for Microencapsulation.

    Science.gov (United States)

    Enck, Kevin; McQuilling, John Patrick; Orlando, Giuseppe; Tamburrini, Riccardo; Sivanandane, Sittadjody; Opara, Emmanuel C

    2017-01-01

    Islet transplantation (IT) has recently been shown to be a promising alternative to pancreas transplantation for reversing diabetes. IT requires the isolation of the islets from the pancreas, and these islets can be used to fabricate a bio-artificial pancreas. Enzymatic digestion is the current gold standard procedure for islet isolation but has lingering concerns. One such concern is that it has been shown to damage the islets due to nonselective tissue digestion. This chapter provides a detailed description of a nonenzymatic method that we are exploring in our lab as an alternative to current enzymatic digestion procedures for islet isolation from human and nonhuman pancreatic tissues. This method is based on selective destruction and protection of specific cell types and has been shown to leave the extracellular matrix (ECM) of islets intact, which may thus enhance islet viability and functionality. We also show that these SOS-isolated islets can be microencapsulated for transplantation.

  4. Selection of Infective Arbuscular Mycorrhizal Fungal Isolates for Field Inoculation

    Directory of Open Access Journals (Sweden)

    Elisa Pellegrino

    2010-09-01

    Full Text Available Arbuscular mycorrhizal (AM fungi play a key role in host plant growth and health, nutrient and water uptake, plant community diversity and dynamics. AM fungi differ in their symbiotic performance, which is the result of the interaction of two fungal characters, infectivity and efficiency. Infectivity is the ability of a fungal isolate to establish rapidly an extensive mycorrhizal symbiosis and is correlated with pre-symbiotic steps of fungal life cycle, such as spore germination and hyphal growth. Here, different AM fungal isolates were tested, with the aim of selecting infective endophytes for field inoculation. Greenhouse and microcosm experiments were performed in order to assess the ability of 12 AM fungal isolates to produce spores, colonize host roots and to perform initial steps of symbiosis establishment, such as spore germination and hyphal growth. AM fungal spore production and root colonization were significantly different among AM fungal isolates. Spore and sporocarp densities ranged from 0.8 to 7.4 and from 0.6 to 2.0 per gram of soil, respectively, whereas root colonization ranged from 2.9 to 72.2%. Percentage of spore or sporocarp germination ranged from 5.8 to 53.3% and hyphal length from 4.7 to 79.8 mm. The ordination analysis (Redundancy Analysis, RDA showed that environmental factors explained about 60% of the whole variance and their effect on fungal infectivity variables was significant (P = 0.002. The biplot clearly showed that variables which might be used to detect infective AM fungal isolates were hyphal length and root colonization. Such analysis may allow the detection of the best parameters to select efficient AM fungal isolates to be used in agriculture.

  5. Casein phosphopeptide-amorphous calcium phosphate and shear bond strength of adhesives to primary teeth enamel.

    Science.gov (United States)

    Farokh Gisovar, Elham; Hedayati, Nassim; Shadman, Niloofar; Shafiee, Leila

    2015-02-01

    CPP-ACP (Phosphopeptide-Amorphous Calcium Phosphate) has an important role in caries prevention in pediatric patients. This study was done, because of the great use of CPP-ACP and the need for restoration for teeth treated with CPP-ACP as well as the importance of shear bond strength of adhesives in the success of restorations. This study aimed to evaluate the effect of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) on shear bond strength of dental adhesives to enamel of primary teeth molars. This in vitro study was conducted on 180 extracted primary molars. They were randomly divided into 6 groups and each group was divided into 2 subgroups (treated with CPP-ACP and untreated). In subgroups with CPP-ACP, enamel was treated with CPP-ACP paste 1 h/d for 5 days. Types of adhesives that were evaluated in this study were Tetric N-Bond, AdheSE, AdheSE One F, single Bond 2, SE Bond, and Adper Prompt L-Pop. Shear bond strength was tested with a universal testing machine and mode of failure was evaluated under stereomicroscope. Data were analyzed by T test, 2-way analysis of variance (ANOVA), Tukey and Fisher exact test using SPSS18. P adhesive systems to enamel of primary teeth treated and untreated with CPP-ACP showed no significant difference (P > 0.05). Mode of failure in all groups regardless of CPP-ACP administration was mainly adhesive type. Our results indicated that CPP-ACP did not affect shear bond strength of studied adhesives to primary teeth enamel. To have a successful and durable composite restoration, having a high strength bonding is essential. Considering the wide use of CPP-ACP in preventing tooth decay and the role of adhesive shear bond strength (SBS) in success of composite restoration, we conducted the present study to evaluate the effect of CPP-ACP on the SBS of adhesives to primary teeth enamel.

  6. Preparation of Affinity Column Based on Zr{sup 4+} Ion for Phosphoproteins Isolation

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Seon Mi; Bae, In Ae; Park, Jung Hyen; Kim, Tae Dong; Choi, Seong Ho [Hannam University, Daejeon (Korea, Republic of)

    2009-06-15

    This paper has described about preparation of Zr{sup 4+} affinity column based on the poly(styreneco- glycidyl methacrylate) prepared by emulsion polymerization of styrene and glycidyl methacrylate in order to isolate phosphopeptide. The Zr{sup 4+} ions were introduced after the phophonation of an epoxy group on polymeric microspheres. The successful preparation of Zr{sup 4+}-immobilized polymeric microsphere stationary phase was confirmed through Fourier transform infrared spectra, optical microscopy, scanning electron microscopy, X-ray photoelectron spectra and inductively coupled plasma-atomic emission spectrometer. The separation efficiency for Zr{sup 4+} affinity column prepared by slurry packing was tested to phosphonated casein and dephosphonated casein. The resolution time (min) of the phosphonated casein was higher than that of dephosphated casein for Zr{sup 4+} affinity polymeric microsphere by liquid chromatography. This Zr{sup 4+} affinity column can be used for isolation of phosphonated casein from casein using liquid chromatography.

  7. Casein phosphopeptides drastically increase the secretion of extracellular proteins in Aspergillus awamori. Proteomics studies reveal changes in the secretory pathway

    OpenAIRE

    Kosalková Katarina; García-Estrada Carlos; Barreiro Carlos; Flórez Martha G; Jami Mohammad S; Paniagua Miguel A; Martín Juan F

    2012-01-01

    Abstract Background The secretion of heterologous animal proteins in filamentous fungi is usually limited by bottlenecks in the vesicle-mediated secretory pathway. Results Using the secretion of bovine chymosin in Aspergillus awamori as a model, we found a drastic increase (40 to 80-fold) in cells grown with casein or casein phosphopeptides (CPPs). CPPs are rich in phosphoserine, but phosphoserine itself did not increase the secretion of chymosin. The stimulatory effect is reduced about 50% u...

  8. Selection of cellulolytic fungi isolated from diverse substrates

    Directory of Open Access Journals (Sweden)

    Mônica Caramez Triches Damaso

    2012-08-01

    Full Text Available The aim of the present work was to select filamentous fungi isolated from diverse substrates to obtain the strains with potential to produce the hydrolytic enzymes. From a total of 215 strains, seven strains from the soils, six from the plants and one from sugarcane bagasse were selected and identified as belonging to the Trichoderma, Penicillium and Aspergillus genera. The best hydrolytic activities obtained by semi-solid fermentation using these strains were approximately: 35; 1; 160; 170 and 120 U/gdm (CMCase, FPase, β-glucosidase, xylanase and polygalacturonase, respectively, demonstrating their potential to synthesize the enzymes compared with the results reported in the literature.

  9. Star formation rates in isolated galaxies selected from the Two-Micron All-Sky Survey

    Science.gov (United States)

    Melnyk, O.; Karachentseva, V.; Karachentsev, I.

    2015-08-01

    We have considered the star formation properties of 1616 isolated galaxies from the 2MASS XSC (Extended Source Catalog) selected sample (2MIG) with the far-ultraviolet GALEX magnitudes. This sample was then compared with corresponding properties of isolated galaxies from the Local Orphan Galaxies (LOG) catalogue and paired galaxies. We found that different selection algorithms define different populations of isolated galaxies. The population of the LOG catalogue, selected from non-clustered galaxies in the Local Supercluster volume, mostly consists of low-mass spiral and late-type galaxies. The specific star formation rate (SSFR) upper limit in isolated and paired galaxies does not exceed the value of ˜dex(-9.4). This is probably common for galaxies of differing activity and environment (at least at z processes is the galaxy mass. However, the environmental influence is notable: paired massive galaxies with logM* > 11.5 have higher (S)SFR than isolated galaxies. Our results suggest that the environment helps to trigger the star formation in the highest mass galaxies. We found that the fraction of AGN in the paired sample is only a little higher than in our isolated galaxy sample. We assume that AGN phenomenon is probably defined by secular galaxy evolution.

  10. Technique for selection of transient radiation-hard junction-isolated integrated circuits

    International Nuclear Information System (INIS)

    Crowley, J.L.; Junga, F.A.; Stultz, T.J.

    1976-01-01

    A technique is presented which demonstrates the feasibility of selecting junction-isolated integrated circuits (JI/ICS) for use in transient radiation environments. The procedure guarantees that all PNPN paths within the integrated circuit are identified and describes the methods used to determine whether the paths represent latchup susceptible structures. Two examples of the latchup analysis are given involving an SSI and an LSI bipolar junction-isolated integrated circuit

  11. Patagonian red wines: selection of Lactobacillus plantarum isolates as potential starter cultures for malolactic fermentation.

    Science.gov (United States)

    Bravo-Ferrada, Bárbara Mercedes; Hollmann, Axel; Delfederico, Lucrecia; Valdés La Hens, Danay; Caballero, Adriana; Semorile, Liliana

    2013-09-01

    The aim of this study was to evaluate fifty-three Lactobacillus plantarum isolates obtained from a Patagonian red wine, molecularly identified and typified using RAPD analysis, in order to select starter cultures for malolactic fermentation (MLF). The results obtained suggest a considerable genetic diversity, taking into account that all L. plantarum isolates were obtained from one cellar and one vintage. Based on the capacity to tolerate a concentration of 14 % ethanol in MRS broth for 2 days, eight isolates were selected for the subsequent analysis. The incidence of various wine stress factors (ethanol, acid pH, lysozyme and sulfur dioxide) on isolates growth was studied. Besides, glucosidase and tannase activities were evaluated, and the presence of genes involved in the synthesis of biogenic amines was examined by PCR. A previously characterized indigenous Oenococcus oeni strain was included with comparative purposes. Differences in technologically relevant characteristics were observed among the eight L. plantarum selected isolates, revealing an isolate-dependent behavior. Detectable glucosidase and tannase activities were found in all isolates. The presence of genes encoding histidine and tyrosine descarboxylases and putrescine carbamoyltransferase was not detected. The ability of L. plantarum isolates to grow and consume L-malic acid in simulated laboratory-scale vinifications revealed that two of them could be considered as possible MLF starter cultures for Patagonian red wines. These isolates will be subjected to further analysis, for a final winery technological characterization.

  12. DEVELOPMENT AND EVALUATION OF A SELECTIVE AND INDICATIVE MEDIUM FOR ISOLATION OF ACTINOBACILLUS-PLEUROPNEUMONIAE FROM TONSILS

    DEFF Research Database (Denmark)

    Jakobsen, Marianne; Nielsen, Jens

    1995-01-01

    In order to isolate ActinobacillIus pleuropneumoniae from mixed bacterial flora a selective and indicative medium was developed. The optimal concentrations of antibiotics were determined for selective chocolate agar (S-TSA) and selective blood agar (S-MBA) using a set of 25 strains of A. pleuropn......In order to isolate ActinobacillIus pleuropneumoniae from mixed bacterial flora a selective and indicative medium was developed. The optimal concentrations of antibiotics were determined for selective chocolate agar (S-TSA) and selective blood agar (S-MBA) using a set of 25 strains of A...

  13. Genetic polymorphism and natural selection of Duffy binding protein of Plasmodium vivax Myanmar isolates

    Science.gov (United States)

    2012-01-01

    Background Plasmodium vivax Duffy binding protein (PvDBP) plays an essential role in erythrocyte invasion and a potential asexual blood stage vaccine candidate antigen against P. vivax. The polymorphic nature of PvDBP, particularly amino terminal cysteine-rich region (PvDBPII), represents a major impediment to the successful design of a protective vaccine against vivax malaria. In this study, the genetic polymorphism and natural selection at PvDBPII among Myanmar P. vivax isolates were analysed. Methods Fifty-four P. vivax infected blood samples collected from patients in Myanmar were used. The region flanking PvDBPII was amplified by PCR, cloned into Escherichia coli, and sequenced. The polymorphic characters and natural selection of the region were analysed using the DnaSP and MEGA4 programs. Results Thirty-two point mutations (28 non-synonymous and four synonymous mutations) were identified in PvDBPII among the Myanmar P. vivax isolates. Sequence analyses revealed that 12 different PvDBPII haplotypes were identified in Myanmar P. vivax isolates and that the region has evolved under positive natural selection. High selective pressure preferentially acted on regions identified as B- and T-cell epitopes of PvDBPII. Recombination may also be played a role in the resulting genetic diversity of PvDBPII. Conclusions PvDBPII of Myanmar P. vivax isolates displays a high level of genetic polymorphism and is under selective pressure. Myanmar P. vivax isolates share distinct types of PvDBPII alleles that are different from those of other geographical areas. These results will be useful for understanding the nature of the P. vivax population in Myanmar and for development of PvDBPII-based vaccine. PMID:22380592

  14. Isolation of cells for selective treatment and analysis using a magnetic microfluidic chip

    KAUST Repository

    Yassine, Omar

    2014-05-01

    This study describes the development and testing of a magnetic microfluidic chip (MMC) for trapping and isolating cells tagged with superparamagnetic beads (SPBs) in a microfluidic environment for selective treatment and analysis. The trapping and isolation are done in two separate steps; first, the trapping of the tagged cells in a main channel is achieved by soft ferromagnetic disks and second, the transportation of the cells into side chambers for isolation is executed by tapered conductive paths made of Gold (Au). Numerical simulations were performed to analyze the magnetic flux and force distributions of the disks and conducting paths, for trapping and transporting SPBs. The MMC was fabricated using standard microfabrication processes. Experiments were performed with E. coli (K12 strand) tagged with 2.8 μm SPBs. The results showed that E. coli can be separated from a sample solution by trapping them at the disk sites, and then isolated into chambers by transporting them along the tapered conducting paths. Once the E. coli was trapped inside the side chambers, two selective treatments were performed. In one chamber, a solution with minimal nutrition content was added and, in another chamber, a solution with essential nutrition was added. The results showed that the growth of bacteria cultured in the second chamber containing nutrient was significantly higher, demonstrating that the E. coli was not affected by the magnetically driven transportation and the feasibility of performing different treatments on selectively isolated cells on a single microfluidic platform.

  15. Glycine-containing selective medium for isolation of Legionellaceae from environmental specimens.

    OpenAIRE

    Wadowsky, R M; Yee, R B

    1981-01-01

    Glycine, at a final concentration of 0.3%, has been shown to be an excellent selective agent for the isolation of Legionellaceae. Stock cultures of Legionella pneumophila were not inhibited on buffered charcoal-yeast extract agar containing the amino acid. Among the other Legionellaceae tested, only one of two strains of L. dumoffii and two of six strains of L. micdadei were appreciably inhibited. This medium permitted the isolation of L. pneumophila from environmental specimens with marked i...

  16. Highly Efficient Single-Step Enrichment of Low Abundance Phosphopeptides from Plant Membrane Preparations

    Directory of Open Access Journals (Sweden)

    Xu Na Wu

    2017-09-01

    Full Text Available Mass spectrometry (MS-based large scale phosphoproteomics has facilitated the investigation of plant phosphorylation dynamics on a system-wide scale. However, generating large scale data sets for membrane phosphoproteins usually requires fractionation of samples and extended hands-on laboratory time. To overcome these limitations, we developed “ShortPhos,” an efficient and simple phosphoproteomics protocol optimized for research on plant membrane proteins. The optimized workflow allows fast and efficient identification and quantification of phosphopeptides, even from small amounts of starting plant materials. “ShortPhos” can produce label-free datasets with a high quantitative reproducibility. In addition, the “ShortPhos” protocol recovered more phosphorylation sites from membrane proteins, especially plasma membrane and vacuolar proteins, when compared to our previous workflow and other membrane-based data in the PhosPhAt 4.0 database. We applied “ShortPhos” to study kinase-substrate relationships within a nitrate-induction experiment on Arabidopsis roots. The “ShortPhos” identified significantly more known kinase-substrate relationships compared to previous phosphoproteomics workflows, producing new insights into nitrate-induced signaling pathways.

  17. Susceptibility of Mycosphaerella fijiensis isolates to the Higromicine B and Carbendazim selection agents

    Directory of Open Access Journals (Sweden)

    Mileidy Cruz-Martín

    2006-10-01

    Full Text Available The  study  of  Musa  -  Mycosphaerella  fijiensis  interactions  supported  by  molecular  tools  such  as  the  genetic transformation could contribute to elucidate new mechanisms related with the pathogenesis and through it to allow  the  development  of  new  selective  fungicides  as  well  as  the  elaboration  of  more  durable  resistance strategies.  This  work  had  as  objective,  to  determine  the  Minimum  Inhibitory  Concentration  (MIC  of  different antimicrobial substances commonly used as selection agents in the genetic transformation study of different species of Mycosphaerella (Hygromycin B and carbendazim. The agar dilution method was used and mycelial suspensions of nine of M. fijiensis isolated by several regions of Villa Clara and Ciego de Ávila was used as inoculums. Concentrations of these substances from 32.0 to 0.015 mg.l l -1  were rehearsed. It was possible to determine  the  MIC  of  both  antimicrobials  substances  against  the  M.fijiensis  isolated  using  this  method.  The MCI of the Hygromycin B for 100% of the isolated was less to 8 μg.ml l -1 and the rehearsed 90% of the isolated ones showed, for the carbendazim, MIC less or similar to 0.125 μg.ml l -1 . CCIBP-Pf-57 isolated did not achieve to be inhibited to the rehearsed concentrations of the Carbendazim. The results obtained in this work evidence the importance of knowing the MIC of the selective agents that are used in the transformation avoiding escapes in the selection. Key words: antimicrobial substance, genetic transformation, Minimal inhibitory concentration, mycelia suspension, Pseudosercospora  fijiensis

  18. Selection of Beauveria bassiana and Metarhizium anisopliae Isolates to Control Triatoma infestans

    Directory of Open Access Journals (Sweden)

    Luz Christian

    1998-01-01

    Full Text Available Twenty three isolates of Beauveria bassiana and 13 isolates of Metarhizium anisopliae were tested on third instar nymphs of Triatoma infestans, a serious vector of Chagas disease. Pathogenicity tests at saturated humidity showed that this insect is very susceptible to fungal infection. At lower relative humidity (50%, conditions expected in the vector microhabitat, virulence was significantly different among isolates. Cumulative mortality 15 days after treatment varied from 17.5 to 97.5%, and estimates of 50% survival time varied from 6 to 11 days. Maintaining lower relative humidity, four B. bassiana and two M. anisopliae isolates were selected for analysis of virulence at different conidial concentrations and temperatures. Lethal concentrations sufficient to kill 50% of insects (LC50 varied from 7.1x105 to 4.3x106 conidia/ml, for a B. bassiana isolate (CG 14 and a M. anisopliae isolate (CG 491 respectively. Most isolates, particularly B. bassiana isolates CG 24 and CG 306, proved to be more virulent at 25 and 30°C, compared to 15 and 20°C. The differential virulence at 50% humidity observed among some B. bassiana isolates was not correlated to phenetic groups in cluster analysis of RAPD markers. In fact, the B. bassiana isolates analyzed presented a high homogeneity (> 73% similarity.

  19. An Improved Test Selection Optimization Model Based on Fault Ambiguity Group Isolation and Chaotic Discrete PSO

    Directory of Open Access Journals (Sweden)

    Xiaofeng Lv

    2018-01-01

    Full Text Available Sensor data-based test selection optimization is the basis for designing a test work, which ensures that the system is tested under the constraint of the conventional indexes such as fault detection rate (FDR and fault isolation rate (FIR. From the perspective of equipment maintenance support, the ambiguity isolation has a significant effect on the result of test selection. In this paper, an improved test selection optimization model is proposed by considering the ambiguity degree of fault isolation. In the new model, the fault test dependency matrix is adopted to model the correlation between the system fault and the test group. The objective function of the proposed model is minimizing the test cost with the constraint of FDR and FIR. The improved chaotic discrete particle swarm optimization (PSO algorithm is adopted to solve the improved test selection optimization model. The new test selection optimization model is more consistent with real complicated engineering systems. The experimental result verifies the effectiveness of the proposed method.

  20. Chemical rationale for selection of isolates for genome sequencing

    DEFF Research Database (Denmark)

    Rank, Christian; Larsen, Thomas Ostenfeld; Frisvad, Jens Christian

    The advances in gene sequencing will in the near future enable researchers to affordably acquire the full genomes of handpicked isolates. We here present a method to evaluate the chemical potential of an entire species and select representatives for genome sequencing. The selection criteria for new...... strains to be sequenced can be manifold, but for studying the functional phenotype, using a metabolome based approach offers a cheap and rapid assessment of critical strains to cover the chemical diversity. We have applied this methodology on the complex A. flavus/A. oryzae group. Though these two species...... are in principal identical, they represent two different phenotypes. This is clearly presented through a correspondence analysis of selected extrolites, in which the subtle chemical differences are visually dispersed. The results points to a handful of strains, which, if sequenced, will likely enhance our...

  1. Selection on plant male function genes identifies candidates for reproductive isolation of yellow monkeyflowers.

    Directory of Open Access Journals (Sweden)

    Jan E Aagaard

    Full Text Available Understanding the genetic basis of reproductive isolation promises insight into speciation and the origins of biological diversity. While progress has been made in identifying genes underlying barriers to reproduction that function after fertilization (post-zygotic isolation, we know much less about earlier acting pre-zygotic barriers. Of particular interest are barriers involved in mating and fertilization that can evolve extremely rapidly under sexual selection, suggesting they may play a prominent role in the initial stages of reproductive isolation. A significant challenge to the field of speciation genetics is developing new approaches for identification of candidate genes underlying these barriers, particularly among non-traditional model systems. We employ powerful proteomic and genomic strategies to study the genetic basis of conspecific pollen precedence, an important component of pre-zygotic reproductive isolation among yellow monkeyflowers (Mimulus spp. resulting from male pollen competition. We use isotopic labeling in combination with shotgun proteomics to identify more than 2,000 male function (pollen tube proteins within maternal reproductive structures (styles of M. guttatus flowers where pollen competition occurs. We then sequence array-captured pollen tube exomes from a large outcrossing population of M. guttatus, and identify those genes with evidence of selective sweeps or balancing selection consistent with their role in pollen competition. We also test for evidence of positive selection on these genes more broadly across yellow monkeyflowers, because a signal of adaptive divergence is a common feature of genes causing reproductive isolation. Together the molecular evolution studies identify 159 pollen tube proteins that are candidate genes for conspecific pollen precedence. Our work demonstrates how powerful proteomic and genomic tools can be readily adapted to non-traditional model systems, allowing for genome-wide screens

  2. Nucleolin (C23), a physiological substrate for casein kinase II

    DEFF Research Database (Denmark)

    Schneider, H R; Issinger, O G

    1988-01-01

    Nucleolin (C23), a 110 kDa phosphoprotein, which is mainly found in the nucleolus has been shown to be a physiological substrate for casein kinase II (CKII). Nucleolin was identified and characterized by immunodetection using an anti-nucleolin antibody. Phosphopeptide patterns from nucleolin...... phosphorylated by purified casein kinase II and of phosphorylated nucleolin which had been isolated from tumor cells grown in the presence of [32P]-o-phosphate, were identical. The partial tryptic digest revealed nine phosphopeptides. Nucleolin isolated from Krebs II mouse ascites cells was phosphorylated...... by purified casein kinase II with about two moles phosphate per one mole of nucleolin....

  3. The molecular basis of FHA domain:phosphopeptide binding specificity and implications for phospho-dependent signaling mechanisms.

    Science.gov (United States)

    Durocher, D; Taylor, I A; Sarbassova, D; Haire, L F; Westcott, S L; Jackson, S P; Smerdon, S J; Yaffe, M B

    2000-11-01

    Forkhead-associated (FHA) domains are a class of ubiquitous signaling modules that appear to function through interactions with phosphorylated target molecules. We have used oriented peptide library screening to determine the optimal phosphopeptide binding motifs recognized by several FHA domains, including those within a number of DNA damage checkpoint kinases, and determined the X-ray structure of Rad53p-FHA1, in complex with a phospho-threonine peptide, at 1.6 A resolution. The structure reveals a striking similarity to the MH2 domains of Smad tumor suppressor proteins and reveals a mode of peptide binding that differs from SH2, 14-3-3, or PTB domain complexes. These results have important implications for DNA damage signaling and CHK2-dependent tumor suppression, and they indicate that FHA domains play important and unsuspected roles in S/T kinase signaling mechanisms in prokaryotes and eukaryotes.

  4. Probing protein phosphatase substrate binding

    DEFF Research Database (Denmark)

    Højlys-Larsen, Kim B.; Sørensen, Kasper Kildegaard; Jensen, Knud Jørgen

    2012-01-01

    Proteomics and high throughput analysis for systems biology can benefit significantly from solid-phase chemical tools for affinity pull-down of proteins from complex mixtures. Here we report the application of solid-phase synthesis of phosphopeptides for pull-down and analysis of the affinity...... profile of the integrin-linked kinase associated phosphatase (ILKAP), a member of the protein phosphatase 2C (PP2C) family. Phosphatases can potentially dephosphorylate these phosphopeptide substrates but, interestingly, performing the binding studies at 4 °C allowed efficient binding to phosphopeptides......, without the need for phosphopeptide mimics or phosphatase inhibitors. As no proven ILKAP substrates were available, we selected phosphopeptide substrates among known PP2Cδ substrates including the protein kinases: p38, ATM, Chk1, Chk2 and RSK2 and synthesized directly on PEGA solid supports through a BAL...

  5. Computational analysis of phosphopeptide binding to the polo-box domain of the mitotic kinase PLK1 using molecular dynamics simulation.

    Directory of Open Access Journals (Sweden)

    David J Huggins

    2010-08-01

    Full Text Available The Polo-Like Kinase 1 (PLK1 acts as a central regulator of mitosis and is over-expressed in a wide range of human tumours where high levels of expression correlate with a poor prognosis. PLK1 comprises two structural elements, a kinase domain and a polo-box domain (PBD. The PBD binds phosphorylated substrates to control substrate phosphorylation by the kinase domain. Although the PBD preferentially binds to phosphopeptides, it has a relatively broad sequence specificity in comparison with other phosphopeptide binding domains. We analysed the molecular determinants of recognition by performing molecular dynamics simulations of the PBD with one of its natural substrates, CDC25c. Predicted binding free energies were calculated using a molecular mechanics, Poisson-Boltzmann surface area approach. We calculated the per-residue contributions to the binding free energy change, showing that the phosphothreonine residue and the mainchain account for the vast majority of the interaction energy. This explains the very broad sequence specificity with respect to other sidechain residues. Finally, we considered the key role of bridging water molecules at the binding interface. We employed inhomogeneous fluid solvation theory to consider the free energy of water molecules on the protein surface with respect to bulk water molecules. Such an analysis highlights binding hotspots created by elimination of water molecules from hydrophobic surfaces. It also predicts that a number of water molecules are stabilized by the presence of the charged phosphate group, and that this will have a significant effect on the binding affinity. Our findings suggest a molecular rationale for the promiscuous binding of the PBD and highlight a role for bridging water molecules at the interface. We expect that this method of analysis will be very useful for probing other protein surfaces to identify binding hotspots for natural binding partners and small molecule inhibitors.

  6. Multivariate fault isolation of batch processes via variable selection in partial least squares discriminant analysis.

    Science.gov (United States)

    Yan, Zhengbing; Kuang, Te-Hui; Yao, Yuan

    2017-09-01

    In recent years, multivariate statistical monitoring of batch processes has become a popular research topic, wherein multivariate fault isolation is an important step aiming at the identification of the faulty variables contributing most to the detected process abnormality. Although contribution plots have been commonly used in statistical fault isolation, such methods suffer from the smearing effect between correlated variables. In particular, in batch process monitoring, the high autocorrelations and cross-correlations that exist in variable trajectories make the smearing effect unavoidable. To address such a problem, a variable selection-based fault isolation method is proposed in this research, which transforms the fault isolation problem into a variable selection problem in partial least squares discriminant analysis and solves it by calculating a sparse partial least squares model. As different from the traditional methods, the proposed method emphasizes the relative importance of each process variable. Such information may help process engineers in conducting root-cause diagnosis. Copyright © 2017 ISA. Published by Elsevier Ltd. All rights reserved.

  7. Surface remineralization potential of casein phosphopeptide-amorphous calcium phosphate on enamel eroded by cola-drinks: An in-situ model study

    Directory of Open Access Journals (Sweden)

    Navneet Grewal

    2013-01-01

    Full Text Available Aim: The aim of this study was to investigate the remineralization potential of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP on enamel eroded by cola drinks. Subjects and Methods: A total of 30 healthy subjects were selected from a random sample of 1200 children and divided into two groups of 15 each wherein calcium and phosphorus analyses and scanning electron microscope (SEM analysis was carried out to investigate the remineralization of enamel surface. A total of 30 non-carious premolar teeth were selected from the human tooth bank (HTB to prepare the in-situ appliance. Three enamel slabs were prepared from the same. One enamel slab was used to obtain baseline values and the other two were embedded into the upper palatal appliances prepared on the subjects′ maxillary working model. The subjects wore the appliance after which 30 ml cola drink exposure was given. After 15 days, the slabs were removed and subjected to respective analysis. Statistical Analysis Used: Means of all the readings of soluble calcium and phosphorous levels at baseline,post cola-drink exposure and post cpp-acp application were subjected to statistical analysis SPSS11.5 version.Comparison within groups and between groups was carried out using ANOVA and F-values at 1% level of significance. Results: Decrease in calcium solubility of enamel in the CPP-ACP application group as compared to post-cola drink exposure group (P < 0.05 was seen. Distinctive change in surface topography of enamel in the post-CPP-ACP application group as compared to post-cola drink exposure group was observed. Conclusion: CPP-ACP significantly promoted remineralization of enamel eroded by cola drinks as revealed by significant morphological changes seen in SEM magnification and spectrophotometric analyses.

  8. Selection of oleuropein-degrading lactic acid bacteria strains isolated from fermenting Moroccan green olives

    Energy Technology Data Exchange (ETDEWEB)

    Ghabbour, N.; Lamzira, Z.; Thonart, P.; Cidalia, P.; Markaouid, M.; Asehraoua, A.

    2011-07-01

    A total of 177 strains of lactic acid bacteria (LAB) were isolated from early-stage Moroccan Picholine green olive fermentation, including Lactobacillus plantarum (44.63%), Lactobacillus pentosus (25.99%), Lactobacillus brevis (9.61%) and Pediococcus pentosaceus (19.77%). All the isolates were screened for their tolerance to olive leaf extract and oleuropein. Most of the isolates (85.3%) were found able to degrade oleuropein, when evaluated by either oleuropein or 5-Bromo-4-chloro-3-indolyl {beta}-D-glucuronide (X-Gluc) as substrates. The biodegradation capacity of the selected strains of each species was confirmed by HPLC analysis. (Author).

  9. Structural and biophysical investigation of the interaction of a mutant Grb2 SH2 domain (W121G) with its cognate phosphopeptide.

    Science.gov (United States)

    Papaioannou, Danai; Geibel, Sebastian; Kunze, Micha B A; Kay, Christopher W M; Waksman, Gabriel

    2016-03-01

    The adaptor protein Grb2 is a key element of mitogenetically important signaling pathways. With its SH2 domain it binds to upstream targets while its SH3 domains bind to downstream proteins thereby relaying signals from the cell membranes to the nucleus. The Grb2 SH2 domain binds to its targets by recognizing a phosphotyrosine (pY) in a pYxNx peptide motif, requiring an Asn at the +2 position C-terminal to the pY with the residue either side of this Asn being hydrophobic. Structural analysis of the Grb2 SH2 domain in complex with its cognate peptide has shown that the peptide adopts a unique β-turn conformation, unlike the extended conformation that phosphopeptides adopt when bound to other SH2 domains. TrpEF1 (W121) is believed to force the peptide into this unusual conformation conferring this unique specificity to the Grb2 SH2 domain. Using X-ray crystallography, electron paramagnetic resonance (EPR) spectroscopy, and isothermal titration calorimetry (ITC), we describe here a series of experiments that explore the role of TrpEF1 in determining the specificity of the Grb2 SH2 domain. Our results demonstrate that the ligand does not adopt a pre-organized structure before binding to the SH2 domain, rather it is the interaction between the two that imposes the hairpin loop to the peptide. Furthermore, we find that the peptide adopts a similar structure when bound to both the wild-type Grb2 SH2 domain and a TrpEF1Gly mutant. This suggests that TrpEF1 is not the determining factor for the conformation of the phosphopeptide. © 2015 The Protein Society.

  10. Population genetic structure and natural selection of apical membrane antigen-1 in Plasmodium vivax Korean isolates.

    Science.gov (United States)

    Kang, Jung-Mi; Lee, Jinyoung; Cho, Pyo-Yun; Moon, Sung-Ung; Ju, Hye-Lim; Ahn, Seong Kyu; Sohn, Woon-Mok; Lee, Hyeong-Woo; Kim, Tong-Soo; Na, Byoung-Kuk

    2015-11-16

    Plasmodium vivax apical membrane antigen-1 (PvAMA-1) is a leading candidate antigen for blood stage malaria vaccine. However, antigenic variation is a major obstacle in the development of an effective vaccine based on this antigen. In this study, the genetic structure and the effect of natural selection of PvAMA-1 among Korean P. vivax isolates were analysed. Blood samples were collected from 66 Korean patients with vivax malaria. The entire PvAMA-1 gene was amplified by polymerase chain reaction and cloned into a TA cloning vector. The PvAMA-1 sequence of each isolate was sequenced and the polymorphic characteristics and effect of natural selection were analysed using the DNASTAR, MEGA4, and DnaSP programs. Thirty haplotypes of PvAMA-1, which were further classified into seven different clusters, were identified in the 66 Korean P. vivax isolates. Domain II was highly conserved among the sequences, but substantial nucleotide diversity was observed in domains I and III. The difference between the rates of non-synonymous and synonymous mutations suggested that the gene has evolved under natural selection. No strong evidence indicating balancing or positive selection on PvAMA-1 was identified. Recombination may also play a role in the resulting genetic diversity of PvAMA-1. This study is the first comprehensive analysis of nucleotide diversity across the entire PvAMA-1 gene using a single population sample from Korea. Korean PvAMA-1 had limited genetic diversity compared to PvAMA-1 in global isolates. The overall pattern of genetic polymorphism of Korean PvAMA-1 differed from other global isolates and novel amino acid changes were also identified in Korean PvAMA-1. Evidences for natural selection and recombination event were observed, which is likely to play an important role in generating genetic diversity across the PvAMA-1. These results provide useful information for the understanding the population structure of P. vivax circulating in Korea and have important

  11. Environmental considerations in the selection of isolation gowns: A life cycle assessment of reusable and disposable alternatives.

    Science.gov (United States)

    Vozzola, Eric; Overcash, Michael; Griffing, Evan

    2018-04-11

    Isolation gowns serve a critical role in infection control by protecting healthcare workers, visitors, and patients from the transfer of microorganisms and body fluids. The decision of whether to use a reusable or disposable garment system is a selection process based on factors including sustainability, barrier effectiveness, cost, and comfort. Environmental sustainability is increasingly being used in the decision-making process. Life cycle assessment is the most comprehensive and widely used tool used to evaluate environmental performance. The environmental impacts of market-representative reusable and disposable isolation gown systems were compared using standard life cycle assessment procedures. The basis of comparison was 1,000 isolation gown uses in a healthcare setting. The scope included the manufacture, use, and end-of-life stages of the gown systems. At the healthcare facility, compared to the disposable gown system, the reusable gown system showed a 28% reduction in energy consumption, a 30% reduction in greenhouse gas emissions, a 41% reduction in blue water consumption, and a 93% reduction in solid waste generation. Selecting reusable garment systems may result in significant environmental benefits compared to selecting disposable garment systems. By selecting reusable isolation gowns, healthcare facilities can add these quantitative benefits directly to their sustainability scorecards. Copyright © 2018 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

  12. In vitro selection and characterization of putative probiotics isolated from the gut of Acipenser baerii (Brandt, 1869)

    NARCIS (Netherlands)

    Geraylou, Z.; Vanhove, M.P.M.; Souffreau, C.; Rurangwa, E.; Buyse, J.; Ollevier, F.

    2014-01-01

    To select and characterize potential probiotic bacteria from the gut microbiota of Siberian sturgeon (Acipenser baerii), 129 strains isolated from the hindgut were screened for antagonistic activity against five fish pathogens. Ten isolates showed antagonism towards three or more pathogens. Nine of

  13. Resistance of Bacteria Isolated from Otamiri River to Heavy Metals and Some Selected Antibiotics

    OpenAIRE

    I.C. Mgbemena; J.C. Nnokwe; L.A. Adjeroh; N.N. Onyemekara

    2012-01-01

    This study is aimed at determining the resistance of bacteria to heavy metals and some antibiotics. The ability of aquatic bacteria isolates from Otamiri River at Ihiagwa in Owerri North, Imo State to tolerate or resist the presence of certain selected heavy metals: Pb+, Zn2+ and Fe2+ and some antibiotics was investigated. Identification tests for the bacteria isolates from Otamiri River revealed them to belong to the genera Pseudomonas, Aeromonas, Bacillus, Escherichia, Micrococcus and Prote...

  14. Tissue specific phosphorylation of mitochondrial proteins isolated from rat liver, heart muscle, and skeletal muscle

    DEFF Research Database (Denmark)

    Bak, Steffen; León, Ileana R; Jensen, Ole Nørregaard

    2013-01-01

    -specific phosphorylation sites were identified in tissue-specific enzymes such as those encoded by HMGCS2, BDH1, PCK2, CPS1, and OTC in liver mitochondria, and CKMT2 and CPT1B in heart and skeletal muscle. Kinase prediction showed an important role for PKA and PKC in all tissues but also for proline-directed kinases......Phosphorylation of mitochondrial proteins in a variety of biological processes is increasingly being recognized and may contribute to the differences in function and energy demands observed in mitochondria from different tissues such as liver, heart, and skeletal muscle. Here, we used a combination...... of TiO2 phosphopeptide-enrichment, HILIC fractionation, and LC-MS/MS on isolated mitochondria to investigate the tissue-specific mitochondrial phosphoproteomes of rat liver, heart, and skeletal muscle. In total, we identified 899 phosphorylation sites in 354 different mitochondrial proteins including...

  15. Selection and characterization of Argentine isolates of Trichoderma harzianum for effective biocontrol of Septoria leaf blotch of wheat.

    Science.gov (United States)

    Stocco, Marina C; Mónaco, Cecilia I; Abramoff, Cecilia; Lampugnani, Gladys; Salerno, Graciela; Kripelz, Natalia; Cordo, Cristina A; Consolo, Verónica F

    2016-03-01

    Species of the genus Trichoderma are economically important as biocontrol agents, serving as a potential alternative to chemical control. The applicability of Trichoderma isolates to different ecozones will depend on the behavior of the strains selected from each zone. The present study was undertaken to isolate biocontrol populations of Trichoderma spp. from the Argentine wheat regions and to select and characterize the best strains of Trichoderma harzianum by means of molecular techniques. A total of 84 out of the 240 strains of Trichoderma were able to reduce the disease severity of the leaf blotch of wheat. Thirty-seven strains were selected for the reduction equal to or greater than 50% of the severity, compared with the control. The percentage values of reduction of the pycnidial coverage ranged between 45 and 80%. The same last strains were confirmed as T. harzianum by polymerase chain reaction amplification of internal transcribed spacers, followed by sequencing. Inter-simple sequence repeat was used to examine the genetic variability among isolates. This resulted in a total of 132 bands. Further numerical analysis revealed 19 haplotypes, grouped in three clusters (I, II, III). Shared strains, with different geographical origins and isolated in different years, were observed within each cluster. The origin of the isolates and the genetic group were partially related. All isolates from Paraná were in cluster I, all isolates from Lobería were in cluster II, and all isolates from Pergamino and Santa Fe were in cluster III. Our results suggest that the 37 native strains of T. harzianum are important in biocontrol programs and could be advantageous for the preparation of biopesticides adapted to the agroecological conditions of wheat culture.

  16. Partial Sequencing of 16S rRNA Gene of Selected Staphylococcus aureus Isolates and its Antibiotic Resistance

    Directory of Open Access Journals (Sweden)

    Harsi Dewantari Kusumaningrum

    2016-08-01

    Full Text Available The choice of primer used in 16S rRNA sequencing for identification of Staphylococcus species found in food is important. This study aimed to characterize Staphylococcus aureus isolates by partial sequencing based on 16S rRNA gene employing primers 16sF, 63F or 1387R. The isolates were isolated from milk, egg dishes and chicken dishes and selected based on the presence of sea gene that responsible for formation of enterotoxin-A. Antibiotic susceptibility of the isolates towards six antibiotics was also tested. The use of 16sF resulted generally in higher identity percentage and query coverage compared to the sequencing by 63F or 1387R. BLAST results of all isolates, sequenced by 16sF, showed 99% homology to complete genome of four S. aureus strains, with different characteristics on enterotoxin production and antibiotic resistance. Considering that all isolates were carrying sea gene, indicated by the occurence of 120 bp amplicon after PCR amplification using primer SEA1/SEA2,  the isolates were most in agreeing to S. aureus subsp. aureus ST288. This study indicated that 4 out of 8 selected isolates were resistant towards streptomycin. The 16S rRNA gene sequencing using 16sF is useful for identification of S. aureus. However, additional analysis such as PCR employing specific gene target, should give a valuable supplementary information, when specific characteristic is expected.

  17. Sexual selection, sexual isolation and pheromones in Drosophila melanogaster strains after long-term maintaining on different diets.

    Science.gov (United States)

    Trajković, Jelena; Miličić, Dragana; Savić, Tatjana; Pavković-Lučić, Sofija

    2017-07-01

    Evolution of reproductive isolation may be a consequence of a variety of signals used in courtship and mate preferences. Pheromones play an important role in both sexual selection and sexual isolation. The abundance of pheromones in Drosophila melanogaster may depend on different environmental factors, including diet. The aim of this study was to ascertain to which degree principal pheromones affect sexual selection in D. melanogaster. We used D. melanogaster strains reared for 14 years on four substrates: standard cornmeal substrate and those containing tomato, banana and carrot. We have previously determined that long-term maintaining of these dietary strains resulted in differences in their cuticular hydrocarbons profile (CHs). In this work, we have tested the level of sexual selection and sexual isolation between aforementioned strains. We found that the high levels of cis-vaccenyl acetate, 7-pentacosene and 7,11-nonacosadiene in the strain reared on a substrate containing carrot affected the individual attractiveness and influenced sexual isolation between flies of this strain and flies reared on a substrate containing banana. Based on these results, long-term different diets, may contribute, to sexual behaviour of D. melanogaster via the effects of principal pheromones. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. In vitro evaluation of casein phosphopeptide-amorphous calcium phosphate effect on the shear bond strength of dental adhesives to enamel.

    Science.gov (United States)

    Shadman, Niloofar; Ebrahimi, Shahram Farzin; Shoul, Maryam Azizi; Sattari, Hasti

    2015-01-01

    Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) is applied for remineralization of early caries lesions or tooth sensitivity conditions and may affect subsequent resin bonding. This in vitro study investigated the effect of CPP-ACP on the shear bond strength of dental adhesives to enamel. Sixty extracted human molar teeth were selected and randomly divided into three groups and six subgroups. Buccal or lingual surfaces of teeth were prepared to create a flat enamel surface. Adhesives used were Tetric N-Bond, AdheSE and AdheSE One F. In three subgroups, before applying adhesives, enamel surfaces were treated with Tooth Mousse CPP-ACP for one hour, rinsed and stored in 37°C temperature with 100% humidity. This procedure was repeated for 5 days and then adhesives were applied and Tetric N-Ceram composite was adhered to the enamel. This procedure was also fulfilled for the other three subgroups without CPP-ACP treatment. After 24 hour water storage, samples were tested for shear bond strength test in a universal testing machine. Failure modes were determined by stereomicroscope. Data were analyzed by t-test and one-way analysis of variance with P enamel only in Tetric N-Bond (P > 0.05). In non-applied CPP-ACP subgroups, there were statistically significant differences among all subgroups. Tetric N-Bond had the highest and AdheSE One F had the lowest shear bond strength. CPP-ACP application reduces the shear bond strength of AdheSE and AdheSE One F to enamel but not Tetric N-Bond.

  19. In vitro evaluation of casein phosphopeptide-amorphous calcium phosphate effect on the shear bond strength of dental adhesives to enamel

    Directory of Open Access Journals (Sweden)

    Niloofar Shadman

    2015-01-01

    Full Text Available Background: Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP is applied for remineralization of early caries lesions or tooth sensitivity conditions and may affect subsequent resin bonding. This in vitro study investigated the effect of CPP-ACP on the shear bond strength of dental adhesives to enamel. Materials and Methods: Sixty extracted human molar teeth were selected and randomly divided into three groups and six subgroups. Buccal or lingual surfaces of teeth were prepared to create a flat enamel surface. Adhesives used were Tetric N-Bond, AdheSE and AdheSE One F. In three subgroups, before applying adhesives, enamel surfaces were treated with Tooth Mousse CPP-ACP for one hour, rinsed and stored in 37°C temperature with 100% humidity. This procedure was repeated for 5 days and then adhesives were applied and Tetric N-Ceram composite was adhered to the enamel. This procedure was also fulfilled for the other three subgroups without CPP-ACP treatment. After 24 hour water storage, samples were tested for shear bond strength test in a universal testing machine. Failure modes were determined by stereomicroscope. Data were analyzed by t-test and one-way analysis of variance with P 0.05. In non-applied CPP-ACP subgroups, there were statistically significant differences among all subgroups. Tetric N-Bond had the highest and AdheSE One F had the lowest shear bond strength. Conclusion: CPP-ACP application reduces the shear bond strength of AdheSE and AdheSE One F to enamel but not Tetric N-Bond.

  20. Facile synthesis of Fe3O4@PDA core-shell microspheres functionalized with various metal ions: A systematic comparison of commonly-used metal ions for IMAC enrichment.

    Science.gov (United States)

    Jiang, Jiebing; Sun, Xueni; Li, Yan; Deng, Chunhui; Duan, Gengli

    2018-02-01

    Metal ions differed greatly in affinity towards phosphopeptides, and thus it is essential to systematically compare the phosphopeptides enrichment ability of different metal ions usually used in the IMAC techniques. In this work, for the first time, eight metal ions, including Nb 5+ , Ti 4+ , Zr 4+ , Ga 3+ , Y 3+ , In 3+ , Ce 4+ , Fe 3+ , were immobilized on the polydopamine (PDA)-coated Fe 3 O 4 (denoted as Fe 3 O 4 @PDA-M n+ ), and systematically compared by the real biosamples, in addition to standard phosphopeptides. Fe 3 O 4 microspheres were synthesized via the solvothermal reaction, followed by self-polymerization of dopamine on the surface. Then through taking advantage of the hydroxyl and amino group of PDA, the eight metal ions were easily adhered to the surface of Fe 3 O 4 @PDA. After characterization, the resultant Fe 3 O 4 @PDA-M n+ microspheres were applied to phosphopeptides enrichment based on the binding affinity between metal ions and phosphopeptides. According to the results, different metal ions presented diverse phosphopeptides enrichment efficiency in terms of selectivity, sensitivity and the enrichment ability from real complex samples, and Fe 3 O 4 @PDA-Nb 5+ and Fe 3 O 4 @PDA-Ti 4+ showed obvious advantages of the phosphopeptides enrichment effect after the comparison. This systematic comparison may provide certain reference for the use and development of IMAC materials in the future. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Mitsunobu mischief: Neighbor-directed histidine N(π)–alkylation provides access to peptides containing selectively functionalized imidazolium heterocycles

    Science.gov (United States)

    Qian, Wen-Jian

    2015-01-01

    There are few methodologies that yield peptides containing His residues with selective N(π), N(π)-bis-alkylated imidazole rings. We have found that, under certain conditions, on-resin Mitsunobu coupling of alcohols with peptides having a N(π)-alkylated His residue results in selective and high-yield alkylation of the imidazole N(π) nitrogen. The reaction requires the presence of a proximal phosphoric, carboxylic or sulfonic acid, and proceeds through an apparent intramolecular mechanism involving Mitsunobu intermediates. These transformations have particular application to phosphopeptides, where “charge masking” of one phosphoryl anionic charge by the cationic histidine imidazolium ion is now possible. This chemistry opens selective access to peptides containing differentially functionalized imidazolium heterocycles, which provide access to new classes of peptides and peptide mimetics. PMID:25739367

  2. Selective cultures for the isolation of biosurfactant producing bacteria: comparison of different combinations of environmental inocula and hydrophobic carbon sources.

    Science.gov (United States)

    Domingues, Patrícia M; Louvado, António; Oliveira, Vanessa; Coelho, Francisco J C R; Almeida, Adelaide; Gomes, Newton C M; Cunha, Angela

    2013-01-01

    The potential of estuarine microniches as reservoirs of biosurfactant-producing bacteria was evaluated by testing different combinations of inocula and hydrophobic carbon sources. Selective cultures using diesel, petroleum, or paraffin as hydrophobic carbon sources were prepared and inoculated with water from the surface microlayer, bulk sediments, and sediment of the rhizosphere of Halimione portulacoides. These inocula were compared regarding the frequency of biosurfactant-producing strains among selected isolates. The community structure of the selective cultures was profiled using denaturing gradient gel electrophoresis (DGGE) of the 16S rRNA gene fragments at the end of the incubation. The DGGE profiles corresponding to the communities established in selective cultures at the end of the incubation revealed that communities were different in terms of structural diversity. The highest diversity was observed in the selective cultures containing paraffin (H (') = 2.5). Isolates were obtained from the selective cultures (66) and tested for biosurfactant production by the atomized oil assay. Biosurfactant production was detected in 17 isolates identified as Microbacterium, Pseudomonas, Rhodococcus, and Serratia. The combination of estuarine surface microlayer (SML) water as inoculum and diesel as carbon source seems promising for the isolation of surfactant-producing bacteria. Supplemental materials are available for this article. Go to the publisher's online edition of Preparative Biochemistry and Biotechnology to view the supplemental file.

  3. The use of naturally occurring selectively isolated bacteria for inhibiting paraffin deposition

    International Nuclear Information System (INIS)

    Lazar, I.; Voicu, A.; Dobrota, S.; Petrisor, I.G.; Stefanescu, M.; Sandulescu, L.; Nicolescu, C.; Mucenica, D.

    1999-01-01

    One of the most severe problems at any oil fields producing paraffinic oils is that of paraffin depositions. Romania which has a long experience in oil production is also faced with this problem in many oil fields. The microbial treatment, based on the activity of naturally occurring, selectively isolated bacteria, is already proved as an effective alternative to conventional methods to prevent and remove paraffin damage. Using such kind of bacterial products, exciting results for inhibiting paraffin depositions have been obtained. In this paper results concerning the naturally occurring bacteria selectively isolated from hydrocarbon polluted sites as well as from paraffinic oils, semi-solid and solid paraffin depositions are presented. After a laboratory screening, 15 bacterial strains (BS 1-15), three bacterial consortia (BC 1-3) and a Special Bacterial Consortium (SBC1) were selected. For the selection of bacterial consortia, the classical enrichment culture method has been used. The Special Bacterial Consortium resulted from a mixture of BS 1-15 and BC 1-3 following the steps of the classical enrichment culture method. The BS 1-15, BC 1-3 and SBC1 have been tested for their performances in producing biosurfactants and biosolvents as well as for hydrocarbon utilisation. The SBC1 has been tested for its ability in degradation of hydrocarbons contained in several types of paraffinic or non-paraffinic oils, and then for inhibiting paraffin deposition on a 'flow equipment' using two types of paraffinic oils. The SBC1 has been also tested for degradation of hydrocarbons contained in semi-solid and solid paraffin depositions. The results obtained could support further applications to prevent and control paraffin depositions

  4. The use of naturally occurring selectively isolated bacteria for inhibiting paraffin deposition

    Energy Technology Data Exchange (ETDEWEB)

    Lazar, I.; Voicu, A.; Dobrota, S.; Petrisor, I.G.; Stefanescu, M.; Sandulescu, L. [Institute of Biology of the Romanian Academy, Spl. Independentei 296, Bucharest (Romania); Nicolescu, C.; Mucenica, D. [PETROSTAR Ploiesti, Bdul Bucuresti 35, Ploiesti (Romania)

    1999-01-01

    One of the most severe problems at any oil fields producing paraffinic oils is that of paraffin depositions. Romania which has a long experience in oil production is also faced with this problem in many oil fields. The microbial treatment, based on the activity of naturally occurring, selectively isolated bacteria, is already proved as an effective alternative to conventional methods to prevent and remove paraffin damage. Using such kind of bacterial products, exciting results for inhibiting paraffin depositions have been obtained. In this paper results concerning the naturally occurring bacteria selectively isolated from hydrocarbon polluted sites as well as from paraffinic oils, semi-solid and solid paraffin depositions are presented. After a laboratory screening, 15 bacterial strains (BS 1-15), three bacterial consortia (BC 1-3) and a Special Bacterial Consortium (SBC1) were selected. For the selection of bacterial consortia, the classical enrichment culture method has been used. The Special Bacterial Consortium resulted from a mixture of BS 1-15 and BC 1-3 following the steps of the classical enrichment culture method. The BS 1-15, BC 1-3 and SBC1 have been tested for their performances in producing biosurfactants and biosolvents as well as for hydrocarbon utilisation. The SBC1 has been tested for its ability in degradation of hydrocarbons contained in several types of paraffinic or non-paraffinic oils, and then for inhibiting paraffin deposition on a `flow equipment` using two types of paraffinic oils. The SBC1 has been also tested for degradation of hydrocarbons contained in semi-solid and solid paraffin depositions. The results obtained could support further applications to prevent and control paraffin depositions

  5. Isolating E.Coli Bacteriophage from Raw Sewage and Determining its Selectivity to the Host Cell

    Directory of Open Access Journals (Sweden)

    SM Imeni

    2016-05-01

    Full Text Available Introduction: Bacteriophages are viruses that infect and destroy prokaryote cells, specifically the bacteria. They act too selective, so as each bacteriophage affects only on specific type of bacteria. Due to their specific features, bacteriophages can be used as an appropriate substitute for antibiotics in infectious diseases treatment. Therefore, this study aimed to isolate E. coli-specific bacteriophage from raw sewage. Methods: Eight samples of raw sewage, each containing approximately 50 ml of raw sewage with 10 minute gap, were prepared from Zargandeh wastewater treatment plant, Tehran, Iran. The sewages were mixed with Brain-heart infusion medium (BHI as a liquid culture medium in order to let the microorganisms grow. Incubation, purification and determination of bacteria were followed repeatedly to isolate the bacteriophage. Then it was tested on E.coli (ATCC 25922, Enterococcus faecalis (ATCC 19433, Staphylococcus aureus (ATCC 2392, and Yersinia enterocolitica (ATCC 9610 in order to determine the bacteriophage selectivity. Results: The E.coli bacteriophages were successfully isolated from all the eight samples, that were completely able to lyse and destroy E.coli bacterial cells, though no effect was observed on other types of bacteria. Conclusion: The study findings revealed that bacteriophages act selectively. Considering the raise of antibiotic resistance in the world, bacteriophages can serve as a good substitute for antibiotics in treating infectious diseases.

  6. Isolation, selection and characterization of root-associated growth promoting bacteria in Brazil Pine (Araucaria angustifolia).

    Science.gov (United States)

    Ribeiro, Carlos Marcelo; Cardoso, Elke Jurandy Bran Nogueira

    2012-01-20

    Araucaria angustifolia, a unique species of this genus that occurs naturally in Brazil, has a high socio-economic and environmental value and is critically endangered of extinction, since it has been submitted to intense predatory exploitation during the last century. Root-associated bacteria from A. angustifolia were isolated, selected and characterized for their biotechnological potential of growth promotion and biocontrol of plant pathogenic fungi. Ninety-seven strains were isolated and subjected to chemical tests. All isolates presented at least one positive feature, characterizing them as potential PGPR. Eighteen isolates produced indole-3-acetic acid (IAA), 27 were able to solubilize inorganic phosphate, 21 isolates were presumable diazotrophs, with pellicle formation in nitrogen-free culture medium, 83 were phosphatases producers, 37 were positive for siderophores and 45 endospore-forming isolates were antagonistic to Fusarium oxysporum, a pathogen of conifers. We also observed the presence of bacterial strains with multiple beneficial mechanisms of action. Analyzing the fatty acid methyl ester (FAME) and partial sequencing of the 16S rRNA gene of these isolates, it was possible to characterize the most effective isolates as belonging to Bacillaceae (9 isolates), Enterobacteriaceae (11) and Pseudomonadaceae (1). As far as we know, this is the first study to include the species Ewingella americana as a PGPR. Copyright © 2011 Elsevier GmbH. All rights reserved.

  7. Selection of Leuconostoc strains isolated from artisanal Serrano Catarinense cheese for use as adjuncts in cheese manufacture.

    Science.gov (United States)

    Seixas, Felipe Nael; Rios, Edson Antônio; Martinez de Oliveira, André Luiz; Beloti, Vanerli; Poveda, Justa Maria

    2018-08-01

    Serrano Catarinense cheese is a raw bovine milk cheese produced in the region of Santa Catarina, Brazil. Twelve representative strains of Leuconostoc isolated from 20 samples of this artisanal cheese were selected and submitted for evaluation of the acidifying, proteolytic, autolytic, aminopeptidase and lipolytic activities, NaCl and acid resistance, production of dextran and biogenic amines and antimicrobial activity. The aim was to genetically and technologically characterize the Leuconostoc strains in order to use them in mixed starter cultures for cheese manufacture. Leuconostoc mesenteroides subsp. mesenteroides was the species that accounted for the largest proportion of isolates of Leuconostoc genus. Two leuconostoc isolates stood out in the acidifying activity, with reduction in pH of 1.12 and 1.04 units. The isolates showed low proteolytic and autolytic activity. Most of the isolates were dextran producers, presented good resistance to the salt and pH conditions of the cheese and showed antimicrobial activity against cheese pathogen bacteria, and none of them produced biogenic amines. These results allowed the selection of five strains (UEL 04, UEL 12, UEL 18, UEL 21 and UEL 28) as good candidates for use as adjunct cultures for cheese manufacture. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

  8. Crystal Structures and Thermodynamic Analysis Reveal Distinct Mechanisms of CD28 Phosphopeptide Binding to the Src Homology 2 (SH2) Domains of Three Adaptor Proteins*

    Science.gov (United States)

    Inaba, Satomi; Numoto, Nobutaka; Ogawa, Shuhei; Morii, Hisayuki; Ikura, Teikichi; Abe, Ryo; Ito, Nobutoshi; Oda, Masayuki

    2017-01-01

    Full activation of T cells and differentiation into effector T cells are essential for many immune responses and require co-stimulatory signaling via the CD28 receptor. Extracellular ligand binding to CD28 recruits protein-tyrosine kinases to its cytoplasmic tail, which contains a YMNM motif. Following phosphorylation of the tyrosine, the proteins growth factor receptor-bound protein 2 (Grb2), Grb2-related adaptor downstream of Shc (Gads), and p85 subunit of phosphoinositide 3-kinase may bind to pYMNM (where pY is phosphotyrosine) via their Src homology 2 (SH2) domains, leading to downstream signaling to distinct immune pathways. These three adaptor proteins bind to the same site on CD28 with variable affinity, and all are important for CD28-mediated co-stimulatory function. However, the mechanism of how these proteins recognize and compete for CD28 is unclear. To visualize their interactions with CD28, we have determined the crystal structures of Gads SH2 and two p85 SH2 domains in complex with a CD28-derived phosphopeptide. The high resolution structures obtained revealed that, whereas the CD28 phosphopeptide bound to Gads SH2 is in a bent conformation similar to that when bound to Grb2 SH2, it adopts a more extended conformation when bound to the N- and C-terminal SH2 domains of p85. These differences observed in the peptide-protein interactions correlated well with the affinity and other thermodynamic parameters for each interaction determined by isothermal titration calorimetry. The detailed insight into these interactions reported here may inform the development of compounds that specifically inhibit the association of CD28 with these adaptor proteins to suppress excessive T cell responses, such as in allergies and autoimmune diseases. PMID:27927989

  9. Crystal Structures and Thermodynamic Analysis Reveal Distinct Mechanisms of CD28 Phosphopeptide Binding to the Src Homology 2 (SH2) Domains of Three Adaptor Proteins.

    Science.gov (United States)

    Inaba, Satomi; Numoto, Nobutaka; Ogawa, Shuhei; Morii, Hisayuki; Ikura, Teikichi; Abe, Ryo; Ito, Nobutoshi; Oda, Masayuki

    2017-01-20

    Full activation of T cells and differentiation into effector T cells are essential for many immune responses and require co-stimulatory signaling via the CD28 receptor. Extracellular ligand binding to CD28 recruits protein-tyrosine kinases to its cytoplasmic tail, which contains a YMNM motif. Following phosphorylation of the tyrosine, the proteins growth factor receptor-bound protein 2 (Grb2), Grb2-related adaptor downstream of Shc (Gads), and p85 subunit of phosphoinositide 3-kinase may bind to pYMNM (where pY is phosphotyrosine) via their Src homology 2 (SH2) domains, leading to downstream signaling to distinct immune pathways. These three adaptor proteins bind to the same site on CD28 with variable affinity, and all are important for CD28-mediated co-stimulatory function. However, the mechanism of how these proteins recognize and compete for CD28 is unclear. To visualize their interactions with CD28, we have determined the crystal structures of Gads SH2 and two p85 SH2 domains in complex with a CD28-derived phosphopeptide. The high resolution structures obtained revealed that, whereas the CD28 phosphopeptide bound to Gads SH2 is in a bent conformation similar to that when bound to Grb2 SH2, it adopts a more extended conformation when bound to the N- and C-terminal SH2 domains of p85. These differences observed in the peptide-protein interactions correlated well with the affinity and other thermodynamic parameters for each interaction determined by isothermal titration calorimetry. The detailed insight into these interactions reported here may inform the development of compounds that specifically inhibit the association of CD28 with these adaptor proteins to suppress excessive T cell responses, such as in allergies and autoimmune diseases. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  10. Isolation, Characterization, and Selection of Molds Associated to Fermented Black Table Olives

    Directory of Open Access Journals (Sweden)

    Simona L. Bavaro

    2017-07-01

    Full Text Available Table olives are one of the most important fermented food in the Mediterranean countries. Apart from lactic acid bacteria and yeasts that mainly conduct the olive fermentation, molds can develop on the brine surface, and can have either deleterious or useful effects on this process. From the food safety point of view, occurring molds could also produce mycotoxins, so, it is important to monitor and control them. In this respect, identification of molds associated to two Italian and two Greek fermented black table olives cultivars, was carried out. Sixty strains were isolated and molecularly identified as Penicillium crustosum (21, P. roqueforti (29, P. paneum (1, P. expansum (6, P. polonicum (2, P. commune (1. A group of 20 selected isolates was subjected to technological (beta-glucosidase, cellulolytic, ligninolytic, pectolytic, and xylanolytic activities; proteolytic enzymes and safety (biogenic amines and secondary metabolites, including mycotoxins characterization. Combining both technological (presence of desired and absence of undesired enzymatic activities and safety aspects (no or low production of biogenic amines and regulated mycotoxins, it was possible to select six strains with biotechnological interest. These are putative candidates for future studies as autochthonous co-starters with yeasts and lactic acid bacteria for black table olive production.

  11. Contributions of natural and sexual selection to the evolution of premating reproductive isolation: a research agenda.

    Science.gov (United States)

    Safran, Rebecca J; Scordato, Elizabeth S C; Symes, Laurel B; Rodríguez, Rafael L; Mendelson, Tamra C

    2013-11-01

    Speciation by divergent natural selection is well supported. However, the role of sexual selection in speciation is less well understood due to disagreement about whether sexual selection is a mechanism of evolution separate from natural selection, as well as confusion about various models and tests of sexual selection. Here, we outline how sexual selection and natural selection are different mechanisms of evolutionary change, and suggest that this distinction is critical when analyzing the role of sexual selection in speciation. Furthermore, we clarify models of sexual selection with respect to their interaction with ecology and natural selection. In doing so, we outline a research agenda for testing hypotheses about the relative significance of divergent sexual and natural selection in the evolution of reproductive isolation. Copyright © 2013 Elsevier Ltd. All rights reserved.

  12. Isolation of two independent allyl alcohol resistant Adh-1 null mutants following selection of pollen and seeds.

    NARCIS (Netherlands)

    Wisman, E.; Ramanna, M.S.; Zabel, P.

    1993-01-01

    The Adh-1 null mutant (B15-1-8) isolated previously was used to establish conditions that allow the selection of ADH-deficient pollen grains and seeds of tomato. New Adh-1 null mutants were then selected among the progenies derived from crosses between the genetically unstable tomato lines Yvms,

  13. Selection of antagonistic bacteria isolated from the Physalis peruviana rhizosphere against Fusarium oxysporum.

    Science.gov (United States)

    Urrea, R; Cabezas, L; Sierra, R; Cárdenas, M; Restrepo, S; Jiménez, P

    2011-09-01

    Cape gooseberries (Physalis peruviana) have become increasingly important in Colombia for both domestic consumption and the international export market. Vascular wilting caused by Fusarium oxysporum is the most damaging disease to P. peruviana crops in Colombia. The control of this pathogen is mainly carried out by chemical and cultural practices, increasing production costs and generating resistance. Therefore, the objectives of this study were to test rhizobacteria isolates from P. peruviana rhizosphere against F. oxysporum under in vitro and in vivo conditions. Over 120 strains were isolated, and five were selected for their high inhibition of F. oxysporum growth and conidia production under in vitro conditions. These strains inhibited growth by 41-58% and reduced three- to fivefold conidia production. In the in vivo assays, all the tested isolates significantly reduced fungal pathogenicity in terms of virulence. Isolate B-3.4 was the most efficient in delaying the onset of the first symptoms. All isolates were identified as belonging to the genus Pseudomonas except for A-19 (Bacillus sp.). Our results confirmed that there are prospective rhizobacteria strains that can be used as biological control agents; some of them being able to inhibit in vitro F. oxysporum growth and sporulation. Incorporating these bacteria into biological control strategies for the disease that causes high economical losses in the second most exported fruit from Colombia would result in a reduced impact on environment and economy. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

  14. Evolution of premating reproductive isolation among conspecific populations of the sea rock-pool beetle Ochthebius urbanelliae driven by reinforcing natural selection.

    Science.gov (United States)

    Porretta, Daniele; Urbanelli, Sandra

    2012-04-01

    How natural selection might be involved in speciation remains a fundamental question in evolutionary biology. When two or more species co-occur in the same areas, natural selection may favor divergence in mating traits. By acting in sympatric but not allopatric populations, natural selection can also affect mate choice within species and ultimately initiate speciation among conspecific populations. Here, we address this potential effect in the sea rock-pool beetles Ochthebius quadricollis and O. urbanelliae. The two species, which inhabit the Mediterranean coasts, co-occurr syntopically in an area along the Italian Tyrrhenian coast and completed reproductive isolation by reinforcement. In this article, through mating trials under laboratory conditions between conspecific populations, we found in O. quadricollis no deviations from random mating. Conversely, in O. urbanelliae, we found a clear pattern of premating isolation between the reinforced populations sympatric with O. quadricollis and those nonreinforced allopatric. This pattern is consistent with the view that natural selection, which completed the reproductive isolation between the two species in sympatry, led incidentally also to partial premating reproductive isolation (I(PSI) estimator from 0.683 to 0.792) between conspecific populations of O. urbanelliae. This case study supports an until recently underappreciated role of natural selection resulting from species interactions in initiating speciation. © 2011 The Author(s). Evolution© 2011 The Society for the Study of Evolution.

  15. Does systemic administration of casein phosphopeptides affect orthodontic movement and root resorption in rats?

    Science.gov (United States)

    Crowther, Lachlan; Shen, Gang; Almuzian, Mohammed; Jones, Allan; Walsh, William; Oliver, Rema; Petocz, Peter; Tarraf, Nour E; Darendeliler, M Ali

    2017-10-01

    To assess the potential effects of casein phosphopeptides (CPPs) on orthodontically induced iatrogenic root resorption (OIIRR) and orthodontic teeth movement. Forty Wistar rats (aged 11 weeks) were randomly divided into experimental group (EG; n = 20) that received a diet supplemented with CPP and control group (CG; n = 20) devoid of diet supplement. A 150 g force was applied using nickel titanium (NiTi) coil that was bonded on maxillary incisors and extended unilaterally to a maxillary first molar. At Day 28, animals in both groups were euthanized. Volumetric assessment of root resorption craters and linear measurement of maxillary first molars movement were blindly examined using a micro-computed tomography scan. Nine rats were excluded from the experiment due to loss during general anesthesia or appliances' failure. Intra-operator reproducibility was high in both volumetric and linear measurements, 92.8 per cent and 98.5-97.6 per cent, respectively. The results reveal that dietary CPP has statistically insignificant effect on the overall OIIRR and orthodontic movement. CPP seems to have statistically insignificant effect on the volume of OIIRR and orthodontic movement in rats. A long-term study with larger sample size using a different concentration of CPP is required to clarify the dentoalveolar effect of CPP. © The Author 2017. Published by Oxford University Press on behalf of the European Orthodontic Society. All rights reserved. For permissions, please email: journals.permissions@oup.com

  16. Phylodynamic analysis of clinical and environmental Vibrio cholerae isolates from Haiti reveals diversification driven by positive selection.

    Science.gov (United States)

    Azarian, Taj; Ali, Afsar; Johnson, Judith A; Mohr, David; Prosperi, Mattia; Veras, Nazle M; Jubair, Mohammed; Strickland, Samantha L; Rashid, Mohammad H; Alam, Meer T; Weppelmann, Thomas A; Katz, Lee S; Tarr, Cheryl L; Colwell, Rita R; Morris, J Glenn; Salemi, Marco

    2014-12-23

    Phylodynamic analysis of genome-wide single-nucleotide polymorphism (SNP) data is a powerful tool to investigate underlying evolutionary processes of bacterial epidemics. The method was applied to investigate a collection of 65 clinical and environmental isolates of Vibrio cholerae from Haiti collected between 2010 and 2012. Characterization of isolates recovered from environmental samples identified a total of four toxigenic V. cholerae O1 isolates, four non-O1/O139 isolates, and a novel nontoxigenic V. cholerae O1 isolate with the classical tcpA gene. Phylogenies of strains were inferred from genome-wide SNPs using coalescent-based demographic models within a Bayesian framework. A close phylogenetic relationship between clinical and environmental toxigenic V. cholerae O1 strains was observed. As cholera spread throughout Haiti between October 2010 and August 2012, the population size initially increased and then fluctuated over time. Selection analysis along internal branches of the phylogeny showed a steady accumulation of synonymous substitutions and a progressive increase of nonsynonymous substitutions over time, suggesting diversification likely was driven by positive selection. Short-term accumulation of nonsynonymous substitutions driven by selection may have significant implications for virulence, transmission dynamics, and even vaccine efficacy. Cholera, a dehydrating diarrheal disease caused by toxigenic strains of the bacterium Vibrio cholerae, emerged in 2010 in Haiti, a country where there were no available records on cholera over the past 100 years. While devastating in terms of morbidity and mortality, the outbreak provided a unique opportunity to study the evolutionary dynamics of V. cholerae and its environmental presence. The present study expands on previous work and provides an in-depth phylodynamic analysis inferred from genome-wide single nucleotide polymorphisms of clinical and environmental strains from dispersed geographic settings in

  17. Crystal Structure of the Human Symplekin-Ssu72-CTD Phosphopeptide Complex

    Energy Technology Data Exchange (ETDEWEB)

    K Xiang; T Nigaike; S Xiang; T Kilic; M Beh; J Manley; L Tong

    2011-12-31

    Symplekin (Pta1 in yeast) is a scaffold in the large protein complex that is required for 3'-end cleavage and polyadenylation of eukaryotic messenger RNA precursors (pre-mRNAs); it also participates in transcription initiation and termination by RNA polymerase II (Pol II). Symplekin mediates interactions between many different proteins in this machinery, although the molecular basis for its function is not known. Here we report the crystal structure at 2.4 {angstrom} resolution of the amino-terminal domain (residues 30-340) of human symplekin in a ternary complex with the Pol II carboxy-terminal domain (CTD) Ser5 phosphatase Ssu72 and a CTD Ser5 phosphopeptide. The N-terminal domain of symplekin has the ARM or HEAT fold, with seven pairs of antiparallel {alpha}-helices arranged in the shape of an arc. The structure of Ssu72 has some similarity to that of low-molecular-mass phosphotyrosine protein phosphatase, although Ssu72 has a unique active-site landscape as well as extra structural features at the C terminus that are important for interaction with symplekin. Ssu72 is bound to the concave face of symplekin, and engineered mutations in this interface can abolish interactions between the two proteins. The CTD peptide is bound in the active site of Ssu72, with the pSer5-Pro6 peptide bond in the cis configuration, which contrasts with all other known CTD peptide conformations. Although the active site of Ssu72 is about 25 {angstrom} from the interface with symplekin, we found that the symplekin N-terminal domain stimulates Ssu72 CTD phosphatase activity in vitro. Furthermore, the N-terminal domain of symplekin inhibits polyadenylation in vitro, but only when coupled to transcription. Because catalytically active Ssu72 overcomes this inhibition, our results show a role for mammalian Ssu72 in transcription-coupled pre-mRNA 3'-end processing.

  18. Comparative evaluation of the effects of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) and xylitol-containing chewing gum on salivary flow rate, pH and buffering capacity in children: An in vivo study.

    Science.gov (United States)

    Hegde, Rahul J; Thakkar, Janhavi B

    2017-01-01

    This study aimed to compare and evaluate the changes in the salivary flow rate, pH, and buffering capacity before and after chewing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) and xylitol-containing chewing gums in children. Sixty children aged between 8 and 12 years were selected for the study. They were randomly divided into Group 1 (CPP-ACP chewing gum) and Group 2 (xylitol-containing chewing gum) comprising thirty children each. Unstimulated and stimulated saliva samples at 15 and 30 min interval were collected from all children. All the saliva samples were estimated for salivary flow rate, pH, and buffering capacity. Significant increase in salivary flow rate, pH, and buffering capacity from baseline to immediately after spitting the chewing gum was found in both the study groups. No significant difference was found between the two study groups with respect to salivary flow rate and pH. Intergroup comparison indicated a significant increase in salivary buffer capacity in Group 1 when compared to Group 2. Chewing gums containing CPP-ACP and xylitol can significantly increase the physiochemical properties of saliva. These physiochemical properties of saliva have a definite relation with caries activity in children.

  19. Urea-modified metal-organic framework of type MIL-101(Cr) for the preconcentration of phosphorylated peptides

    International Nuclear Information System (INIS)

    Yang, Xiaoqing; Xia, Yan

    2016-01-01

    Mass spectrometry (MS) is the most powerful tool in phosphoproteomics research. However, phosphopeptides usually are present in low concentrations and their preconcentration therefore is highly desired. We describe a two-step method for the synthesis of a metal organic framework of the type MIL-101(Cr) that is modified with urea (then designated as MIL-101(Cr)-UR 2 ). It possesses large surface area, good solvent stability and high affinity for some phosphates. Due to the presence of modified urea functions, this material allows for selective and effective enrichment of phosphorylated peptides. It was successfully applied to the enrichment of phosphopeptides from non-fat-milk. The method was applied to the detection of phosphopeptides in a tryptic digest of β-casein where is showed a detection sensitivity as low as 10 −10 M. (author)

  20. Functional investigation of β-adrenoceptors in human isolated detrusor focusing on the novel selective β3-adrenoceptor agonist KUC-7322

    NARCIS (Netherlands)

    Igawa, Yasuhiko; Schneider, Tim; Yamazaki, Yoshinobu; Tatemichi, Satoshi; Homma, Yukio; Nishizawa, Osamu; Michel, Martin C.

    2012-01-01

    This study aimed to characterize the β-adrenoceptor (β-AR) subtype mediating relaxation of isolated human bladder strips and to explore relaxation by the novel β3-AR-selective agonist KUC-7322 for its relaxant effect on the human isolated detrusor and for its effect on the carbachol (CCh)-induced

  1. Population genetic structure and natural selection of Plasmodium falciparum apical membrane antigen-1 in Myanmar isolates.

    Science.gov (United States)

    Kang, Jung-Mi; Lee, Jinyoung; Moe, Mya; Jun, Hojong; Lê, Hương Giang; Kim, Tae Im; Thái, Thị Lam; Sohn, Woon-Mok; Myint, Moe Kyaw; Lin, Khin; Shin, Ho-Joon; Kim, Tong-Soo; Na, Byoung-Kuk

    2018-02-07

    Plasmodium falciparum apical membrane antigen-1 (PfAMA-1) is one of leading blood stage malaria vaccine candidates. However, genetic variation and antigenic diversity identified in global PfAMA-1 are major hurdles in the development of an effective vaccine based on this antigen. In this study, genetic structure and the effect of natural selection of PfAMA-1 among Myanmar P. falciparum isolates were analysed. Blood samples were collected from 58 Myanmar patients with falciparum malaria. Full-length PfAMA-1 gene was amplified by polymerase chain reaction and cloned into a TA cloning vector. PfAMA-1 sequence of each isolate was sequenced. Polymorphic characteristics and effect of natural selection were analysed with using DNASTAR, MEGA4, and DnaSP programs. Polymorphic nature and natural selection in 459 global PfAMA-1 were also analysed. Thirty-seven different haplotypes of PfAMA-1 were identified in 58 Myanmar P. falciparum isolates. Most amino acid changes identified in Myanmar PfAMA-1 were found in domains I and III. Overall patterns of amino acid changes in Myanmar PfAMA-1 were similar to those in global PfAMA-1. However, frequencies of amino acid changes differed by country. Novel amino acid changes in Myanmar PfAMA-1 were also identified. Evidences for natural selection and recombination event were observed in global PfAMA-1. Among 51 commonly identified amino acid changes in global PfAMA-1 sequences, 43 were found in predicted RBC-binding sites, B-cell epitopes, or IUR regions. Myanmar PfAMA-1 showed similar patterns of nucleotide diversity and amino acid polymorphisms compared to those of global PfAMA-1. Balancing natural selection and intragenic recombination across PfAMA-1 are likely to play major roles in generating genetic diversity in global PfAMA-1. Most common amino acid changes in global PfAMA-1 were located in predicted B-cell epitopes where high levels of nucleotide diversity and balancing natural selection were found. These results highlight the

  2. [Effect of casein phosphopeptide-amorphouscalcium phosphate (CPP-ACP) treatment on the shear bond strength of orthodontic brackets after tooth bleaching].

    Science.gov (United States)

    Lu, Jing; Ding, Xiao-jun; Yu, Xiao-ping; Gong, Yi-ming

    2015-10-01

    To evaluate the effect of casein phosphopeptide-amorphouscalcium phosphate (CPP-ACP) treatment on the shear bond strength of orthodontic brackets after tooth bleaching. One hundred extracted human premolars were randomly divided and treated according to 5 groups (n=20) : (1) no treatment; (2) 10% carbamide peroxide bleaching; (3) 38% hydrogen peroxide bleaching; (4)10% carbamide peroxide bleaching and CPP-ACP paste; (5)38% hydrogen peroxide bleaching and CPP-ACP paste. In all groups, the brackets were bonded using a conventional acid-etch and bond system (Transbond XT, 3M Unitek, Monrovia, Calif). The shear bond strength adhesive remnant index (ARI) of the brackets were determined and the data was analyzed by ANOVA and Bonferroni test using SPSS13.0 software package. The use of 10% carbamide peroxide and 38% hydrogen peroxide bleaching significantly decreased the shear bond strength of orthodontic brackets when compared with untreated group (P0.05). The ARI did not show any significant difference before and after CPP-ACP treatment. After tooth bleaching, CPP-ACP treatment have little influence on the shear bond strength of orthodontic brackets.

  3. Quantitative Phosphoproteomics of Tomato Mounting a Hypersensitive Response Reveals a Swift Suppression of Photosynthetic Activity and a Differential Role for Hsp90 Isoforms

    DEFF Research Database (Denmark)

    Stulemeijer, Iris J E; Joosten, Matthieu H A J; Jensen, Ole N

    2009-01-01

    secreted by the pathogenic fungus Cladosporium fulvum. Phosphopeptides were isolated from tomato seedlings expressing both Cf-4 and Avr4 using titanium dioxide columns and LC-MS/MS analysis led to the identification of 50 phosphoproteins, most of which have not been described in tomato before...

  4. Fourier transform IR studies on the interaction of selected chemicals with isolated cuticles

    International Nuclear Information System (INIS)

    Luque, P.; Ramirez, F.J.; Heredia, A.; Bukovac, M.J.

    1994-01-01

    It is known that the plant cuticle represents the first barrier that must be overcome by any chemical reaching the plant surface from the atmosphere before entering the plant. Because of the importance of the cuticle as a barrier to penetration of a wide variety of compounds, its morphology, chemistry, and permeability have been extensively studied. However, only limited information is available on the nature of functional chemical groups present and their interaction and role during the penetration process. The usefulness of in situ Fourier transform infrared spectroscopy studies in identifying functional groups present in isolated cuticles is described and their relationships to the structure of the cuticular membrane are discussed. Applications of infrared spectroscopy on the presence and role of phenolics in the cuticle structure and during the cuticle development, nitrogen oxide binding to isolated cuticles, and the interactions between selected chemical probes during sorption by the cuticle are also described. (orig.)

  5. Combining casein phosphopeptide-amorphous calcium phosphate with fluoride: synergistic remineralization potential of artificially demineralized enamel or not?

    Science.gov (United States)

    Elsayad, Iman; Sakr, Amal; Badr, Yahia

    2009-07-01

    Recaldent is a product of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP). The remineralizing potential of CPP-ACP per se, or when combined with 0.22% Fl gel on artificially demineralized enamel using laser florescence, is investigated. Mesial surfaces of 15 sound human molars are tested using a He-Cd laser beam at 441.5 nm with 18-mW power as an excitation source on a suitable setup based on a Spex 750-M monochromator provided with a photomultiplier tube (PMT) for detection of collected autofluorescence from sound enamel. Mesial surfaces are subjected to demineralization for ten days. The spectra from demineralized enamel are measured. Teeth are divided into three groups according to the remineralizing regimen: group 1 Recaldent per se, group 2 Recaldent combined with fluoride gel and ACP, and group 3 artificial saliva as a positive control. After following these protocols for three weeks, the spectra from the remineralized enamel are measured. The spectra of enamel autofluorescence are recorded and normalized to peak intensity at about 540 nm to compare spectra from sound, demineralized, and remineralized enamel surfaces. A slight red shift occurred in spectra from demineralized enamel, while a blue shift may occur in remineralized enamel. Group 2 shows the highest remineralizing potential. Combining fluoride and ACP with CPP-ACP can give a synergistic effect on enamel remineralization.

  6. Site selection and evaluation studies of the Waste Isolation Pilot Plant (WIPP), Los Medanos, Eddy County, NM

    International Nuclear Information System (INIS)

    Griswold, G.B.

    1977-12-01

    Bedded-salt deposits of the Salado Formation have been selected for evaluation for a proposed Waste Isolation Pilot Plant (WIPP) to be located in Eddy County, NM, approximately 26 mi east of Carlsbad. Site selection and evaluation studies that included geologic mapping, geophysical surveys, drilling, and resource appraisal were conducted over and under the prospective location. The lower portion of the Salado meets essential criteria for waste isolation. Beds chosen for waste storage lie 2074 to 2730 ft below the surface. High-purity salt exists at these depths, and the geologic structure revealed by geophysical surveys indicates that these beds are essentially flat. Additional geophysical surveys are now under way. The initial interpretation of the new data indicates that more structure may exist in the salt beds in the northern portion of the site area. Full evaluation of potentially commercial deposits of potash and natural gas within the WIPP site will be reported by separate studies, as will be the hydrologic details of the region

  7. The Effectiveness of Seismic Isolation System for Nuclear Equipment

    International Nuclear Information System (INIS)

    Kim, Min-Kyu; Choun, Young-Sun; Seo, Jeong-Moon

    2005-04-01

    In this study, the Emergency Diesel Generator and Off-site Transformer were selected for isolation. For the selection of the most suitable base isolation system, the literature review and the numerical analysis were performed. For the decision of the parameter of isolation system, the sensitivity analysis was performed. Finally the conceptual design of each equipment was performed. In case of EDG, the Coil Spring and Viscous Damper system was selected for isolation system and 45% isolation effect was determined. For the OST, the FPS was selected and 69% isolation effect was determined

  8. The Effectiveness of Seismic Isolation System for Nuclear Equipment

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Min-Kyu; Choun, Young-Sun; Seo, Jeong-Moon

    2005-04-15

    In this study, the Emergency Diesel Generator and Off-site Transformer were selected for isolation. For the selection of the most suitable base isolation system, the literature review and the numerical analysis were performed. For the decision of the parameter of isolation system, the sensitivity analysis was performed. Finally the conceptual design of each equipment was performed. In case of EDG, the Coil Spring and Viscous Damper system was selected for isolation system and 45% isolation effect was determined. For the OST, the FPS was selected and 69% isolation effect was determined.

  9. Large-scale analysis of in Vivo phosphorylated membrane proteins by immobilized metal ion affinity chromatography and mass spectrometry

    DEFF Research Database (Denmark)

    Nühse, Thomas S; Stensballe, Allan; Jensen, Ole N

    2003-01-01

    specificity. We investigated the potential of IMAC in combination with capillary liquid chromatography coupled to tandem mass spectrometry for the identification of plasma membrane phosphoproteins of Arabidopsis. Without chemical modification of peptides, over 75% pure phosphopeptides were isolated from...... plasma membrane digests and detected and sequenced by mass spectrometry. We present a scheme for two-dimensional peptide separation using strong anion exchange chromatography prior to IMAC that both decreases the complexity of IMAC-purified phosphopeptides and yields a far greater coverage...... of monophosphorylated peptides. Among the identified sequences, six originated from different isoforms of the plasma membrane H(+)-ATPase and defined two previously unknown phosphorylation sites at the regulatory C terminus. The potential for large-scale identification of phosphorylation sites on plasma membrane...

  10. Comparative evaluation of the effects of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP and xylitol-containing chewing gum on salivary flow rate, pH and buffering capacity in children: An in vivo study

    Directory of Open Access Journals (Sweden)

    Rahul J Hegde

    2017-01-01

    Full Text Available Aim: This study aimed to compare and evaluate the changes in the salivary flow rate, pH, and buffering capacity before and after chewing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP and xylitol-containing chewing gums in children. Materials and Methods: Sixty children aged between 8 and 12 years were selected for the study. They were randomly divided into Group 1 (CPP-ACP chewing gum and Group 2 (xylitol-containing chewing gum comprising thirty children each. Unstimulated and stimulated saliva samples at 15 and 30 min interval were collected from all children. All the saliva samples were estimated for salivary flow rate, pH, and buffering capacity. Results: Significant increase in salivary flow rate, pH, and buffering capacity from baseline to immediately after spitting the chewing gum was found in both the study groups. No significant difference was found between the two study groups with respect to salivary flow rate and pH. Intergroup comparison indicated a significant increase in salivary buffer capacity in Group 1 when compared to Group 2. Conclusion: Chewing gums containing CPP-ACP and xylitol can significantly increase the physiochemical properties of saliva. These physiochemical properties of saliva have a definite relation with caries activity in children.

  11. Isolation and Fatty Acid Profile of Selected Microalgae Strains from the Red Sea for Biofuel Production

    Directory of Open Access Journals (Sweden)

    Khalid M. Abu-Salah

    2013-05-01

    Full Text Available The isolation of lipid-rich autochthonous strains of microalgae is a crucial stage for the development of a microalgae-based biofuel production plant, as these microalgae already have the necessary adaptations to withstand competition, predation and the temperatures observed at each production site. This is particularly important in extreme climates such as in Saudi Arabia. Resorting to fluorescence activated cell sorting (FACS we screened for and isolated several microalgal strains from samples collected from the Red Sea. Relying on the fluorescence of BODIPY 505/515 (4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diazasindacene and growth performance, four promising candidates were identified and the total lipid content and fatty acid profile was assessed for biofuels production. Selected isolates were classified as chlorophytes, belonging to three different genera: Picochlorum, Nannochloris and Desmochloris. The lipid contents were assessed microscopically by means of BODIPY 505/515-associated fluorescence to detect intracellular lipid bodies, which revealed several lipid drops in all selected strains. This result was confirmed by lipid gravimetric determination, which demonstrated that all strains under study presented inner cell lipid contents ranging from 20% to 25% of the biomass dry weight. Furthermore, the fatty acid methyl esters profile of all strains seems ideal for biodiesel production due to a low degree of polyunsaturated fatty acid methyl esters and high amount of palmitic and oleic acids.

  12. Structure-based design of an osteoclast-selective, nonpeptide Src homology 2 inhibitor with in vivo antiresorptive activity

    Science.gov (United States)

    Shakespeare, William; Yang, Michael; Bohacek, Regine; Cerasoli, Franklin; Stebbins, Karin; Sundaramoorthi, Raji; Azimioara, Mihai; Vu, Chi; Pradeepan, Selvi; Metcalf, Chester; Haraldson, Chad; Merry, Taylor; Dalgarno, David; Narula, Surinder; Hatada, Marcos; Lu, Xiaode; van Schravendijk, Marie Rose; Adams, Susan; Violette, Shelia; Smith, Jeremy; Guan, Wei; Bartlett, Catherine; Herson, Jay; Iuliucci, John; Weigele, Manfred; Sawyer, Tomi

    2000-01-01

    Targeted disruption of the pp60src (Src) gene has implicated this tyrosine kinase in osteoclast-mediated bone resorption and as a therapeutic target for the treatment of osteoporosis and other bone-related diseases. Herein we describe the discovery of a nonpeptide inhibitor (AP22408) of Src that demonstrates in vivo antiresorptive activity. Based on a cocrystal structure of the noncatalytic Src homology 2 (SH2) domain of Src complexed with citrate [in the phosphotyrosine (pTyr) binding pocket], we designed 3′,4′-diphosphonophenylalanine (Dpp) as a pTyr mimic. In addition to its design to bind Src SH2, the Dpp moiety exhibits bone-targeting properties that confer osteoclast selectivity, hence minimizing possible undesired effects on other cells that have Src-dependent activities. The chemical structure AP22408 also illustrates a bicyclic template to replace the post-pTyr sequence of cognate Src SH2 phosphopeptides such as Ac-pTyr-Glu-Glu-Ile (1). An x-ray structure of AP22408 complexed with Lck (S164C) SH2 confirmed molecular interactions of both the Dpp and bicyclic template of AP22408 as predicted from molecular modeling. Relative to the cognate phosphopeptide, AP22408 exhibits significantly increased Src SH2 binding affinity (IC50 = 0.30 μM for AP22408 and 5.5 μM for 1). Furthermore, AP22408 inhibits rabbit osteoclast-mediated resorption of dentine in a cellular assay, exhibits bone-targeting properties based on a hydroxyapatite adsorption assay, and demonstrates in vivo antiresorptive activity in a parathyroid hormone-induced rat model. PMID:10944210

  13. Isolation, characterization, and evaluation of wild isolates of Lactobacillus reuteri from pig feces.

    Science.gov (United States)

    Lee, Deog Yong; Seo, Yeon-Soo; Rayamajhi, Nabin; Kang, Mi Lan; Lee, Su In; Yoo, Han Sang

    2009-12-01

    Lactic acid bacteria (LAB) are a well-used probiotics for health improvements in both humans and animals. Despite of several benefits, non-host-specific LAB showed poor probiotics effects due to difficulty in colonization and competition with normal flora. Therefore, the feasibility of porcine LAB isolates was evaluated as a probiotics. Ten of 49 Lactobacillus spp. isolates harbored 2 approximately 10 kb plasmid DNA. Seven strains were selected based on the safety test, such as hemolytic activity, ammonia, indole, and phenylalanine production. After safety test, five strains were selected again by several tests, such as epithelial adherence, antimicrobial activity, tolerance against acid, bile, heat, and cold-drying, and production of acid and hydrogen peroxide. Then, enzyme profiles (ZYM test) and antibiotics resistance were analyzed for further characterization. Five Lactobacillus reuteri isolates from pig feces were selected by safety and functional tests. The plasmid DNA which was able to develop vector system was detected in the isolates. Together with these approaches, pig-specific Lactobacillus spp. originated from pigs were selected. These strains may be useful tools to develop oral delivery system.

  14. Antimicrobial and antibiofilm effects of selected food preservatives against Salmonella spp. isolated from chicken samples.

    Science.gov (United States)

    Er, Buket; Demirhan, Burak; Onurdag, Fatma Kaynak; Ozgacar, Selda Özgen; Oktem, Aysel Bayhan

    2014-03-01

    Salmonella spp. are widespread foodborne pathogens that contaminate egg and poultry meats. Attachment, colonization, as well as biofilm formation capacity of Salmonella spp. on food and contact surfaces of food may cause continuous contamination. Biofilm may play a crucial role in the survival of salmonellae under unfavorable environmental conditions, such as in animal slaughterhouses and processing plants. This could serve as a reservoir compromising food safety and human health. Addition of antimicrobial preservatives extends shelf lives of food products, but even when products are supplemented with adequate amounts of preservatives, it is not always possible to inhibit the microorganisms in a biofilm community. In this study, our aims were i) to determine the minimum inhibitory concentrations (MIC) and minimum biofilm inhibitory concentrations (MBIC) of selected preservatives against planktonic and biofilm forms of Salmonella spp. isolated from chicken samples and Salmonella Typhimurium SL1344 standard strain, ii) to show the differences in the susceptibility patterns of same strains versus the planktonic and biofilm forms to the same preservative agent, and iii) to determine and compare antimicrobial and antibiofilm effects of selected food preservatives against Salmonella spp. For this purpose, Salmonella Typhimurium SL1344 standard strain and 4 Salmonella spp. strains isolated from chicken samples were used. Investigation of antimicrobial and antibiofilm effects of selected food preservatives against Salmonella spp. was done according to Clinical and Laboratory Standards Institute M100-S18 guidelines and BioTimer assay, respectively. As preservative agents, pure ciprofloxacin, sodium nitrite, potassium sorbate, sodium benzoate, methyl paraben, and propyl paraben were selected. As a result, it was determined that MBIC values are greater than the MIC values of the preservatives. This result verified the resistance seen in a biofilm community to food

  15. Selection of hyperproduction of AmpC and SME-1 in a carbapenem-resistant Serratia marcescens isolate during antibiotic therapy.

    Science.gov (United States)

    Hemarajata, Peera; Amick, Thomas; Yang, Shangxin; Gregson, Aric; Holzmeyer, Cameron; Bush, Karen; Humphries, Romney M

    2018-02-19

    Antibiotic selective pressure may result in changes to antimicrobial susceptibility throughout the course of infection, especially for organisms that harbour chromosomally encoded AmpC β-lactamases, notably Enterobacter spp., in which hyperexpression of ampC may be induced following treatment with cephalosporins. In this study, we document a case of bacteraemia caused by a blaSME-1-harbouring Serratia marcescens that subsequently developed resistance to expanded-spectrum cephalosporins, piperacillin/tazobactam and fluoroquinolones, over the course of several months of treatment with piperacillin/tazobactam and ciprofloxacin. Susceptibility testing and WGS were performed on three S. marcescens isolates from the patient. β-Lactamase activity in the presence or absence of induction by imipenem was measured by nitrocefin hydrolysis assays. Expression of ampC and blaSME-1 under the same conditions was determined by real-time PCR. WGS demonstrated accumulation of missense and nonsense mutations in ampD associated with stable derepression of AmpC. Gene expression and β-lactamase activity of both AmpC and SME-1 were inducible in the initial susceptible isolate, but were constitutively high in the resistant isolate, in which total β-lactamase activity was increased by 128-fold. Although development of such in vitro resistance due to selective pressure imposed by antibiotics is reportedly low in S. marcescens, our findings highlight the need to evaluate isolates on a regular basis during long-term antibiotic therapy.

  16. Selection, assessment of virulence to Alphitobius diaperinus, and Pr1 enzyme production of Beauveria bassiana (Bals. Vuill. isolates cultured at stress temperatures

    Directory of Open Access Journals (Sweden)

    Kelly Christiane Constanski

    2015-12-01

    Full Text Available The entomopathogenic fungus Beauveria bassiana is a promising agent for use in insect control. Its pathogenic activity, as well as other factors such as temperature that can interfere with its development, should be assessed, thus, establishing the foundations for B. bassiana use in biological control programs. The objective of this study was to select and induce tolerance of B. bassiana isolates to high and low temperatures and to assess their virulence before and after exposure to those temperatures. A pre-selection test was performed, in which the tolerance of isolates to stress temperatures was tested and compared to the ideal growth temperature of 25 °C for this organism. For the isolates/temperature combinations resulting in growth, conidia germination and colony-forming units (CFUs were assessed. The isolates Unioeste 4 and Unioeste 40 exhibited >95% germinated conidia at 16 and 31 °C. Thereafter, they underwent four consecutive passages at maximum and minimum tolerated temperatures (10 and 37 °C. A significant difference in germination was observed between the two isolates at all temperatures tested. More CFUs were observed for Unioeste 4 compared to Unioeste 40 at all temperatures, and in the case of the latter, there was no difference in CFU formation at 10 and 25 °C. For both isolates, decreased vegetative growth was observed at 37 °C. Recovery of virulence was observed in both isolates, as determined by insect mortality. No relationship was observed between production of the enzyme Pr1 and the virulence of the isolates.

  17. Compost as a source of microbial isolates for the bioremediation of heavy metals: In vitro selection

    International Nuclear Information System (INIS)

    Vargas-García, María del Carmen; López, María José; Suárez-Estrella, Francisca; Moreno, Joaquín

    2012-01-01

    Heavy metal pollution has become a major environmental concern nowadays and the bioremediation of polluted habitats is an increasingly popular strategy due to both its efficiency and safety. A screening and selection protocol based on different composting processes was designed in order to isolate heavy metal-resistant microorganisms. A collection of 51 microorganisms was obtained and most of them showed the capability to tolerate heavy metals in multi-polluted aqueous systems (Cd(II), Cr(VI), Ni, Pb, Zn(II)), as well as to remove them. The highest detoxification ratios were observed for Pb. Some of the isolates detoxifying more than a 90% of this metal, while the other metals were removed in a range between 20% and 60%. The best isolates (Graphium putredinis, Fusarium solani, Fusarium sp. and Penicillium chrysogenum) were further assayed in order to determine the predominant removal mechanism and the potential use of their dead biomass as a biosorbent. Intracellular accumulation was the prevalent mechanism for most isolates and metals, with the exception of Ni. In this case, the proportion removed by extracellular adsorption was similar or even higher than that removed by intracellular accumulation. Thus, the efficiency of living cells was higher than that of dead biomass except in the case of Ni. - Highlights: ► Composting is a good reservoir for the isolation of HM-resistant microorganisms. ► Pb was the most removed heavy metal in multi-polluted aqueous systems. ► Intracellular accumulation was the predominant mechanism for heavy metal removal. ► Graphium putredinis, which detoxifies organic pollutants, was the most efficient isolate.

  18. Optimization of Culture Parameters for Maximum Polyhydroxybutyrate Production by Selected Bacterial Strains Isolated from Rhizospheric Soils.

    Science.gov (United States)

    Lathwal, Priyanka; Nehra, Kiran; Singh, Manpreet; Jamdagni, Pragati; Rana, Jogender S

    2015-01-01

    The enormous applications of conventional non-biodegradable plastics have led towards their increased usage and accumulation in the environment. This has become one of the major causes of global environmental concern in the present century. Polyhydroxybutyrate (PHB), a biodegradable plastic is known to have properties similar to conventional plastics, thus exhibiting a potential for replacing conventional non-degradable plastics. In the present study, a total of 303 different bacterial isolates were obtained from soil samples collected from the rhizospheric area of three crops, viz., wheat, mustard and sugarcane. All the isolates were screened for PHB (Poly-3-hydroxy butyric acid) production using Sudan Black staining method, and 194 isolates were found to be PHB positive. Based upon the amount of PHB produced, the isolates were divided into three categories: high, medium and low producers. Representative isolates from each category were selected for biochemical characterization; and for optimization of various culture parameters (carbon source, nitrogen source, C/N ratio, different pH, temperature and incubation time periods) for maximizing PHB accumulation. The highest PHB yield was obtained when the culture medium was supplemented with glucose as the carbon source, ammonium sulphate at a concentration of 1.0 g/l as the nitrogen source, and by maintaining the C/N ratio of the medium as 20:1. The physical growth parameters which supported maximum PHB accumulation included a pH of 7.0, and an incubation temperature of 30 degrees C for a period of 48 h. A few isolates exhibited high PHB accumulation under optimized conditions, thus showing a potential for their industrial exploitation.

  19. Cluster Head Selection in a Homogeneous Wireless Sensor Network Ensuring Full Connectivity with Minimum Isolated Nodes

    Directory of Open Access Journals (Sweden)

    Tapan Kumar Jain

    2014-01-01

    Full Text Available The research work proposes a cluster head selection algorithm for a wireless sensor network. A node can be a cluster head if it is connected to at least one unique neighbor node where the unique neighbor is the one that is not connected to any other node. If there is no connected unique node then the CH is selected on the basis of residual energy and the number of neighbor nodes. With the increase in number of clusters, the processing energy of the network increases; hence, this algorithm proposes minimum number of clusters which further leads to increased network lifetime. The major novel contribution of the proposed work is an algorithm that ensures a completely connected network with minimum number of isolated nodes. An isolated node will remain only if it is not within the transmission range of any other node. With the maximum connectivity, the coverage of the network is automatically maximized. The superiority of the proposed design is verified by simulation results done in MATLAB, where it clearly depicts that the total numbers of rounds before the network dies out are maximum compared to other existing protocols.

  20. Isolation and selection of new biosurfactant producing bacteria from degraded palm kernel cake under liquid state fermentation.

    Science.gov (United States)

    Jamal, Parveen; Mir, Shajrat; Alam, Md Zahangir; Wan Nawawi, Wan M Fazli

    2014-01-01

    Biosurfactants are surface-active compounds produced by different microorganisms. The aim of this study was to introduce palm kernel cake (PKC) as a novel substrate for biosurfactant production using a potent bacterial strain under liquid state fermentation. This study was primarily based on the isolation and identification of biosurfactant-producing bacteria that could utilize palm kernel cake as a new major substrate. Potential bacterial strains were isolated from degraded PKC and screened for biosurfactant production with the help of the drop collapse assay and by analyzing the surface tension activity. From the screened isolates, a new strain, SM03, showed the best and most consistent results, and was therefore selected as the most potent biosurfactant-producing bacterial strain. The new strain was identified as Providencia alcalifaciens SM03 using the Gen III MicroPlate Biolog Microbial Identification System. The yield of the produced biosurfactant was 8.3 g/L.

  1. Isolation of human simple repeat loci by hybridization selection.

    Science.gov (United States)

    Armour, J A; Neumann, R; Gobert, S; Jeffreys, A J

    1994-04-01

    We have isolated short tandem repeat arrays from the human genome, using a rapid method involving filter hybridization to enrich for tri- or tetranucleotide tandem repeats. About 30% of clones from the enriched library cross-hybridize with probes containing trimeric or tetrameric tandem arrays, facilitating the rapid isolation of large numbers of clones. In an initial analysis of 54 clones, 46 different tandem arrays were identified. Analysis of these tandem repeat loci by PCR showed that 24 were polymorphic in length; substantially higher levels of polymorphism were displayed by the tetrameric repeat loci isolated than by the trimeric repeats. Primary mapping of these loci by linkage analysis showed that they derive from 17 chromosomes, including the X chromosome. We anticipate the use of this strategy for the efficient isolation of tandem repeats from other sources of genomic DNA, including DNA from flow-sorted chromosomes, and from other species.

  2. Isolating GaSb Membranes Grown Metamorphically on GaAs Substrates Using Highly Selective Substrate Removal Etch Processes

    Science.gov (United States)

    Renteria, E. J.; Muniz, A. J.; Addamane, S. J.; Shima, D. M.; Hains, C. P.; Balakrishnan, G.

    2015-05-01

    The etch rates of NH4OH:H2O2 and C6H8O7:H2O2 for GaAs and GaSb have been investigated to develop a selective etch for GaAs substrates and to isolate GaSb epilayers grown on GaAs. The NH4OH:H2O2 solution has a greater etch rate differential for the GaSb/GaAs material system than C6H8O7:H2O2 solution. The selectivity of NH4OH:H2O2 for GaAs/GaSb under optimized etch conditions has been observed to be as high as 11,000 ± 2000, whereas that of C6H8O7:H2O2 has been measured up to 143 ± 2. The etch contrast has been verified by isolating 2- μm-thick GaSb epilayers that were grown on GaAs substrates. GaSb membranes were tested and characterized with high- resolution x-ray diffraction and atomic force microscopy.

  3. Selective isolation of gonyautoxins 1,4 from the dinoflagellate Alexandrium minutum based on molecularly imprinted solid-phase extraction.

    Science.gov (United States)

    Lian, Ziru; Wang, Jiangtao

    2017-09-15

    Gonyautoxins 1,4 (GTX1,4) from Alexandrium minutum samples were isolated selectively and recognized specifically by an innovative and effective extraction procedure based on molecular imprinting technology. Novel molecularly imprinted polymer microspheres (MIPMs) were prepared by double-templated imprinting strategy using caffeine and pentoxifylline as dummy templates. The synthesized polymers displayed good affinity to GTX1,4 and were applied as sorbents. Further, an off-line molecularly imprinted solid-phase extraction (MISPE) protocol was optimized and an effective approach based on the MISPE coupled with HPLC-FLD was developed for selective isolation of GTX1,4 from the cultured A. minutum samples. The separation method showed good extraction efficiency (73.2-81.5%) for GTX1,4 and efficient removal of interferences matrices was also achieved after the MISPE process for the microalgal samples. The outcome demonstrated the superiority and great potential of the MISPE procedure for direct separation of GTX1,4 from marine microalgal extracts. Copyright © 2017. Published by Elsevier Ltd.

  4. Isolated Calyx Mistaken for a Cyst: Inappropriately Performed Catheter-Directed Sclerotherapy and Safe Removal of the Catheter After Selective Embolization

    Energy Technology Data Exchange (ETDEWEB)

    Gwak, Jng Won, E-mail: jjungwonie@hanmail.net; Lee, Seung Hwa, E-mail: gareureung@daum.net; Chung, Hwan Hoon, E-mail: chungmic@korea.ac.kr; Je, Bo Kyung, E-mail: purity21@hanmail.net; Yeom, Suk kyu, E-mail: pagoda20@hanmail.net [Korea University College of Medicine, Department of Radiology, Ansan Hospital (Korea, Republic of); Sung, Deuk Jae, E-mail: urora@korea.ac.kr [Korea University College of Medicine, Department of Radiology, Anam Hospital (Korea, Republic of)

    2015-02-15

    We present a case of isolated calyx that was mistaken for a large cyst. A 47-year-old woman was referred for sclerotherapy of a large cystic lesion on her left kidney. Computed tomography (CT) and ultrasound showed that the cystic lesion was a large cyst. We noticed that the cystic lesion was not a typical simple cyst, even after two sessions of catheter-mediated sclerotherapy. Isolated calyx was presumed by medical history review and was confirmed by aspirated fluid analysis and far delayed-phase CT after intravenous contrast injection. We performed meticulous selective arterial embolization for an isolated calyx and inserted a catheter that could be removed without complication.

  5. Nickel nanoparticle decorated graphene for highly selective isolation of polyhistidine-tagged proteins

    Science.gov (United States)

    Liu, Jia-Wei; Yang, Ting; Ma, Lin-Yu; Chen, Xu-Wei; Wang, Jian-Hua

    2013-12-01

    Nickel nanoparticle decorated graphene (GP-Ni) is prepared by one-pot hydrothermal reduction of graphene oxide and nickel cations by hydrazine hydrate in the presence of poly(sodium-p-styrenesulfonate) (PSS). The GP-Ni hybrid is characterized by XRD, TEM, SEM, XPS, Raman and FT-IR spectra, demonstrating the formation of poly-dispersed nickel nanoparticles with an average size of 83 nm attached on the surface of graphene sheets. The GP-Ni hybrid exhibits ferromagnetic behavior with a magnetization saturation of 31.1 emu g-1 at 10 000 Oersted (Oe). The GP-Ni also possesses favorable stability in aqueous medium and rapid magnetic response to an external magnetic field. These make it a novel magnetic adsorbent for the separation/isolation of His6-tagged recombinant proteins from a complex sample matrix (cell lysate). The targeted protein species is captured onto the surface of the GP-Ni hybrid via specific metal affinity force between polyhistidine groups and nickel nanoparticles. The SDS-PAGE assay indicates highly selective separation of His6-tagged Smt A from cell lysate. The GP-Ni hybrid displays favorable performance on the separation/isolation of His6-tagged recombinant proteins with respect to the commercial NTA-Ni2+ column.

  6. Clinical evaluation of remineralization potential of casein phosphopeptide amorphous calcium phosphate nanocomplexes for enamel decalcification in orthodontics.

    Science.gov (United States)

    Wang, Jun-xiang; Yan, Yan; Wang, Xiu-jing

    2012-11-01

    Enamel decalcification in orthodontics is a concern for dentists and methods to remineralize these lesions are the focus of intense research. The aim of this study was to evaluate the remineralizing effect of casein phosphopeptide amorphous calcium phosphate (CPP-ACP) nanocomplexes on enamel decalcification in orthodontics. Twenty orthodontic patients with decalcified enamel lesions during fixed orthodontic therapy were recruited to this study as test group and twenty orthodontic patients with the similar condition as control group. GC Tooth Mousse, the main component of which is CPP-ACP, was used by each patient of test group every night after tooth-brushing for six months. For control group, each patient was asked to brush teeth with toothpaste containing 1100 parts per million (ppm) of fluoride twice a day. Standardized intraoral images were taken for all patients and the extent of enamel decalcification was evaluated before and after treatment over this study period. Measurements were statistically compared by t test. After using CPP-ACP for six months, the enamel decalcification index (EDI) of all patients had decreased; the mean EDI before using CPP-ACP was 0.191 ± 0.025 and that after using CPP-ACP was 0.183 ± 0.023, the difference was significant (t = 5.169, P 0.05). CPP-ACP can effectively improve the demineralized enamel lesions during orthodontic treatment, so it has some remineralization potential for enamel decalcification in orthodontics.

  7. Environmental carbonate chemistry selects for phenotype of recently isolated strains of Emiliania huxleyi

    Science.gov (United States)

    Rickaby, Rosalind E. M.; Hermoso, Michaël; Lee, Renee B. Y.; Rae, Benjamin D.; Heureux, Ana M. C.; Balestreri, Cecilia; Chakravarti, Leela; Schroeder, Declan C.; Brownlee, Colin

    2016-05-01

    Coccolithophorid algae, particularly Emiliania huxleyi, are prolific biomineralisers that, under many conditions, dominate communities of marine eukaryotic plankton. Their ability to photosynthesise and form calcified scales (coccoliths) has placed them in a unique position in the global carbon cycle. Contrasting reports have been made with regards to the response of E. huxleyi to ocean acidification. Therefore, there is a pressing need to further determine the fate of this key organism in a rising CO2 world. In this paper, we investigate the phenotype of newly isolated, genetically diverse, strains of E. huxleyi from UK Ocean Acidification Research Programme (UKOA) cruises around the British Isles, the Arctic, and the Southern Ocean. We find a continuum of diversity amongst the physiological and photosynthetic parameters of different strains of E. huxleyi morphotype A under uniform, ambient conditions imposed in the laboratory. This physiology is best explained by adaptation to carbonate chemistry in the former habitat rather than being prescribed by genetic fingerprints such as the coccolithophore morphology motif (CMM). To a first order, the photosynthetic capacity of each strain is a function of both aqueous CO2 availability, and calcification rate, suggestive of a link between carbon concentrating ability and calcification. The calcification rate of each strain is related linearly to the natural environmental [CO32-] at the site of isolation, but a few exceptional strains display low calcification rates at the highest [CO32-] when calcification is limited by low CO2 availability and/or a lack of a carbon concentrating mechanism. We present O2-electrode measurements alongside coccolith oxygen isotopic composition and the uronic acid content (UAC) of the coccolith associated polysaccharide (CAP), that act as indirect tools to show the differing carbon concentrating ability of the strains. The environmental selection revealed amongst our recently isolated strain

  8. Selective isolation and screening of fungi with herbicidal potential ...

    African Journals Online (AJOL)

    The respective fungal isolates were cultivated in modified Fries media under standard condition. The mycelia and the filtrate were extracted with ethyl acetate and the concentrated extract was evaluated for the herbicidal activity adopting leaf necrosis assay. Among the different isolates, extract prepared from A. alternata and ...

  9. Compost as a source of microbial isolates for the bioremediation of heavy metals: In vitro selection

    Energy Technology Data Exchange (ETDEWEB)

    Vargas-Garcia, Maria del Carmen, E-mail: mcvargas@ual.es; Lopez, Maria Jose, E-mail: mllopez@ual.es; Suarez-Estrella, Francisca, E-mail: fsuarez@ual.es; Moreno, Joaquin, E-mail: jcasco@ual.es

    2012-08-01

    Heavy metal pollution has become a major environmental concern nowadays and the bioremediation of polluted habitats is an increasingly popular strategy due to both its efficiency and safety. A screening and selection protocol based on different composting processes was designed in order to isolate heavy metal-resistant microorganisms. A collection of 51 microorganisms was obtained and most of them showed the capability to tolerate heavy metals in multi-polluted aqueous systems (Cd(II), Cr(VI), Ni, Pb, Zn(II)), as well as to remove them. The highest detoxification ratios were observed for Pb. Some of the isolates detoxifying more than a 90% of this metal, while the other metals were removed in a range between 20% and 60%. The best isolates (Graphium putredinis, Fusarium solani, Fusarium sp. and Penicillium chrysogenum) were further assayed in order to determine the predominant removal mechanism and the potential use of their dead biomass as a biosorbent. Intracellular accumulation was the prevalent mechanism for most isolates and metals, with the exception of Ni. In this case, the proportion removed by extracellular adsorption was similar or even higher than that removed by intracellular accumulation. Thus, the efficiency of living cells was higher than that of dead biomass except in the case of Ni. - Highlights: Black-Right-Pointing-Pointer Composting is a good reservoir for the isolation of HM-resistant microorganisms. Black-Right-Pointing-Pointer Pb was the most removed heavy metal in multi-polluted aqueous systems. Black-Right-Pointing-Pointer Intracellular accumulation was the predominant mechanism for heavy metal removal. Black-Right-Pointing-Pointer Graphium putredinis, which detoxifies organic pollutants, was the most efficient isolate.

  10. Detection of Extended Spectrum Beta-Lactamases Resistance Genes among Bacteria Isolated from Selected Drinking Water Distribution Channels in Southwestern Nigeria.

    Science.gov (United States)

    Adesoji, Ayodele T; Ogunjobi, Adeniyi A

    2016-01-01

    Extended Spectrum Beta-Lactamases (ESBL) provide high level resistance to beta-lactam antibiotics among bacteria. In this study, previously described multidrug resistant bacteria from raw, treated, and municipal taps of DWDS from selected dams in southwestern Nigeria were assessed for the presence of ESBL resistance genes which include bla TEM, bla SHV, and bla CTX by PCR amplification. A total of 164 bacteria spread across treated (33), raw (66), and municipal taps (68), belonging to α-Proteobacteria, β-Proteobacteria, γ-Proteobacteria, Flavobacteriia, Bacilli, and Actinobacteria group, were selected for this study. Among these bacteria, the most commonly observed resistance was for ampicillin and amoxicillin/clavulanic acid (61 isolates). Sixty-one isolates carried at least one of the targeted ESBL genes with bla TEM being the most abundant (50/61) and bla CTX being detected least (3/61). Klebsiella was the most frequently identified genus (18.03%) to harbour ESBL gene followed by Proteus (14.75%). Moreover, combinations of two ESBL genes, bla SHV + bla TEM or bla CTX + bla TEM, were observed in 11 and 1 isolate, respectively. In conclusion, classic bla TEM ESBL gene was present in multiple bacterial strains that were isolated from DWDS sources in Nigeria. These environments may serve as foci exchange of genetic traits in a diversity of Gram-negative bacteria.

  11. Selected Lactobacillus strains isolated from sugary and milk kefir reduce Salmonella infection of epithelial cells in vitro.

    Science.gov (United States)

    Zavala, L; Golowczyc, M A; van Hoorde, K; Medrano, M; Huys, G; Vandamme, P; Abraham, A G

    2016-09-01

    The isolation of potentially probiotic strains and the subsequent study of their properties are very important steps to gain insight in the health benefits ascribed to sugary and milk kefir. The aim of the present study was to characterise fifteen Lactobacillus strains isolated from these beverages by determining some surface properties and their ability to antagonise enterocyte cell damage after Salmonella infection in vitro. Lactobacillus surface properties were determined by hydrophobicity, autoaggregation, and coaggregation assays with Salmonella. In addition, lactobacilli adhesion to Caco-2/TC-7 cells and the effect on Salmonella invasion were evaluated. Finally, the disassembly of F-actin cytoskeleton on intestinal epithelial cells was assayed in vitro when Salmonella infection was performed in the presence of selected Lactobacillus strains. Ten out of the 15 strains showed a high adhesion capacity to Caco-2/TC-7 cells. Most of the strains were hydrophilic and non-autoaggregating. Strains isolated from sugary kefir were non-coaggregating with Salmonella, while strains Lactobacillus paracasei CIDCA 83120, 83121, 83123, 83124, 8339, 83102 isolated from milk kefir were able to coaggregate after 1 h. L. paracasei CIDCA 8339 and Lactobacillus kefiri CIDCA 83102 were able to diminish Salmonella invasion to the enterocytes. An antagonistic effect on cytoskeleton disruption elicited by the pathogen was also demonstrated. Our results suggest that both strains isolated from milk kefir could be considered as appropriate probiotic candidates.

  12. CHROMagar COL-APSE: a selective bacterial culture medium for the isolation and differentiation of colistin-resistant Gram-negative pathogens

    DEFF Research Database (Denmark)

    Abdul Momin, Muhd Haziq F; Bean, David C; Hendriksen, Rene S.

    2017-01-01

    Purpose. A selective chromogenic culture medium for the laboratory isolation and differentiation of colistin resistant Acinetobacter, Pseudomonas, Stenotrophomonas and Enterobacteriaceae spp. (CHROMagar COL-APSE) was developed, evaluated and compared to an existing selective bacterial culture......-resistant non-fermentative bacteria (Acinetobacter, Pseudomonas and Stenotrophomonas). CHROMagar COL-APSE was also more sensitive in supporting the growth of Enterobacteriaceae with COL resistance associated with the carriage of mcr-1. Conclusion. CHROMagar COL-APSE is a sensitive and specific medium...

  13. Influence of selected Rhizoctonia solani isolates on sugar beet seedlings

    Directory of Open Access Journals (Sweden)

    Skonieczek Paweł

    2016-04-01

    Full Text Available From 2008 to 2010 the levels of sugar beet seedlings infection caused by Rhizoctonia solani were compared in laboratory tests. Seven sugar beet lines were tested: H56, H66, S2, S3, S4, S5 and S6 as well as three control cultivars: Carlos, Esperanza and Janosik. Sugar beet lines with tolerance to rhizoctoniosis and cultivars without tolerance were infected artificially by R. solani isolates: R1, R28a and R28b. These isolates belong to the second anastomosis group (AG, which is usually highly pathogenic to beet roots. The aim of the experiment was to test whether the tolerance of sugar beet genotypes to R. solani AG 2 prevents both root rot, and damping-off of seedlings, induced by the pathogen. Sugar beet lines tolerant to brown root rot in laboratory tests were significantly less sensitive to infection of the seedlings by R. solani AG 2 isolates in comparison to control cultivars. Rhizoctonia solani AG 2 isolates demonstrated considerable differences in pathogenicity against seedlings of sugar beet lines and cultivars. The strongest infection of sugar beet seedlings occurred with the isolate R28b. The greatest tolerance to infection by AG 2 isolates was found for the S5 and S3 breeding lines.

  14. Summary of computational support and general documentation for computer code (GENTREE) used in Office of Nuclear Waste Isolation Pilot Salt Site Selection Project

    International Nuclear Information System (INIS)

    Beatty, J.A.; Younker, J.L.; Rousseau, W.F.; Elayat, H.A.

    1983-01-01

    A Decision Tree Computer Model was adapted for the purposes of a Pilot Salt Site Selection Project conducted by the Office of Nuclear Waste Isolation (ONWI). A deterministic computer model was developed to structure the site selection problem with submodels reflecting the five major outcome categories (Cost, Safety, Delay, Environment, Community Impact) to be evaluated in the decision process. Time-saving modifications were made in the tree code as part of the effort. In addition, format changes allowed retention of information items which are valuable in directing future research and in isolation of key variabilities in the Site Selection Decision Model. The deterministic code was linked to the modified tree code and the entire program was transferred to the ONWI-VAX computer for future use by the ONWI project

  15. Selection of bacteria with hydrocarbon degrading capacity isolated from Colombian Caribbean sediments

    International Nuclear Information System (INIS)

    Narvaez Florez, Silvia; Gomez, Martha L; Martinez Maria M

    2008-01-01

    Thirty one bacterial isolations in minimal salts supplemented medium with hydrocarbons (ACPM or crude oil) as sole carbon source were isolated from sediment samples from the Colombian Caribbean. Bacterial strains underwent selection tests in different concentrations of hydrocarbons; 11 tolerant crude oil and ACPM strains in a range of 1-8%v/v were chosen. A mixed bacterial culture was created and assessed its ability to degrade hydrocarbons in a laboratory-scale test, with a concentration of 2% v/v of ACPM over a period of 21 days. Measurements of biomass in Colony Forming Units (CFU)/mL were used to develop the growth curve of the mixed culture. Hydrocarbons remotion was measured by mass chromatography. The mixed culture was able to degrade the 68.6% of aliphatic hydrocarbons in preference of long chain n- alkenes (C12- C31), reaching a maximum growth of 3.13 x 10 9 UFC / mL. Degradation of aromatic hydrocarbons was not evidenced under the observation time. Nine of the eleven strains were identified using the biochemical systems BBL and API 50 CHB/E; they belonged to the genus Klebsiella, Chromobacterium, Flavimonas, Enterobacter,Pseudomonas, and Bacillus. The evaluated strains have enzymatic potential to degrade hydrocarbons and it is necessary to characterize them at molecular level in order to develop and effective consortium for field application

  16. Selection and geographic isolation influence hummingbird speciation: genetic, acoustic and morphological divergence in the wedge-tailed sabrewing (Campylopterus curvipennis

    Directory of Open Access Journals (Sweden)

    Ornelas Juan

    2011-02-01

    Full Text Available Abstract Background Mesoamerica is one of the most threatened biodiversity hotspots in the world, yet we are far from understanding the geologic history and the processes driving population divergence and speciation for most endemic taxa. In species with highly differentiated populations selective and/or neutral factors can induce rapid changes to traits involved in mate choice, promoting reproductive isolation between allopatric populations that can eventually lead to speciation. We present the results of genetic differentiation, and explore drift and selection effects in promoting acoustic and morphological divergence among populations of Campylopterus curvipennis, a lekking hummingbird with an extraordinary vocal variability across Mesoamerica. Results Analyses of two mitochondrial genes and ten microsatellite loci genotyped for 160 individuals revealed the presence of three lineages with no contemporary gene flow: C. c. curvipennis, C. c. excellens, and C. c. pampa disjunctly distributed in the Sierra Madre Oriental, the Tuxtlas region and the Yucatan Peninsula, respectively. Sequence mtDNA and microsatellite data were congruent with two diversification events: an old vicariance event at the Isthmus of Tehuantepec (c. 1.4 Ma, and a more recent Pleistocene split, isolating populations in the Tuxtlas region. Hummingbirds of the excellens group were larger, and those of the pampa group had shorter bills, and lineages that have been isolated the longest shared fewer syllables and differed in spectral and temporal traits of a shared syllable. Coalescent simulations showed that fixation of song types has occurred faster than expected under neutrality but the null hypothesis that morphological divergence resulted from drift was not rejected. Conclusions Our phylogeographic analyses uncovered the presence of three Mesoamerican wedge-tailed sabrewing lineages, which diverged at different time scales. These results highlight the importance of the

  17. Genotypic characterization of Azotobacteria isolated from Argentinean soils and plant-growth-promoting traits of selected strains with prospects for biofertilizer production.

    Science.gov (United States)

    Rubio, Esteban Julián; Montecchia, Marcela Susana; Tosi, Micaela; Cassán, Fabricio Darío; Perticari, Alejandro; Correa, Olga Susana

    2013-01-01

    The genetic diversity among 31 putative Azotobacter isolates obtained from agricultural and non-agricultural soils was assessed using rep-PCR genomic fingerprinting and identified to species level by ARDRA and partial 16S rRNA gene sequence analysis. High diversity was found among the isolates, identified as A. chroococcum, A. salinestris, and A. armeniacus. Selected isolates were characterized on the basis of phytohormone biosynthesis, nitrogenase activity, siderophore production, and phosphate solubilization. Indole-3 acetic-acid (IAA), gibberellin (GA3) and zeatin (Z) biosynthesis, nitrogenase activity, and siderophore production were found in all evaluated strains, with variation among them, but no phosphate solubilization was detected. Phytohormones excreted to the culture medium ranged in the following concentrations: 2.2-18.2 μ g IAA mL(-1), 0.3-0.7 μ g GA3 mL(-1), and 0.5-1.2 μ g Z mL(-1). Seed inoculations with further selected Azotobacter strains and treatments with their cell-free cultures increased the number of seminal roots and root hairs in wheat seedlings. This latter effect was mimicked by treatments with IAA-pure solutions, but it was not related to bacterial root colonization. Our survey constitutes a first approach to the knowledge of Azotobacter species inhabiting Argentinean soils in three contrasting geographical regions. Moreover, this phenotypic characterization constitutes an important contribution to the selection of Azotobacter strains for biofertilizer formulations.

  18. Natural selection and type 2 diabetes-associated mortality in an isolated indigenous community in the valley of Oaxaca, southern Mexico.

    Science.gov (United States)

    Little, Bertis B; Peña Reyes, Maria Eugenia; Malina, Robert M

    2017-03-01

    This study tests the hypothesis that natural selection is associated with type 2 diabetes (T2D)-associated mortality and fertility in a rural isolated Zapotec community in the Valley of Oaxaca, southern Mexico. Mortality data and related demographic and genealogic information were linked with data for fertility, prereproductive mortality and family history of mortality attributed to T2D. Physician verified T2D mortality (n = 27) between 1980 and 2009 and imputed T2D (n = 70) from cardiovascular mortality (68% random sample) and renal failure (44% random sample). Bootstrapping was used to obtain a robust variance estimate in survival analysis and multivariate analysis of variance. Estimated maximum natural selection by Crow's Index occurred circa 1930 and was relaxed after this time in the study population. Cox-regression survival analysis of T2D mortality with covariates (family history of T2D, cardiovascular disease, renal failure) indicated a significant hazard ratio (HR = 5.95, 95% CI: 1.38-25.67, p natural selection decreased, and favored offspring survival of non-T2D descedants. The results indicated statistically significant directional selection against T2D and imputed T2D to this population isolate. © 2016 Wiley Periodicals, Inc.

  19. Mechanisms responsible for imipenem resistance among Pseudomonas aeruginosa clinical isolates exposed to imipenem concentrations within the mutant selection window.

    Science.gov (United States)

    Vassilara, Foula; Galani, Irene; Souli, Maria; Papanikolaou, Konstantinos; Giamarellou, Helen; Papadopoulos, Antonios

    2017-07-01

    The aim of this study was to determine the propensities of imipenem to select for resistant Pseudomonas aeruginosa mutants by determining the mutant prevention concentrations (MPCs) for 9 unrelated clinical isolates and the accession of any relationship with mechanisms of resistance development. The MPC/MIC ratios ranged from 4 to 16. Detection of resistance mechanisms in the mutant derivatives of the nine isolates mainly revealed inactivating mutations in the gene coding for outer membrane protein OprD. Point mutations leading to premature stop codons or amino acid substitution S278P, ≥1bp deletion leading to frameshift mutations and interruption of the oprD by an insertion sequence, were observed. MPC and mutant selection window (MSW) are unique parameters that may guide the implementation of antimicrobial treatment, providing useful information about the necessary imipenem concentration needed in the infection area, in order to avoid the emergence of resistance, especially in clinical situations with high bacterial load. Copyright © 2017 Elsevier Inc. All rights reserved.

  20. Ureaplasma parvum undergoes selection in utero resulting in genetically diverse isolates colonizing the chorioamnion of fetal sheep.

    Science.gov (United States)

    Dando, Samantha J; Nitsos, Ilias; Polglase, Graeme R; Newnham, John P; Jobe, Alan H; Knox, Christine L

    2014-02-01

    Ureaplasmas are the microorganisms most frequently isolated from the amniotic fluid of pregnant women and can cause chronic intrauterine infections. These tiny bacteria are thought to undergo rapid evolution and exhibit a hypermutatable phenotype; however, little is known about how ureaplasmas respond to selective pressures in utero. Using an ovine model of chronic intraamniotic infection, we investigated if exposure of ureaplasmas to subinhibitory concentrations of erythromycin could induce phenotypic or genetic indicators of macrolide resistance. At 55 days gestation, 12 pregnant ewes received an intraamniotic injection of a nonclonal, clinical Ureaplasma parvum strain followed by (i) erythromycin treatment (intramuscularly, 30 mg/kg/day, n = 6) or (ii) saline (intramuscularly, n = 6) at 100 days gestation. Fetuses were then delivered surgically at 125 days gestation. Despite injecting the same inoculum into all the ewes, significant differences between amniotic fluid and chorioamnion ureaplasmas were detected following chronic intraamniotic infection. Numerous polymorphisms were observed in domain V of the 23S rRNA gene of ureaplasmas isolated from the chorioamnion (but not the amniotic fluid), resulting in a mosaiclike sequence. Chorioamnion isolates also harbored the macrolide resistance genes erm(B) and msr(D) and were associated with variable roxithromycin minimum inhibitory concentrations. Remarkably, this variability occurred independently of exposure of ureaplasmas to erythromycin, suggesting that low-level erythromycin exposure does not induce ureaplasmal macrolide resistance in utero. Rather, the significant differences observed between amniotic fluid and chorioamnion ureaplasmas suggest that different anatomical sites may select for ureaplasma subtypes within nonclonal, clinical strains. This may have implications for the treatment of intrauterine ureaplasma infections.

  1. Casein Phosphopeptide-Amorphous Calcium Phosphate Reduces Streptococcus mutans Biofilm Development on Glass Ionomer Cement and Disrupts Established Biofilms.

    Science.gov (United States)

    Dashper, Stuart G; Catmull, Deanne V; Liu, Sze-Wei; Myroforidis, Helen; Zalizniak, Ilya; Palamara, Joseph E A; Huq, N Laila; Reynolds, Eric C

    2016-01-01

    Glass ionomer cements (GIC) are dental restorative materials that are suitable for modification to help prevent dental plaque (biofilm) formation. The aim of this study was to determine the effects of incorporating casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) into a GIC on the colonisation and establishment of Streptococcus mutans biofilms and the effects of aqueous CPP-ACP on established S mutans biofilms. S. mutans biofilms were either established in flow cells before a single ten min exposure to 1% w/v CPP-ACP treatment or cultured in static wells or flow cells with either GIC or GIC containing 3% w/w CPP-ACP as the substratum. The biofilms were then visualised using confocal laser scanning microscopy after BacLight LIVE/DEAD staining. A significant decrease in biovolume and average thickness of S. mutans biofilms was observed in both static and flow cell assays when 3% CPP-ACP was incorporated into the GIC substratum. A single ten min treatment with aqueous 1% CPP-ACP resulted in a 58% decrease in biofilm biomass and thickness of established S. mutans biofilms grown in a flow cell. The treatment also significantly altered the structure of these biofilms compared with controls. The incorporation of 3% CPP-ACP into GIC significantly reduced S. mutans biofilm development indicating another potential anticariogenic mechanism of this material. Additionally aqueous CPP-ACP disrupted established S. mutans biofilms. The use of CPP-ACP containing GIC combined with regular CPP-ACP treatment may lower S. mutans challenge.

  2. Selection for autochthonous bifidobacteial isolates adapted to simulated gastrointestinal fluid

    Directory of Open Access Journals (Sweden)

    H Jamalifar

    2010-03-01

    Full Text Available "nBackground and the purpose of the study: Bifidobacterial strains are excessively sensitive to acidic conditions and this can affect their living ability in the stomach and fermented foods, and as a result, restrict their use as live probiotic cultures. The aim of the present study was to obtain bifidobacterial isolates with augmented tolerance to simulated gastrointestinal condition using cross-protection method. "nMethods: Individual bifidobacterial strains were treated in acidic environment and also in media containing bile salts and NaCl. Viability of the acid and acid-bile-NaCl tolerant isolates was further examined in simulated gastric and small intestine by subsequent incubation of the probiotic bacteria in the corresponding media for 120 min. Antipathogenic activities of the adapted isolates were compared with those of the original strains. "nResults and major conclusion: The acid and acid-bile-NaCl adapted isolates showed improved viabilities significantly (p<0.05 in simulated gastric fluid compared to their parent strains. The levels of reduction in bacterial count (Log cfu/ml of the acid and acid-bile-NaCl adapted isolates obtained in simulated gastric fluid ranged from 0.64-3.06 and 0.36-2.43 logarithmic units after 120 min of incubation. There was no significant difference between the viability of the acid-bile-NaCl-tolerant isolates and the original strains in simulated small intestinal condition except for Bifidobacterium adolescentis (p<0.05. The presence of 15 ml of supernatants of acid-bile-NaCl-adapted isolates and also those of the initial Bifidobacterium strains inhibited pathogenic bacterial growth for 24 hrs. Probiotic bacteria with improved ability to survive in harsh gastrointestinal environment could be obtained by subsequent treatment of the strains in acid, bile salts and NaCl environments.

  3. SENSITIVITY OF MOLDS ISOLATED FROM WAREHOUSES OF FOOD PRODUCTION FACILITY ON SELECTED ESSENTIAL OILS

    Directory of Open Access Journals (Sweden)

    Łukasz Kręcidło

    2015-07-01

    Full Text Available Storage of raw materials is one of steps in food production chain. The aim of this study was to estimate the influence of selected essential oils on the growth of four fungal strains: Trichoderma viride, Rhizomucor miehei, Penicillium chrysogenum, Penicillium janthinellum. Strains were isolated from warehouses of the food production facility. Selected essential oils: thyme oil, rosewood oil and rosemary oil were used to assess antifungal activity. Chemical composition of essential oils was determined by Gas Chromatography-Mass Spectroscopy (GC-MS. Antifungal activity of essential oils was estimated in relative to peracetic acid (PAA and sterile water with Tween 80 (0,5%. The influence of essential oils on fungal growth was carried by medium poisoning method. Increment of fungal mycelium was measured every day by 10 days. The thyme essential oils totally inhibited fungal growth in the lowest concentration of 1 mm3·cm-3. The most resistant strain was Penicillium janthinellum.

  4. The Implementation of a Hospital-wide Practice for the Selective Use of Carbapenems Based on the Monitoring of Susceptibility of Pseudomonas aeruginosa Isolates.

    Science.gov (United States)

    Ohshima, Toshio; Asai, Satomi; Miyazawa, Miki; Yamamoto, Yukari; Hisada, Akifumi; Kumazawa, Chie; Hashimoto, Masayoshi; Fukawa, Katsuji; Iwashita, Hideo; Umezawa, Kazuo; Yamada, Sanetoshi; Yamamoto, Yoshiro; Miyachi, Hayato

    2017-12-20

    To control carbapenem-resistant Pseudomonas aeruginosa, we implemented a hospital-wide policy concerning the selective use of carbapenems based on the monitoring of P. aeruginosa isolates for susceptibility to five carbapenems using a customized dry plate method. In this study, we retrospectively investigated the outcome of our measures to control carbapenem-resistant P. aeruginosa. To select effective carbapenems, 100 clinical isolates were collected, and the minimum inhibitory concentration (MIC) to 5 carbapenems (IPM/CS, MEPM, DRPM, BIPM and PAPM/BP) was monitored using a customized dry plate method from 2006 to 2013. Carbapenems, which were associated with a high rate of drug resistance in P. aeruginosa, were restricted from use during our intervention study. The antimicrobial use density per 100 bed-days (AUD 100 ) of carbapenems and the detection rates of carbapenem (IPM/CS and MEPM)-resistant P. aeruginosa were determined during the period of the intervention. The isolates consistently showed higher rates of drug-resistant P. aeruginosa in IPM/CS and PAPM/BP. Thus, DRPM, MEPM and BIPM were adopted for hospital-wide use. The detection rates of all IPM/Cs and MEPM-resistant P. aeruginosa significantly decreased. Meanwhile, the consumption of carbapenems showed an increasing trend. The outcome of the hospital-wide implementation of the selective use of carbapenems based on periodic monitoring of the susceptibility of P. aeruginosa isolates was retrospectively studied. Implementation of this measure might have contributed in part to the control of carbapenem-resistant P. aeruginosa in our hospital.

  5. Serological comparison of selected isolates of Aeromonas salmonicida ssp. Salmonicida

    Science.gov (United States)

    Hahnel, G.B.; Gould, R.W.; Boatman, E.S.

    1983-01-01

    Eight isolates of Acronionus salmonicida ssp. salmonicida were collected during furunculosis epizootics in North American Pacific coast states and provinces. Both virulent and avirulent forms of each isolate, confirmed by challenge and electron microscopy, were examined. Serological comparisons by cross-absorption agglutination tests revealed no serological differences between isolates. Using the double diffusion precipitin test, a single band was observed when antigen from a sonicated virulent strain was reacted with antiserum against a sonicated, virulent strain absorbed with homologous, avirulent strain. The presence of the single band was eliminated by excess sonication.

  6. Insulin increases phosphorylation of mitochondrial proteins in human skeletal muscle in vivo

    DEFF Research Database (Denmark)

    Zhao, Xiaolu; Bak, Steffen; Pedersen, Andreas James Thestrup

    2014-01-01

    , we investigated the effect of insulin on the phosphorylation of mitochondrial proteins in human skeletal muscle in vivo. Using a combination of TiO2 phosphopeptide-enrichment, HILIC fractionation, and LC−MS/MS, we compared the phosphoproteomes of isolated mitochondria from skeletal muscle samples...... obtained from healthy individuals before and after 4 h of insulin infusion. In total, we identified 207 phosphorylation sites in 95 mitochondrial proteins. Of these phosphorylation sites, 45% were identified in both basal and insulin-stimulated samples. Insulin caused a 2-fold increase in the number...... of different mitochondrial phosphopeptides (87 ± 7 vs 40 ± 7, p = 0.015) and phosphoproteins (46 ± 2 vs 26 ± 3, p = 0.005) identified in each mitochondrial preparation. Almost half of the mitochondrial phosphorylation sites (n = 94) were exclusively identified in the insulin-stimulated state and included...

  7. Isolation and selection of plant growth-promoting bacteria associated with sugarcane

    Directory of Open Access Journals (Sweden)

    Ariana Alves Rodrigues

    2016-06-01

    Full Text Available Microorganisms play a vital role in maintaining soil fertility and plant health. They can act as biofertilizers and increase the resistance to biotic and abiotic stress. This study aimed at isolating and characterizing plant growth-promoting bacteria associated with sugarcane, as well as assessing their ability to promote plant growth. Endophytic bacteria from leaf, stem, root and rhizosphere were isolated from the RB 867515 commercial sugarcane variety and screened for indole acetic acid (IAA production, ability to solubilize phosphate, fix nitrogen and produce hydrogen cyanide (HCN, ammonia and the enzymes pectinase, cellulase and chitinase. A total of 136 bacteria were isolated, with 83 of them presenting some plant growth mechanism: 47 % phosphate solubilizers, 26 % nitrogen fixers and 57 % producing IAA, 0.7 % HCN and chitinase, 45 % ammonia, 30 % cellulose and 8 % pectinase. The seven best isolates were tested for their ability to promote plant growth in maize. The isolates tested for plant growth promotion belong to the Enterobacteriaceae family and the Klebsiella, Enterobacter and Pantoea genera. Five isolates promoted plant growth in greenhouse experiments, showing potential as biofertilizers.

  8. The impact of the increased use of piperacillin/tazobactam on the selection of antibiotic resistance among invasive Escherichia coli and Klebsiella pneumoniae isolates.

    Science.gov (United States)

    Lee, Jina; Oh, Chi Eun; Choi, Eun Hwa; Lee, Hoan Jong

    2013-08-01

    Increasing antimicrobial resistance is related to the selective pressure exerted by antibiotic usage. This study evaluated the impact of the increased use of piperacillin/tazobactam (TZP) on the selection of antibiotic resistance. From 1999 to 2010, Escherichia coli and Klebsiella pneumoniae invasive isolates obtained from hospitalized Korean children were analyzed in antibiotic susceptibility tests and subjected to characterization of the β-lactamase types. Antibiotic consumption data were also analyzed. Between January 1999 and December 2010, 409 invasive isolates of E. coli (n=170) and K. pneumoniae (n=239) were obtained. A rebound of extended-spectrum β-lactamase (ESBL) prevalence with an increase in total antibiotics was noted. Non-susceptibility to TZP was determined in 7.6% of E. coli isolates and 20.9% of K. pneumoniae isolates. Despite the increase in TZP usage, the overall prevalence of TZP resistance did not significantly increase over time, especially in E. coli. The mechanisms for TZP resistance included the presence of AmpC producers, possible TEM-1 hyperproducers, and multiple β-lactamases in individual organisms of a given isolate. Replacement of only the antibiotic class appears to be insufficient to control antibiotic resistance, and continued efforts to decrease overall antibiotic pressure are needed, especially in highly endemic situations. Copyright © 2013 International Society for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  9. A Discovery Strategy for Selective Inhibitors of c-Src in Complex with the Focal Adhesion Kinase SH3/SH2-binding Region.

    Science.gov (United States)

    Moroco, Jamie A; Baumgartner, Matthew P; Rust, Heather L; Choi, Hwan Geun; Hur, Wooyoung; Gray, Nathanael S; Camacho, Carlos J; Smithgall, Thomas E

    2015-08-01

    The c-Src tyrosine kinase co-operates with the focal adhesion kinase to regulate cell adhesion and motility. Focal adhesion kinase engages the regulatory SH3 and SH2 domains of c-Src, resulting in localized kinase activation that contributes to tumor cell metastasis. Using assay conditions where c-Src kinase activity required binding to a tyrosine phosphopeptide based on the focal adhesion kinase SH3-SH2 docking sequence, we screened a kinase-biased library for selective inhibitors of the Src/focal adhesion kinase peptide complex versus c-Src alone. This approach identified an aminopyrimidinyl carbamate compound, WH-4-124-2, with nanomolar inhibitory potency and fivefold selectivity for c-Src when bound to the phospho-focal adhesion kinase peptide. Molecular docking studies indicate that WH-4-124-2 may preferentially inhibit the 'DFG-out' conformation of the kinase active site. These findings suggest that interaction of c-Src with focal adhesion kinase induces a unique kinase domain conformation amenable to selective inhibition. © 2014 John Wiley & Sons A/S.

  10. Genotypic Characterization of Azotobacteria Isolated from Argentinean Soils and Plant-Growth-Promoting Traits of Selected Strains with Prospects for Biofertilizer Production

    Directory of Open Access Journals (Sweden)

    Esteban Julián Rubio

    2013-01-01

    Full Text Available The genetic diversity among 31 putative Azotobacter isolates obtained from agricultural and non-agricultural soils was assessed using rep-PCR genomic fingerprinting and identified to species level by ARDRA and partial 16S rRNA gene sequence analysis. High diversity was found among the isolates, identified as A. chroococcum, A. salinestris, and A. armeniacus. Selected isolates were characterized on the basis of phytohormone biosynthesis, nitrogenase activity, siderophore production, and phosphate solubilization. Indole-3 acetic-acid (IAA, gibberellin (GA3 and zeatin (Z biosynthesis, nitrogenase activity, and siderophore production were found in all evaluated strains, with variation among them, but no phosphate solubilization was detected. Phytohormones excreted to the culture medium ranged in the following concentrations: 2.2–18.2 μg IAA mL−1, 0.3–0.7 μg GA3 mL−1, and 0.5–1.2 μg Z mL−1. Seed inoculations with further selected Azotobacter strains and treatments with their cell-free cultures increased the number of seminal roots and root hairs in wheat seedlings. This latter effect was mimicked by treatments with IAA-pure solutions, but it was not related to bacterial root colonization. Our survey constitutes a first approach to the knowledge of Azotobacter species inhabiting Argentinean soils in three contrasting geographical regions. Moreover, this phenotypic characterization constitutes an important contribution to the selection of Azotobacter strains for biofertilizer formulations.

  11. Determination of the minimum inhibitory concentration (MIC and mutant prevention concentration (MPC of selected antimicrobials in bovine and swine Pasteurella multocida, Escherichia coli, and Staphylococcus aureus isolates

    Directory of Open Access Journals (Sweden)

    Kateřina Nedbalcová

    2015-01-01

    Full Text Available We compared the values of the minimum inhibitory concentration (MIC and mutant prevention concentration (MPC values ​​of three antimicrobial agents for 72 bovine isolates of Pasteurella multocida, 80 swine isolates of P. multocida, 80 bovine isolates of Escherichia coli, 80 swine isolates of E. coli, and 80 isolates of Staphylococcus aureus from bovine mastitis. The ratio of MIC90​​/MPC90 which limited mutant selection window (MSW was ≤ 0.12/4 mg/l for enrofloxacin, 0.5/≥ 64 mg/l for florfenicol and 4/≥ 128 mg/l for tulathromycin in bovine P. multocida isolates, ≤ 0.12/2 mg/l for enrofloxacin, 0.5/≥ 64 mg/l for florfenicol and 4/≥ 128 mg/l for tulathromycin in swine P. multocida isolates, 1/16 mg/l for enrofloxacin, 8/≥ 64 mg/l for florfenicol and 8/≥ 128 mg/l for tulathromycin in bovine E. coli isolates, 0.5/16 mg/l for enrofloxacin, ≥ 64/≥ 64 mg/l for florfenicol and 8/≥ 128 mg/l for tulathromycin in swine E. coli isolates, and 0.25/16 mg/l for enrofloxacin, 4/≥ 64 mg/l for florfenicol and 4/≥ 128 mg/l for tulathromycin in S. aureus isolates. These findings indicate that the dosage of antimicrobial agents to achieve serum concentration equal to or higher than MPC could reduce selection of resistant bacterial subpopulation.

  12. In vitro screening of selected probiotic properties of Lactobacillus strains isolated from traditional fermented cabbage and cucumber.

    Science.gov (United States)

    Zielińska, Dorota; Rzepkowska, Anna; Radawska, Anna; Zieliński, Konrad

    2015-02-01

    Most important during probiotic selection are gastric acid and bile tolerance, the adhesion to the luminal epithelium to colonize the lower gastrointestinal tract of a human and safety for human consumption. The aim of this study was to evaluate the selected probiotic in vitro properties of Lactobacillus spp. Strains isolated from traditional fermented food. A total 38 strains were isolated from the pickled samples and 14 were identified as Lactobacillus spp. The survival of almost all strains after incubation at pH 2.5 did not change markedly, and remained at above 90 % (10(9) CFU/mL). The strains also exhibited a high survival rate at pH 3.5 (>90 %), whereas pH 1.5 all were died. Just four strains could survive 90 min. at pH 1.5 (survival rates of 81-94 % after 24 h, whereas after incubation in 2 and 4 % bile salt solution it was 59-94 %. All tested strains showed very good and good resistance to 0.4 % phenol addition, however only Lb. johnsonii K4 was able to multiply. The hydrophobic nature of the cell surface of the tested strains was moderated recording hydrophobicity of Lb. johnsonii K4 and Lb. rhamnosus K3 above 60 %. Safety evaluation excluded four of tested strains as candidate probiotics, according to antibiotic resistance patterns and certain metabolic activities. On the basis on the results 10 of the selected Lactobacillus strains are safe and can survive under gastrointestinal conditions, which requires them to future in vitro and in vivo probiotic studies.

  13. Effects of conditioners on microshear bond strength to enamel after carbamide peroxide bleaching and/or casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) treatment.

    Science.gov (United States)

    Adebayo, O A; Burrow, M F; Tyas, M J

    2007-11-01

    To evaluate (a) the enamel microshear bond strength (MSBS) of a universal adhesive and (b) the effects of conditioning with a self-etching primer adhesive with/without prior bleaching and/or casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) application. Thirty-five molars were cut into four sections, assigned randomly to four groups (no treatment; 16% carbamide peroxide bleaching; CPP-ACP-containing paste (Tooth Mousse, TM); bleaching and TM) and treated accordingly. Specimens were divided into two for bonding with either a self-etching primer (Clearfil SE Bond, CSE) or a total-etch adhesive (Single Bond, SB). Specimens for CSE bonding were subdivided for one of four preconditioning treatments (no conditioning; 30-40% phosphoric acid (PA); 15% EDTA; 20% polyacrylic acid conditioner (Cavity conditioner, CC) and treated. The adhesives were applied and resin composite bonded to the enamel using microtubes (internal diameter 0.75mm). Bonds were stressed in shear until failure, mean MSBS calculated and data analysed using ANOVA with Tukey's HSD test (alpha=0.05). The modes of bond failure were assessed and classified. Two-way ANOVA revealed significant differences between treatments (Padhesive system on treated enamel may significantly improve bond strengths.

  14. Different Patterns of pfcrt and pfmdr1 Polymorphisms in P. falciparum Isolates from Nigeria and Brazil: The Potential Role of Antimalarial Drug Selection Pressure

    Science.gov (United States)

    Gbotosho, Grace O.; Folarin, Onikepe A.; Bustamante, Carolina; Pereira da Silva, Luis Hildebrando; Mesquita, Elieth; Sowunmi, Akintunde; Zalis, Mariano G.; Oduola, Ayoade M. J.; Happi, Christian T.

    2012-01-01

    The effect of antimalarial drug selection on pfcrt and pfmdr1 polymorphisms in Plasmodium falciparum isolates from two distinct geographical locations was determined in 70 and 18 P. falciparum isolates from Nigeria and Brazil, respectively, using nested polymerase chain reaction and direct DNA sequencing approaches. All isolates from Brazil and 72% from Nigeria harbored the mutant SVMNT and CVIET pfcrt haplotype, respectively. The pfcrt CVMNT haplotype was also observed in (7%) of the Nigerian samples. One hundred percent (100%) and 54% of the parasites from Brazil and Nigeria, respectively, harbored wild-type pfmdr1Asn86. We provide first evidence of emergence of the CVMNT haplotype in West Africa. The high prevalence of pfcrt CVIET and SVMNT haplotypes in Nigeria and Brazil, respectively, is indicative of different selective pressure by chloroquine and amodiaquine. Continuous monitoring of pfcrt SVMNT haplotype is required in endemic areas of Africa, where artesunate-amodiaquine combination is used for treatment of acute uncomplicated malaria. PMID:22302850

  15. Molecular mechanisms of membrane impermeability in clinical isolates of Enterobacteriaceae exposed to imipenem selective pressure.

    Science.gov (United States)

    Pavez, Monica; Vieira, Camila; de Araujo, Maria Rita; Cerda, Alvaro; de Almeida, Lara Mendes; Lincopan, Nilton; Mamizuka, Elsa Masae

    2016-07-01

    Intrinsic mechanisms leading to carbapenem-induced membrane impermeability and multidrug resistance are poorly understood in clinical isolates of Enterobacteriaceae. In this study, molecular behaviours during the establishment of membrane impermeability in members of the Enterobacteriaceae family under imipenem selective pressure were investigated. Clinical isolates (n = 22) exhibiting susceptibility to multiple antibiotics or characterised as extended-spectrum β-lactamase (ESBL)- or AmpC-producers were submitted to progressive passages on Mueller-Hinton agar plates containing subclinical concentrations of imipenem [0.5 × the minimum inhibitory concentration (MIC)]. Changes in outer membrane permeability were evaluated by determination of antimicrobial MICs, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), and gene expression analysis related to membrane permeability (i.e. omp35-like, omp36-like and acrA) and regulatory mechanisms (i.e. marA and ompR) by quantitative reverse transcription PCR. Following imipenem induction, 73% of isolates showed increased carbapenem MICs by ≥2 doubling dilutions. At an early stage of treatment, imipenem modulated the expression of porins and efflux pump genes, represented by a reduction of 78% in omp36-like and a two-fold increase in acrA expression. Transcriptional factors marA and ompR were also affected by imipenem induction, increasing mRNA expression by 14- and 4-fold, respectively. High marA expression levels were associated with higher values of acrA expression. These results suggest that imipenem is an important factor in the development of an adaptive response to carbapenems by regulating key genes involved in the control of efflux pumps and porins, which could lead to a multidrug-resistant profile in clinical isolates, contributing to possible treatment failure. Copyright © 2016 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.

  16. Casein Phosphopeptide-Amorphous Calcium Phosphate Reduces Streptococcus mutans Biofilm Development on Glass Ionomer Cement and Disrupts Established Biofilms.

    Directory of Open Access Journals (Sweden)

    Stuart G Dashper

    Full Text Available Glass ionomer cements (GIC are dental restorative materials that are suitable for modification to help prevent dental plaque (biofilm formation. The aim of this study was to determine the effects of incorporating casein phosphopeptide-amorphous calcium phosphate (CPP-ACP into a GIC on the colonisation and establishment of Streptococcus mutans biofilms and the effects of aqueous CPP-ACP on established S mutans biofilms. S. mutans biofilms were either established in flow cells before a single ten min exposure to 1% w/v CPP-ACP treatment or cultured in static wells or flow cells with either GIC or GIC containing 3% w/w CPP-ACP as the substratum. The biofilms were then visualised using confocal laser scanning microscopy after BacLight LIVE/DEAD staining. A significant decrease in biovolume and average thickness of S. mutans biofilms was observed in both static and flow cell assays when 3% CPP-ACP was incorporated into the GIC substratum. A single ten min treatment with aqueous 1% CPP-ACP resulted in a 58% decrease in biofilm biomass and thickness of established S. mutans biofilms grown in a flow cell. The treatment also significantly altered the structure of these biofilms compared with controls. The incorporation of 3% CPP-ACP into GIC significantly reduced S. mutans biofilm development indicating another potential anticariogenic mechanism of this material. Additionally aqueous CPP-ACP disrupted established S. mutans biofilms. The use of CPP-ACP containing GIC combined with regular CPP-ACP treatment may lower S. mutans challenge.

  17. Selection and identification of fungi isolated from sugarcane bagasse and their application for phenanthrene removal from soil.

    Science.gov (United States)

    Cortés-Espinosa, D V; Fernández-Perrino, F J; Arana-Cuenca, A; Esparza-García, F; Loera, O; Rodríguez-Vázquez, R

    2006-01-01

    This work investigated the identification and selection of fungi isolated from sugarcane bagasse and their application for phenanthrene (Phe) removal from soil. Fungi were identified by PCR amplification of ITS regions as Aspergillus terrus, Aspergillus fumigatus and Aspergillus niger, Penicillium glabrum and Cladosporium cladosporioides. A primary selection of fungi was accomplished in plate, considering Phe tolerance of every strain in two different media: potato dextrose agar (PDA) and mineral medium (MM). The radial extension rate (r(r)) in PDA exhibited significant differences (p<0.05) at 200 and 400 ppm of Phe. A secondary selection of A. niger, C. cladosporoides, and P. glabrum sp. was achieved based on their tolerance to 200, 400, 600 and 800 ppm of Phe, in solid culture at a sugarcane bagasse/contaminated soil ratio of 95:5, in Toyamas, Czapeck and Wunder media. Under these conditions, a maximum (70%) Phe removal by A. niger was obtained. In addition C. cladosporioides and A. niger were able to remove high (800 ppm) Phe concentrations.

  18. Optimal selective renewal policy for systems subject to propagated failures with global effect and failure isolation phenomena

    International Nuclear Information System (INIS)

    Maaroufi, Ghofrane; Chelbi, Anis; Rezg, Nidhal

    2013-01-01

    This paper considers a selective maintenance policy for multi-component systems for which a minimum level of reliability is required for each mission. Such systems need to be maintained between consecutive missions. The proposed strategy aims at selecting the components to be maintained (renewed) after the completion of each mission such that a required reliability level is warranted up to the next stop with the minimum cost, taking into account the time period allotted for maintenance between missions and the possibility to extend it while paying a penalty cost. This strategy is applied to binary-state systems subject to propagated failures with global effect, and failure isolation phenomena. A set of rules to reduce the solutions space for such complex systems is developed. A numerical example is presented to illustrate the modeling approach and the use of the reduction rules. Finally, the Monte-Carlo simulation is used in combination with the selective maintenance optimization model to deal with a number of successive missions

  19. An analysis of the binding characteristics of a panel of recently selected ICAM-1 binding Plasmodium falciparum patient isolates

    DEFF Research Database (Denmark)

    Madkhali, Aymen M; Alkurbi, Mohammed O; Szestak, Tadge

    2014-01-01

    patterns of lab-adapted patient isolates after selecting on ICAM-1. We investigated the binding phenotypes using variant ICAM-1 proteins including ICAM-1Ref, ICAM-1Kilifi, ICAM-1S22/A, ICAM-1L42/A and ICAM-1L44/A using static assays. The study also examined ICAM-1 blocking by four anti-ICAM-1 monoclonal...

  20. Isolating GaSb membranes grown metamorphically on GaAs substrates using highly selective substrate removal etch processes

    Energy Technology Data Exchange (ETDEWEB)

    Lavrova, Olga [Univ. of New Mexico, Albuquerque, NM (United States). Dept. of Electrical and Computer Engineering. Center for High Technology Materials; Balakrishnan, Ganesh [Univ. of New Mexico, Albuquerque, NM (United States). Dept. of Electrical and Computer Engineering. Center for High Technology Materials

    2017-02-24

    The etch rates of NH4OH:H2O2 and C6H8O7:H2O2 for GaAs and GaSb have been investigated to develop a selective etch for GaAs substrates and to isolate GaSb epilayers grown on GaAs. The NH4OH:H2O2 solution has a greater etch rate differential for the GaSb/GaAs material system than C6H8O7:H2O2 solution. The selectivity of NH4OH:H2O2 for GaAs/GaSb under optimized etch conditions has been observed to be as high as 11471 ± 1691 whereas that of C6H8O7:H2O2 has been measured up to 143 ± 2. The etch contrast has been verified by isolating 2 μm thick GaSb epi-layers that were grown on GaAs substrates. GaSb membranes were tested and characterized with high-resolution X-Ray diffraction (HR-XRD) and atomic force microscopy (AFM).

  1. Isolation Effect in Immediate and Delayed Recall

    Science.gov (United States)

    Bellezza, Francis S.; Cheney, Terry L.

    1973-01-01

    If the hypothesis of selective rehearsal is used to account for the isolation effect, then the recall of isolated items will depend both on the serial position of the isolated item and on whether recall is immediate or delayed. (Author)

  2. Pooled-DNA sequencing identifies genomic regions of selection in Nigerian isolates of Plasmodium falciparum.

    Science.gov (United States)

    Oyebola, Kolapo M; Idowu, Emmanuel T; Olukosi, Yetunde A; Awolola, Taiwo S; Amambua-Ngwa, Alfred

    2017-06-29

    The burden of falciparum malaria is especially high in sub-Saharan Africa. Differences in pressure from host immunity and antimalarial drugs lead to adaptive changes responsible for high level of genetic variations within and between the parasite populations. Population-specific genetic studies to survey for genes under positive or balancing selection resulting from drug pressure or host immunity will allow for refinement of interventions. We performed a pooled sequencing (pool-seq) of the genomes of 100 Plasmodium falciparum isolates from Nigeria. We explored allele-frequency based neutrality test (Tajima's D) and integrated haplotype score (iHS) to identify genes under selection. Fourteen shared iHS regions that had at least 2 SNPs with a score > 2.5 were identified. These regions code for genes that were likely to have been under strong directional selection. Two of these genes were the chloroquine resistance transporter (CRT) on chromosome 7 and the multidrug resistance 1 (MDR1) on chromosome 5. There was a weak signature of selection in the dihydrofolate reductase (DHFR) gene on chromosome 4 and MDR5 genes on chromosome 13, with only 2 and 3 SNPs respectively identified within the iHS window. We observed strong selection pressure attributable to continued chloroquine and sulfadoxine-pyrimethamine use despite their official proscription for the treatment of uncomplicated malaria. There was also a major selective sweep on chromosome 6 which had 32 SNPs within the shared iHS region. Tajima's D of circumsporozoite protein (CSP), erythrocyte-binding antigen (EBA-175), merozoite surface proteins - MSP3 and MSP7, merozoite surface protein duffy binding-like (MSPDBL2) and serine repeat antigen (SERA-5) were 1.38, 1.29, 0.73, 0.84 and 0.21, respectively. We have demonstrated the use of pool-seq to understand genomic patterns of selection and variability in P. falciparum from Nigeria, which bears the highest burden of infections. This investigation identified known

  3. Selection of Lactic Acid Bacteria with Probiotic Potential Isolated from the Fermentation Process of "Cupuaçu" (Theobroma grandiflorum).

    Science.gov (United States)

    Ornellas, Roberta Maria Santos; Santos, Tiza Teles; Arcucio, Leonardo Borges; Sandes, Sávio Henrique Cicco; Oliveira, Mayara Messias; Dias, Cristiano Villela; de Carvalho Silva, Samuel; Uetanabaro, Ana Paula Trovatti; Vinderola, Gabriel; Nicoli, Jacques Robert

    2017-01-01

    In the present study, nine lactic acid bacteria isolated from the fermentation process of "cupuaçu" (Theobroma grandiflorum) were selected for probiotic use. In vitro (resistance to gastrointestinal environment, in vitro antagonism and co-aggregation with pathogens) and in vivo (intestinal colonization and ex vivo antagonism in germ-free mice, cumulative mortality, translocation to liver and spleen, histopathological examination of liver and ileum and mRNA cytokine gene expression during an experimental infection with S. Typhimurium) assays were used. Among the nine Lactobacillus strains isolated from the "cupuaçu" fermentation, L. plantarum 81 and L. plantarum 90 were selected as potential probiotics based on better results obtained in in vitro evaluations (production of diffusible inhibitory compounds and co-aggregation) as well as in vivo experiments (resistance to gastrointestinal environment, ex vivo antagonism, higher survival after enteropathogen challenge, lower hepatic translocation of enteropathogen, lower histopathological lesions in ileum and liver and anti-inflammatory pattern of immunological response). Concluding, L. plantarum 81 and L. plantarum 90 showed in vitro and in vivo capacities for probiotic use through different mechanisms of protection and its origin would allow an easier adaptation in an alimentary matrix for its administration.

  4. Screening and characterization of phosphate solubilizing bacteria from isolate of thermophilic bacteria

    Science.gov (United States)

    Yulianti, Evy; Rakhmawati, Anna

    2017-08-01

    The aims of this study were to select bacteria that has the ability to dissolve phosphate from thermophilic bacteria isolates after the Merapi eruption. Five isolates of selected bacteria was characterized and continued with identification. Selection was done by using a pikovskaya selective medium. Bacterial isolates were grown in selective medium and incubated for 48 hours at temperature of 55 ° C. Characterization was done by looking at the cell and colony morphology, physiological and biochemical properties. Identification was done with the Profile Matching method based on the reference genus Oscillospira traced through Bergey's Manual of Determinative Bacteriology. Dendogram was created based on similarity index SSM. The results showed there were 14 isolates of bacteria that were able to dissolve phosphate indicated by a clear zone surrounding the bacterial colony on selective media. Five isolates were selected with the largest clear zone. Isolates D79, D92, D110a, D135 and D75 have different characters. The result of phenotypic characters identification with Genus Oscillospira profile has a percentage of 100% similarity to isolate D92 and D110a; 92.31% for isolates D79, and 84.6% for isolates D75 and D135. Dendogram generated from average linkage algorithm / UPGMA using the Simple Matching Coefficient (SSM) algorithms showed, isolate thermophilic bacteria D75 and D135 are combined together to form cluster 1. D110a and D92 form a sub cluster A. Sub cluster A and D79 form cluster 2

  5. Selective isolation and noninvasive analysis of circulating cancer stem cells through Raman imaging.

    Science.gov (United States)

    Cho, Hyeon-Yeol; Hossain, Md Khaled; Lee, Jin-Ho; Han, Jiyou; Lee, Hun Joo; Kim, Kyeong-Jun; Kim, Jong-Hoon; Lee, Ki-Bum; Choi, Jeong-Woo

    2018-04-15

    Circulating cancer stem cells (CCSCs), a rare circulating tumor cell (CTC) type, recently arose as a useful resource for monitoring and characterizing both cancers and their metastatic derivatives. However, due to the scarcity of CCSCs among hematologic cells in the blood and the complexity of the phenotype confirmation process, CCSC research can be extremely challenging. Hence, we report a nanoparticle-mediated Raman imaging method for CCSC characterization which profiles CCSCs based on their surface marker expression phenotypes. We have developed an integrated combinatorial Raman-Active Nanoprobe (RAN) system combined with a microfluidic chip to successfully process complete blood samples. CCSCs and CTCs were detected (90% efficiency) and classified in accordance with their respective surface marker expression via completely distinct Raman signals of RANs. Selectively isolated CCSCs (93% accuracy) were employed for both in vitro and in vivo tumor phenotyping to identify the tumorigenicity of the CCSCs. We utilized our new method to predict metastasis by screening blood samples from xenograft models, showing that upon CCSC detection, all subjects exhibited liver metastasis. Having highly efficient detection and noninvasive isolation capabilities, we have demonstrated that our RAN-based Raman imaging method will be valuable for predicting cancer metastasis and relapse via CCSC detection. Moreover, the exclusion of peak overlapping in CCSC analysis with our Raman imaging method will allow to expand the RAN families for various cancer types, therefore, increasing therapeutic efficacy by providing detailed molecular features of tumor subtypes. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Functional characterization of autophosphorylation sites of the activated insulin receptor-tyrosine kinase

    International Nuclear Information System (INIS)

    Flores-Riveros, J.R.; Lane, M.D.

    1987-01-01

    Insulin receptor, solubilized from 3T3-L1 cellular membranes and then purified, was autophosphorylated with [γ- 32 P]ATP in the absence or presence of insulin. Specific phosphopeptides generated by trypsin digestion of the 32 P-labeled β-subunit were identified and separated by reverse phase HPLC. In the absence of insulin, radioactivity of the phosphopeptides is evenly distributed among four major peaks designated as sites I, II, III and IV, according to their order of elution. This pattern is maintained for at least the first 30 min of autophosphorylation. When the reaction is carried out in the presence of insulin, > 50% of the total 32 P radioactivity is found in site I and the rate of 32 P incorporation into this site is markedly higher than into sites II, III and IV. Maximal activation of tyrosine kinase activity, as estimated by substrate phosphorylation, is coincident with the nearly complete phosphorylation of site I. Delayed activation of previously autophosphorylated receptor by insulin, but not by EGF or IGF-I, produced a similar pattern where phosphorylated site I predominates. These observations indicate that one major insulin-regulated autophosphorylation site in the β-subunit is responsible for activation of the insulin receptor tyrosine kinase. The isolation of this phosphopeptide on a preparative scale and its characterization are now in progress

  7. The Effect of Casein Phosphopeptide-amorphous Calcium Fluoride Phosphate on the Remineralization of Artificial Caries Lesions: An In Vitro Study

    Directory of Open Access Journals (Sweden)

    Ngoc Vo Truong Nhu

    2017-08-01

    Full Text Available The studies on electron microstructure of the effect of the use of products that contain casein phosphopeptide-amorphous calcium fluoride phosphate (CPP-ACPF on enamel remineralization are still needed. It is important method to observe of the morphological changes of teeth in different conditions. Objective: To evaluate the remineralization potential of paste on enamel lesions using scanning electron microscopy (SEM. Methods: Sixty enamel specimens were prepared from extracted human premolars. The specimens were placed in a demineralizing solution for four days to produce artificial carious lesions. The specimens were then randomly assigned to two study groups: group A (control group and group B. Group B was incubated in remineralizing paste (CPP-ACPF for 30 minutes per day for 10 days. The control group received no intervention with remineralizing paste. All 60 specimens were stored in artificial saliva at 370C. After remineralization, the samples were observed using SEM. Results: The statistical analysis showed a decrease in the lesion area between the demineralized and remineralized samples, but no significant difference was observed in the lesion depth for group B. There was a significant increase observed in both the lesion depth and lesion area for group A (p = 0.03. Conclusion: The results showed the capacity of CPP-ACPF in supplying calcium and phosphate to the enamel, decreasing the dissolution of the enamel surface and increasing the remineralization of the enamel surface. 

  8. Selective isolation of potentially phosphate-mobilizing, biosurfactant-producing and biodegradative bacteria associated with a sub-Arctic, terricolous lichen, Peltigera membranacea.

    Science.gov (United States)

    Sigurbjörnsdóttir, Margrét Auður; Vilhelmsson, Oddur

    2016-06-01

    Lichens are the symbiotic association of fungi and a photosynthetic partner. However, non-phototrophic bacteria are also present and thought to comprise an essential part of the lichen symbiosis, although their roles in the symbiosis are still poorly understood. In this study, we isolated and characterized 110 non-phototrophic bacterial lichen associates from thalli of the terricolous lichen Peltigera membranacea The biodegradative and other nutrient-scavenging properties studied among selected isolates were phosphate mobilization, biosurfactant production and degradation of napthalene and several biopolymers, suggesting organic and inorganic nutrient scavenging as roles for bacteria in the lichen symbiotic association. Identification by partial 16S rRNA gene sequencing revealed that the isolates comprised 18 genera within the Proteobacteria, Actinobacteria, Bacteroidetes and Firmicutes, many with high similarities with bacteria typically associated with the plant and rhizosphere environments, could suggest that plants may be important sources of terricolous lichen-associated bacteria, or vice versa. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  9. Molecular characterization and evaluation of mycorrhizal capacity of Suillus isolates from central Spain for the selection of fungal inoculants.

    Science.gov (United States)

    Ruiz-Díez, Beatriz; Rincón, Ana M; de Felipe, María R; Fernández-Pascual, Mercedes

    2006-10-01

    Suillus fungal specimens of pine forests from a Mediterranean area of central Spain (Madrid region) were studied based on molecular and physiological analysis of sporocarps to obtain fungal native inocula to produce mycorrhizal Pinus halepensis Miller in nursery. Variation within the internal transcribed spacer (ITS) region of the ribosomal RNA genes of Suillus was examined by restriction fragment length polymorphism (RFLP) and direct sequencing of polymerase chain reaction products. Ribosomal DNA (rDNA) spacers were amplified from pure cultures obtained from fruit bodies of a range of Suillus species: Suillus bellinii (Inzenga) Watling, Suillus bovinus (Pers.) Kuntze, Suillus collinitus (Fr.) Kuntze, Suillus granulatus (L.) Snell, Suillus mediterraneensis (Jacquet. & Blum) Redeuil, Suillus luteus L. (Gray), and Suillus variegatus (Sw.) Kuntze. Interspecific variation in the length and number of restriction sites of the amplified ITS region was observed. This variation was confirmed by sequencing, which allowed us to identify some isolates. This is the first time that the ITS sequence of S. mediterraneensis is completely described. No intraspecific rDNA variation was observed within isolates of S. collinitus, S. mediterraneensis, and S. luteus. The phylogenetic analysis established the close relationship among these Mediterranean fungal species. As a further step to characterize the different isolates and to understand the relation between genetic and functional diversity, some physiological variables were evaluated. Intraspecific variation in axenic fungal growth and in mycorrhizal capacities was detected, especially within S. collinitus isolates. The fungal isolates stimulated the growth of P. halepensis in different rates. These studies indicated that ITS analysis, in conjunction with mycorrhizal tests, provides suitable combined tools for the analysis of Suillus spp. in a small geographic area for selecting isolates with final afforestation purposes.

  10. An Innovative Approach to Treat Incisors Hypomineralization (MIH): A Combined Use of Casein Phosphopeptide-Amorphous Calcium Phosphate and Hydrogen Peroxide-A Case Report.

    Science.gov (United States)

    Mastroberardino, Stefano; Campus, Guglielmo; Strohmenger, Laura; Villa, Alessandro; Cagetti, Maria Grazia

    2012-01-01

    Molar Incisor Hypomineralization (MIH) is characterized by a developmentally derived deficiency in mineral enamel. Affected teeth present demarcated enamel opacities, ranging from white to brown; also hypoplasia can be associated. Patient frequently claims aesthetic discomfort if anterior teeth are involved. This problem leads patients to request a bleaching treatment to improve aestheticconditions.Nevertheless, hydrogen peroxide can produce serious side-effects, resulting from further mineral loss. Microabrasion and/or a composite restoration are the treatments of choice in teeth with mild/moderate MIH, but they also need enamel loss. Recently, a new remineralizing agent based on Casein Phosphopeptide-Amorphous Calcium Phosphate (CPP-ACP) has been proposed to be effective in hypomineralized enamel, improving also aesthetic conditions. The present paper presents a case report of a young man with white opacities on incisors treated with a combined use of CPP-ACP mousse and hydrogen peroxide gel to correct the aesthetic defect. The patient was instructed to use CPP-ACP for two hours per day for three months in order to obtain enamel remineralization followed by a combined use of CPP-ACP and bleaching agent for further two months. At the end of this five-month treatment, a noticeable aesthetic improvement of the opacities was observed.

  11. An Innovative Approach to Treat Incisors Hypomineralization (MIH: A Combined Use of Casein Phosphopeptide-Amorphous Calcium Phosphate and Hydrogen Peroxide—A Case Report

    Directory of Open Access Journals (Sweden)

    Stefano Mastroberardino

    2012-01-01

    Full Text Available Molar Incisor Hypomineralization (MIH is characterized by a developmentally derived deficiency in mineral enamel. Affected teeth present demarcated enamel opacities, ranging from white to brown; also hypoplasia can be associated. Patient frequently claims aesthetic discomfort if anterior teeth are involved. This problem leads patients to request a bleaching treatment to improve aestheticconditions.Nevertheless, hydrogen peroxide can produce serious side-effects, resulting from further mineral loss. Microabrasion and/or a composite restoration are the treatments of choice in teeth with mild/moderate MIH, but they also need enamel loss. Recently, a new remineralizing agent based on Casein Phosphopeptide-Amorphous Calcium Phosphate (CPP-ACP has been proposed to be effective in hypomineralized enamel, improving also aesthetic conditions. The present paper presents a case report of a young man with white opacities on incisors treated with a combined use of CPP-ACP mousse and hydrogen peroxide gel to correct the aesthetic defect. The patient was instructed to use CPP-ACP for two hours per day for three months in order to obtain enamel remineralization followed by a combined use of CPP-ACP and bleaching agent for further two months. At the end of this five-month treatment, a noticeable aesthetic improvement of the opacities was observed.

  12. Schizosaccharomyces isolation method

    Directory of Open Access Journals (Sweden)

    Benito Santiago

    2014-01-01

    Full Text Available This study discusses the optimization of a selective and differential medium which would facilitate the isolation of Schizosaccharomyces (a genus with a low incidence compared to other microorganisms to select individuals from this genus for industrial purposes, especially in light of the recent recommendation of the use of yeasts from this genus in the wine industry by the International Organisation of Vine and Wine, or to detect the presence of such yeasts, for those many authors who consider them food spoilers. To this end, we studied various selective differential agents based on the main physiological characteristics of these species, such as their high resistances to high concentrations of sugar, sulfur dioxide, sorbic acid, benzoic acid, acetic acid or malo ethanolic fermentation. This selective medium is based on the genus resistance to the antibiotic actidione and its high resistance to inhibitory agents such as benzoic acid. Malic acid was used as a differential factor due to the ability of this genus to metabolise it to ethanol, which allows detecting of the degradation of this compound. Lastly, the medium was successfully used to isolate strains of Schizosaccharomyces pombe from honey and honeycombs.

  13. Approaches for enhancing in situ detection of enterocin genes in thermized milk, and selective isolation of enterocin-producing Enterococcus faecium from Baird-Parker agar.

    Science.gov (United States)

    Vandera, Elpiniki; Tsirka, Georgia; Kakouri, Athanasia; Koukkou, Anna-Irini; Samelis, John

    2018-05-21

    Enterococci are naturally selected for growth in thermized ewes'/goats' milk mixtures used for traditional cooked hard cheese processing in Greece. A culture-independent PCR-based approach was applied to detect the presence of enterocin-encoding genes in naturally culture-enriched thermized milk (TM). Portions of TM (63 °C, 30 s) collected from a commercial cheese plant before addition of starters were fermented at 37 °C for 48 h to facilitate growth of indigenous enterococci. The multiple enterocin-producing (m-Ent+) Enterococcus faecium KE82 and the nisin A-producing Lactococcus lactis subsp. cremoris M104 served as bacteriocin-positive inocula in separate TM treatments. The PCR results revealed a constant presence of the enterocin A, B and P genes in TM fermented naturally at 37 °C. Eleven out of 42 (26.2%) lactic isolates from the enriched TM cultures without inoculation were Ent+ E. faecium assigned to three biotypes. Biotype I (4 isolates) included single entA possessors, whereas biotype II (5 isolates) and biotype III (2 isolates) were m-Ent+ variants profiling entA-entB-entP and entA-entB genes, respectively. Biotype II displayed the strongest antilisterial activity in vitro. Surprisingly, 85.7% (6/7) of the m-Ent+ E. faecium were selectively isolated from Baird-Parker agar, reflecting their natural resistance to 0.01% tellurite contained in the egg yolk supplement. No cytolysin-positive E. faecalis or other Ent+ Enterococcus spp. were isolated. In conclusion, commercially thermized Greek milk is a natural pool or 'reservoir' of antagonistic Ent+ or m-Ent+ E. faecium strains that can be easily detected and recovered by applying this PCR-based approach to naturally fermented milks or cheese products. Copyright © 2018 Elsevier B.V. All rights reserved.

  14. Pepino mosaic virus isolates and differential symptomatology in tomato

    NARCIS (Netherlands)

    Hanssen, I.M.; Paeleman, A.; Vandewoestijne, E.; Bergen, Van L.; Bragard, C.; Lievens, B.; Vanachter, A.C.R.C.; Thomma, B.P.H.J.

    2009-01-01

    Based on a survey conducted in commercial tomato production in Belgium in 2006, four Pepino mosaic virus (PepMV) isolates that differed in symptom expression in the crop of origin were selected for greenhouse trials. The selected isolates were inoculated onto tomato plants grown in four separate

  15. Effects of the addition of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) on mechanical properties of luting and lining glass ionomer cement

    Science.gov (United States)

    Heravi, Farzin; Bagheri, Hossein; Rangrazi, Abdolrasoul; Mojtaba Zebarjad, Seyed

    2016-07-01

    Recently, the addition of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) into glass ionomer cements (GICs) has attracted interest due to its remineralization of teeth and its antibacterial effects. However, it should be investigated to ensure that the incorporation of CPP-ACP does not have significant adverse effects on its mechanical properties. The purpose of this study was to evaluate the effects of the addition of CPP-ACP on the mechanical properties of luting and lining GIC. The first step was to synthesize the CPP-ACP. Then the CPP-ACP at concentrations of 1%, 1.56% and 2% of CPP-ACP was added into a luting and lining GIC. GIC without CPP-ACP was used as a control group. The results revealed that the incorporation of CPP-ACP up to 1.56%(w/w) increased the flexural strength (29%), diametral tensile strength (36%) and microhardness (18%), followed by a reduction in these mechanical properties at 2%(w/w) CPP-ACP. The wear rate was significantly decreased (23%) in 1.56%(w/w) concentration of CPP-ACP and it was increased in 2%(w/w). Accordingly, the addition of 1.56%(w/w) CPP-ACP into luting and lining GIC had no adverse effect on the mechanical properties of luting and lining GIC and could be used in clinical practice.

  16. Antibiotic content of selective culture media for isolation of Capnocytophaga species from oral polymicrobial samples.

    Science.gov (United States)

    Ehrmann, E; Jolivet-Gougeon, A; Bonnaure-Mallet, M; Fosse, T

    2013-10-01

    In oral microbiome, because of the abundance of commensal competitive flora, selective media with antibiotics are necessary for the recovery of fastidious Capnocytophaga species. The performances of six culture media (blood agar, chocolate blood agar, VCAT medium, CAPE medium, bacitracin chocolate blood agar and VK medium) were compared with literature data concerning five other media (FAA, LB, TSBV, CapR and TBBP media). To understand variable growth on selective media, the MICs of each antimicrobial agent contained in this different media (colistin, kanamycin, trimethoprim, trimethoprim-sulfamethoxazole, vancomycin, aztreonam and bacitracin) were determined for all Capnocytophaga species. Overall, VCAT medium (Columbia, 10% cooked horse blood, polyvitaminic supplement, 3·75 mg l(-1) of colistin, 1·5 mg l(-1) of trimethoprim, 1 mg l(-1) of vancomycin and 0·5 mg l(-1) of amphotericin B, Oxoid, France) was the more efficient selective medium, with regard to the detection of Capnocytophaga species from oral samples (P culture, a simple blood agar allowed the growth of all Capnocytophaga species. Nonetheless, in oral samples, because of the abundance of commensal competitive flora, selective media with antibiotics are necessary for the recovery of Capnocytophaga species. The demonstrated superiority of VCAT medium made its use essential for the optimal detection of this bacterial genus. This work showed that extreme caution should be exercised when reporting the isolation of Capnocytophaga species from oral polymicrobial samples, because the culture medium is a determining factor. © 2013 The Society for Applied Microbiology.

  17. The supply solutions for isolated rural consumers

    International Nuclear Information System (INIS)

    Hazi, Gheorghe; Solomon, Petre; Hazi, Aneta

    2004-01-01

    This paper establishes the supply optimal solutions for isolated rural consumers. A complex technical-economical calculation method is developed for selection of the best solutions. This analysis is based on the minimization of the net present value, NPV, criterion. Using the results of this calculation, one can select easily the supply solution for a given active power and for a given distance separating the power source and the isolated consumer. (authors)

  18. Comparison of seismic isolation concepts for FBR

    International Nuclear Information System (INIS)

    Shiojiri, H.; Mazda, T.; Kasai, H.; Kanda, J.N.; Kubo, T.; Madokoro, M.; Shimomura, T.; Nojima, O.

    1989-01-01

    This paper seeks to verify the reliability and effectiveness of seismic isolation for FBR. Some results of the preliminary study of the program are described. Seismic isolation concepts and corresponding seismic isolation devices were selected. Three kinds of seismically-isolated FBR plant concepts were developed by applying promising seismic isolation concepts to the non-isolated FBR plant, and by developing plant component layout plans and building structural designs. Each plant was subjected to seismic response analysis and reduction in the amount of material of components and buildings were estimated for each seismic isolation concepts. Research and development items were evaluated

  19. Antimicrobial susceptibility survey of pathogens isolated from selected patients in Northern Italy

    Directory of Open Access Journals (Sweden)

    Elisabetta Maioli

    2005-03-01

    Full Text Available The Clinical Microbiology Laboratory of the University of Genoa participated, during the year 2003, in an international antimicrobial surveillance program.The collection of isolates was done according to the site of infection and/or type of patient. Four hundred twenty (420 clinical isolates were analyzed during this year and the frequencies of the different pathogens were investigated. A reference centre carried out susceptibility tests. Oxacillin-resistant Staphylococcus aureus represented 47.6% of all S. aureus isolates from blood stream infections and 33.3% of all S. aureus isolated from skin and soft tissue infections in hospitalised patients.These strains showed resistance to most of the antimicrobial agents evaluated, except vancomycin, teicoplanin, quinupristin/dalfopristin and linezolid which registered 100% of susceptibility. Some isolates from blood stream infections such as E. coli demonstrated resistance to ciprofloxacin (23.3%, levofloxacin (20%, and gatifloxacin (16.6%, and Klebsiella pneumoniae was resistant (18% to all fluoroquinolones tested. Pseudomonas aeruginosa manifested resistance to ciprofloxacin (16.6%, while 27.7% of these strains were resistant both to levofloxacin and gatifloxacin. All the Enterobacter cloacae isolated from blood were susceptible to ciprofloxacin, levofloxacin and gatifloxacin. Haemophilus influenzae and Moraxella catarrhalis collected from community-acquired respiratory tract infections were all inhibited by ciprofloxacin, levofloxacin and gatifloxacin. E. coli isolated from urinary tract infections in hospitalised patients were resistant to ciprofloxacin, levofloxacin and gatifloxacin (2.7%. All Salmonella spp. collected from samples of patients affected by infections of the gastro-intestinal tract were susceptible to all fluoroquinolones. Penicillin resistance in Streptococcus pneumoniae was found in 21.4% of isolates from patients with respiratory tract infections. Fluoroquinolone resistance was

  20. Selected fault testing of electronic isolation devices used in nuclear power plant operation

    International Nuclear Information System (INIS)

    Villaran, M.; Hillman, K.; Taylor, J.; Lara, J.; Wilhelm, W.

    1994-05-01

    Electronic isolation devices are used in nuclear power plants to provide electrical separation between safety and non-safety circuits and systems. Major fault testing in an earlier program indicated that some energy may pass through an isolation device when a fault at the maximum credible potential is applied in the transverse mode to its output terminals. During subsequent field qualification testing of isolators, concerns were raised that the worst case fault, that is, the maximum credible fault (MCF), may not occur with a fault at the maximum credible potential, but rather at some lower potential. The present test program investigates whether problems can arise when fault levels up to the MCF potential are applied to the output terminals of an isolator. The fault energy passed through an isolated device during a fault was measured to determine whether the levels are great enough to potentially damage or degrade performance of equipment on the input (Class 1E) side of the isolator

  1. MOX Fabrication Isolation Considerations

    Energy Technology Data Exchange (ETDEWEB)

    Eric L. Shaber; Bradley J Schrader

    2005-08-01

    This document provides a technical position on the preferred level of isolation to fabricate demonstration quantities of mixed oxide transmutation fuels. The Advanced Fuel Cycle Initiative should design and construct automated glovebox fabrication lines for this purpose. This level of isolation adequately protects the health and safety of workers and the general public for all mixed oxide (and other transmutation fuel) manufacturing efforts while retaining flexibility, allowing parallel development and setup, and minimizing capital expense. The basis regulations, issues, and advantages/disadvantages of five potential forms of isolation are summarized here as justification for selection of the preferred technical position.

  2. Phosphoproteins in extracellular vesicles as candidate markers for breast cancer

    OpenAIRE

    Chen, I-Hsuan; Xue, Liang; Hsu, Chuan-Chih; Paez, Juan Sebastian Paez; Pan, Li; Andaluz, Hillary; Wendt, Michael K.; Iliuk, Anton B.; Zhu, Jian-Kang; Tao, W. Andy

    2017-01-01

    Protein phosphorylation is a major regulatory mechanism for many cellular functions, but no phosphoprotein in biofluids has been developed for disease diagnosis because of the presence of active phosphatases. This study presents a general strategy to isolate and identify phosphoproteins in extracellular vesicles (EVs) from human plasma as potential markers to differentiate disease from healthy states. We identified close to 10,000 unique phosphopeptides in EVs from small volumes of plasma sam...

  3. Isolation of saprophytic Leptospira spp. from a selected environmental water source of Argentina.

    Science.gov (United States)

    Scialfa, Exequiel; Grune, Sylvia; Brihuega, Bibiana; Aguirre, Pablo; Rivero, Marina

    2017-11-29

    Ten Leptospira spp. strains were isolated from water samples from Nievas stream, Olavarría, Buenos Aires province (Argentina). The isolates showed the typical motility and morphology of the genus Leptospira under dark field microscopy, developing in liquid EMJH medium after eight days of incubation at 13°C and 30°C. All isolates were negative by the Multiple Locus Variable Number Tandem Repeat Analysis (MLVA). Molecular identification by 16S rRNA gene sequencing identified all isolates as nonpathogenic leptospires. Four isolates showed a genetic profile identical to that of the reference strain Leptospira biflexa serovar Patoc, and six isolates revealed sequence similarities within the 97-98% range, closely related to Leptospira yanagawae and Leptospira meyeri, respectively. Strains ScialfaASA42, ScialfaASA45, ScialfaASA44, ScialfaASA47, ScialfaASA49, ScialfaASA50 and ScialfaASA51 possibly represent a novel species of the genus Leptospira. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  4. Isolation of peat swamp forest foliar endophyte fungi as biofertilizer

    Directory of Open Access Journals (Sweden)

    Safinah Surya Hakim

    2017-01-01

    Full Text Available Peatland restoration activity is facing many obstacles, particularly in planting techniques and poor nutrient in peat soil. Naturally, endophytic fungi are abundant and have great potential as biofertilizer. This research investigates the potential endophytic fungi isolated from leaves of peat swamp tree species for biofertilizer. Research activities include: exploration, in vitro test to examine the phosphate solubilization and identification. Result showed that there were 360 leave segments collected from 4 sampling locations. The colonization percentage of 222 isolates ranged from 52.17% - 60.17%. Fifty seven morphospecies were selected from 222 isolates. Twelve isolates demonstrated ability to produce clear zones and ten isolates were selected for identification. It is concluded that twelve isolated demonstrated potential ability to produce clear zone and Penicillum citrinum isolate P3.10 was identified as an isolate that show the highest potential ability as a biofertilizer

  5. Comparação da eficiência dos caldos de enriquecimento seletivo no isolamento de Salmonella Dublin Comparison of the efficiency of selective enrichment broths for Salmonella Dublin isolation

    Directory of Open Access Journals (Sweden)

    D.G. Silva

    2008-06-01

    Full Text Available The aim of this study was to compare three different selective enrichment broths: Rappaport-Vassiliadis (RV, selenite cystine (SC and Muller-Kauffmann tetrathionate (MKT for Salmonella Dublin isolation from faecal samples of calf experimentally infected. The bacteriological procedure involved pre-enrichment stages in Hajna-GN broth (only for the samples inoculated in RV broth, selective enrichment, culture in modified brilliant green agar (BGA, presumptive biochemistry tests (using triple-sugar-iron agar and lysine-agar and slide agglutination test with poli-O and poli-H Salmonella antiserum. The effects of enrichment temperatures using RV broth were also evaluated (37ºC and 42ºC. SC broth was significantly more efficient in the isolation of Salmonella Dublin (P<0,05, whereas RV broth incubated at 42ºC had a lower efficiency in the microbiological isolation.

  6. Antibacterial Activities of Lactic Acid Bacteria Isolated from Selected ...

    African Journals Online (AJOL)

    Members of lactic acid bacteria (LAB) are known probiotics and have been reported to have antimicrobial properties. Although various researchers have documented the isolation of these bacteria from fruits and vegetables, studies on LAB associated with lettuce, cucumber and cabbage are limited and non-existing in ...

  7. Isolation and characterization of iron chelators from turmeric (Curcuma longa): selective metal binding by curcuminoids.

    Science.gov (United States)

    Messner, Donald J; Surrago, Christine; Fiordalisi, Celia; Chung, Wing Yin; Kowdley, Kris V

    2017-10-01

    Iron overload disorders may be treated by chelation therapy. This study describes a novel method for isolating iron chelators from complex mixtures including plant extracts. We demonstrate the one-step isolation of curcuminoids from turmeric, the medicinal food spice derived from Curcuma longa. The method uses iron-nitrilotriacetic acid (NTA)-agarose, to which curcumin binds rapidly, specifically, and reversibly. Curcumin, demethoxycurcumin, and bisdemethoxycurcumin each bound iron-NTA-agarose with comparable affinities and a stoichiometry near 1. Analyses of binding efficiencies and purity demonstrated that curcuminoids comprise the primary iron binding compounds recovered from a crude turmeric extract. Competition of curcuminoid binding to the iron resin was used to characterize the metal binding site on curcumin and to detect iron binding by added chelators. Curcumin-Iron-NTA-agarose binding was inhibited by other metals with relative potency: (>90% inhibition) Cu 2+  ~ Al 3+  > Zn 2+  ≥ Ca 2+  ~ Mg 2+  ~ Mn 2+ (80% by addition of iron to the media; uptake was completely restored by desferoxamine. Ranking of metals by relative potencies for blocking curcumin uptake agreed with their relative potencies in blocking curcumin binding to iron-NTA-agarose. We conclude that curcumin can selectively bind toxic metals including iron in a physiological setting, and propose inhibition of curcumin binding to iron-NTA-agarose for iron chelator screening.

  8. Evaluation of isolation methods for pathogenic Yersinia enterocolitica from pig intestinal content.

    Science.gov (United States)

    Laukkanen, R; Hakkinen, M; Lundén, J; Fredriksson-Ahomaa, M; Johansson, T; Korkeala, H

    2010-03-01

    The aim of this study was to evaluate the efficiency of four isolation methods for the detection of pathogenic Yersinia enterocolitica from pig intestinal content. The four methods comprised of 15 isolation steps using selective enrichments (irgasan-ticarcillin-potassium chlorate and modified Rappaport broth) and mildly selective enrichments at 4 or 25 degrees C. Salmonella-Shigella-desoxycholate-calcium chloride agar, cefsulodin-irgasan-novobiocin agar were used as plating media. The most sensitive method detected 78% (53/68) of the positive samples. Individual isolation steps using cold enrichment as the only enrichment or as a pre-enrichment step with further selective enrichment showed the highest sensitivities (55-66%). All isolation methods resulted in high numbers of suspected colonies not confirmed as pathogenic Y. enterocolitica. Cold enrichment should be used in the detection of pathogenic Y. enterocolitica from pig intestinal contents. In addition, more than one parallel isolation step is needed. The study shows that depending on the isolation method used for Y. enterocolitica, the detected prevalence of Y. enterocolitica in pig intestinal contents varies greatly. More selective and sensitive isolation methods need to be developed for pathogenic Y. enterocolitica.

  9. Atomic force microscopic comparison of remineralization with casein-phosphopeptide amorphous calcium phosphate paste, acidulated phosphate fluoride gel and iron supplement in primary and permanent teeth: An in-vitro study

    Directory of Open Access Journals (Sweden)

    Nikita Agrawal

    2014-01-01

    Full Text Available Context: Demineralization of tooth by erosion is caused by frequent contact between the tooth surface and acids present in soft drinks. Aim: The present study objective was to evaluate the remineralization potential of casein-phosphopeptide-amorphous calcium phosphate (CPP-ACP paste, 1.23% acidulated phosphate fluoride (APF gel and iron supplement on dental erosion by soft drinks in human primary and permanent enamel using atomic force microscopy (AFM. Materials and Methods: Specimens were made from extracted 15 primary and 15 permanent teeth which were randomly divided into three treatment groups: CPP-ACP paste, APF gel and iron supplement. AFM was used for baseline readings followed by demineralization and remineralization cycle. Results and Statistics: Almost all group of samples showed remineralization that is a reduction in surface roughness which was higher with CPP-ACP paste. Statistical analysis was performed using by one-way ANOVA and Mann-Whitney U-test with P < 0.05. Conclusions: It can be concluded that the application of CPP-ACP paste is effective on preventing dental erosion from soft drinks.

  10. Selection of Beauveria bassiana sensu lato and Metarhizium anisopliae sensu lato isolates as microbial control agents against the boll weevil (Anthonomus grandis) in Argentina.

    Science.gov (United States)

    Nussenbaum, A L; Lecuona, R E

    2012-05-01

    The boll weevil (Anthonomus grandis) is the main pest of cotton in the Americas. The aim of this work was to evaluate isolates of the entomopathogenic fungi Beauveria bassiana sensu lato and Metarhizium anisopliae sensu lato virulent against A. grandis. Screening was performed to evaluate the pathogenicity of 28 isolates of M. anisopliae s.l. and 66 isolates of B. bassiana s.l. against boll weevil adults. To select the isolates, LC(50) values of the most virulent isolates were calculated, and compatibility between the fungi and insecticides was studied. In addition, the effects of these isolates on the feeding behavior of the adults were evaluated. Isolates Ma 50 and Ma 20 were the most virulent against A. grandis and their LC(50) values were 1.13×10(7) and 1.20×10(7) conidia/ml, respectively. In addition, these isolates were compatible with pyrethroid insecticides, but none with endosulfan. On the other hand, infected females reduced the damage caused by feeding on the cotton squares and their weight gain. This shows that entomopathogenic fungi cause mortality in the insects, but also these fungi could influence the feeding behavior of the females. In summary, these results indicate the possibility of the use of M. anisopliae s.l. as a microbiological control agent against boll weevils. Also, this species could be included in an Integrated Pest Management program. Copyright © 2012 Elsevier Inc. All rights reserved.

  11. Isolated trochlear nerve palsy with midbrain hemorrhage

    Directory of Open Access Journals (Sweden)

    Raghavendra S

    2010-01-01

    Full Text Available Midbrain hemorrhage causing isolated fourth nerve palsy is extremely rare. Idiopathic, traumatic and congenital abnormalities are the most common causes of fourth nerve palsy. We report acute isolated fourth nerve palsy in an 18-year-old lady due to a midbrain hemorrhage probably due to a midbrain cavernoma. The case highlights the need for neuroimaging in selected cases of isolated trochlear nerve palsy.

  12. Molecular detection of genes encoding AcrAB , Qep A efflux pumps in Klebsiella pneumoniae strains isolated from hospitalized patients in selected hospitals in Tehran

    Directory of Open Access Journals (Sweden)

    Mohsen Heidary

    2017-03-01

    Full Text Available Abstract Background and Objectives: Increasing emergence of fluoroquinolone resistance among clinical isolates of Klebsiella pneumoniae  (K. pneumoniae, has limited the treatment options for treatment of infections caused by these bacteria. The aim of this study was to investigate the dissemination of genes encoding AcrAB and QepA efflux pumps among K. pneumoniae strains. Methods: This study was carried out on 117 K. pneumoniae strains isolated from patients hospitalized in selected hospitals in Tehran city, 2015-2016, Iran. Antimicrobial susceptibility tests were performed using disk diffusion method (based on CLSI guidelines and identification of acr A, acr B and qep A genes using PCR assay. Results: In this study, colistin and tigecycline had the best effect against clinical isolates of K. pneumoniae. According to PCR results, 110 (94% isolates had acrA gene and 102 (87% isolates had acrB gene, respectively. The qepA gene was not found in any of the K. pneumoniae strains. Conclusion: According to the results of the present study, dissemination of the genes encoding AcrAB efflux pumps among K. pneumoniae strains, which cause resistance to fluoroquinolones, is a matter of concern. Therefore, infection control and prevention of the spread of drug-resistant bacteria requires careful management in drug prescription and identification of resistant isolates.

  13. HIV-1 isolation from infected peripheral blood mononuclear cells

    NARCIS (Netherlands)

    Dispinseri, Stefania; Saba, Elisa; Vicenzi, Elisa; Kootstra, Neeltje A.; Schuitemaker, Hanneke; Scarlatti, Gabriella

    2014-01-01

    Human immunodeficiency virus 1 (HIV-1) isolation from peripheral blood mononuclear cells (PBMCs) allows retrieval of replication-competent viral variants. In order to impose the smallest possible selective pressure on the viral isolates, isolation must be carried out in primary cultures of cells and

  14. Patterns of reproductive isolation in the Drosophila subquinaria complex: can reinforced premating isolation cascade to other species?

    Science.gov (United States)

    Humphreys, Devon P.; Rundle, Howard D.; Dyer, Kelly A.

    2016-01-01

    Abstract The reinforcement of premating barriers due to reduced hybrid fitness in sympatry may cause secondary sexual isolation within a species as a by-product. Consistent with this, in the fly Drosophila subquinaria, females that are sympatric with D. recens mate at very low rates not only with D. recens, but also with conspecific D. subquinaria males from allopatry. Here, we ask if these effects of reinforcement cascade more broadly to affect sexual isolation with other closely related species. We assay reproductive isolation of these species with D. transversa and find that choosy D. subquinaria females from the region sympatric with D. recens discriminate strongly against male D. transversa, whereas D. subquinaria from the allopatric region do not. This increased sexual isolation cannot be explained by natural selection to avoid mating with this species, as they are allopatric in geographic range and we do not identify any intrinsic postzygotic isolation between D. subquinaria and D. transversa. Variation in epicuticular hydrocarbons, which are used as mating signals in D. subquinaria, follow patterns of premating isolation: D. transversa and allopatric D. subquinaria are most similar to each other and differ from sympatric D. subquinaria, and those of D. recens are distinct from the other two species. We suggest that the secondary effects of reinforcement may cascade to strengthen reproductive isolation with other species that were not a target of selection. These effects may enhance the divergence that occurs in allopatry to help explain why some species are already sexually isolated upon secondary contact. PMID:29491905

  15. Development of novel agar media for isolating guaiacol producing Alicyclobacillus spp.

    Science.gov (United States)

    Chang, S S; Park, S H; Kang, D H

    2013-06-03

    The purpose of this study is to develop a selective and differential medium (SK2 agar) for isolating guaiacol producing Alicyclobacillus. Forty-one selected dyes and vanillic acid were incorporated in SK agar for screening selective and differential agents. Two guaiacol producing (1016, 1101) and two non-guaiacol producing (19220, C-GD 1-1) Alicyclobacillus isolates were streaked onto media and color differentiation of the isolates was assessed. Among 41 tested dyes, Chrome Azurol S (CAS) allowed color differentiation of the two types of Alicyclobacillus. Colonies of guaiacol producing Alicyclobacillus isolates appeared as dark purple to royal blue color with yellow background, whereas non-guaiacol producing Alicyclobacillus isolates produced cream colored colonies with yellow background. Vanillic acid not only served as a precursor for guaiacol formation but also inhibited non-guaiacol producing Alicyclobacillus. Non-guaiacol producing isolates did not grow on SK agar containing more than 70 ppm vanillic acid, whereas the recovery of guaiacol producing isolates was unaffected. When compared with other Alicyclobacillus isolation media, not only was SK2 agar capable of selectively recovering guaiacol-producing Alicyclobacillus, the degree of growth was also approximately equal if not better than orange serum agar, potato dextrose agar, and K agar. The development of SK2 agar provides the fruit juice industry with an inexpensive, simple to use alternative for the detection of guaiacol producing Alicyclobacillus. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. Frequency of isolation and antibiotic resistance patterns of bacterial isolates from wound infections

    Directory of Open Access Journals (Sweden)

    Stojanović-Radić, Z.

    2016-12-01

    Full Text Available Six hundred and thirteen bacterial strains were isolated from wound swabs and the isolates were identified on the basis of growth on differential and selective media. In order to test the sensitivity of isolated strains to different antibiotics, the disc diffusion method, according to EUCAST protocol v 5.0 was used. The most common species isolated from wound swabs was Staphylococcus epidermidis (18.4%, followed by Staphylococcus aureus, Pseudomonas aeruginosa and Enterococcus faecalis (16.8%, 12.7% and 10.4%, respectively. The maximum resistance of Gram-positive cocci was observed to penicillin and the lowest to linezolid. Gram-negative bacteria showed the highest resistance to tetracyclines, while the same strains demonstrated the highest sensitivity to polypeptide antibiotics. Comparison of the resistance patterns of Gramnegative and Gram-positive bacterial strains showed significant difference in the tetracycline efficiency.

  17. Selection of rhizosphere local microbial as bioactive inoculant based on irradiated compost

    International Nuclear Information System (INIS)

    Dadang Sudrajat; Nana Mulyana; Arief Adhari

    2014-01-01

    One of the main components of carrier based on irradiation compost for bio organic fertilizer is a potential microbial isolates role in nutrient supply and growth hormone. This research was conducted to obtain microbial isolates from plant root zone (rhizosphere), further isolation and selection in order to obtain potential isolates capable of nitrogen fixation (N 2 ), resulting in growth hormone (Indole Acetic Acid), and phosphate solubilizing. Selected potential isolates used as bioactive microbial inoculants formulation in irradiation compost based. Forty eight (48) rhizosphere samples were collected from different areas of West and Central Java. One hundred sixteen (116) isolates have been characterized for their morphological, cultural, staining and biochemical characteristics. Isolates have been selected for further screening of PGPR traits. Parameters assessed were Indole Acetic Acid (IAA) content analysis with colorimetric methods, dinitrogen fixation using gas chromatography, phosphate solubility test qualitatively (in the media pikovskaya) and quantitative assay of dissolved phosphate (spectrophotometry). Evaluation of the ability of selected isolates on the growth of corn plants were done in pots. The isolates will be used as inoculant consortium base on compost irradiation. The selection obtained eight (8) bacterial isolates identified as Bacillus circulans (3 isolates), Bacillus stearothermophilus (1 isolate), Azotobacter sp (3 isolates), Pseudomonas diminuta (1 isolate). The highest phosphate released (91,21 mg/l) was by BD2 isolate (Bacillus circulan) with a holo zone size (1.32 cm) on Pikovskaya agar medium. Isolate of Pseudomonas diminuta (KACI) was capable to produce the highest IAA hormone (74.34 μg/ml). The highest nitrogen (N 2 ) fixation activity was shown by Azotobacter sp isolates (KDB2) at a rate of 235.05 nmol/hour. The viability test showed that all selected isolates in compost irradiation carrier slightly decreased after 3 months of

  18. Divergent Selection and Then What Not: The Conundrum of Missing Reproductive Isolation in Misty Lake and Stream Stickleback

    Directory of Open Access Journals (Sweden)

    Katja Räsänen

    2012-01-01

    Full Text Available In ecological speciation, reproductive isolation evolves as a consequence of adaptation to different selective environments. A frequent contributor to this process is the evolution of positive assortative mate choice between ecotypes. We tested this expectation for lake and inlet stream threespine stickleback (Gasterosteus aculeatus from the Misty system (Vancouver Island, Canada, which show strong genetically based adaptive divergence and little genetic exchange in nature. This, and work on other stickleback systems, led us to expect positive assortative mating. Yet, our standard “no-choice” laboratory experiment on common-garden fish revealed no evidence for this—despite divergence in traits typically mediating assortative mating in stickleback. These results remind us that divergent natural selection may not inevitably lead to the evolution of positive assortative mate choice. The apparent lack of strong and symmetric reproductive barriers in this system presents a conundrum: why are such barriers not evident despite strong adaptive divergence and low gene flow in nature?

  19. Fermentation of Apple Juice with a Selected Yeast Strain Isolated from the Fermented Foods of Himalayan Regions and Its Organoleptic Properties

    OpenAIRE

    Kanwar, S. S.; Keshani,

    2016-01-01

    Twenty-three Saccharomyces cerevisiae strains isolated from different fermented foods of Western Himalayas have been studied for strain level and functional diversity in our department. Among these 23 strains, 10 S. cerevisiae strains on the basis of variation in their brewing traits were selected to study their organoleptic effect at gene level by targeting ATF1 gene, which is responsible for ester synthesis during fermentation. Significant variation was observed in ATF1 gene sequences, sugg...

  20. Isolation of Salmonella and Shigella species from house flies ...

    African Journals Online (AJOL)

    Salmonella and Shigella species were isolated from House flies (Musca domestica L.) from various sampling sites using selective media. Out of 34 pooled samples Shigella species were isolated in all (100%) of the samples while Salmonella species were isolated in 21 (61.7%) of the samples. The flies pooled from the ...

  1. A select bibliography with abstracts of reports related to Waste Isolation Pilot Plant geotechnical studies (1972--1990)

    Energy Technology Data Exchange (ETDEWEB)

    Powers, D.W. [Powers (Dennis W.), Anthony, TX (United States); Martin, M.L. [International Technology, Inc., Las Vegas, NV (United States)

    1993-08-01

    This select bibliography contains 941 entries. Each bibliographic entry contains the citation of a report, conference paper, or journal article containing geotechnical information about the Waste Isolation Pilot Plant (WIPP). The entries cover the period from 1972, when investigation began for a WIPP Site in southeastern New Mexico, through December 1990. Each entry is followed by an abstract. If an abstract or suitable summary existed, it has been included; 316 abstracts were written for other documents. For some entries, an annotation has been provided to clarify the abstract, comment on the setting and significance of the document, or guide the reader to related reports. An index of key words/phrases is included for all entries.

  2. A select bibliography with abstracts of reports related to Waste Isolation Pilot Plant geotechnical studies (1972--1990)

    International Nuclear Information System (INIS)

    Powers, D.W.; Martin, M.L.

    1993-08-01

    This select bibliography contains 941 entries. Each bibliographic entry contains the citation of a report, conference paper, or journal article containing geotechnical information about the Waste Isolation Pilot Plant (WIPP). The entries cover the period from 1972, when investigation began for a WIPP Site in southeastern New Mexico, through December 1990. Each entry is followed by an abstract. If an abstract or suitable summary existed, it has been included; 316 abstracts were written for other documents. For some entries, an annotation has been provided to clarify the abstract, comment on the setting and significance of the document, or guide the reader to related reports. An index of key words/phrases is included for all entries

  3. Invitro Evaluation Of Antibacterial Activity Of Lactic Acid Bacteria Isolated From Ergo And Qotchqotcha Ethiopian Traditional Fermented Foods Against Some Selected Food Borne Pathogens

    Directory of Open Access Journals (Sweden)

    Lamenew Fenta

    2017-09-01

    Full Text Available The prevention of pathogenic bacteria by lactic acid bacteria LAB isolated directly from foods is an innovative approach. With the aim of determining the anti-bacterial activity of Lactic acid bacteria isolated from ergo and qotchqotcha Ethiopian fermented food 12 samples of each were taken from the 4 different kebeles of Assosa town. Isolation of LAB from the selected samples were carried out using MRS media. The different set of isolates were characterized using primary biochemical tests. Isolates which were gram positive catalase negative and KOH negative were considered to be presumptive LAB and further characterized by using different biochemical tests for further identification. 16 isolates from ergo samples were isolated. Based on Bergeys manual of determinative bacteriology the 16 isolates belonged to four 4 LAB species namely Lactobacillus acidophilus 18.75 Lactobacillus casei 31.25 Streptococcus thermophiles 25 and Lactobacillus bulgaricus 25. Likewise 5 isolates were isolated from Qotchqotcha and the five isolates were found to be Lactobacillus acidophilus 80 and Pediococcus acidilactici 20. Cell free solution from MRS broth culture of theses LAB was prepared and tested against Escherichia coli O157H7 and Staphylococcus aureus using agar-well diffusion method. Of the 16 isolates isolated from ergo 9 of them show antimicrobial activity against E. coli O157H7 with a largest inhibition zone measured about 7.331.20mm by EK0101 and 12 of them show antimicrobial activity against S. aureus with a largest inhibition zone measured about 11.66 0.88mm by EK0201. On the basis of morphological and biochemical test done EK0101 was found to be presumptive Lactobacillus acidophilus and EK0201 to be Streptococcus thermophiles. All of the isolates isolated from Qotchqotcha showed antimicrobial activity against the tested organisms though there was a significant difference in their activity P0.05. The isolate QK0201 showed an inhibition zone of about 6

  4. Selected lactic acid-producing bacterial isolates with the capacity to reduce Salmonella translocation and virulence gene expression in chickens.

    Directory of Open Access Journals (Sweden)

    Xiaojian Yang

    Full Text Available BACKGROUND: Probiotics have been used to control Salmonella colonization/infection in chickens. Yet the mechanisms of probiotic effects are not fully understood. This study has characterized our previously-selected lactic acid-producing bacterial (LAB isolates for controlling Salmonella infection in chickens, particularly the mechanism underlying the control. METHODOLOGY/PRINCIPAL FINDINGS: In vitro studies were conducted to characterize 14 LAB isolates for their tolerance to low pH (2.0 and high bile salt (0.3-1.5% and susceptibility to antibiotics. Three chicken infection trials were subsequently carried out to evaluate four of the isolates for reducing the burden of Salmonella enterica serovar Typhimurium in the broiler cecum. Chicks were gavaged with LAB cultures (10(6-7 CFU/chick or phosphate-buffered saline (PBS at 1 day of age followed by Salmonella challenge (10(4 CFU/chick next day. Samples of cecal digesta, spleen, and liver were examined for Salmonella counts on days 1, 3, or 4 post-challenge. Salmonella in the cecum from Trial 3 was also assessed for the expression of ten virulence genes located in its pathogenicity island-1 (SPI-1. These genes play a role in Salmonella intestinal invasion. Tested LAB isolates (individuals or mixed cultures were unable to lower Salmonella burden in the chicken cecum, but able to attenuate Salmonella infection in the spleen and liver. The LAB treatments also reduced almost all SPI-1 virulence gene expression (9 out of 10 in the chicken cecum, particularly at the low dose. In vitro treatment with the extracellular culture fluid from a LAB culture also down-regulated most SPI-1 virulence gene expression. CONCLUSIONS/SIGNIFICANCE: The possible correlation between attenuation of Salmonella infection in the chicken spleen and liver and reduction of Salmonella SPI-1 virulence gene expression in the chicken cecum by LAB isolates is a new observation. Suppression of Salmonella virulence gene expression in

  5. Incidence and Antibiotic Susceptibility Pattern of Bacterial Isolates ...

    African Journals Online (AJOL)

    bacterial isolates. The isolates were characterized and identified by standard microbiological methods. Antibiotic susceptibility testing was carried out ... Pure cultures were characterized using morphological appearances on selective and .... J Nepal Health Res Council. 2007; 5(1): 22-26. 4. Taiwo SS, Okesina AB, Onile BA.

  6. Ecological Divergence and the Origins of Intrinsic Postmating Isolation with Gene Flow

    Directory of Open Access Journals (Sweden)

    Aneil F. Agrawal

    2011-01-01

    Full Text Available The evolution of intrinsic postmating isolation has received much attention, both historically and in recent studies of speciation genes. Intrinsic isolation often stems from between-locus genetic incompatibilities, where alleles that function well within species are incompatible with one another when brought together in the genome of a hybrid. It can be difficult for such incompatibilities to originate when populations diverge with gene flow, because deleterious genotypic combinations will be created and then purged by selection. However, it has been argued that if genes underlying incompatibilities are themselves subject to divergent selection, then they might overcome gene flow to diverge between populations, resulting in the origin of incompatibilities. Nonetheless, there has been little explicit mathematical exploration of such scenarios for the origin of intrinsic incompatibilities during ecological speciation with gene flow. Here we explore theoretical models for the origin of intrinsic isolation where genes subject to divergent natural selection also affect intrinsic isolation, either directly or via linkage disequilibrium with other loci. Such genes indeed overcome gene flow, diverge between populations, and thus result in the evolution of intrinsic isolation. We also examine barriers to neutral gene flow. Surprisingly, we find that intrinsic isolation sometimes weakens this barrier, by impeding differentiation via ecologically based divergent selection.

  7. Isolasi, Seleksi Dan Opttmasi Produksi Protease Daribeberapaisolat Bakteri*(isolation, Selection and Optimalization of Protease Production of Some Bacterial Isolates)

    OpenAIRE

    Naiola, Elidar; Widhyastuti, Nunuk

    2002-01-01

    Thirty-seven out of sixty-one bacterial isolates from various sources of samples were screened for protease production. The isolate of ISO PL3 could produce the highest enzyme activity, and it was used as a standard bacterial strain in this observation. For any reason,we implemented ISO PL2 to study the optimum condition for producing bacterial protease. Result shows that the maximum protease activity was obtained in a medium containing 100 gram of rice brand in a liter tofu liquid waste. The...

  8. Batesian mimicry promotes pre- and postmating isolation in a snake mimicry complex.

    Science.gov (United States)

    Pfennig, David W; Akcali, Christopher K; Kikuchi, David W

    2015-04-01

    We evaluated whether Batesian mimicry promotes early-stage reproductive isolation. Many Batesian mimics occur not only in sympatry with their model (as expected), but also in allopatry. As a consequence of local adaptation within both sympatry (where mimetic traits are favored) and allopatry (where nonmimetic traits are favored), divergent, predator-mediated natural selection should disfavor immigrants between these selective environments as well as any between-environment hybrids. This selection might form the basis for both pre- and postmating isolation, respectively. We tested for such selection in a snake mimicry complex by placing clay replicas of sympatric, allopatric, or hybrid phenotypes in both sympatry and allopatry and measuring predation attempts. As predicted, replicas with immigrant phenotypes were disfavored in both selective environments. Replicas with hybrid phenotypes were also disfavored, but only in a region of sympatry where previous studies have detected strong selection favoring precise mimicry. By fostering immigrant inviability and ecologically dependent selection against hybrids (at least in some habitats), Batesian mimicry might therefore promote reproductive isolation. Thus, although Batesian mimicry has long been viewed as a mechanism for convergent evolution, it might play an underappreciated role in fueling divergent evolution and possibly even the evolution of reproductive isolation and speciation. © 2015 The Author(s).

  9. Impact of social isolation on behavioral health in elderly: Systematic review.

    Science.gov (United States)

    Choi, Hanbyul; Irwin, Michael R; Cho, Hyong Jin

    2015-12-22

    To examine and compare the effects of subjective and objective social isolation on behavioral health in elderly adults. A systematic search of PubMed was performed for original research articles from peer-reviewed journals examining one of the following topics: "Social isolation and sleep disturbance", "social isolation and depression", or "social isolation and fatigue in older adults". Studies were selected following the criteria established based on the aim of this review. Data were extracted from the articles by two independent reviewers. Due to the heterogeneity in study designs and outcome measures of the included studies, qualitative and narrative analyses were conducted. The set criteria were used to select a total of 16 studies for the review. Of the 16, 13 were cross-sectional studies. The characteristics of study populations were identified as follows. A total of 12 studies randomly selected subjects irrespective of pre-existing health conditions. Consequently, an unspecified number of the study subjects had chronic diseases in the studies compared. In addition, cultural and ethnic backgrounds of studies in this review were diverse, and included subjects living in North America, South America, Asia, Europe, and Oceania. Both subjective and objective types of social isolation increased behavioral symptoms, such as sleep disturbance, depressive symptoms, and fatigue in older adults. Furthermore, a few recent studies reported stronger effects of subjective social isolation than objective social isolation on sleep disturbance and depressive symptoms. Social isolation affects behavioral health in older adults. Compared to the objective social isolation, subjective social isolation contributes more significantly to sleep disturbance and depression.

  10. The Study of Isolated Bacteria Application for Bioremediation Agent of Uranium Radionuclide in the Environment

    International Nuclear Information System (INIS)

    Yazid, Mochd

    2007-01-01

    Application of the isolated bacteria on the Low Level Uranium Waste as uranium bioremediation agent in the environment has been studied. The objective of this research is to study the possibility of isolated bacteria to be used on uranium remediation process. The isolation of uranium resistance bacteria was carried out on the selective medium SBS containing 10 mg/l uranium, incubated at 37°C until the growth was visible. Selection of binding uranium bacteria was carried out based on their ability to grow on liquid medium containing various concentration of uranium that shacked on 120 rpm speed. The isolated bacteria with the highest specific growth rate constant (μ) were selected for biochemical characterization and identification by matching profile method. The result of this research showed that three selected isolate bacteria were able to grow well on liquid SBS medium until 100 mg/l uranium concentration. The identification results showed that two of them were suspected belong to the genus Pseudomonas and one isolates belong to the genus of Bacillus. The uranium reduction studied was performed by growing up the isolated bacteria on the SBS liquid medium that containing 40 mg/l uranium. Bacterial growth were measured by weighted of bacterial biomass and uranium concentration were measured by spectrophotometer. The research result showed that the selected isolates bacteria may applicable for bioremediation agent because of their ability to grow well on liquid SBS medium and their ability on uranium concentration reduction. The efficiency of reduction by Pseudomonas in the isolated bacteria one were 78.51 % and in the isolated bacteria three were 91.47 % , and Bacillus in the isolate bacteria six were 52.73%. (author)

  11. Isolation of gallic acid-producing microorganisms and their use in ...

    African Journals Online (AJOL)

    A total number of eighty gallic acid producing strains were isolated from forest soil or plant samples. Among these strains, thirteen isolates were selected for gallic acid production and these isolates were Aspergillus niger 1, A. niger 2, A. niger 3, Penicillium canescens (3), P. frequentans (2), P. spinulosum (2), ...

  12. WIPP: Waste Isolation Pilot Plant

    International Nuclear Information System (INIS)

    1984-01-01

    The following aspects of the Waste Isolation Pilot Plant are discussed briefly: history and site selection; salt as a disposal medium; transporting waste materials; early key events; impacts on New Mexico; project organization; and site certification profile

  13. O-naphthoquinone isolated from Capraria biflora L. induces selective cytotoxicity in tumor cell lines.

    Science.gov (United States)

    de S Wisintainer, G G N; Scola, G; Moura, S; Lemos, T L G; Pessoa, C; de Moraes, M O; Souza, L G S; Roesch-Ely, M; Henriques, J A P

    2015-12-21

    Biflorin is an o-naphthoquinone isolated from the roots of the plant Capraria biflora L. (Scrophulariaceae). In this study, the cytotoxic effects of biflorin were verified, and late apoptosis was detected in various cancer cell lines by in situ analysis. The cytotoxicity was further evaluated exclusively for 48 h of treatment in different tumor and non-tumor cell lines (Hep-2, HeLa, HT-29, A-375, and A-549, and HEK-293, respectively). The results indicated that biflorin induced selective cytotoxicity in tumor cells. HeLa cells were more susceptible to biflorin, followed by HT-29, A-549, A-375, and Hep-2 at all concentrations (range 5-50 μg/mL), and the highest half-maximal inhibitory concentration IC50 (56.01 ± 1.17 μg/mL) was observed in HEK-293 cells. Late apoptotic/necrotic events, observed by in situ immunostaining with Annexin V, varied with each cell line; an increase in late apoptotic events was observed corresponding to the increase in biflorin dosage. Hep-2 cells showed a greater percentage of late apoptotic events among the tumor cell lines when treated with higher concentrations of biflorin (69.63 ± 2.28%). The non-tumor HEK-293 line showed greater resistance to late apoptotic events, as well as a lower level of cytotoxicity (77.69 ± 6.68%) than the tested tumor lines. The data presented indicate that biflorin showed an important, possibly selective, cytotoxicity against tumor cell lines, thereby revealing a promising novel substance with potential anticancer activity for tumor therapy.

  14. Mass-selective isolation of ions stored in a quadrupole ion trap. A simulation study

    Science.gov (United States)

    March, Raymond E.; Londry, Frank A.; Alfred, Roland L.; Franklin, Anthony M.; Todd, John F. J.

    1992-01-01

    Trajectories of single ions stored in the quadrupole ion trap have been calculated using a simulation program described as the specific program for quadrupolar resonance (SPQR). Previously, the program has been used for the investigation of quadrupolar resonance excitation of ions with a static working point (or co-ordinates) in the stability diagram. The program has been modified to accommodate continuous d.c. and/or r.f. voltage ramps so as to permit calculation of ion trajectories while the working point is being changed. The modified program has been applied to the calculation of ion trajectories during ion isolation, or mass-selective storage, in the ion trap. The quadrupolar resonance excitation aspect of SPQR was not used in this study. Trajectories are displayed as temporal variations of ion kinetic energy, and axial and radial excursions from the centre of the ion trap. The working points of three ion species (m/z 144, 146 and 148), located initially on the qz, axis with qz [approximate] 0.12, were moved to the vicinity of the upper apex by a combination of r.f. and d.c. voltages applied in succession. Stable trajectories were maintained only for the ion species of m/z 146 for which the working point lay within this apex; the other ion species were ejected either radially or axially. The d.c. voltage was then reduced to zero so as to restore the working point of the isolated ion species to the qz axis. The amplitude of the r.f voltage was reduced to its initial value so as to retrieve the initial working point for m/z 146. The process extended over a real time of 2.9 ms, and was collision-free. The trajectory of the isolated ion was stable during this process; the ion species with m/z value lower than that of the target ion, that is, m/z 144, was ejected axially at the [beta]z = 1 boundary, while that with higher m/z value, that is, m/z 148, was ejected radially at the [beta]r = 0 boundary, as expected. The moderating effects of buffer gas were not taken

  15. Isolation of putative probionts from cod rearing environment

    DEFF Research Database (Denmark)

    Lauzon, H.L.; Gudmundsdottir, S.; Pedersen, M.H.

    2008-01-01

    , metabolite production and adhesion to fish cell lines. Our study demonstrated that 14% of screened bacteria (n = 188) had antagonistic properties towards fish pathogens. The majority of these isolates were Gram-positive (81%), belonging to Firmicutes (69.2%) and Actinobacteria (11.5%) phyla based on 16S r...... was designed to search for new probiotics to target this critical period in cod rearing. Potential probionts were selected from the naturalmicrobiota of cod aquacultural environment. The selection was based on several criteria: pathogen inhibition potential, growth characteristics, strain identification......RNA gene sequencing. Only 6 (3.2%) of 188 isolates could inhibit all three pathogens tested: Vibrio anguillarum, Aeromonas salmonicida subsp. achromogenes and Vibrio salmonicida. Differences observed in activity intensity and spectrum among inhibitory isolates emphasise the need to develop probiotic...

  16. Selected, annotated bibliography of studies relevant to the isolation of nuclear wastes

    International Nuclear Information System (INIS)

    Hyder, L.K.; Fore, C.S.; Vaughan, N.D.; Faust, R.A.

    1980-09-01

    This annotated bibliography of 705 references represents the first in a series to be published by the Ecological Sciences Information Center containing scientific, technical, economic, and regulatory information relevant to nuclear waste isolation. Most references discuss deep geologic disposal, with fewer studies of deep seabed disposal; space disposal is also included. The publication covers both domestic and foreign literature for the period 1954 to 1980. Major chapters selected are Chemical and Physical Aspects; Container Design and Performance; Disposal Site; Envirnmental Transport; General Studies and Reviews; Geology, Hydrology and Site Resources; Regulatory and Economic Aspects; Repository Design and Engineering; Transportation Technology; Waste Production; and Waste Treatment. Specialized data fields have been incorporated to improve the ease and accuracy of locating pertinent references. Specific radionuclides for which data are presented are listed in the Measured Radionuclides field, and specific parameters which affect the migration of these radionuclides are presented in the Measured Parameters field. The references within each chapter are arranged alphabetically by leading author, corporate affiliation, or title of the document. When the author is not given, the corporate affiliation appears first. If these two levels of authorship are not given, the title of the document is used as the identifying level. Indexes are provided for author(s), keywords, subject category, title, geographic location, measured parameters, measured radionuclides, and publication description

  17. Diagnostic properties of three conventional selective plating media for selection of Bacillus cereus, B. thuringiensis and B. weihenstephanensis

    DEFF Research Database (Denmark)

    Hendriksen, Niels Bohse; Hansen, Bjarne Munk

    2011-01-01

    The aim of this study was to assess the diagnostic properties of the two selective plating media and a chromogenic medium for identification of Bacillus cereus. The 324 isolates were B. cereus (37%), Bacillus weihenstephanensis (45%) or Bacillus thuringiensis (18%), as identified by a new...... combination of techniques. All isolates were growing on mannitol–egg yolk–polymyxin agar (MYP), and they did not form acid from mannitol. However, a significant lower number of B. thuringiensis isolates did not show lecithinase activity. All isolates were also growing on polymyxin–egg yolk...... recommended selective plating media MYP and PEMBA for detection of B. cereus group bacteria both have their limitations for identification of some B. cereus, B. weihenstephanensis or B. thuringiensis. However, MYP is preferable compared to PEMBA. The chromogenic medium has its own advantages and limitations...

  18. Isolation and characterization of a cellulolytic actinomycete Microbispora bispora

    Energy Technology Data Exchange (ETDEWEB)

    Waldron, Jr, C R; Becker-Vallone, C A; Eveleigh, D E

    1986-09-01

    Protocols for the isolation of cellulolytic actinomycetes are described, and their use illustrated in the selection of thermophilic bacteria from soil. One isolate, Microbispora bispora, was selected for further study. It grew readily at 55/sup 0/C, produced an extracellular cellulase in good yield (endoglucanase, 5.9 U/ml) that had a broad pH range (pH 5.5 - 7.2) and was thermally stable. Its aryl-..beta..-glucosidase was cell-associated and was relatively resistant to end-product inhibition.

  19. Isolation and identification of radiation resistant yeasts from sea water

    International Nuclear Information System (INIS)

    Park, Jong Cheon; Jeong, Yong Uk; Kim, Du Hong; Jo, Eun A

    2011-12-01

    This study was conducted to isolate radiation-resistant yeasts from sea water for development of application technology of radiation-resistant microorganism. · Isolation of 656 yeasts from sea water and selection of 2 radiation-resistant yeasts (D 10 value >3) · Identification of isolated yeasts as Filobasidium elegans sharing 99% sequence similarity · Characterization of isolated yeast with ability to repair of the DNA damage and membrane integrity to irradiation

  20. Sequential elution from IMAC (SIMAC)

    DEFF Research Database (Denmark)

    Thingholm, Tine E.; Larsen, Martin R.

    2016-01-01

    Phosphoproteomics relies on methods for efficient purification and sequencing of phosphopeptides from highly complex biological systems, especially when using low amounts of starting material. Current methods for phosphopeptide enrichment, e.g., Immobilized Metal ion Affinity Chromatography and t...

  1. Enrichment and separation of mono- and multiply phosphorylated peptides using sequential elution from IMAC prior to mass spectrometric analysis

    DEFF Research Database (Denmark)

    Thingholm, Tine E; Jensen, Ole N; Larsen, Martin R

    2009-01-01

    Phospho-proteomics relies on methods for efficient purification and sequencing of phosphopeptides from highly complex biological systems using low amounts of starting material. Current methods for phosphopeptide enrichment, e.g., immobilized metal affinity chromatography and titanium dioxide chro...

  2. Effect of isolates of fibre degrading bacteria on body weight changes ...

    African Journals Online (AJOL)

    tc

    2013-05-22

    May 22, 2013 ... evaluate the effect of supplementation of fibrolytic bac- terial isolate on body weight changes, milk production, nutrient intake and nutrient utilization in lactating Murrah buffaloes. This study was conducted to evaluate the digestibility of lignocellulosic feeds by some selected bacterial isolates isolated from ...

  3. Identification at Biovar Level of Brucella Isolates Causing Abortion in Small Ruminants of Iran

    Directory of Open Access Journals (Sweden)

    Ali Mohammad Behroozikhah

    2012-01-01

    Full Text Available To determine the most prevalent biovar responsible for brucellosis in sheep and goat populations of Iran, a cross-sectional study was carried out over 2 years in six provinces selected based on geography and disease prevalence. Specimens obtained from referred aborted sheep and goat fetuses were cultured on Brucella selective media for microbiological isolation. Brucellae were isolated from 265 fetuses and examined for biovar identification using standard microbiological methods. Results showed that 246 isolates (92.8% were B. melitensis biovar 1, 18 isolates (6.8% were B. melitensis biovar 2, and, interestingly, one isolate (0.4% obtained from Mazandaran province was B. abortus biovar 3. In this study, B. melitensis biovar 3 was isolated in none of the selected provinces, and all isolates from 3 provinces (i.e., Chehar-mahal Bakhtiari, Markazi, and Ilam were identified only as B. melitensis biovar 1. In conclusion, we found that B. melitensis biovar 1 remains the most prevalent cause of small ruminant brucellosis in various provinces of Iran.

  4. Quantitative phosphoproteomics applied to the yeast pheromone signaling pathway

    DEFF Research Database (Denmark)

    Gruhler, Albrecht; Olsen, Jesper Velgaard; Mohammed, Shabaz

    2005-01-01

    of a detailed molecular view of complex biological processes. We present a quantitative modification-specific proteomic approach that combines stable isotope labeling by amino acids in cell culture (SILAC) for quantitation with IMAC for phosphopeptide enrichment and three stages of mass spectrometry (MS....... Phosphopeptide fractions were analyzed by LC-MS using a linear ion trap-Fourier transform ion cyclotron resonance mass spectrometer. MS/MS and neutral loss-directed MS/MS/MS analysis allowed detection and sequencing of phosphopeptides with exceptional accuracy and specificity. Of more than 700 identified...

  5. Isolation, selection and evaluation of yeasts for use in fermentation of coffee beans by the wet process.

    Science.gov (United States)

    de Melo Pereira, Gilberto Vinícius; Soccol, Vanete Thomaz; Pandey, Ashok; Medeiros, Adriane Bianchi Pedroni; Andrade Lara, João Marcos Rodrigues; Gollo, André Luiz; Soccol, Carlos Ricardo

    2014-10-01

    During wet processing of coffee, the ripe cherries are pulped, then fermented and dried. This study reports an experimental approach for target identification and selection of indigenous coffee yeasts and their potential use as starter cultures during the fermentation step of wet processing. A total of 144 yeast isolates originating from spontaneously fermenting coffee beans were identified by molecular approaches and screened for their capacity to grow under coffee-associated stress conditions. According to ITS-rRNA gene sequencing, Pichia fermentans and Pichia kluyveri were the most frequent isolates, followed by Candida Candida glabrata, quercitrusa, Saccharomyces sp., Pichia guilliermondii, Pichia caribbica and Hanseniaspora opuntiae. Nine stress-tolerant yeast strains were evaluated for their ability to produce aromatic compounds in a coffee pulp simulation medium and for their pectinolytic activity. P. fermentans YC5.2 produced the highest concentrations of flavor-active ester compounds (viz., ethyl acetate and isoamyl acetate), while Saccharomyces sp. YC9.15 was the best pectinase-producing strain. The potential impact of these selected yeast strains to promote flavor development in coffee beverages was investigated for inoculating coffee beans during wet fermentation trials at laboratory scale. Inoculation of a single culture of P. fermentans YC5.2 and co-culture of P. fermentans YC5.2 and Saccharomyces sp. YC9.15 enhanced significantly the formation of volatile aroma compounds during the fermentation process compared to un-inoculated control. The sensory analysis indicated that the flavor of coffee beverages was influenced by the starter cultures, being rated as having the higher sensory scores for fruity, buttery and fermented aroma. This demonstrates a complementary role of yeasts associated with coffee quality through the synthesis of yeast-specific volatile constituents. The yeast strains P. fermentans YC5.2 and Saccharomyces sp. YC9.15 have a great

  6. Isolation, Identification, and Evaluation of Novel Probiotic Strains Isolated from Feces of Breast-Fed Infants.

    Science.gov (United States)

    Panya, Marutpong; Lulitanond, Viraphong; Rattanachaikunsopon, Pongsak; Srivoramas, Thanyakarn; Chaiwong, Tarinee

    2016-01-01

    To isolate, identify, and evaluate the probiotic properties of lactic acid bacteria (LAB) isolated from the feces of breast-fed infants. The probiotic tests included investigation of hemolysis activity, survival in simulated gastrointestinal tract conditions (acid and bile salt tolerance), susceptibility to antibiotics, and ability to inhibit selected bacterial pathogens (Escherichia coli O157:H7, Vibrio cholerae and Salmonella enterica subsp enterica serovar Typhimurium). The bacterial species identification was performed by both carbohydrate utilization and partial 16S ribosomal RNA sequencing. Five of fifty LAB isolates (UBU-03, UBU-06, UBU-09, UBU-34, and UBU-37) showed good probiotic properties. These five isolates showed non-hemolysis type (gamma-hemolysis), susceptibility to all antibiotics tested except for vancomycin, ability to survive in the simulated gastrointestinal conditions of both acid and bile salt solution, and ability to inhibit growth of E. coli O157: H7 and V. cholerae. Bacterial species identification revealed that all five isolates were firmly identified as Lactobacillus rhamnosus species. The L. rhamnosus strains that were isolated and characterized in this study could be considered as probiotic strains, and then used for further probiotic characterization in human cell cultures or animal models.

  7. Feather wastes digestion by new isolated strains Bacillus sp. in ...

    African Journals Online (AJOL)

    Feather wastes digestion by new isolated strains Bacillus sp. in Morocco. ... The most efficient isolated strain selected was compared with Bacillus subtilis ATCC 6633. Results showed ... African Journal of Biotechnology Vol.3(1) 2004: 67-70 ...

  8. Radioactive waste isolation in salt: peer review of Office of Nuclear Waste Isolation's Socioeconomic Program Plan

    International Nuclear Information System (INIS)

    Winter, R.; Fenster, D.; O'Hare, M.; Zillman, D.; Harrison, W.; Tisue, M.

    1984-07-01

    The following recommendations have been abstracted from the body of this report. The Office of Nuclear Waste Isolation's Socioeconomic Program Plan for the Establishment of Mined Geologic Repositories to Isolate Nuclear Waste should be modified to: (1) encourage active public participation in the decision-making processes leading to repository site selection; (2) clearly define mechanisms for incorporating the concerns of local residents, state and local governments, and other potentially interested parties into the early stages of the site selection process. In addition, the Office of Nuclear Waste Isolation should carefully review the overall role that these persons and groups, including local pressure groups organized in the face of potential repository development, will play in the siting process; (3) place significantly greater emphasis on using primary socioeconomic data during the site selection process, reversing the current overemphasis on secondary data collection, description of socioeconomic conditions at potential locations, and development of analytical methodologies; (4) include additional approaches to solving socioeconomic problems. For example, a reluctance to acknowledge that solutions to socioeconomic problems need to be found jointly with interested parties is evident in the plan; (5) recognize that mitigation mechanisms other than compensation and incentives may be effective; (6) as soon as potential sites are identified, the US Department of Energy (DOE) should begin discussing impact mitigation agreements with local officials and other interested parties; and (7) comply fully with the pertinent provisions of NWPA

  9. Isolation of thermotolerant, halotolerant, facultative biosurfactant-producing bacteria.

    Science.gov (United States)

    Ghojavand, H; Vahabzadeh, F; Mehranian, M; Radmehr, M; Shahraki, Kh A; Zolfagharian, F; Emadi, M A; Roayaei, E

    2008-10-01

    Several facultative bacterial strains tolerant to high temperature and salinity were isolated from the oil reservoir brines of an Iranian oil field (Masjed-I Soleyman). Some of these isolates were able to grow up to 60 degrees C and at high concentration of NaCl (15% w/v). One of the isolates grew at 40 degrees C, while it was able to grow at 15% w/v NaCl. Tolerances to NaCl levels decreased as the growth temperatures were increased. Surfactant production ability was detected in some of these isolates. The use of biosurfactant is considered as an effective mechanism in microbial-enhanced oil recovery processes detected in some of these isolates. The surfactant producers were able to grow at high temperatures and salinities to about 55 degrees C and 10% w/v, respectively. These isolates exhibited morphological and physiological characteristics of the Bacillus genus. The partial sequencing of the 16S ribosomal deoxyribonucleic acid gene of the selected isolates was assigned them to Bacillus subtilis group. The biosurfactant produced by these isolates caused a substantial decrease in the surface tension of the culture media to 26.7 mN/m. By the use of thin-layer chromatography technique, the presence of the three compounds was detected in the tested biosurfactant. Infrared spectroscopy and (1)H nuclear magnetic resonance analysis were used, and the partial structural characterization of the biosurfactant mixture of the three compounds was found to be lipopeptidic in nature. The possibility of use of the selected bacterial strains reported, in the present study, in different sectors of the petroleum industry has been addressed.

  10. COMPARATIVE EVALUATION OF CULTURE MEDIA FOR PATHOGEN ISOLATION OF PURULENT BACTERIAL MENINGITIS

    Directory of Open Access Journals (Sweden)

    Ya. V. Podkopaev

    2016-01-01

    Full Text Available The State Research Center for Applied Microbiology and Biotechnology has designed two nutrient media — chocolate agar and PBM-agar to isolate pathogens of purulent bacterial meningitis (PBM. In our previous research using collected microbial strains the media were shown to be highly susceptible and to provide the growth of Neisseria meningiti-dis, Streptococcus pneumoniae and Haemophilus influenzae strains, when inoculated with microbial suspensions containing single cells. When isolating Haemophilus influenzae, meningococci, and pneumococci the use of selective additives in both media assures selective isolation of required microorganisms, inhibiting contaminants. The objective of this research was to assess the media in bacteriological tests of clinical samples collected from the upper and lower respiratory tract in humans. The bacteriological plating of throat smear specimens (n = 90 from children and adults at the age of 0 to 66 with disorder of the upper respiratory tract on chocolate agar, PBM-agar and on a control medium in the absence of selective additives resulted in the equal amount of microbial cultures isolated. Of 154 isolated cultures 2, 23 and 9 were attributed to Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae, respectively. The plating of throat smears (n = 10 from healthy people at the age of 30 to 55 on the analyzable and control media in the presence of additives allowed us to selectively isolate Haemophilus influenzae and Streptococcus pneumoniae cultures without a quantitative loss, with contaminants inhibited. By their growth characteristics chocolate agar and PBM-agar were highly competitive with reference media being used in clinical practice for isolating main causative agents of purulent bacterial meningitis.

  11. Using selective chromogenic plates to optimize isolation of group B Streptococcus in pregnant women

    Directory of Open Access Journals (Sweden)

    Romano Mattei

    2014-03-01

    Full Text Available Group B Streptococcus (GBS remains the leading cause of severe bacterial infections (sepsis, meningitis, pneumonia in neonates. We compared the detection of GBS from recto-vaginal swabs on blood agar and two chromogenic media and evaluated their antibiotic susceptibility. A total of 1351 swabs were taken from pregnant women at 35-37 weeks of gestation. Following enrichment in Todd Hewitt broth + nalidixic acid and colistin, the samples were plated on Columbia CNA agar (CNA, chromID Strepto B agar (STRB and Granada Agar (GRAN, respectively. GBS were found in 22.4% of recto-vaginal swabs from pregnant women. Sensitivity, specificity, positive and negative predictive values of GBS detection were 88%, 88%, 81% and 96% for CNA, 99%, 97%, 90% and 99% for STRB and 94%, 99%, 98% e 99% for GRAN; Cohen’s k index concordances for CNA, STREB and GRAN were 0.68, 0.92 and 0.96, respectively. All isolates were susceptible to penicillin, whereas resistances of erythromycin and clindamycin were 40% and 42%, respectively. To conclude, selective broth enrichment combined with chromogenic plates is recommended for GBS screening in pregnant women.

  12. Probiotic features of two oral Lactobacillus isolates

    OpenAIRE

    Zavisic, Gordana; Petricevic, Sasa; Radulovic, Zeljka; Begovic, Jelena; Golic, Natasa; Topisirovic, Ljubisa; Strahinic, Ivana

    2012-01-01

    In this study, we checked lactobacilli strains of human origin for their potential as probiotic. Samples were collected from oral mucosa of 16 healthy individuals, out of which twenty isolates were obtained and two of them were selected and identified as Lactobacillus plantarum (G1) and L. casei (G3). Both isolates exhibited antagonistic action towards pathogenic microorganisms such as Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Salmonella abony, and Clostridium sporogene...

  13. Enrichment and desalting of tryptic protein digests and the protein depletion using boron nitride

    Energy Technology Data Exchange (ETDEWEB)

    Fischnaller, Martin; Köck, Rainer; Bakry, Rania, E-mail: rania.bakry@uibk.ac.at; Bonn, Günther K.

    2014-05-01

    Highlights: • Protein tryptic digests were desalted and enriched utilizing hexagonal boron nitride. • Phosphopeptides were desalted with high recovery rates. • Boron nitride exhibits high wettability allowing fast sample preparation. • Boron nitride shows protein depletion capability applied for peptide purification. - Abstract: Sample preparation still remains a great challenge in modern bioanalysis and the interest in new efficient solid phase extraction (SPE) materials still remains high. In this work, hexagonal boron nitride (h-BN) is introduced as a new SPE material for the isolation and enrichment of peptides. The h-BN is isoelectronic and structurally similar to graphite. It has remarkable properties including good thermal conductivity, excellent thermal and chemical stability and a better oxidation resistance than graphite. BN attracts increasing interest because of its wide range of applicability. In the present work, the great potential of h-BN, as a new SPE-material, on the enrichment, preconcentration and desalting of tryptic digest of model proteins is demonstrated. A special attention was dedicated to the efficient enrichment of hydrophilic phosphopeptides. Two elution protocols were developed for the enrichment of peptides compatible for subsequent MALDI-MS and ESI-MS analysis. In addition, the recoveries of 5 peptides and 3 phosphopeptides with wide range of pI values utilizing h-BN materials with different surface areas were investigated. 84–106% recovery rate could be achieved using h-BN materials. The results were compared with those obtained using graphite and silica C18 under the same elution conditions, and lower recoveries were obtained. In addition, h-BN was found to have a capability of protein depletion, which is requisite for the peptide profiling.

  14. Enrichment and desalting of tryptic protein digests and the protein depletion using boron nitride

    International Nuclear Information System (INIS)

    Fischnaller, Martin; Köck, Rainer; Bakry, Rania; Bonn, Günther K.

    2014-01-01

    Highlights: • Protein tryptic digests were desalted and enriched utilizing hexagonal boron nitride. • Phosphopeptides were desalted with high recovery rates. • Boron nitride exhibits high wettability allowing fast sample preparation. • Boron nitride shows protein depletion capability applied for peptide purification. - Abstract: Sample preparation still remains a great challenge in modern bioanalysis and the interest in new efficient solid phase extraction (SPE) materials still remains high. In this work, hexagonal boron nitride (h-BN) is introduced as a new SPE material for the isolation and enrichment of peptides. The h-BN is isoelectronic and structurally similar to graphite. It has remarkable properties including good thermal conductivity, excellent thermal and chemical stability and a better oxidation resistance than graphite. BN attracts increasing interest because of its wide range of applicability. In the present work, the great potential of h-BN, as a new SPE-material, on the enrichment, preconcentration and desalting of tryptic digest of model proteins is demonstrated. A special attention was dedicated to the efficient enrichment of hydrophilic phosphopeptides. Two elution protocols were developed for the enrichment of peptides compatible for subsequent MALDI-MS and ESI-MS analysis. In addition, the recoveries of 5 peptides and 3 phosphopeptides with wide range of pI values utilizing h-BN materials with different surface areas were investigated. 84–106% recovery rate could be achieved using h-BN materials. The results were compared with those obtained using graphite and silica C18 under the same elution conditions, and lower recoveries were obtained. In addition, h-BN was found to have a capability of protein depletion, which is requisite for the peptide profiling

  15. Heat resistance of thermoduric enterococci isolated from milk.

    Science.gov (United States)

    McAuley, Catherine M; Gobius, Kari S; Britz, Margaret L; Craven, Heather M

    2012-03-15

    Enterococci are reported to survive pasteurisation but the extent of their survival is unclear. Sixty-one thermoduric enterococci isolates were selected from laboratory pasteurised milk obtained from silos in six dairy factories. The isolates were screened to determine log(10) reductions incurred after pasteurisation (63°C/30 min) and ranked from highest to lowest log(10) reduction. Two isolates each of Enterococcus faecalis, Enterococcus faecium, Enterococcus durans and Enterococcus hirae, exhibiting the median and the greatest heat resistance, as well as E. faecalis ATCC 19433, were selected for further heat resistance determinations using an immersed coil apparatus. D values were calculated from survival curves plotted from viable counts obtained after heating isolates in Brain Heart Infusion Broth at 63, 69, 72, 75 and 78°C followed by rapid cooling. At 72°C, the temperature employed for High Temperature Short Time (HTST) pasteurisation (72°C/15s), the D values extended from 0.3 min to 5.1 min, depending on the isolate and species. These data were used to calculate z values, which ranged from 5.0 to 9.8°C. The most heat sensitive isolates were E. faecalis (z values 5.0, 5.7 and 7.5°C), while the most heat resistant isolates were E. durans (z values 8.7 and 8.8°C), E. faecium (z value 9.0°C) and E. hirae (z values 8.5 and 9.8°C). The data show that heat resistance in enterococci is highly variable. Copyright © 2011 Elsevier B.V. All rights reserved.

  16. Isolation and identification of a bacterium from marine shrimp digestive tract: A new degrader of starch and protein

    Science.gov (United States)

    Li, Jiqiu; Tan, Beiping; Mai, Kangsen

    2011-09-01

    It is a practical approach to select candidate probiotic bacterial stains on the basis of their special traits. Production of digestive enzyme was used as a trait to select a candidate probiotic bacterial strain in this study. In order to select a bacterium with the ability to degrade both starch and protein, an ideal bacterial strain STE was isolated from marine shrimp ( Litopenaeus vannamei) intestines by using multiple selective media. The selected isolate STE was identified on the basis of its morphological, physiological, and biochemical characteristics as well as molecular analyses. Results of degradation experiments confirmed the ability of the selected isolate to degrade both starch and casein. The isolate STE was aerobic, Gram-negative, rod-shaped, motile and non-spore-forming, and had catalase and oxidase activities but no glucose fermentation activity. Among the tested carbon/nitrogen sources, only Tween40, alanyl-glycine, aspartyl-glycine, and glycyl-l-glutamic acid were utilized by the isolate STE. Results of homology comparison analyses of the 16S rDNA sequences showed that the isolate STE had a high similarity to several Pseudoalteromonas species and, in the phylogenetic tree, grouped with P. ruthenica with maximum bootstrap support (100%). In conclusion, the isolate STE was characterized as a novel strain belonging to the genus Pseudoalteromonas. This study provides a further example of a probiotic bacterial strain with specific characteristics isolated from the host gastrointestinal tract.

  17. Fermentation of Apple Juice with a Selected Yeast Strain Isolated from the Fermented Foods of Himalayan Regions and Its Organoleptic Properties.

    Science.gov (United States)

    Kanwar, S S; Keshani

    2016-01-01

    Twenty-three Saccharomyces cerevisiae strains isolated from different fermented foods of Western Himalayas have been studied for strain level and functional diversity in our department. Among these 23 strains, 10 S. cerevisiae strains on the basis of variation in their brewing traits were selected to study their organoleptic effect at gene level by targeting ATF1 gene, which is responsible for ester synthesis during fermentation. Significant variation was observed in ATF1 gene sequences, suggesting differences in aroma and flavor of their brewing products. Apple is a predominant fruit in Himachal Pradesh and apple cider is one of the most popular drinks all around the world hence, it was chosen for sensory evaluation of six selected yeast strains. Organoleptic studies and sensory analysis suggested Sc21 and Sc01 as best indigenous strains for soft and hard cider, respectively, indicating their potential in enriching the local products with enhanced quality.

  18. New approach for isolation of VNTR markers.

    OpenAIRE

    Nakamura, Y; Carlson, M; Krapcho, K; Kanamori, M; White, R

    1988-01-01

    Elsewhere we have reported an efficient method for isolating VNTR (Variable Number of Tandem Repeats) markers. Several of the VNTR markers isolated in those experiments were sequenced, and a DNA sequence of 9 bp (GNNGTGGG) emerged as an apparent consensus sequence for VNTR markers. To confirm this result and to develop more VNTR markers, we synthesized nine different 18-base-long oligonucleotides whose sequences each included GNNGTGGG. When 102 cosmid clones selected by these oligonucleotides...

  19. Isolation and characterization of two malathiondegrading ...

    African Journals Online (AJOL)

    Pseudomonas aeruginosa and Pseudomonas mendocina degrading malathion were studied. Morphological, biochemical and 16S rRNA genes for bacterial identification were selected. Biodegradation of some organophosphorus compounds with the 2 bacterial isolates was determined by high performance liquid ...

  20. Search for unknown isotopes using the TIARA-ISOL and the JAERI-ISOL

    Energy Technology Data Exchange (ETDEWEB)

    Asai, Masato [Nagoya Univ. (Japan). School of Engineering

    1997-07-01

    The new neutron-deficient isotopes {sup 127}Pr and {sup 125}Pr, and the new neutron-rich isotopes {sup 166}Tb, {sup 165}Gd and {sup 161}Sm were identified using the TIARA-ISOL with {sup 36}Ar + {sup 92,94}Mo reactions, and the gas-jet coupled JAERI-ISOL with a proton-induced fission of {sup 238}U, respectively. The element-selective oxidation technique was used to reduce large contamination from isobars and molecular ions with production cross sections about one or two orders of magnitude as large as those of the new isotopes. The good signal-to-noise ratios achieved in the present measurements were essential to observe and identify weak X/{gamma} rays from the new isotopes. (author)

  1. Selection, isolation and growth kinetic study of a bacterial consortium obtained from the Potengi mangrove in the presence of crude oil

    Energy Technology Data Exchange (ETDEWEB)

    Costa, C.C.; Vaz, M.R.F.; Santos, E.S.; Macedo, G.R. [Universidade Federal do Rio Grande do Norte (UFRN), Natal, RN (Brazil). Dept. de Engenharia Quimica], E-mail: natcintia@gmail.com; Costa, J.G. da [Universidade Federal do Amazonas (UFAM), Coari, AM (Brazil). Inst. de Saude e Biotecnologia

    2011-10-15

    The selection, isolation and kinetic study of a bacterial consortium obtained from a sample of soil from the Potengi mangrove, located in the city of Natal, Rio Grande do Norte, Brazil, has been carried out using the enrichment culture technique to observe aspects such as the evaluation of main growth parameters. The kinetic study used a rotary incubator shaker at 150rpm, under 30 deg C. The bacterial consortium isolated from the estuary of the Potengi River showed a good acclimation in minimum mineral medium with 1% (v/v) of oil. The cell concentration reached 2.55 g/L at 16h of cultivation and surface tension dropped. The maximum productivity in cells obtained was of 0.3 g/L.h, the specific velocity of growth was of 0.075h{sup -1}, with a generation time (tg) of 9.24h. This study seeks to demonstrate that the consortium can be used as inoculants in biological treatments, capable of reducing the waste's degradation time. (author)

  2. Poly-γ-Glutamic Acids Contribute to Biofilm Formation and Plant Root Colonization in Selected Environmental Isolates of Bacillus subtilis

    Science.gov (United States)

    Yu, Yiyang; Yan, Fang; Chen, Yun; Jin, Christopher; Guo, Jian-Hua; Chai, Yunrong

    2016-01-01

    Bacillus subtilis is long known to produce poly-γ-glutamic acids (γ-PGA) as one of the major secreted polymeric substances. In B. subtilis, the regulation of γ-PGA production and its physiological role are still unclear. B. subtilis is also capable of forming structurally complex multicellular communities, or biofilms, in which an extracellular matrix consisting of secreted proteins and polysaccharides holds individual cells together. Biofilms were shown to facilitate B. subtilis–plant interactions. In this study, we show that different environmental isolates of B. subtilis, all capable of forming biofilms, vary significantly in γ-PGA production. This is possibly due to differential regulation of γ-PGA biosynthesis genes. In many of those environmental isolates, γ-PGA seems to contribute to robustness and complex morphology of the colony biofilms, suggesting a role of γ-PGA in biofilm formation. Our evidence further shows that in selected B. subtilis strains, γ-PGA also plays a role in root colonization by the bacteria, pinpointing a possible function of γ-PGA in B. subtilis–plant interactions. Finally, we found that several pathways co-regulate both γ-PGA biosynthesis genes and genes for the biofilm matrix in B. subtilis, but in an opposing fashion. We discussed potential biological significance of that. PMID:27891125

  3. Biodegradation of propargite by Pseudomonas putida, isolated from tea rhizosphere.

    Science.gov (United States)

    Sarkar, Soumik; Seenivasan, Subbiah; Asir, Robert Premkumar Samuel

    2010-02-15

    Biodegradation of miticide propargite was carried out in vitro by selected Pseudomonas strains isolated from tea rhizosphere. A total number of 13 strains were isolated and further screened based on their tolerance level to different concentrations of propargite. Five best strains were selected and further tested for their nutritional requirements. Among the different carbon sources tested glucose exhibited the highest growth promoting capacity and among nitrogen sources ammonium nitrate supported the growth to the maximum. The five selected Pseudomonas strain exhibited a range of degradation capabilities. Mineral salts medium (MSM) amended with glucose provided better environment for degradation with the highest degradation potential in strain SPR 13 followed by SPR 8 (71.9% and 69.0% respectively).

  4. Casein phosphopeptides and CaCl2 increase penicillin production and cause an increment in microbody/peroxisome proteins in Penicillium chrysogenum.

    Science.gov (United States)

    Domínguez-Santos, Rebeca; Kosalková, Katarina; García-Estrada, Carlos; Barreiro, Carlos; Ibáñez, Ana; Morales, Alejandro; Martín, Juan-Francisco

    2017-03-06

    Transport of penicillin intermediates and penicillin secretion are still poorly characterized in Penicillium chrysogenum (re-identified as Penicillium rubens). Calcium (Ca 2+ ) plays an important role in the metabolism of filamentous fungi, and casein phosphopeptides (CPP) are involved in Ca 2+ internalization. In this study we observe that the effect of CaCl 2 and CPP is additive and promotes an increase in penicillin production of up to 10-12 fold. Combination of CaCl 2 and CPP greatly promotes expression of the three penicillin biosynthetic genes. Comparative proteomic analysis by 2D-DIGE, identified 39 proteins differentially represented in P. chrysogenum Wisconsin 54-1255 after CPP/CaCl 2 addition. The most interesting group of overrepresented proteins were a peroxisomal catalase, three proteins of the methylcitrate cycle, two aminotransferases and cystationine β-synthase, which are directly or indirectly related to the formation of penicillin amino acid precursors. Importantly, two of the enzymes of the penicillin pathway (isopenicillin N synthase and isopenicillin N acyltransferase) are clearly induced after CPP/CaCl 2 addition. Most of these overrepresented proteins are either authentic peroxisomal proteins or microbody-associated proteins. This evidence suggests that addition of CPP/CaCl 2 promotes the formation of penicillin precursors and the penicillin biosynthetic enzymes in peroxisomes and vesicles, which may be involved in transport and secretion of penicillin. Penicillin biosynthesis in Penicillium chrysogenum is one of the best characterized secondary metabolism processes. However, the mechanism by which penicillin is secreted still remains to be elucidated. Taking into account the role played by Ca 2+ and CPP in the secretory pathway and considering the positive effect that Ca 2+ exerts on penicillin production, the analysis of global protein changes produced after CPP/CaCl 2 addition is very helpful to decipher the processes related to the

  5. Characterization of Petroleum Hydrocarbon Decomposing Fungi Isolated from Mangrove Rhizosphere

    Directory of Open Access Journals (Sweden)

    Nuni Gofar

    2011-01-01

    Full Text Available The research was done to obtain the isolates of soil borne fungi isolated from mangrove rhizosphere which were capable of degrading petroleum hydrocarbon compounds. The soil samples were collected from South Sumatra mangrove forest which was contaminated by petroleum. The isolates obtained were selected based on their ability to survive, to grow and to degrade polycyclic aromatic hydrocarbons in medium containing petroleum residue. There were 3 isolates of soil borne hydrocarbonoclastic fungi which were able to degrade petroleum in vitro. The 3 isolates were identified as Aspergillus fumigates, A. parasiticus, and Chrysonilia sitophila. C. sitophila was the best isolate to decrease total petroleum hydrocarbon (TPH from medium containing 5-20% petroleum residue.

  6. Prevalence and antibiogram profiles of Escherichia coli O157:H7 isolates recovered from three selected dairy farms in the Eastern Cape Province, South Africa

    Directory of Open Access Journals (Sweden)

    Luyanda Msolo

    2016-12-01

    Full Text Available Objective: To investigate the occurrence and antibiotics susceptibility of Escherichia coli (E. coli O157:H7 isolates from raw milk, cattle udder, milking machines and worker’s hand swabs from three selected commercial dairy farms in the Amathole District Municipality, Eastern Cape Province, South Africa. Methods: Raw milk samples were collected from bulk storage tanks and swab samples were collected from milking machines, cattle udders and worker’s hands fortnightly over a sixmonth sampling regime between June and November 2014. A standard culture-based method was used for the enumeration and isolation of E. coli O157:H7, presumptive identification using sorbitol MacConkey agar (supplemented with cefixime (50 µg/L and potassium tellurite (25 mg/L. A serological confirmation of the presumptive E. coli O157:H7 isolates was conducted using the O157 latex agglutination test kit. Results: A total of 252 E. coli O157:H7 isolates were further subjected to PCR amplification of rfbEO157 and flCH7 genes of which 27(11% of the isolates were confirmed positive E. coli O157:H7. The percentage antibiotic resistance of the 27 E. coli O157:H7 isolates from the dairy farms revealed penicillin [23 (85%], tetracycline [22 (81%], erythromycin [19 (70%], streptomycin [14 (52%] and chloramphenicol [12 (45%]. The highest resistances were penicillin [23 (85%] and tetracycline [22 (81%]. Conclusions: These findings revealed that the dairy farms are potential reservoirs of E. coli O157:H7 serotype, and harbor antibiotic-resistant determinants, a concern to public and environmental health.

  7. Ohcratoxin producing Aspergillus spp. Isolated from tropical soils in Sarawak, Malaysia

    Directory of Open Access Journals (Sweden)

    J.S.S. Seelan

    2010-03-01

    Full Text Available Aspergillus strains isolated from tropical soils were selected for additional characterization and for ochratoxin analysis, which was determined by ELISA method and high performance liquid chromatography (HPLC profiles. Because of its great morphological variability and mycotoxin production availability, 18 isolates of Aspergillus species were selected for this study. Only two isolates of these tropical soils, A. sulphureus and A. carbonarius, showed positive results for ohcratoxin (OA in lower concentration (0.05-0.10 µg/ml. Ochratoxin production by these species was confirmed by high performance liquid chromatography. HPLC analysis for ochratoxin producing A. sulphureus and A. carbonarius showed retention time, Rt value = 4.417 and Rt value = 4.081 respectively.

  8. Effectively designed molecularly imprinted polymers for selective isolation of the antidiabetic drug metformin and its transformation product guanylurea from aqueous media

    Energy Technology Data Exchange (ETDEWEB)

    Kyzas, George Z.; Nanaki, Stavroula G. [Laboratory of Polymer Chemistry and Technology, Department of Chemistry, Aristotle University of Thessaloniki, GR-541 24 Thessaloniki (Greece); Koltsakidou, Anastasia; Papageorgiou, Myrsini; Kechagia, Maria [Laboratory of Environmental Pollution Control, Department of Chemistry, Aristotle University of Thessaloniki, GR-541 24 Thessaloniki (Greece); Bikiaris, Dimitrios N. [Laboratory of Polymer Chemistry and Technology, Department of Chemistry, Aristotle University of Thessaloniki, GR-541 24 Thessaloniki (Greece); Lambropoulou, Dimitra A., E-mail: dlambro@chem.auth.gr [Laboratory of Environmental Pollution Control, Department of Chemistry, Aristotle University of Thessaloniki, GR-541 24 Thessaloniki (Greece)

    2015-03-25

    Highlights: • Preparation of novel MIPs with remarkable recognition properties. • Selective isolation and removal of metformin and guanylurea from aqueous media. • Detailed adsorbent characterization and adsorption studies. • Successful application of synthesized MIPs as dispersive SPE sorbents. • Estimation of expanded uncertainty following a bottom-up approach. - Abstract: In the present study, two novel molecularly imprinted polymers (MIPs) with remarkable recognition properties for metformin and its transformation product, guanylurea, have been prepared for their selective, enrichment, isolation and removal from aqueous media. The prepared adsorbents were characterized by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and swelling experiments. The performance of the prepared MIPs was evaluated by various parameters including the influence of pH, contact time, temperature and initial compound concentration. The effects on the adsorption behavior of the removal process parameters were studied and the equilibrium data were fitted by the Langmuir and Freundlich models. Due to the imprinting effect, adsorption performance of MIPs was always superior to its corresponding NIP (non-imprinted polymer), with maximum adsorption capacity ∼80 mg g{sup −1} for both MIPs. Stability and reusability of the MIPs up to the 5th cycle meant that they could be applied repeatedly without losing substantial removal ability. In the next step, the prepared MIP nanoparticles were evaluated as sorbents in a dispersive solid phase extraction (D-SPE) configuration for selective enrichment and determination of metformin and guanylurea in different aqueous matrices. Under the working extraction conditions, the D-SPE method showed good linearity in the range of 50–1000 ng L{sup −1}, repeatability of the extractions (RSD 2.1–5.1%, n = 3), and low limits of detection (1.5–3.4 ng L{sup −1}). The expanded uncertainty of the data obtained

  9. cDNA fingerprinting of osteoprogenitor cells to isolate differentiation stage-specific genes.

    OpenAIRE

    Candeliere, G A; Rao, Y; Floh, A; Sandler, S D; Aubin, J E

    1999-01-01

    A cDNA fingerprinting strategy was developed to identify genes based on their differential expression pattern during osteoblast development. Preliminary biological and molecular staging of cDNA pools prepared by global amplification PCR allowed discrim-inating choices to be made in selection of expressed sequence tags (ESTs) to be isolated. Sequencing of selected ESTs confirmed that both known and novel genes can be isolated from any developmental stage of interest, e.g. from primitive progen...

  10. Isolation and characterization of mutated alcohol oxidases from the yeast Hansenula polymorpha with decreased affinity toward substrates and their use as selective elements of an amperometric biosensor

    Directory of Open Access Journals (Sweden)

    Schuhmann Wolfgang

    2007-06-01

    Full Text Available Abstract Background Accurate, rapid, and economic on-line analysis of ethanol is very desirable. However, available biosensors achieve saturation at very low ethanol concentrations and thus demand the time and labour consuming procedure of sample dilution. Results Hansenula polymorpha (Pichia angusta mutant strains resistant to allyl alcohol in methanol medium were selected. Such strains possessed decreased affinity of alcohol oxidase (AOX towards methanol: the KM values for AOX of wild type and mutant strains CA2 and CA4 are shown to be 0.62, 2.48 and 1.10 mM, respectively, whereas Vmax values are increased or remain unaffected. The mutant AOX alleles from H. polymorpha mutants CA2 and CA4 were isolated and sequenced. Several point mutations in the AOX gene, mostly different between the two mutant alleles, have been identified. Mutant AOX forms were isolated and purified, and some of their biochemical properties were studied. An amperometric biosensor based on the mutated form of AOX from the strain CA2 was constructed and revealed an extended linear response to the target analytes, ethanol and formaldehyde, as compared to the sensor based on the native AOX. Conclusion The described selection methodology opens up the possibility of isolating modified forms of AOX with further decreased affinity toward substrates without reduction of the maximal velocity of reaction. It can help in creation of improved ethanol biosensors with a prolonged linear response towards ethanol in real samples of wines, beers or fermentation liquids.

  11. IDENTIFICATION AND BIOLOGICAL ACTIVITY OF POTENTIAL PROBIOTIC BACTERIA ISOLATED FROM POECILIA RETICULATA (GUPPY

    Directory of Open Access Journals (Sweden)

    Aparna Balakrishna

    2014-08-01

    Full Text Available Antagonistic activities against candidate indicator strains, adhesion to mucus and biofilm formation of potential probiotic strains isolated from Poecilia reticulata were evaluated. Four isolated strains (MBTU_PB1, MBTU_PB2, MBTU_PB3 and MBTU_PB4 showed moderate to strong antagonistic activities against the tested five indicator strains (Aeromonas hydrophila1739, Vibrio cholera 3906, Flavobacterium 2495, Acinetobacter 1271 and Alcaligenes 1424 and these isolates were further identified using biochemical tests and 16S rDNA gene sequence analysis. Except the whole cell product, the other three cellular components, namely, heat-killed whole cell product, intracellular product and extracellular product of all the four selected isolates were equally effective, as revealed by the zone of inhibitions to the tested indicator strains. The in vitro adhesion property or the ability of colonization is often considered as a selection criteria for probiotics. All the selected four strains had higher adhesion abilities than the indicator strains. Further, these four strains had the ability to form biofilms on polystyrene surfaces. The in vitro characterization of these four strains suggests possibility of using the isolates, as individual strain or in combination, for probiotic therapy in aquaculture.

  12. Mites as selective fungal carriers in stored grain habitats.

    Science.gov (United States)

    Hubert, Jan; Stejskal, Václav; Kubátová, Alena; Munzbergová, Zuzana; Vánová, Marie; Zd'árková, Eva

    2003-01-01

    Mites are well documented as vectors of micromycetes in stored products. Since their vectoring capacity is low due to their small size, they can be serious vectors only where there is selective transfer of a high load of specific fungal species. Therefore the aim of our work was to find out whether the transfer of fungi is selective. Four kinds of stored seeds (wheat, poppy, lettuce, mustard) infested by storage mites were subjected to mycological analysis. We compared the spectrum of micromycete species isolated from different species of mites (Acarus siro, Lepidoglyphus destructor, Tyrophagus putrescentiae, Caloglyphus rhizoglyphoides and Cheyletus malaccensis) and various kinds of stored seeds. Fungi were separately isolated from (a) the surface of mites, (b) the mites' digestive tract (= faeces), and (c) stored seeds and were then cultivated and determined. The fungal transport via mites is selective. This conclusion is supported by (i) lower numbers of isolated fungal species from mites than from seeds; (ii) lower Shannon-Weaver diversity index in the fungal communities isolated from mites than from seeds; (iii) significant effect of mites/seeds as environmental variables on fungal presence in a redundancy analysis (RDA); (iv) differences in composition of isolated fungi between mite species shown by RDA. The results of our work support the hypothesis that mite-fungal interactions are dependent on mite species. The fungi attractive to mites seem to be dispersed more than others. The selectivity of fungal transport via mites enhances their pest importance.

  13. Antimicrobial resistence of Shigella species isolated during 2004 and 2005 from selected sites in Zimbabwe.

    Science.gov (United States)

    Ndlovu, N; Tarupiwa, A; Mudzori, J T

    2006-01-01

    To determine the predominant serotype and antibiotic sensitivity pattern of Shigella isolates during 2004 and 2005 in Zimbabwe. Cross sectional study. National Microbiology Reference Laboratory (NMRL), Harare, Zimbabwe. 259 clinical isolates of Shigella species isolated during 2004 and 2005 in Zimbabwe were studied. These samples had been referred to the NMRL for further testing. Serotype and antibiotic sensitivity pattern of Shigella species. Of the 259 clinical isolates of Shigella tested the following species were serotyped; 141 (54.4%) were S. flexneri; 70 (27%) S. sonnei; 38 (14.7%) S. dysenteriae and 10 (3.9%) S. boydii. About 4% of all Shigella isolates tested showed full sensitivity to commonly used antibiotics, 20.8% were resistant to one antibiotic only while 75.3% were resistant to at least two antibiotics. The most common resistance among Shigella species was to cotrimoxazole (89%), tetracycline (73%), ampicillin (49%) and chloramphenicol (41%). High susceptibility among Shigella species was observed to nalidixic acid (86%), ciprofloxacin (99%) and ceftazidine (99%). There was a low drug resistance of Shigella species to nalidixic acid, a drug of choice in Zimbabwe, except among Shigella dysenteriae type 1 strains. Continuous monitoring of the susceptibility patterns of Shigella species is important in order to detect the emergence of drug resistance and to update guidelines for antibiotic treatment in shigellosis.

  14. HYPOVIRULENT ISOLATES OF FUSARIUM COLLECTED FROM CHILI CROPS IN BOYOLALI REGENCY, CENTRAL JAVA, INDONESIA

    Directory of Open Access Journals (Sweden)

    Supyani

    2015-02-01

    Full Text Available Fusarium, a genus of filamentous fungi, has many species which serving as important pathogens to many diseases in crops. Till today, there have not been effective and efficient control methods for such fungi. Recently, scientists agree that application of biological agents is a tactful choice. Development of hypovirulent strains of fungus as biocontrol agents is very limited. This research was aimed to find hypovirulent isolates of Fusarium from field as biological agents. A hundred isolates of Fusarium from chili were collected in Boyolali, Central Java. Morphological characterization revealed that isolates performed varied colony phenotypes. Based on colony phenotype pattern, isolates were classified into five groups. From each group, one hypovirulent isolate was selected based on colony growth rate on potato dextrose agar media. The selected hypovirulent isolates were used for virulence assay in apple. The result showed that there were four hypovirulent isolates i.e.: B6, C15, D19, and E20 isolates. Total RNA extraction of the identified hypovirulent isolates revealed the existence of viral RNA in C15 isolate. Based on the existence of viral RNA in C15 isolate, the hypovirulent traits were due to mycoviral infection, whereas the hypovirulent traits performed by the other three were due to genetic factors.

  15. A new approach to isolating siderophore-producing actinobacteria.

    Science.gov (United States)

    Nakouti, I; Sihanonth, P; Hobbs, G

    2012-07-01

    This study was conducted to investigate the application of 2,2'-dipyridyl as a new approach to isolating siderophore-producing actinobacteria. Isolation of actinobacteria from soil was conducted by a soil dilution plate technique using starch-casein agar. Iron starvation was fostered by the incorporation of the iron chelator 2,2'-dipyridyl in the isolation medium. Pretreatment of the samples at an elevated temperature (40°C) ensured that the majority of nonsporulating bacteria were excluded. The survivors of this treatment were largely actinobacteria. Of the viable cultures grown in the presence of 2,2'-dipyridyl, more than 78-88% (average of three separate studies) were reported to produce siderophore-like compounds compared to 13-18% (average of three separate studies) when grown on the basic media in the absence of the chelating agent. The most prolific producers as assessed by the chrome azurol sulphate (CAS) assay were further characterized and found to belong to the genus Streptomyces. Selective pressure using 2,2'-dipyridyl as an iron-chelating agent in starch-casein media increased the isolation of siderophore-producing actinobacteria compared to the unamended medium. The study described represents a new approach to the isolation of siderophore-producing actinobacteria using a novel procedure that places a selection on cell population based upon the incorporation of a chelating agent in the medium. © 2012 The Authors. Letters in Applied Microbiology © 2012 The Society for Applied Microbiology.

  16. Synthesis of Optimal Isolation Systems of Hand-Transmitted Vibration

    Directory of Open Access Journals (Sweden)

    Marek Książek

    1997-01-01

    Full Text Available In this article a procedure is presented for the analytical synthesis of optimal vibration isolation for a hand-arm system subjected to stochastic excitation. A general approach is discussed for a selected vibration isolation criterion. The general procedure is illustrated by analytical examples for different hand-arm systems described by their driving-point impedances. The influence of particular forms of excitation and the structure of the vibroisolated hand-arm systems on the resultant vibration isolation is then discussed. Some numerical examples illustrating the procedure have also been included.

  17. Isolation of Individual Egg Cells and Zygotes in Alstroemeria Followed by Manual Selection with a Microcapillary-connected Micropump

    Science.gov (United States)

    HOSHINO, YOICHIRO; MURATA, NAHO; SHINODA, KOICHI

    2006-01-01

    • Aims To develop a procedure for isolating living egg cells and zygotes from Alstroemeria ovules. • Scope An attempt was made to isolate egg cells and zygotes from the ovules of Alstroemeria aurea. The ovules were histologically observed using a clearing procedure which revealed the localization and sizes of the embryo sacs and egg apparatus within the ovules. For the isolation of egg cells, ovules were cut into sections with a surgical blade and treated with an enzyme solution. Subsequently, these ovule sections were dissected using a glass needle under an inverted microscope. Egg cells successfully isolated by this procedure were collected using microcapillaries connected to a micropump. For zygote isolation, ovules were excised from ovaries 24 h after self-pollination. By treating excised ovules with an enzyme solution and subsequently dissecting them using a glass needle, zygotes were successfully isolated from the ovules and collected with a microcapillary. The isolated zygotes were associated with pollen tubes and one of the synergids. Egg cells and zygotes were viable for up to 2 h following isolation, as determined by fluorescein diacetate staining. • Conclusions The procedures for isolating egg cells and zygotes in Alstroemeria were established, and each egg cell and zygote was captured with a microcapillary. PMID:16621859

  18. The Effect of Casein Phosphopeptide-Amorf Calcium Phosphate and Acidulated Phosphate Fluoride Gel on Dental Erosion in Primary Teeth: An in Vitro Study.

    Science.gov (United States)

    Maden, Eda Arat; Acar, Özge; Altun, Ceyhan; Polat, Günseli Güven

    This study aimed to investigate the effect of acidulated phosphate fluoride (APF) gel and casein phosphopeptide/amorphous calciumphosphate (CPP-ACP) on the dental erosion produced by carbonated soft drink in primary teeth. This study evaluated by an in vitro model the effect of APF gel and CPP-ACP on the dental enamel previously subjected to erosive challenge with carbonated soft drink. Sixty sound human primary molars were prepared by embedding the crown sections in acrylic resin blocks leaving the enamel surfaces exposed. The surface roughness of the enamel was measured with prophilometry at baseline. Specimens were randomly divided into three treatment groups (n:20): artificial saliva, CPP-ACP, 1.23% APF gel. All specimens were then exposed to an erosive challenge of carbonated soft drink and artificial saliva for 20 cycles of 20 seconds each. Demineralization-remineralization cycles was repeated twice at eight-hour intervals and roughness values were measured. Enamel samples were treated with artificial saliva, CPP-ACP, 1.23% APF gel applied for 10 min after erosive challenge. The arithmetic average roughness (Ra) readings were recorded after remineralization agents were applied. The mean surface roughness in all groups increased significantly after erosion process and decreased after remineralization treatment. After treatment, the mean surface roughness of the 1.23% APF gel group was significantly less than the other groups and the mean surface roughness of the artificial saliva group was significantly more than the other groups. 1.23% APF gel showed the highest protective effect against erosive enamel loss. Under the conditions of this study, artificial saliva, CPP-ACP and 1.23% APF treatments were able to reduce erosive enamel loss produced by carbonated soft drink in primary teeth. However, 1.23% APF gel showed the highest protective effect against erosive enamel loss.

  19. Live Cell Genomics: RNA Exon-Specific RNA-Binding Protein Isolation.

    Science.gov (United States)

    Bell, Thomas J; Eberwine, James

    2015-01-01

    RNA-binding proteins (RBPs) are essential regulatory proteins that control all modes of RNA processing and regulation. New experimental approaches to isolate these indispensable proteins under in vivo conditions are needed to advance the field of RBP biology. Historically, in vitro biochemical approaches to isolate RBP complexes have been useful and productive, but biological relevance of the identified RBP complexes can be imprecise or erroneous. Here we review an inventive experimental to isolate RBPs under the in vivo conditions. The method is called peptide nucleic acid (PNA)-assisted identification of RBP (PAIR) technology and it uses cell-penetrating peptides (CPPs) to deliver photo-activatible RBP-capture molecule to the cytoplasm of the live cells. The PAIR methodology provides two significant advantages over the most commonly used approaches: (1) it overcomes the in vitro limitation of standard biochemical approaches and (2) the PAIR RBP-capture molecule is highly selective and adaptable which allows investigators to isolate exon-specific RBP complexes. Most importantly, the in vivo capture conditions and selectivity of the RBP-capture molecule yield biologically accurate and relevant RBP data.

  20. Measuring of -1,3 Glucanase Activity in Trichoderma Virens Isolates and Selection of the Best Isolates for Biological Control of Cucumber Root Rot

    Directory of Open Access Journals (Sweden)

    Fatemeh Zavvari

    2013-02-01

    Full Text Available In this study four isolates of  Trichoderma virens (T.virens 414, T.virens 414.8, T.virens 304 and  T.virens 404.4 were compared based on extra cellular β-1,3 glucanase production and biological control of root rot cucumber (Phytophthora drerchsleri in laboratory and green house experiments.The in vitro potential of those isolates were evaluated against  P.drechsleri  through production of volatile and dual culture. The pH (3, 5, 7, 8, 9 and temperature effects(5, 15, 20, 25, 30, 40˚C on Trichoderma mycelial growth were also evaluated. Colony growth rate of thease isolates were also studied in water agar culture medium containing carboxy methyle cellulose (CMC and showed direct correlation with β-1,3 glucanase secretion. According to results T. virens 304 and T. virens 414.8  were the best isolates in biocontrol of . drechsleri. Result also showed direct correlation between β-1,3 glucanase secretion and inhibition growth of phytophthora  mycelium and reduction of phytophthora infection.

  1. Progress in ISOL target-ion source systems

    Energy Technology Data Exchange (ETDEWEB)

    Koester, U. [Institut Laue Langevin, 6 Rue Jules Horowitz, F-38042 Grenoble Cedex 9 (France); ISOLDE, CERN, CH-1211 Geneve 23 (Switzerland)], E-mail: koester@ill.fr; Arndt, O. [HGF VISTARS and Institut fuer Kernchemie, Johannes-Gutenberg Universitaet Mainz, D-55128 Mainz (Germany); Bouquerel, E.; Fedoseyev, V.N. [ISOLDE, CERN, CH-1211 Geneve 23 (Switzerland); Franberg, H. [ISOLDE, CERN, CH-1211 Geneve 23 (Switzerland); Laboratory for Radio- and Environmental Chemistry, Paul Scherrer Institut, CH-5232 Villigen PSI (Switzerland); Joinet, A. [ISOLDE, CERN, CH-1211 Geneve 23 (Switzerland); Centre d' Etude Spatiale des Rayonnements, 9 Av. du Colonel Roche, F-31028 Toulouse Cedex 4 (France); Jost, C. [HGF VISTARS and Institut fuer Kernchemie, Johannes-Gutenberg Universitaet Mainz, D-55128 Mainz (Germany); Kerkines, I.S.K. [Laboratory of Physical Chemistry, National and Kapodistrian University of Athens, Department of Chemistry, Zografou 157 71, GR (Greece); Cherry L. Emerson Center for Scientific Computation and Department of Chemistry, Emory University, Atlanta, GA 30322 (United States); Kirchner, R. [Gesellschaft fuer Schwerionenforschung, Planckstr. 1, D-64291 Darmstadt (Germany)

    2008-10-15

    The heart of every ISOL (isotope separation on-line) facility is its target and ion source system. Its efficiency, selectivity and rapidity is decisive for the production of intense and pure ion beams of short-lived isotopes. Recent progress in ISOL target and ion source technology is discussed at the examples of radioactive ion beams of exotic zinc and tin isotopes that were purified by isothermal chromatography and molecular sideband separation respectively. An outlook is given to which other elements these purification methods are applicable.

  2. Progress in ISOL target-ion source systems

    International Nuclear Information System (INIS)

    Koester, U.; Arndt, O.; Bouquerel, E.; Fedoseyev, V.N.; Franberg, H.; Joinet, A.; Jost, C.; Kerkines, I.S.K.; Kirchner, R.

    2008-01-01

    The heart of every ISOL (isotope separation on-line) facility is its target and ion source system. Its efficiency, selectivity and rapidity is decisive for the production of intense and pure ion beams of short-lived isotopes. Recent progress in ISOL target and ion source technology is discussed at the examples of radioactive ion beams of exotic zinc and tin isotopes that were purified by isothermal chromatography and molecular sideband separation respectively. An outlook is given to which other elements these purification methods are applicable.

  3. Site selection report basalt waste isolation program near-surface test facility

    International Nuclear Information System (INIS)

    Sharpe, S.D.

    1978-01-01

    A site selection committee was established to review the information gathered on potential sites and to select a site for the Near-Surface Test Facility Phase I. A decision was made to use a site on the north face of Gable Mountain located on the Hanford Site. This site provided convenient access to the Pomona Basalt Flow. This flow was selected for use at this site because it exhibited the characteristics established in the primary criteria. These criteria were: the flows thickness; its dryness; its nearness to the surface; and, its similarities to basalt units which are candidates for the repository. After the selection of the Near-Surface Test Facility Phase I Site, the need arose for an additional facility to demonstrate safe handling, storage techniques, and the physical effects of radioactive materials on an in situ basalt formation. The committee reviewed the sites selected for Phase I and chose the same site for locating Phase II of the Near-Surface Test Facility

  4. BanglaLekha-Isolated: A multi-purpose comprehensive dataset of Handwritten Bangla Isolated characters

    Directory of Open Access Journals (Sweden)

    Mithun Biswas

    2017-06-01

    Full Text Available BanglaLekha-Isolated, a Bangla handwritten isolated character dataset is presented in this article. This dataset contains 84 different characters comprising of 50 Bangla basic characters, 10 Bangla numerals and 24 selected compound characters. 2000 handwriting samples for each of the 84 characters were collected, digitized and pre-processed. After discarding mistakes and scribbles, 1,66,105 handwritten character images were included in the final dataset. The dataset also includes labels indicating the age and the gender of the subjects from whom the samples were collected. This dataset could be used not only for optical handwriting recognition research but also to explore the influence of gender and age on handwriting. The dataset is publicly available at https://data.mendeley.com/datasets/hf6sf8zrkc/2.

  5. Selective medium for aerobic incubation of Campylobacter

    Science.gov (United States)

    Studies were conducted on the formulation of a selective medium that could be used to isolate Campylobacter from mixed bacterial cultures using aerobic incubation. A non-selective, basal broth medium was prepared and supplemented with Bolton, Cefex, or Skirrow antibiotic mixtures. The ability of pur...

  6. Selected, annotated bibliography of studies relevant to the isolation of nuclear wastes. [705 references

    Energy Technology Data Exchange (ETDEWEB)

    Hyder, L.K.; Fore, C.S.; Vaughan, N.D.; Faust, R.A.

    1980-09-01

    This annotated bibliography of 705 references represents the first in a series to be published by the Ecological Sciences Information Center containing scientific, technical, economic, and regulatory information relevant to nuclear waste isolation. Most references discuss deep geologic disposal, with fewer studies of deep seabed disposal; space disposal is also included. The publication covers both domestic and foreign literature for the period 1954 to 1980. Major chapters selected are Chemical and Physical Aspects; Container Design and Performance; Disposal Site; Envirnmental Transport; General Studies and Reviews; Geology, Hydrology and Site Resources; Regulatory and Economic Aspects; Repository Design and Engineering; Transportation Technology; Waste Production; and Waste Treatment. Specialized data fields have been incorporated to improve the ease and accuracy of locating pertinent references. Specific radionuclides for which data are presented are listed in the Measured Radionuclides field, and specific parameters which affect the migration of these radionuclides are presented in the Measured Parameters field. The references within each chapter are arranged alphabetically by leading author, corporate affiliation, or title of the document. When the author is not given, the corporate affiliation appears first. If these two levels of authorship are not given, the title of the document is used as the identifying level. Indexes are provided for author(s), keywords, subject category, title, geographic location, measured parameters, measured radionuclides, and publication description.

  7. Genomic characterization of Burkholderia pseudomallei isolates selected for medical countermeasures testing: comparative genomics associated with differential virulence.

    Directory of Open Access Journals (Sweden)

    Jason W Sahl

    Full Text Available Burkholderia pseudomallei is the causative agent of melioidosis and a potential bioterrorism agent. In the development of medical countermeasures against B. pseudomallei infection, the US Food and Drug Administration (FDA animal Rule recommends using well-characterized strains in animal challenge studies. In this study, whole genome sequence data were generated for 6 B. pseudomallei isolates previously identified as candidates for animal challenge studies; an additional 5 isolates were sequenced that were associated with human inhalational melioidosis. A core genome single nucleotide polymorphism (SNP phylogeny inferred from a concatenated SNP alignment from the 11 isolates sequenced in this study and a diverse global collection of isolates demonstrated the diversity of the proposed Animal Rule isolates. To understand the genomic composition of each isolate, a large-scale blast score ratio (LS-BSR analysis was performed on the entire pan-genome; this demonstrated the variable composition of genes across the panel and also helped to identify genes unique to individual isolates. In addition, a set of ~550 genes associated with pathogenesis in B. pseudomallei were screened against the 11 sequenced genomes with LS-BSR. Differential gene distribution for 54 virulence-associated genes was observed between genomes and three of these genes were correlated with differential virulence observed in animal challenge studies using BALB/c mice. Differentially conserved genes and SNPs associated with disease severity were identified and could be the basis for future studies investigating the pathogenesis of B. pseudomallei. Overall, the genetic characterization of the 11 proposed Animal Rule isolates provides context for future studies involving B. pseudomallei pathogenesis, differential virulence, and efficacy to therapeutics.

  8. Francisella tularensis subsp. novicida isolated from a human in Arizona

    Directory of Open Access Journals (Sweden)

    Birdsell Dawn N

    2009-11-01

    Full Text Available Abstract Background Francisella tularensis is the etiologic agent of tularemia and is classified as a select agent by the Centers for Disease Control and Prevention. Currently four known subspecies of F. tularensis that differ in virulence and geographical distribution are recognized:tularensis (type A, holarctica (type B, mediasiatica, and novicida. Because of the Select Agent status and differences in virulence and geographical location, the molecular analysis of any clinical case of tularemia is of particular interest. We analyzed an unusual Francisella clinical isolate from a human infection in Arizona using multiple DNA-based approaches. Findings We report that the isolate is F. tularensis subsp. novicida, a subspecies that is rarely isolated. Conclusion The rarity of this novicida subspecies in clinical settings makes each case study important for our understanding of its role in disease and its genetic relationship with other F. tularensis subspecies.

  9. Isolation and identification of lactic acid bacteria from abalone (Haliotis asinina as a potential candidate of probiotic

    Directory of Open Access Journals (Sweden)

    YAYAN SOFYAN

    2010-01-01

    Full Text Available Sarkono, Faturrahman, Sofyan Y. 2010. Isolation and identification of lactic acid bacteria from abalone (Haliotis asinina as a potential candidate of probiotic. Nusantara Bioscience 2: 38-42. The purpose of this study was to isolate, select and characterize lactic acid bacteria (LAB from abalone as a potential candidate probiotic in abalone cultivation system. Selective isolation of LAB performed using de Man Rogosa Sharpe medium. LAB isolate that potential as probiotics was screened. Selection was based on its ability to suppress the growth of pathogenic bacteria, bacterial resistance to acidic conditions and bacterial resistance to bile salts (bile. Further characterization and identification conducted to determine the species. The results showed that two of the ten isolates potential to be developed as probiotic bacteria that have the ability to inhibit several pathogenic bacteria such as Eschericia coli, Bacillus cereus dan Staphylococus aureus, able to grow at acidic condition and bile tolerance during the incubation for 24 hour. Based on the API test kit, the both of isolate identified as members of the species Lactobacillus paracasei ssp. paracasei.

  10. Isolation, screening and identification of novel isolates of Actinomycetes from India for antimicrobial applications

    Directory of Open Access Journals (Sweden)

    Vineeta Singh

    2016-12-01

    Full Text Available The search for novel bioactive compounds from the natural environment has been rapidly increased with the increase in multi-drug resistant (MDR pathogens. In the present study, the antimicrobial potential of novel actinomycetes has been evaluated by initial screening of six soil samples. Primary and secondary screening was performed against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Candida albicans, Candida tropicalis, Trichophyton rubrum, and other MDR bacterial and fungal test strains, and at the end thirteen active isolates were selected for further study. Microbial strains were identified on the basis of growth conditions and other biochemical characters. Five most active microbial strains were identified using 16S rRNA sequence homology and designated as Streptomyces xanthophaeus MTCC 11938, Streptomyces variabilis MTCC 12266, Streptomyces xanthochromogenes MTCC 11937, Streptomyces levis EU 124569 and Streptomyces sp. NCIM 5500. Four antibacterial and three antifungal compounds isolated from the above five isolates were purified and partially characterized using UV absorption and IR spectra. Two antibacterial metabolites, belong to chromone and peptide antibiotic, respectively. The antifungal compounds were found to be of non-polyene nature. In conclusion, we study the isolation of novel bacterial strains of actinomycetes for producing novel compounds having antibacterial and antifungal activities from the unexplored agro-ecological niches of India. Also, this study paves the way for further characterization of these isolates of Streptomyces sp. for their optimum utilization for antimicrobial purposes.

  11. An optimized method for mouse liver sinusoidal endothelial cell isolation

    Energy Technology Data Exchange (ETDEWEB)

    Meyer, Jeremy, E-mail: jeremy.meyer@hcuge.ch [Division of Digestive and Transplantation Surgery, University Hospitals of Geneva, Rue Gabrielle-Perret-Gentil 4, 1211 Genève 14 (Switzerland); Unit of Surgical Research, University of Geneva, Rue Michel-Servet 1, 1206 Genève (Switzerland); Lacotte, Stéphanie, E-mail: stephanie.lacotte@unige.ch [Unit of Surgical Research, University of Geneva, Rue Michel-Servet 1, 1206 Genève (Switzerland); Morel, Philippe, E-mail: philippe.morel@hcuge.ch [Division of Digestive and Transplantation Surgery, University Hospitals of Geneva, Rue Gabrielle-Perret-Gentil 4, 1211 Genève 14 (Switzerland); Unit of Surgical Research, University of Geneva, Rue Michel-Servet 1, 1206 Genève (Switzerland); Gonelle-Gispert, Carmen, E-mail: carmen.gonelle@unige.ch [Unit of Surgical Research, University of Geneva, Rue Michel-Servet 1, 1206 Genève (Switzerland); Bühler, Léo, E-mail: leo.buhler@hcuge.ch [Division of Digestive and Transplantation Surgery, University Hospitals of Geneva, Rue Gabrielle-Perret-Gentil 4, 1211 Genève 14 (Switzerland); Unit of Surgical Research, University of Geneva, Rue Michel-Servet 1, 1206 Genève (Switzerland)

    2016-12-10

    The objective of the present study was to develop an accurate and reproducible method for liver sinusoidal endothelial cell (LSEC) isolation in mice. Non-parenchymal cells were isolated using a modified two-step collagenase digestion combined with Optiprep density gradient centrifugation. LSEC were further purified using two prevalent methods, short-term selective adherence and CD146+ magnetic-activated cell sorting (MACS), and compared in terms of cell yield, viability and purity to our purification technique using CD11b cell depletion combined with long-term selective adherence. LSEC purification using our technique allowed to obtain 7.07±3.80 million LSEC per liver, while CD146+ MACS and short-term selective adherence yielded 2.94±1.28 and 0.99±0.66 million LSEC, respectively. Purity of the final cell preparation reached 95.10±2.58% when using our method. In contrast, CD146+ MACS and short-term selective adherence gave purities of 86.75±3.26% and 47.95±9.82%, respectively. Similarly, contamination by non-LSEC was the lowest when purification was performed using our technique, with a proportion of contaminating macrophages of only 1.87±0.77%. Further, isolated cells analysed by scanning electron microscopy presented typical LSEC fenestrations organized in sieve plates, demonstrating that the technique allowed to isolate bona fide LSEC. In conclusion, we described a reliable and reproducible technique for the isolation of high yields of pure LSEC in mice. This protocol provides an efficient method to prepare LSEC for studying their biological functions. - Highlights: • This protocol provides an efficient method to prepare primary mouse LSEC for studying their biological functions. • The liver cell dispersion step was improved by performing a retrograde cannulation of the liver. • The cell yield and the purity obtained were higher than comparative techniques in mice. • Contaminating macrophages were removed by introducing a CD11b- magnetic

  12. An optimized method for mouse liver sinusoidal endothelial cell isolation

    International Nuclear Information System (INIS)

    Meyer, Jeremy; Lacotte, Stéphanie; Morel, Philippe; Gonelle-Gispert, Carmen; Bühler, Léo

    2016-01-01

    The objective of the present study was to develop an accurate and reproducible method for liver sinusoidal endothelial cell (LSEC) isolation in mice. Non-parenchymal cells were isolated using a modified two-step collagenase digestion combined with Optiprep density gradient centrifugation. LSEC were further purified using two prevalent methods, short-term selective adherence and CD146+ magnetic-activated cell sorting (MACS), and compared in terms of cell yield, viability and purity to our purification technique using CD11b cell depletion combined with long-term selective adherence. LSEC purification using our technique allowed to obtain 7.07±3.80 million LSEC per liver, while CD146+ MACS and short-term selective adherence yielded 2.94±1.28 and 0.99±0.66 million LSEC, respectively. Purity of the final cell preparation reached 95.10±2.58% when using our method. In contrast, CD146+ MACS and short-term selective adherence gave purities of 86.75±3.26% and 47.95±9.82%, respectively. Similarly, contamination by non-LSEC was the lowest when purification was performed using our technique, with a proportion of contaminating macrophages of only 1.87±0.77%. Further, isolated cells analysed by scanning electron microscopy presented typical LSEC fenestrations organized in sieve plates, demonstrating that the technique allowed to isolate bona fide LSEC. In conclusion, we described a reliable and reproducible technique for the isolation of high yields of pure LSEC in mice. This protocol provides an efficient method to prepare LSEC for studying their biological functions. - Highlights: • This protocol provides an efficient method to prepare primary mouse LSEC for studying their biological functions. • The liver cell dispersion step was improved by performing a retrograde cannulation of the liver. • The cell yield and the purity obtained were higher than comparative techniques in mice. • Contaminating macrophages were removed by introducing a CD11b- magnetic

  13. Isolation of Acremonium species producing cephalosporine C (CPC ...

    African Journals Online (AJOL)

    SERVER

    2007-11-19

    Nov 19, 2007 ... selective media and Acremonium sp. were isolated. The presence of CPC antibiotic in the fermentation broths of these species was ... Penicillium sp. 28%. Alternaria sp. .... On-line high performance liquid chromatography for ...

  14. Isolation, identification and characterization of regional indigenous Saccharomyces cerevisiae strains

    Directory of Open Access Journals (Sweden)

    Hana Šuranská

    2016-03-01

    Full Text Available Abstract In the present work we isolated and identified various indigenous Saccharomyces cerevisiae strains and screened them for the selected oenological properties. These S. cerevisiae strains were isolated from berries and spontaneously fermented musts. The grape berries (Sauvignon blanc and Pinot noir were grown under the integrated and organic mode of farming in the South Moravia (Czech Republic wine region. Modern genotyping techniques such as PCR-fingerprinting and interdelta PCR typing were employed to differentiate among indigenous S. cerevisiae strains. This combination of the methods provides a rapid and relatively simple approach for identification of yeast of S. cerevisiae at strain level. In total, 120 isolates were identified and grouped by molecular approaches and 45 of the representative strains were tested for selected important oenological properties including ethanol, sulfur dioxide and osmotic stress tolerance, intensity of flocculation and desirable enzymatic activities. Their ability to produce and utilize acetic/malic acid was examined as well; in addition, H2S production as an undesirable property was screened. The oenological characteristics of indigenous isolates were compared to a commercially available S. cerevisiae BS6 strain, which is commonly used as the starter culture. Finally, some indigenous strains coming from organically treated grape berries were chosen for their promising oenological properties and these strains will be used as the starter culture, because application of a selected indigenous S. cerevisiae strain can enhance the regional character of the wines.

  15. Genomic diversity of Escherichia isolates from diverse habitats.

    Directory of Open Access Journals (Sweden)

    Seungdae Oh

    Full Text Available Our understanding of the Escherichia genus is heavily biased toward pathogenic or commensal isolates from human or animal hosts. Recent studies have recovered Escherichia isolates that persist, and even grow, outside these hosts. Although the environmental isolates are typically phylogenetically distinct, they are highly related to and phenotypically indistinguishable from their human counterparts, including for the coliform test. To gain insights into the genomic diversity of Escherichia isolates from diverse habitats, including freshwater, soil, animal, and human sources, we carried out comparative DNA-DNA hybridizations using a multi-genome E. coli DNA microarray. The microarray was validated based on hybridizations with selected strains whose genome sequences were available and used to assess the frequency of microarray false positive and negative signals. Our results showed that human fecal isolates share two sets of genes (n>90 that are rarely found among environmental isolates, including genes presumably important for evading host immune mechanisms (e.g., a multi-drug transporter for acids and antimicrobials and adhering to epithelial cells (e.g., hemolysin E and fimbrial-like adhesin protein. These results imply that environmental isolates are characterized by decreased ability to colonize host cells relative to human isolates. Our study also provides gene markers that can distinguish human isolates from those of warm-blooded animal and environmental origins, and thus can be used to more reliably assess fecal contamination in natural ecosystems.

  16. Morphological, Physiological, and Taxonomic Characterization of Actinobacterial Isolates Living as Endophytes of Cacao Pods and Cacao Seeds.

    Science.gov (United States)

    Tchinda, Romaric Armel Mouafo; Boudjeko, Thaddée; Simao-Beaunoir, Anne-Marie; Lerat, Sylvain; Tsala, Éric; Monga, Ernest; Beaulieu, Carole

    2016-01-01

    Vascular plants are commonly colonized by endophytic actinobacteria. However, very little is known about the relationship between these microorganisms and cacao fruits. In order to determine the physiological and taxonomic relationships between the members of this community, actinobacteria were isolated from cacao fruits and seeds. Among the 49 isolates recovered, 11 morphologically distinct isolates were selected for further characterization. Sequencing of the 16S rRNA gene allowed the partition of the selected isolates into three phylogenetic clades. Most of the selected endophytic isolates belonged to the Streptomyces violaceusniger clade. Physiological characterization was carried out and a similarity index was used to cluster the isolates. However, clustering based on physiological properties did not match phylogenetic lineages. Isolates were also characterized for traits commonly associated with plant growth-promoting bacteria, including antibiosis and auxin biosynthesis. All isolates exhibited resistance to geldanamycin, whereas only two isolates were shown to produce this antibiotic. Endophytes were inoculated on radish seedlings and most isolates were found to possess plant growth-promoting abilities. These endophytic actinobacteria inhibited the growth of various plant pathogenic fungi and/or bacteria. The present study showed that S. violaceusniger clade members represent a significant part of the actinobacterial community living as endophytes in cacao fruits and seeds. While several members of this clade are known to be geldanamycin producers and efficient biocontrol agents of plant diseases, we herein established the endophytic lifestyle of some of these microorganisms, demonstrating their potential as plant health agents.

  17. Reliability analysis of containment isolation systems

    International Nuclear Information System (INIS)

    Pelto, P.J.; Counts, C.A.

    1984-06-01

    The Pacific Northwest Laboratory (PNL) is reviewing available information on containment systems design, operating experience, and related research as part of a project being conducted by the Division of Systems Integration, US Nuclear Regulatory Commission. The basic objective of this work is to collect and consolidate data relevant to assessing the functional performance of containment isolation systems and to use this data to the extent possible to characterize containment isolation system reliability for selected reference designs. This paper summarizes the results from initial efforts which focused on collection of data from available documents and briefly describes detailed review and analysis efforts which commenced recently. 5 references

  18. Selection and optimization of extracellular lipase production using ...

    African Journals Online (AJOL)

    The aim of this study was to isolate and select lipase-producing microorganisms originated from different substrates, as well as to optimize the production of microbial lipase by submerged fermentation under different nutrient conditions. Of the 40 microorganisms isolated, 39 showed a halo around the colonies and 4 were ...

  19. The Use of Variable Q1 Isolation Windows Improves Selectivity in LC-SWATH-MS Acquisition.

    Science.gov (United States)

    Zhang, Ying; Bilbao, Aivett; Bruderer, Tobias; Luban, Jeremy; Strambio-De-Castillia, Caterina; Lisacek, Frédérique; Hopfgartner, Gérard; Varesio, Emmanuel

    2015-10-02

    As tryptic peptides and metabolites are not equally distributed along the mass range, the probability of cross fragment ion interference is higher in certain windows when fixed Q1 SWATH windows are applied. We evaluated the benefits of utilizing variable Q1 SWATH windows with regards to selectivity improvement. Variable windows based on equalizing the distribution of either the precursor ion population (PIP) or the total ion current (TIC) within each window were generated by an in-house software, swathTUNER. These two variable Q1 SWATH window strategies outperformed, with respect to quantification and identification, the basic approach using a fixed window width (FIX) for proteomic profiling of human monocyte-derived dendritic cells (MDDCs). Thus, 13.8 and 8.4% additional peptide precursors, which resulted in 13.1 and 10.0% more proteins, were confidently identified by SWATH using the strategy PIP and TIC, respectively, in the MDDC proteomic sample. On the basis of the spectral library purity score, some improvement warranted by variable Q1 windows was also observed, albeit to a lesser extent, in the metabolomic profiling of human urine. We show that the novel concept of "scheduled SWATH" proposed here, which incorporates (i) variable isolation windows and (ii) precursor retention time segmentation further improves both peptide and metabolite identifications.

  20. Antioxidant Activity of Whey from Milk Fermented with Lactobacillus Species Isolated from Nigerian Fermented Foods

    Directory of Open Access Journals (Sweden)

    Ifeoma Korie

    2010-01-01

    Full Text Available Eight Lactobacillus isolates obtained from five indigenous fermented foods (ogi, ogi baba, wara, kunnu and ugba were investigated. Wara is a dairy-based food while the others are not dairy-based. The bacteria were isolated on MRS agar and purified by successive streaking on the same medium. The whey fraction of skimmed milk fermented with each isolate was assayed for radical scavenging effects using 2,2-diphenyl-1-picrylhydrazyl (DPPH radical. All the whey fractions showed radical scavenging activities. The five isolates with the highest activities were selected. On the basis of Gram stain reaction, cellular morphology, biochemical tests and carbohydrate utilization profiles they were identified as strains of Lactobacillus brevis, L. fermentum, L. plantarum, L. casei and L. delbrueckii. The antioxidant activities of whey fractions from 24-hour fermentations with the selected organisms were investigated using both radical scavenging effects and lipid peroxidation inhibitory activity. The radical scavenging activity was generally higher than the lipid peroxidation inhibition, except in the L. plantarum strain, which did not show any significant difference in both activities. The probiotic potential of the isolates was evaluated by pH and bile tolerance. None of the selected isolates showed any growth at pH=2.0 but L. casei and L. delbrueckii survived at this pH. Four of the five selected isolates were able to grow in 0.5 % dehydrated bile, with L. casei strain showing the highest level of growth, followed by L. delbrueckii. L. plantarum strain was not bile tolerant. The ability of L. casei and L. delbrueckii strains to survive at pH=2 and grow in the presence of bile indicates that the isolates may be able to colonize the gastrointestinal tract. The findings of this study indicate that Lactobacillus strains isolated from indigenous Nigerian fermented foods could be useful as starter cultures to provide antioxidants in food and that fermented milk

  1. Isolation and identification of culturable extremely halophilic archaea of Inche-Boroun wetland

    Directory of Open Access Journals (Sweden)

    Mehrnoosh Rasooli

    2013-11-01

    Full Text Available Haloarchaeal diversity of Inche-Boroun wetland in north of Iran in Golestan province was investigated by using culture-dependent methods. Sampling was carried out in May and September 2010. In each sampling, 4 distinct region of wetland were analyzed by using complex media like MGM, JCM168, MH1 and an alkaliphilic medium containing 23% salts. After incubation at 40ºC, a total of 406 isolates were prepared and 2.1×106 CFU/ml were obtained in culture media. Among all isolates, 361 isolates were obtained from MGM and 39 isolates from JCM 168, 3 isolates from MH1 and 3 isolate from alkaliphilic media. Initial morphological, biochemical and physiological tests were performed. According to the results, 45 isolates were selected and phylogenetic analysis of 16S rRNA was performed for them. Among selected strains, 40 isolates belonged to Halobacteriacaea and were related to Haloarcula, Halorubrum, Haloferax, Halobellus, Halogeometricum, Halobacterium, Halolamina, Halorhabdus and Halostagnicola (respectively 30, 27.5, 17.5, 10, 5.2, 2.6, 2.6, 2.6 and 2.6 percent of Haloarchaeal strains. A total of 5 strains belonged to the kingdom of Bacteria and were related to Rhodovibrio, Pseudomonas and Salicola (respectively 40, 40 and 20 percent of bacterial strains. According to our results and the limited numbers of haloarchaeal genera that having been discovered until now, it seemed that the culturable prokaryotic populations in this hypersaline environment was diverse.

  2. Laser Ion Source Development for ISOL Systems at RIA

    CERN Document Server

    Liu, Yuan; Beene, James R; Bilheux, Hassina Z; Brueck, Kim; Geppert, Christopher; Havener, Charles; Kessler, Thomas; Krause, Herbert F; Schultz, David R; Stracener, Dan; Vane, C R; Wendt, Klaus

    2005-01-01

    The isobaric purity of radioactive ion beams (RIBs) is of crucial importance to many experiments. Laser ion sources based on resonant photoionization have already proved to be of great value at existing ISOL RIB facilities. In these ion sources, ions of a selected isotope are produced by laser radiation via stepwise atomic resonant excitations followed by ionization in the last transition. Because each element has its own unique atomic energy levels, the resonant photoionization process can provide elemental selectivity of nearly 100%. We have initiated a research effort to develop a prototype laser ion source with the potential to achieve the high selectivity and high efficiency required for research with ISOL-generated RIBs at the Rare Isotope Accelerator (RIA). A pilot experiment has been conducted to demonstrate resonant photoionization of three atomic species using all-solid-state tunable Ti:Sapphire lasers. Three Ti:Sapphire lasers were provided by the University of Mainz and used in the experiment for ...

  3. Isolation, identification and screening of potential cellulase-free ...

    African Journals Online (AJOL)

    compaq

    2012-09-05

    Sep 5, 2012 ... Eight fungal species were selected for further study based on clearing zones ... lignolytic enzymes, instead of chlorine, are used to break the xylan, cellulose, lignin ... purchased from Sigma chemicals Co., USA. Isolation and ...

  4. Primary isolation strain determines both phage type and receptors recognised by Campylobacter jejuni bacteriophages.

    Directory of Open Access Journals (Sweden)

    Martine C Holst Sørensen

    Full Text Available In this study we isolated novel bacteriophages, infecting the zoonotic bacterium Campylobacter jejuni. These phages may be used in phage therapy of C. jejuni colonized poultry to prevent spreading of the bacteria to meat products causing disease in humans. Many C. jejuni phages have been isolated using NCTC12662 as the indicator strain, which may have biased the selection of phages. A large group of C. jejuni phages rely on the highly diverse capsular polysaccharide (CPS for infection and recent work identified the O-methyl phosphoramidate modification (MeOPN of CPS as a phage receptor. We therefore chose seven C. jejuni strains each expressing different CPS structures as indicator strains in a large screening for phages in samples collected from free-range poultry farms. Forty-three phages were isolated using C. jejuni NCTC12658, NCTC12662 and RM1221 as host strains and 20 distinct phages were identified based on host range analysis and genome restriction profiles. Most phages were isolated using C. jejuni strains NCTC12662 and RM1221 and interestingly phage genome size (140 kb vs. 190 kb, host range and morphological appearance correlated with the isolation strain. Thus, according to C. jejuni phage grouping, NCTC12662 and NCTC12658 selected for CP81-type phages, while RM1221 selected for CP220-type phages. Furthermore, using acapsular ∆kpsM mutants we demonstrated that phages isolated on NCTC12658 and NCTC12662 were dependent on the capsule for infection. In contrast, CP220-type phages isolated on RM1221 were unable to infect non-motile ∆motA mutants, hence requiring motility for successful infection. Hence, the primary phage isolation strain determines both phage type (CP81 or CP220 as well as receptors (CPS or flagella recognised by the isolated phages.

  5. Bioremediation capability and characterization of bacteria isolated from petroleum contaminated soils in Iran

    Directory of Open Access Journals (Sweden)

    Golafarin Ghoreishi

    2017-07-01

    Full Text Available This study was carried out to isolate bacteria for bioremediation of petroleum polluted soils. Five samples were used for isolation in this study. They were four soil samples in addition to one kerosene sample. The soil samples including soils contaminated by crude oil and gas oil and two soil samples with no outward contamination which were collected from Shiraz Oil Refinery sites. Seven strains were selected among the isolated colonies for further experiments. The selected isolates were cultured in standard succinate medium (SSM minimal medium in which 2.5% v/v kerosene was used as carbon source. In another bacterial SSM culture, carbon, sulfur or nitrogen source was removed and 20% v/v kerosene added to check the ability of isolates to utilizekerosene as sole source for C, N and S. Finally, cultures of four strains with higher growth in modified SSM cultures were selected for GC analysis. In this study they were named C2 and C4 which were isolated from crude oil contaminated soil and SI1 and SI2 isolated from soils with no outward contamination. GC analysis showed that C2 could degrade 69% of 5% v/v kerosene in 7 d, while C4 and SI1 degraded 48% and 42% of 5% v/v kerosene during this 7-d period respectively, and the degradation ability of SI2 was 38% after 7 d. Analysis of 16S rRNA gene showed that C2 was close to Citrobacter sedlakii, C4 and SI1 were related to Entrobacter hormeachei and SI2 was close to Entrobacter cloacae, respectively.

  6. Isolation and characterization of Faecalibacterium prausnitzii from calves and piglets.

    Directory of Open Access Journals (Sweden)

    Carla Foditsch

    Full Text Available The goal of our study was to isolate and characterize Faecalibacterium prausnitzii from fecal samples of healthy calves and piglets, in order to develop a novel probiotic for livestock animals. We identified 203 isolates of Faecalibacterium sp., which were clustered in 40 genetically distinct groups. One representative isolate from each cluster was selected for further characterization. The concentrations of the short chain fatty acids (SCFA acetate, butyrate, propionate and isobutyrate in the culture media were measured by gas chromatography. We observed reduction in the concentration of acetate followed by concomitant increase in the concentration of butyrate, suggesting that the isolates were consuming acetate present in the media and producing butyrate. Butyrate production correlated positively with bacterial growth. Since butyrate has many benefits to the colonic epithelial cells, the selection of strains that produce higher amounts of butyrate is extremely important for the development of this potential probiotic. The effect of pH and concentration of bile salts on bacterial growth was also evaluated in order to mimic the conditions encountered by F. prausnitzii in vivo. The optimal pH for growth ranged between 5.5 and 6.7, while most isolates were inhibited by of the lowest concentration of bile salts tested (0.1%. Antimicrobial resistance profile showed that most isolates of Faecalibacterium sp. were resistant against ciprofloxacin and sulfamethoxazole-trimethoprim. More than 50% of the isolates were resistant to tetracycline, amikacin, cefepime and cefoxitin. A total of 19 different combinations of multidrug resistance were observed among the isolates. Our results provide new insights into the cultural and physiological characteristics of Faecalibacterium prausnitzii illustrating large variability in short chain fatty acid production, in vitro growth, sensitivity to bile salts, and antibiotic resistance and suggesting that future

  7. Permanent isolation surface barrier development plan

    International Nuclear Information System (INIS)

    Wing, N.R.

    1994-01-01

    The exhumation and treatment of wastes may not always be the preferred alternative in the remediation of a waste site. In-place disposal alternatives, under certain circumstances, may be the most desirable alternatives to use in the protection of human health and the environment. The implementation of an in-place disposal alternative will likely require some type of protective covering that will provide long-term isolation of the wastes from the accessible environment. Even if the wastes are exhumed and treated, a long-term barrier may still be needed to adequately dispose of the treated wastes or any remaining waste residuals. Currently, no open-quotes provenclose quotes long-term barrier is available. The Hanford Site Permanent Isolation Surface Barrier Development Program (BDP) was organized to develop the technology needed to provide a long-term surface barrier capability for the Hanford Site. The permanent isolation barrier technology also could be used at other sites. Permanent isolation barriers use engineered layers of natural materials to create an integrated structure with redundant protective features. Drawings of conceptual permanent isolation surface barriers are shown. The natural construction materials (e.g., fine soil, sand, gravel, riprap, asphalt) have been selected to optimize barrier performance and longevity. The objective of current designs is to use natural materials to develop a maintenance-free permanent isolation surface barrier that isolates wastes for a minimum of 1,000 years by limiting water drainage to near-zero amounts; reducing the likelihood of plant, animal, and human intrusion; controlling the exhalation of noxious gases; and minimizing erosion-related problems

  8. Permanent isolation surface barrier development plan

    Energy Technology Data Exchange (ETDEWEB)

    Wing, N.R.

    1994-01-01

    The exhumation and treatment of wastes may not always be the preferred alternative in the remediation of a waste site. In-place disposal alternatives, under certain circumstances, may be the most desirable alternatives to use in the protection of human health and the environment. The implementation of an in-place disposal alternative will likely require some type of protective covering that will provide long-term isolation of the wastes from the accessible environment. Even if the wastes are exhumed and treated, a long-term barrier may still be needed to adequately dispose of the treated wastes or any remaining waste residuals. Currently, no {open_quotes}proven{close_quotes} long-term barrier is available. The Hanford Site Permanent Isolation Surface Barrier Development Program (BDP) was organized to develop the technology needed to provide a long-term surface barrier capability for the Hanford Site. The permanent isolation barrier technology also could be used at other sites. Permanent isolation barriers use engineered layers of natural materials to create an integrated structure with redundant protective features. Drawings of conceptual permanent isolation surface barriers are shown. The natural construction materials (e.g., fine soil, sand, gravel, riprap, asphalt) have been selected to optimize barrier performance and longevity. The objective of current designs is to use natural materials to develop a maintenance-free permanent isolation surface barrier that isolates wastes for a minimum of 1,000 years by limiting water drainage to near-zero amounts; reducing the likelihood of plant, animal, and human intrusion; controlling the exhalation of noxious gases; and minimizing erosion-related problems.

  9. Isolation and Characterization of melanized yeast form of Aureobasidium pullulans and physiological studies on the melanization process

    International Nuclear Information System (INIS)

    El-Gamal, M.S.; El-Bialy, H.A.; Elsayed, M.A.; Khalifa, M.A.

    2017-01-01

    Melanin pigments have immense application potentials in the fields of agriculture , cosmetics and pharmaceutical industries . Isolation and characterization of melanin producer from local resources is the main aim of the present study . Six melanized yeast isolates were selected from seventeen isolation resources including natural sources (Eucalyptus globules plant), industrial wastes (Tomato paste processing) and surface contaminants (Bathrooms). The selected yeasts were screened for melanin production in an individual experiment , results revealed the ability of yeast isolate selected from wastes of tomato paste processing (W 3 ) to produce melanin at 300 mM concentration . The selected melanized yeast was identified as Aureobasidium pullulans according to the morphological characteristics and 18S rRNA . Melanin production by the selected yeast was maximized by optimization of the cultural conditions and nutritional parameters by nearly two folds ( 550 mM ). The highest melanin yield was achieved in the fermentation medium contains 30 gl -1 sucrose , 35 gl -1 peptone and 1 mM L - DOPA concentration after nine days of incubation at 30°C and its pH value is adjusted at 7.0. Melanin characterization by FTIR and NMR resemble aliphatic and aromatic composition of A . pullulans ’ s melanin . In conclusion, the selected yeast is a promising candidate for melanin production in a semipilot scale

  10. Isolation and Screening of Thermo-Stable Cellulase Enzyme Fungal Producer at Different Temperature

    International Nuclear Information System (INIS)

    Noor Ashiqin Jamroo; Noor Azrimi Umor; Kamsani

    2015-01-01

    Thermo stable cellulase from fungi has high potential for industrial application. In this study, wild -type of fungal were isolate from different sources such as hot spring water, sea water, soft wood, rice straw and cow dung. The isolates were characterized by cultural and morphological observation. Based on morphological characteristics, the genera of all fungal cultures were identified namely Aspergillus fumigatus. The screening for thermo stable cellulase were done using 2 % carboxymethyl cellulose and congo red as an indicator at temperature 30, 37, 45 and 50 degree Celsius respectively. Out of 26 fungal isolates, only eight isolates were selected for further screening and showed the abilities to secrete cellulases by forming distinct halo zones on selective agar plate. The maximum halo zone ranging from 32 mm to 35 mm were obtained after 72 hour incubation at 50 degree Celsius by H2, SW1 and C1 isolates. As contrary other isolates showed halo zone range from 22 mm to 29 mm at same temperature. All the isolates showed the abilities to secrete cellulase enzyme at other temperature but lower when compared to 50 degree Celsius referred to the halo zone obtained. The SW1 isolates showed highest cellulolytic index which was 2.93 measured at 37 degree Celsius and 2.67 at 50 degree Celsius respectively. (author)

  11. Halotolerant ability and α-amylase activity of some saltwater fungal isolates

    NARCIS (Netherlands)

    Niknejad, F.; Moshfegh, M.; Najafzadeh, M.J.; Houbraken, J.; Rezaei, S.; Zarrini, G.; Faramarzi, M.A.; Nafissi-Varcheh, N.

    2013-01-01

    Four halotolerant fungal isolates originating from the saltwater Lake Urmia in Iran were selected during a screening program for salt resistance and α-amylase activity. The isolates were identified based on sequencing the ITS region and a part of the β-tubulin gene, as Penicillium chrysogenum

  12. HIV-1 isolation from infected peripheral blood mononuclear cells.

    Science.gov (United States)

    Dispinseri, Stefania; Saba, Elisa; Vicenzi, Elisa; Kootstra, Neeltje A; Schuitemaker, Hanneke; Scarlatti, Gabriella

    2014-01-01

    Human immunodeficiency virus 1 (HIV-1) isolation from peripheral blood mononuclear cells (PBMCs) allows retrieval of replication-competent viral variants. In order to impose the smallest possible selective pressure on the viral isolates, isolation must be carried out in primary cultures of cells and not in tumor derived cell lines. The procedure involves culture of PBMCs from an infected patient with phytohemagglutinin (PHA)-stimulated PBMC from seronegative donors, which provide susceptible target cells for HIV replication. HIV can be isolated from the bulk population of PBMCs or after cloning of the cells to obtain viral biological clones. Viral production is determined with p24 antigen (Ag) detection assays or with reverse transcriptase (RT) activity assay. Once isolated, HIV-1 can be propagated by infecting PHA-stimulated PBMCs from healthy donors. Aliquots from culture with a high production of virus are stored for later use.

  13. Evaluation of a Chromogenic Culture Medium for Isolation of Clostridium difficile within 24 Hours ▿

    Science.gov (United States)

    Perry, John D.; Asir, Kerry; Halimi, Diane; Orenga, Sylvain; Dale, Joanne; Payne, Michelle; Carlton, Ruth; Evans, Jim; Gould, F. Kate

    2010-01-01

    Rapid and effective methods for the isolation of Clostridium difficile from stool samples are desirable to obtain isolates for typing or to facilitate accurate diagnosis of C. difficile-associated diarrhea. We report on the evaluation of a prototype chromogenic medium (ID C. difficile prototype [IDCd]) for isolation of C. difficile. The chromogenic medium was compared using (i) 368 untreated stool samples that were also inoculated onto CLO medium, (ii) 339 stool samples that were subjected to alcohol shock and also inoculated onto five distinct selective agars, and (iii) standardized suspensions of 10 C. difficile ribotypes (untreated and alcohol treated) that were also inoculated onto five distinct selective agars. Two hundred thirty-six isolates of C. difficile were recovered from 368 untreated stool samples, and all but 1 of these strains (99.6%) were recovered on IDCd within 24 h, whereas 74.6% of isolates were recovered on CLO medium after 48 h. Of 339 alcohol-treated stool samples cultured onto IDCd and five other selective agars, C. difficile was recovered from 218 samples using a combination of all media. The use of IDCd allowed recovery of 96.3% of isolates within 24 h, whereas 51 to 83% of isolates were recovered within 24 h using the five other media. Finally, when they were challenged with pure cultures, all 10 ribotypes of C. difficile generated higher colony counts on IDCd irrespective of alcohol pretreatment or duration of incubation. We conclude that IDCd is an effective medium for isolation of C. difficile from stool samples within 24 h. PMID:20739493

  14. Polyclonal antibodies of Ganoderma boninense isolated from ...

    African Journals Online (AJOL)

    Polyclonal antibodies of Ganoderma boninense isolated from Malaysian oil palm for detection of basal stem rot disease. ... ELISA-PAb shows better detection as compared to cultural-based method, Ganoderma selective medium (GSM) with an improvement of 18% at nursery trial. The present study also demonstrates ...

  15. Selection and evaluation of Debaryomyces hansenii isolates as potential bioprotective agents against toxigenic penicillia in dry-fermented sausages.

    Science.gov (United States)

    Núñez, Félix; Lara, María S; Peromingo, Belén; Delgado, Josué; Sánchez-Montero, Lourdes; Andrade, María J

    2015-04-01

    Biocontrol using autochthonous Debaryomyces hansenii isolates is a potentially suitable strategy for inhibiting toxigenic moulds in dry-cured meat products. The antifungal activity of 280 D. hansenii isolated from dry-cured meat products as well as the mode of action of the most active isolates against toxigenic penicillia were evaluated in this work. A 13.9% of the D. hansenii isolates showed inhibitory activity in a radial inhibition assay. The effects on penicillia growth of both the cell-free culture filtrate and volatile compounds from active yeast isolates were analysed. Penicillia growth inhibition by D. hansenii was probably based on additive or synergistic effects of several inhibiting factors such as competition for nutrient and space, and production of soluble or volatile compounds. When four D. hansenii isolates were tested on dry-fermented sausage, two of them produced a significantly growth reduction of the ochratoxigenic Penicillium verrucosum, keeping its counts under the level considered as hazardous for the mycotoxin presence. Therefore, the use of these two D. hansenii isolates during the processing of dry-fermented meat product could be a promising tool to control toxigenic moulds in the meat industry. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. Design and test of aircraft engine isolators for reduced interior noise

    Science.gov (United States)

    Unruh, J. F.; Scheidt, D. C.

    1982-01-01

    Improved engine vibration isolation was proposed to be the most weight and cost efficient retrofit structure-borne noise control measure for single engine general aviation aircraft. A study was carried out the objectives: (1) to develop an engine isolator design specification for reduced interior noise transmission, (2) select/design candidate isolators to meet a 15 dB noise reduction design goal, and (3) carry out a proof of concept evaluation test. Analytical model of the engine, vibration isolators and engine mount structure were coupled to an empirical model of the fuselage for noise transmission evaluation. The model was used to develop engine isolator dynamic properties design specification for reduced noise transmission. Candidate isolators ere chosen from available product literature and retrofit to a test aircraft. A laboratory based test procedure was then developed to simulate engine induced noise transmission in the aircraft for a proof of concept evaluation test. Three candidate isolator configurations were evaluated for reduced structure-borne noise transmission relative to the original equipment isolators.

  17. Isolation and application of hydrocarbon degradation of indigenous microbial from oil contaminated soil

    International Nuclear Information System (INIS)

    Dadang Sudrajat; Nana Mulyana; Tri Retno DL

    2015-01-01

    The aims of this research are to obtain indigenous potential microbes from bacterial and fungal groups which have capable of degrading hydrocarbon from crude oil contaminated soil. The research carried out were isolation, selection, and identification potential microbial isolates capable of degrading hydrocarbon from oil contaminated soil located at Cepu East Java. The isolates were tested for their growth and ability to degrades crude oil. Each isolate was inoculated unto minimum mineral salt medium (MSM) contained 1% crude oil. Viability and stability test of selected isolates were carried out on irradiated compost carrier materials contained 5% crude oil. The fours series microbial s consortium consists of microbial consortium I, II, III, and IV were tested for the in vitro biodegradability of hydrocarbon. The results shows there sixty two (62) isolates are obtained, among them 42 bacteria and 20 molds. From 42 bacterial isolates, only 8 strains were potent hydrocarbon degraders. Three of these isolates are identified Bacillus cereus (BMC2), Bacillus sp (BMC4), and Pseudomonas sp (BMC6). Whereas from 20 fungal isolates, only 4 strains were potent hydrocarbon degraders. Two of these isolates are identified Aspergillus fumigatus (FMC2) and Aspergillus niger (FMC6). All isolates show good growth in mineral salt medium contained crude oil with decrease in pH. The ability of decrease of TPH content by the bacterial and fungal isolates were 54, 61, 67, 74, and 78% respectively at day 30. The viability and stability of microbial isolates show considerable good viability on irradiated compost carrier materials after 14 days storage. From the fours series microbial consortium, the highest TPH degradation rates is found in microbial consortium III (BMC6, BMC2, and FMC6) with 89,1% in 5 weeks. (author)

  18. Isolation, speciation, and antibiogram of clinically relevant non-diphtherial Corynebacteria (Diphtheroids

    Directory of Open Access Journals (Sweden)

    B S Reddy

    2012-01-01

    Full Text Available Purpose: Coryneform or the non-diphtherial Corynebacterium species largely remains a neglected group with the traditional consideration of these organisms as contaminants. This concept, however, is slowly changing in the light of recent observations. This study has been done to find out the species distribution and antibiogram of various members of the clinically relevant Coryneform group, isolated from various clinical materials. Materials and Methods: One hundred and fourteen non-duplicate isolates of diphtheroids from various clinical isolates were selected for the study. The isolates were identified to the species level by using a battery of tests; and antimicrobial susceptibility was tested by using a combination of Clinical and Laboratory Standards Institute (CLSI and the British Society for Antimicrobial Chemotherapy (BSAC guidelines, in the absence of definitive CLSI guidelines. Results: Corynebacterium amycolatum was the predominant species (35.9% in our series followed by the CDC Group G organisms (15.7%. Each of the remaining 19 species comprised of less than 10% of the isolates. More than half the total isolates were resistant to the penicillins, erythromycin, and clindamycin; while excellent activity (all the strains being susceptible was shown by vancomycin, linezolid, and tigecycline. Chloramphenicol and tetracycline also had good activity in inhibiting more than 80% of the isolates. Multiply drug resistance was exhibited by all the species. Conclusion: This study was an attempt to establish the clinical significance of coryneform organisms. The high level of resistance shown by this group to some of the common antibacterial agents highlights the importance of processing these isolates in select conditions to guide the clinicians towards an appropriate therapy.

  19. A CMOS-compatible silicon substrate optimization technique and its application in radio frequency crosstalk isolation

    International Nuclear Information System (INIS)

    Li Chen; Liao Huailin; Huang Ru; Wang Yangyuan

    2008-01-01

    In this paper, a complementary metal-oxide semiconductor (CMOS)-compatible silicon substrate optimization technique is proposed to achieve effective isolation. The selective growth of porous silicon is used to effectively suppress the substrate crosstalk. The isolation structures are fabricated in standard CMOS process and then this post-CMOS substrate optimization technique is carried out to greatly improve the performances of crosstalk isolation. Three-dimensional electro-magnetic simulation is implemented to verify the obvious effect of our substrate optimization technique. The morphologies and growth condition of porous silicon fabricated have been investigated in detail. Furthermore, a thick selectively grown porous silicon (SGPS) trench for crosstalk isolation has been formed and about 20dB improvement in substrate isolation is achieved. These results demonstrate that our post-CMOS SGPS technique is very promising for RF IC applications. (cross-disciplinary physics and related areas of science and technology)

  20. Antibiotic profiles of bacteria isolated on selective campylobacter media

    Science.gov (United States)

    The objective of this study was to determine antibiotic profiles of non-Campylobacter bacteria recovered on selective Campylobacter media. Broiler carcasses were obtained from a processing facility, and whole-carcass rinses were performed by shaking carcasses in plastic bags with 200 mL of distilled...

  1. The determination of minimum inhibitory concentrations of selected antimicrobials for porcine Haemophilus parasuis isolates from the Czech Republic

    Directory of Open Access Journals (Sweden)

    Kateřina Nedbalcová

    2017-01-01

    Full Text Available Haemophilus parasuis isolates obtained from pigs in the Czech Republic were tested for their susceptibility to amoxicillin, penicillin, ceftiofur, enrofloxacin, tetracycline, and tulathromycin by determination of minimum inhibitory concentrations using the broth microdilution method. The H. parasuis isolates were mostly susceptible to majority of tested antimicrobials (amoxicillin 90%, penicillin 73.3%, enrofloxacin 83.3%, and tulathromycin 83.3%. All isolates were susceptible to ceftiofur. On the other hand, no isolate was susceptible to tetracycline, 30% of tested isolates were intermediately susceptible, and 70% were resistant. These findings indicate that tested antimicrobials with the exception of tetracycline should be the preferred option used for the treatment of infection caused by H. parasuis but due to the potential transmission of resistance from animals to humans, the use of ceftiofur is considered as a last resort option in antimicrobial treatment of animals.

  2. Lysogenic Streptococcus suis isolate SS2-4 containing prophage SMP showed increased mortality in zebra fish compared to the wild-type isolate.

    Directory of Open Access Journals (Sweden)

    Fang Tang

    Full Text Available Streptococcus suis (S. suis infection is considered to be a major problem in the swine industry worldwide. Based on the capsular type, 33 serotypes of S. suis have been described, with serotype 2 (SS2 being the most frequently isolated from diseased piglets. Little is known, however, about the pathogenesis and virulence factors of S. suis. Research on bacteriophages highlights a new area in S. suis research. A S. suis serotype 2 bacteriophage, designated SMP, has been previously isolated in our laboratory. Here, we selected a lysogenic isolate in which the SMP phage was integrated into the chromosome of strain SS2-4. Compared to the wild-type isolate, the lysogenic strain showed increased mortality in zebra fish. Moreover the sensitivity of the lysogenic strain to lysozyme was seven times higher than that of the wild-type.

  3. Laboratory invesitgations of variability of Ascochyta fabae Speg. isolates obtained from horse bean

    Directory of Open Access Journals (Sweden)

    Antoni Józef Filipowicz

    2014-08-01

    Full Text Available Fifty five isolates of Ascochyta fabae Speg. were investigated. They were selected from 1650 isolates of this fungus obtained from horse bean seeds in 1974-1976. All the isolates grew and sporulated on Potato Dextrose Agar, Malt Agar and Horse Bean Agar. The rate of their growth amounted to 1-4 mm per 24 hours. The variability of isolates in size of pycnidia and conidia and number of sepia was noticed. A few spores with untypical shapes were observed as well.

  4. Hydrocarbon degradation and plant colonization of selected bacterial strains isolated from the rhizsophere and plant interior of Italian ryegrass and Birdsfoot trefoil

    Science.gov (United States)

    Sohail, Y.; Andria, V.; Reichenauer, T. G.; Sessitsch, A.

    2009-04-01

    Hydrocarbon-degrading strains were isolated from the rhizosphere, root and shoot interior of Italian ryegrass (Lolium multiflorum var. Taurus), Birdsfoot trefoil (Lotus corniculatus var. Leo) grown in a soil contaminated with petroleum oil. Strains were tested regarding their phylogeny and their degradation efficiency. The most efficient strains were tested regarding their suitability to be applied for phytoremediation of diesel oils. Sterilized and non-sterilized agricultural soil, with and with out compost, were spiked with diesel and used for planting Italian ryegrass and birdsfoot trefoil. Four selected strains with high degradation activities, derived from the rhizosphere and plant interior, were selected for individual inoculation. Plants were harvested at flowering stage and plant biomass and hydrocarbon degradation was determined. Furthermore, it was investigated to which extent the inoculant strains were able to survive and colonize plants. Microbial community structures were analysed by 16S rRNA and alkB gene analysis. Results showed efficient colonization by the inoculant strains and improved degradation by the application of compost combined with inoculation as well as on microbial community structures will be presented.

  5. TiSH - a robust and sensitive global phosphoproteomics strategy employing a combination of TiO(2), SIMAC, and HILIC

    DEFF Research Database (Denmark)

    Engholm-Keller, Kasper; Birck, Pernille; Størling, Joachim

    2012-01-01

    losses. We demonstrate the capability of this strategy by quantitative investigation of early interferon-γ signaling in low quantities of insulinoma cells. We identified ~6600 unique phosphopeptides from 300μg of peptides/condition (22 unique phosphopeptides/μg) in a duplex dimethyl labeling experiment....... This strategy thus shows great potential for interrogating signaling networks from low amounts of sample with high sensitivity and specificity....

  6. Phosphoproteins in extracellular vesicles as candidate markers for breast cancer.

    Science.gov (United States)

    Chen, I-Hsuan; Xue, Liang; Hsu, Chuan-Chih; Paez, Juan Sebastian Paez; Pan, Li; Andaluz, Hillary; Wendt, Michael K; Iliuk, Anton B; Zhu, Jian-Kang; Tao, W Andy

    2017-03-21

    The state of protein phosphorylation can be a key determinant of cellular physiology such as early-stage cancer, but the development of phosphoproteins in biofluids for disease diagnosis remains elusive. Here we demonstrate a strategy to isolate and identify phosphoproteins in extracellular vesicles (EVs) from human plasma as potential markers to differentiate disease from healthy states. We identified close to 10,000 unique phosphopeptides in EVs isolated from small volumes of plasma samples. Using label-free quantitative phosphoproteomics, we identified 144 phosphoproteins in plasma EVs that are significantly higher in patients diagnosed with breast cancer compared with healthy controls. Several biomarkers were validated in individual patients using paralleled reaction monitoring for targeted quantitation. This study demonstrates that the development of phosphoproteins in plasma EV as disease biomarkers is highly feasible and may transform cancer screening and monitoring.

  7. Isolation and characterization of biosurfactant producing bacteria from Persian Gulf (Bushehr provenance).

    Science.gov (United States)

    Hassanshahian, Mehdi

    2014-09-15

    Biosurfactants are surface active materials that are produced by some microorganisms. These molecules increase biodegradation of insoluble pollutants. In this study sediments and seawater samples were collected from the coastline of Bushehr provenance in the Persian Gulf and their biosurfactant producing bacteria were isolated. Biosurfactant producing bacteria were isolated by using an enrichment method in Bushnell-Hass medium with diesel oil as the sole carbon source. Five screening tests were used for selection of Biosurfactant producing bacteria: hemolysis in blood agar, oil spreading, drop collapse, emulsification activity and Bacterial Adhesion to Hydrocarbon test (BATH). These bacteria were identified using biochemical and molecular methods. Eighty different colonies were isolated from the collected samples. The most biosurfactant producing isolates related to petrochemical plants of Khark Island. Fourteen biosurfactant producing bacteria were selected between these isolates and 7 isolates were screened as these were predominant producers that belong to Shewanella alga, Shewanella upenei, Vibrio furnissii, Gallaecimonas pentaromativorans, Brevibacterium epidermidis, Psychrobacter namhaensis and Pseudomonas fluorescens. The largest clear zone diameters in oil spreading were observed for G. pentaromativorans strain O15. Also, this strain has the best emulsification activity and reduction of surface tension, suggesting it is the best of thee isolated strains. The results of this study confirmed that there is high diversity of biosurfactant producing bacteria in marine ecosystem of Iran and by application of these bacteria in petrochemical waste water environmental problems can be assisted. Copyright © 2014 Elsevier Ltd. All rights reserved.

  8. Susceptibility of peach GF 305 seedlings and selected herbaceous plants to plum pox virus isolates from western Slovakia.

    Science.gov (United States)

    Glasa, M; Matisová, J; Hricovský, I; Kúdela, O

    1997-12-01

    The susceptibility of peach GF 305 seedlings and herbaceous plants to five plum pox virus (PPV) isolates from orchards of western Slovakia was investigated. PPV was isolated from diseased plum, apricot and peach trees, and transmitted by chip-budding to peach GF 305. The herbaceous plants were infected by mechanical inoculation. The transmission was analysed by symptomatology and double sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Infected peaches developed leaf distortion, tissue clearing along the veins and small chlorotic spots (isolate BOR-3). With exception of BOR-3, the PPV isolates transmitted from peach caused local chlorotic spots on Chenopodium foetidum. The character of symptoms changed when a sap from PPV-infected Nicotiana benthamiana was used as virus inoculum. From N. benthamiana, the PPV isolates could be transmitted to Pisum sativum, cv. Colmo (light green mosaic), N. clevelandii and N. clevelandii x N. glutinosa hybrid (latent infection or chlorotic spots).

  9. A protocol for large scale genomic DNA isolation for cacao genetics ...

    African Journals Online (AJOL)

    Advances in DNA technology, such as marker assisted selection, detection of quantitative trait loci and genomic selection also require the isolation of DNA from a large number of samples and the preservation of tissue samples for future use in cacao genome studies. The present study proposes a method for the ...

  10. A Seismic Isolation Application Using Rubber Bearings; Hangar Project in Turkey

    International Nuclear Information System (INIS)

    Sesigur, Haluk; Cili, Feridun

    2008-01-01

    Seismic isolation is an effective design strategy to mitigate the seismic hazard wherein the structure and its contents are protected from the damaging effects of an earthquake. This paper presents the Hangar Project in Sabiha Goekcen Airport which is located in Istanbul, Turkey. Seismic isolation system where the isolation layer arranged at the top of the columns is selected. The seismic hazard analysis, superstructure design, isolator design and testing were based on the Uniform Building Code (1997) and met all requirements of the Turkish Earthquake Code (2007). The substructure which has the steel vertical trusses on facades and RC H shaped columns in the middle axis of the building was designed with an R factor limited to 2.0 in accordance with Turkish Earthquake Code. In order to verify the effectiveness of the isolation system, nonlinear static and dynamic analyses are performed. The analysis revealed that isolated building has lower base shear (approximately 1/4) against the non-isolated structure

  11. Potential of Glycosidase from Non-Saccharomyces Isolates for Enhancement of Wine Aroma.

    Science.gov (United States)

    Hu, Kai; Qin, Yi; Tao, Yong-Sheng; Zhu, Xiao-Lin; Peng, Chuan-Tao; Ullah, Niamat

    2016-04-01

    The aim of this work was to rapidly screen indigenous yeasts with high levels of β-glucosidase activity and assess the potential of glycosidase extracts for aroma enhancement in winemaking. A semiquantitative colorimetric assay was applied using 96-well plates to screen yeasts from 3 different regions of China. Isolates with high β-glucosidase activity were confirmed by the commonly used pNP assay. Among 493 non-Saccharomyces isolates belonging to 8 generas, 3 isolates were selected for their high levels of β-glucosidase activity and were identified as Hanseniaspora uvarum, Pichia membranifaciens, and Rhodotorula mucilaginosa by sequence analysis of the 26S rDNA D1/D2 domain. β-Glucosidase in the glycosidase extract from H. uvarum strain showed the highest activity in winemaking conditions among the selected isolates. For aroma enhancement in winemaking, the glycosidase extract from H. uvarum strain exhibited catalytic specificity for aromatic glycosides of C13 -norisoprenoids and some terpenes, enhancing fresh floral, sweet, berry, and nutty aroma characteristics in wine. © 2016 Institute of Food Technologists®

  12. Mycoremediation of Textile Dyes: Application of Novel Autochthonous Fungal Isolates

    Directory of Open Access Journals (Sweden)

    Sweety

    2017-07-01

    Full Text Available Four fungal isolates Trichoderma virens, Phlebiopsis cf. ravenelii, Talaromyces stipitatus, Aspergillus niger originally isolated from the textile dye contaminated soil of Meerut (U.P. India. They were used for the decolorization studies of selected textile azo dyes under laboratory conditions. Out of total 74 isolates, selected four fungal strains were picked on the basis of primary screening carried out using agar layer decolorization method. Decolorization efficiency of textile dyes was studied at an interval of 3, 5, 7 and 9 days at temperatures 20, 25, 30 and 40°C using five synthetic dyes viz. Xylene cynol FF, Brilliant blue R, Aniline Blue, Orange G II and Crystal violet. Decolorization study was carried out under shaking and stationary conditions at pH 4.0, 5.4, 6.5, and 8.0. The results obtained showed that Trichoderma virens and Aspergillus niger were more efficient then Phlebiopsis cf. ravenelii and Talaromyces stipitatus. Highest biodegradation activities of dyes by these aboriginal fungal isolates were observed at pH 5.4 after 9 days of incubation. Maximum decolorization 99.84 % was achieved by Aspergillus niger, followed by Trichoderma virens. This is the first report where the bioremediation aspects of Phlebiopsis cf. ravenelii and Talaromyces stipitatus has been revealed.

  13. Identification of potential local isolated for biosurfactant production

    Science.gov (United States)

    Shafiei, Zahra; Yusoff, Wan Mohtar Wan; Hamid, Aidil Abdul; Moazami, Nasrin; Hamzah, Ainon; Fooladi, Taybeh

    2013-11-01

    Biosurfactant are amphiphilic molecule that have received increasing attention in recent years because of their role in the growth of microorganisms on water-insoluble hydrophobic materials such as hydrocarbons as well as their commercial potential in the cosmetics, food, oil recovery and agricultural industries. In this study a potential biosurfactant producing strain was isolated from several soil samples of Terengganu oil refinery, Malaysia and selected during preliminary screening using hemolytic activity, oil spreading and drop collapsed technique. Isolates with at least more than one positive response to these three methods were subjected to complementary screening by measuring surface tension reduction as well as emulsification capacity. The biosurfactant produced by isolated 5M was able to reduced surface tension of culture medium from 60 mN/m to30mN/m. The biochemical and morphological characterization, 16SrRNA gene sequencing showed that the isolated 5M belongs to bacillus groups. The maximum production of biosurfactant by Bacillus 5M was observed after 48 h of incubation.

  14. The isolation of Moellerella wisconsensis from stool samples in the U.K.

    Science.gov (United States)

    Marshall, A R; Al-Jumaili, I J; Bint, A J

    1986-01-01

    Three strains of Moellerella wisconsensis were isolated from a total of 400 stool specimens screened for this organism by means of a new selective medium developed in this laboratory. This is the first report of the isolation of this organism in the U.K. The exact role of M. wisconsensis in causing diarrhoea remains to be elucidated.

  15. Radioactive-waste isolation pilot plant

    International Nuclear Information System (INIS)

    Weart, W.D.

    1977-01-01

    The objective of the Waste Isolation Pilot Plant (WIPP) program is to demonstrate the suitability of bedded salt, specifically, the bedded salt deposits in the Los Medanos area of southeastern New Mexico, as a disposal medium for radioactive wastes. Our program responsibilities include site selection considerations, all aspects of design and development, technical guidance of facility operation, environmental impact assessment, and technical support to ERDA for developing public understanding of the facility

  16. Evidence of adaptive tolerance to nickel in isolates of Cenococcum geophilum from serpentine soils.

    Science.gov (United States)

    Gonçalves, Susana C; Martins-Loução, M Amélia; Freitas, Helena

    2009-04-01

    Selection for metal-tolerant ecotypes of ectomycorrhizal (ECM) fungi has been reported in instances of metal contamination of soils as a result of human activities. However, no study has yet provided evidence that natural metalliferous soils, such as serpentine soils, can drive the evolution of metal tolerance in ECM fungi. We examined in vitro Ni tolerance in isolates of Cenococcum geophilum from serpentine and non-serpentine soils to assess whether isolates from serpentine soils exhibited patterns consistent with adaptation to elevated levels of Ni, a typical feature of serpentine. A second objective was to investigate the relationship between Ni tolerance and specific growth rates (micro) among isolates to increase our understanding of possible tolerance/growth trade-offs. Isolates from both soil types were screened for Ni tolerance by measuring biomass production in liquid media with increasing Ni concentrations, so that the effective concentration of Ni inhibiting fungal growth by 50% (EC(50)) could be determined. Isolates of C. geophilum from serpentine soils exhibited significantly higher tolerance to Ni than non-serpentine isolates. The mean Ni EC(50) value for serpentine isolates (23.4 microg ml(-1)) was approximately seven times higher than the estimated value for non-serpentine isolates (3.38 microg ml(-1)). Although there was still a considerable variation in Ni sensitivity among the isolates, none of the serpentine isolates had EC(50) values for Ni within the range found for non-serpentine isolates. We found a negative correlation between EC(50) and micro values among isolates (r = -0.555). This trend, albeit only marginally significant (P = 0.06), indicates a potential trade-off between tolerance and growth, in agreement with selection against Ni tolerance in "normal" habitats. Overall, these results suggest that Ni tolerance arose among serpentine isolates of C. geophilum as an adaptive response to Ni exposure in serpentine soils.

  17. Isolation of cells for selective treatment and analysis using a magnetic microfluidic chip

    KAUST Repository

    Yassine, Omar; Gooneratne, Chinthaka Pasan; Abu Smara, D.; Li, F.; Mohammed, H.; Merzaban, Jasmeen; Kosel, Jü rgen

    2014-01-01

    . Experiments were performed with E. coli (K12 strand) tagged with 2.8 μm SPBs. The results showed that E. coli can be separated from a sample solution by trapping them at the disk sites, and then isolated into chambers by transporting them along the tapered

  18. Sexual imprinting on ecologically divergent traits leads to sexual isolation in sticklebacks.

    Science.gov (United States)

    Kozak, Genevieve M; Head, Megan L; Boughman, Janette W

    2011-09-07

    During sexual imprinting, offspring learn parental phenotypes and then select mates who are similar to their parents. Imprinting has been thought to contribute to the process of speciation in only a few rare cases; this is despite imprinting's potential to generate assortative mating and solve the problem of recombination in ecological speciation. If offspring imprint on parental traits under divergent selection, these traits will then be involved in both adaptation and mate preference. Such 'magic traits' easily generate sexual isolation and facilitate speciation. In this study, we show that imprinting occurs in two ecologically divergent stickleback species (benthics and limnetics: Gasterosteus spp.). Cross-fostered females preferred mates of their foster father's species. Furthermore, imprinting is essential for sexual isolation between species; isolation was reduced when females were raised without fathers. Daughters imprinted on father odour and colour during a critical period early in development. These traits have diverged between the species owing to differences in ecology. Therefore, we provide the first evidence that imprinting links ecological adaptation to sexual isolation between species. Our results suggest that imprinting may facilitate the evolution of sexual isolation during ecological speciation, may be especially important in cases of rapid diversification, and thus play an integral role in the generation of biodiversity.

  19. Screening of Azotobacter isolates for PGP properties and antifungal activity

    Directory of Open Access Journals (Sweden)

    Bjelić Dragana Đ.

    2015-01-01

    Full Text Available Аmong 50 bacterial isolates obtained from maize rhizospherе, 13 isolates belonged to the genus Azotobacter. Isolates were biochemically characterized and estimated for pH and halo tolerance ability and antibiotic resistance. According to characterization, the six representative isolates were selected and further screened in vitro for plant growth promoting properties: production of indole-3-acetic acid (IAA, siderophores, hydrogen cyanide (HCN, exopolysaccharides, phosphate solubilization and antifungal activity (vs. Helminthosporium sp., Macrophomina sp., Fusarium sp.. Beside HCN production, PGP properties were detected for all isolates except Azt7. All isolates produced IAA in the medium without L-tryptophan and the amount of produced IAA increased with concentration of precursor in medium. The highest amount of IAA was produced by isolates Azt4 (37.69 and 45.86 μg ml-1 and Azt5 (29.44 and 50.38 μg ml-1 in the medium with addition of L-tryptophan (2.5 and 5 mM. The isolates showed the highest antifungal activity against Helminthosporium sp. and the smallest antagonistic effect on Macrophomina sp. Radial Growth Inhibition (RGI obtained by the confrontation of isolates with tested phytopathogenic fungi, ranged from 10 to 48%. [Projekat Ministarstva nauke Republike Srbije, br. TR 31073

  20. Isolation and characterization of biosurfactant producing bacteria from Persian Gulf (Bushehr provenance)

    International Nuclear Information System (INIS)

    Hassanshahian, Mehdi

    2014-01-01

    Highlights: • Biosurfactant producing bacteria were isolated from Persian Gulf. • There is high diversity of biosurfactant producing bacteria in the Persian Gulf. • These bacteria are very useful for management of oil pollution in the sea. - Abstract: Biosurfactants are surface active materials that are produced by some microorganisms. These molecules increase biodegradation of insoluble pollutants. In this study sediments and seawater samples were collected from the coastline of Bushehr provenance in the Persian Gulf and their biosurfactant producing bacteria were isolated. Biosurfactant producing bacteria were isolated by using an enrichment method in Bushnell-Hass medium with diesel oil as the sole carbon source. Five screening tests were used for selection of Biosurfactant producing bacteria: hemolysis in blood agar, oil spreading, drop collapse, emulsification activity and Bacterial Adhesion to Hydrocarbon test (BATH). These bacteria were identified using biochemical and molecular methods. Eighty different colonies were isolated from the collected samples. The most biosurfactant producing isolates related to petrochemical plants of Khark Island. Fourteen biosurfactant producing bacteria were selected between these isolates and 7 isolates were screened as these were predominant producers that belong to Shewanella alga, Shewanella upenei, Vibrio furnissii, Gallaecimonas pentaromativorans, Brevibacterium epidermidis, Psychrobacter namhaensis and Pseudomonas fluorescens. The largest clear zone diameters in oil spreading were observed for G. pentaromativorans strain O15. Also, this strain has the best emulsification activity and reduction of surface tension, suggesting it is the best of thee isolated strains. The results of this study confirmed that there is high diversity of biosurfactant producing bacteria in marine ecosystem of Iran and by application of these bacteria in petrochemical waste water environmental problems can be assisted

  1. Selective vs. nonselective media and direct plating vs. enrichment technique in isolation of Vibrio cholerae: recommendations for clinical laboratories.

    Science.gov (United States)

    Rennels, M B; Levine, M M; Daya, V; Angle, P; Young, C

    1980-09-01

    The occurrence of human cholera along the Gulf of Mexico and the isolation of Vibrio cholerae O1 from the Gulf and Chesapeake Bay make it imperative that microbiology laboratories along estuaries develop the capabilities to culture for these pathogens. In attempts to devise a simplified but efficient culture procedure, a selective medium, thiosulfate-citrate-bile salts-sucrose (TCBS) agar, was compared with a nonselective medium, gelatin agar (GA), and the utility of enrichment was examined. TCBS agar detected 99% of the stools found to be positive by all techniques combined, whereas GA identified only 80%. Of acute diarrheal stools, 96% were positive on direct plating, whereas only 66% of formed stools containing V. cholerae were detected by direct plating. Stools from patients with acute diarrhea can be plated directly into TCBS agar alone; stools from persons shedding low numbers of organisms (such as contacts, carriers, or patients receiving antibiotics) should be incubated first in an enrichment broth and then on TCBS agar.

  2. Genetic Diversity of a Natural Population of Apple stem pitting virus Isolated from Apple in Korea

    Directory of Open Access Journals (Sweden)

    Ju Yeon Yoon

    2014-06-01

    Full Text Available Apple stem pitting virus (ASPV, of the Foveavirus genus in the family Betaflexiviridae, is one of the most common viruses of apple and pear trees. To examine variability of the coat protein (CP gene from ASPV, eight isolates originating from 251 apple trees, which were collected from 22 apple orchards located in intensive apple growing areas of the North Gyeongsang and North Jeolla Provinces in Korea, were sequenced and compared. The nucleotide sequence identity of the CP gene of eight ASPV isolates ranged from 77.0 to 97.0%, while the amino acid sequence identity ranged from 87.7 to 98.5%. The N-terminal region of the viral CP gene was highly variable, whereas the C-terminal region was conserved. Genetic algorithm recombination detection (GARD and single breakpoint recombination (SBP analyses identified base substitutions between eight ASPV isolates at positions 54 and 57 and position 771, respectively. GABranch analysis was used to determine whether the eight isolates evolved due to positive selection. All values in the GABranch analysis showed a ratio of substitution rates at non-synonymous and synonymous sites (dNS/dS below 1, suggestive of strong negative selection forces during ASPV CP history. Although negative selection dominated CP evolution in the eight ASPV isolates, SLAC and FEL tests identified four possible positive selection sites at codons 10, 22, 102, and 158. This is the first study of the ASPV genome in Korea.

  3. Isolation and molecular characterization of phytase producing ...

    African Journals Online (AJOL)

    Isolation and molecular characterization of phytase producing bacteria from Malaysia hot springs. ... The strains were further analyzed in broth culture using sodium phytate as substrate. Among them, strain L3 was selected as the best producer (0.16 U/ml after 72 h of culture). This phytase showed optimal activity at 37 °C ...

  4. [Isolation of actinobacteria with antibiotic associated with soft coral Nephthea sp].

    Science.gov (United States)

    Ma, Liang; Zhang, Wenjun; Zhu, Yiguang; Wu, Zhengchao; Saurav, Kumar; Hang, Hui; Zhang, Changsheng

    2013-10-04

    The present study aims to isolate and identify actinobacteria associated with the soft coral Nephthea sp., and to isolate natural products from these actinobacteria under the guidance of PCR screening for polyketides synthase (PKS) genes. Eleven selective media were used to isolate actinobacteria associated with the soft coral Nephthea sp. collected from Yongxin Island. The isolated actinobacteria were classified on the basis of phylogenetic tree analysis of their 16S rRNA genes. Degenerated primers targeted on conserved KS (ketoacyl-synthase) domain of type I PKS genes were used to screen for potential isolates. The positive isolates were cultured in three different media to check their producing profiles. One bioactive strain that is rich in metabolites was subjected to larger scale fermentation for isolating bioactive natural products. A total of 20 strains were isolated from Nephthea sp., and were categorized into 3 genera including Streptomyces, Dietzia and Salinospora, among which 18 strains were positive in screening with type I PKS genes. Two bioactive compounds rifamycin S and rifamycin W were isolated and identified from Salinospora arenicola SH04. This is the first report of isolating indigenous marine actinobacteria Salinospora from the soft coral Nephthea sp. It provides an example of isolating bioactive secondary metabolites from cultivable actinobacteria associated with Nephthea sp. by PCR screening.

  5. Possible bioremediation of arsenic toxicity by isolating indigenous bacteria from the middle Gangetic plain of Bihar, India

    Directory of Open Access Journals (Sweden)

    Ghanshyam Kumar Satyapal

    2018-03-01

    Full Text Available In middle Gangetic plain, high arsenic concentration is present in water, which causes a significant health risk. Total 48 morphologically distinct arsenite resistant bacteria were isolated from middle Gangetic plain. The minimum inhibitory concentration (MIC values of arsenite varied widely in the range 1–15 mM of the isolates. On the basis of their MIC, two isolates, AK1 (KY569423 and AK9 (KY569424 were selected. The analysis of the 16S rRNA gene sequence of selected isolates revealed that they are belong to the genus Pseudomonas. The AgNO3 test based microplate method revealed that isolates, AK1 and AK9, have potential in transformation of arsenic species. Further, the presence of aoxR, aoxB and aoxC genes in the both isolated strain AK1 and AK9 was confirmed, which play an important role in arsenic bioremediation by arsenite oxidation. Isolated strains also showed heavy metal resistance against Cr(IV, Ni(II, Co(II, Pb(II, Cu(II, Hg(II, Ag(I and Cd(II.

  6. Geological problems in radioactive waste isolation

    International Nuclear Information System (INIS)

    Witherspoon, P.A.

    1991-01-01

    The problem of isolating radioactive wastes from the biosphere presents specialists in the fields of earth sciences with some of the most complicated problems they have ever encountered. This is especially true for high level waste (HLW) which must be isolated in the underground and away from the biosphere for thousands of years. Essentially every country that is generating electricity in nuclear power plants is faced with the problem of isolating the radioactive wastes that are produced. The general consensus is that this can be accomplished by selecting an appropriate geologic setting and carefully designing the rock repository. Much new technology is being developed to solve the problems that have been raised and there is a continuing need to publish the results of new developments for the benefit of all concerned. The 28th International Geologic Congress that was held July 9--19, 1989 in Washington, DC provided an opportunity for earth scientists to gather for detailed discussions on these problems. Workshop W3B on the subject, ''Geological Problems in Radioactive Waste Isolation -- A World Wide Review'' was organized by Paul A Witherspoon and Ghislain de Marsily and convened July 15--16, 1989 Reports from 19 countries have been gathered for this publication. Individual papers have been cataloged separately

  7. Geological problems in radioactive waste isolation

    Energy Technology Data Exchange (ETDEWEB)

    Witherspoon, P.A. (ed.)

    1991-01-01

    The problem of isolating radioactive wastes from the biosphere presents specialists in the fields of earth sciences with some of the most complicated problems they have ever encountered. This is especially true for high level waste (HLW) which must be isolated in the underground and away from the biosphere for thousands of years. Essentially every country that is generating electricity in nuclear power plants is faced with the problem of isolating the radioactive wastes that are produced. The general consensus is that this can be accomplished by selecting an appropriate geologic setting and carefully designing the rock repository. Much new technology is being developed to solve the problems that have been raised and there is a continuing need to publish the results of new developments for the benefit of all concerned. The 28th International Geologic Congress that was held July 9--19, 1989 in Washington, DC provided an opportunity for earth scientists to gather for detailed discussions on these problems. Workshop W3B on the subject, Geological Problems in Radioactive Waste Isolation -- A World Wide Review'' was organized by Paul A Witherspoon and Ghislain de Marsily and convened July 15--16, 1989 Reports from 19 countries have been gathered for this publication. Individual papers have been cataloged separately.

  8. Heavy metal and antibiotic resistance of Acinetobacter spp. isolated from diesel fuel polluted

    Directory of Open Access Journals (Sweden)

    Kais Kassim Ghaima

    2018-04-01

    Full Text Available Heavy metals pollution of soil and wastewater is a global problem that threatens the environment as they are not degraded or removed and the potential threat to human health comes from the multiple resistances to heavy metals and antibiotics among bacterial populations. The present study was aimed to isolate and identify multiple metal/antibiotic resistant Acinetobacter spp. from diesel fuel polluted soil of Al-Dora, Baghdad, Iraq. Initially, a total of 24 bacterial cultures (coded KNZ–1 to KNZ–24 were isolated and identified up to genus level as Acinetobacter by morphological, physiological and biochemical characteristics. Screening of heavy metals resistant Acinetobacter were conducted by streaking the isolates on nutrient agar plates supplemented with different concentrations: 10, 25, 50 and 100mg/L of the three heavy metals; Hg2+, Cd2+ and Pb2+. Out of 24 isolates, 6 (25% isolates (KNZ–3, KNZ–5, KNZ–8, KNZ–12, KNZ–16 and KNZ–21 were selected as a multiple heavy metal resistant (MHMR Acinetobacter with maximum tolerable concentrations (MTCs; 100–200mg/L for Hg2+, 300-600mg/L for Cd2+ and 100–300mg/L for Pb2+. Antibiotic resistance pattern of the selected MHMR isolates was determined by Kirby-Bauer disc diffusion method against 12 different antibiotics belonging to 7 classes. Out of 6 isolates, 4 isolates were multidrug resistance (MDR with varying degrees. Among them isolate, KNZ–16 showed a wide range of resistance to all tested antibiotics except Levofloxacin and Imipenem. It was concluded that dual resistant Acinetobacter is useful in the bioremediation of environments polluted with heavy metals especially the biodegradation of organic pollutants.

  9. Selection and Characterization of Endophytic Bacteria as Biocontrol Agents of Tomato Bacterial Wilt Disease

    Directory of Open Access Journals (Sweden)

    ABDJAD ASIH NAWANGSIH

    2011-06-01

    Full Text Available Biological control of bacterial wilt pathogen (Ralstonia solanacearum of tomato using endophytic bacteria is one of the alternative control methods to support sustainable agriculture. This study was conducted to select and characterize endophytic bacteria isolated from healthy tomato stems and to test their ability to promote plant growth and suppress bacterial wilt disease. Among 49 isolates successfully isolated, 41 were non-plant pathogenic. Green house test on six selected isolates based on antagonistic effect on R. solanacearum or ability to suppress R. solanacearum population in dual culture assays obtained BC4 and BL10 isolates as promising biocontrol agents. At six weeks after transplanting, plants treated with BC4 isolate showed significantly lower disease incidence (33% than that of control (83%. Plants height was not significantly affected by endophytic bacterial treatments. Based on 16S rRNA sequence, BC4 isolate had 97% similarity with Staphylococcus epidermidis (accession number EU834240.1, while isolate BL10 had 98% similarity with Bacillus amyloliquefaciens strain JK-SD002 (accession number AB547229.1.

  10. Isolation and characterization of actinomycete antagonists of a fungal root pathogen.

    Science.gov (United States)

    Crawford, D L; Lynch, J M; Whipps, J M; Ousley, M A

    1993-11-01

    By use of selective media, 267 actinomycete strains were isolated from four rhizosphere-associated and four non-rhizosphere-associated British soils. Organic media with low nutrient concentrations were found to be best for isolating diverse actinomycetes while avoiding contamination and overgrowth of isolation media by eubacteria and fungi. While all isolates grew well at pHs 6.5 to 8.0, a few were unable to grow at pH 6.0 and a significant number failed to grow at pH 5.5. Eighty-two selected isolates were screened for in vitro antagonism towards Pythium ultimum by use of a Difco cornmeal agar assay procedure. Five isolates were very strong antagonists of the fungus, four were strong antagonists, and ten others were weakly antagonistic. The remaining isolates showed no antagonism by this assay. Additional studies showed that several of the P. ultimum antagonists also strongly inhibited growth of other root-pathogenic fungi. Twelve isolates showing antifungal activity in the in vitro assay were also tested for their effects on the germination and short-term growth of lettuce plants in glasshouse pot studies in the absence of pathogens. None of the actinomycetes prevented seed germination, although half of the isolates retarded seed germination and outgrowth of the plants by 1 to 3 days. During 18-day growth experiments, biomass yields of some actinomycete-inoculated plants were reduced in comparison with untreated control plants, although all plants appeared healthy and well rooted. None of the actinomycetes significantly enhanced plant growth over these short-term experiments. For some, but not all, actinomycetes, some correlations between delayed seed germination and reduced 18-day plant biomass yields were seen. For others, plant biomass yields were not reduced despite an actinomycete-associated delay in seed germination and plant outgrowth. Preliminary glasshouse experiments indicated that some of the actinomycetes protect germinating lettuce seeds against

  11. Isolation of glutathione-deficient mutants of the yeast Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Kistler, M.; Eckardt, F.; Summer, K.-H.

    1986-01-01

    Glutathione-deficient (gsh - ) mutants of the yeast Saccharomyces cerevisiae were isolated after UV treatment using MNNG as selective agent. For genetic and biochemical characterization 5 mutant strains were chosen which exhibited considerably decreased residual GSH contents varying from 2 to 6% of the wild-type levels. All 5 isolates showed a 2:2 segregation of the gsh - :GSH + phenotypes alluding to a monogenic recessive mutation. Complementation analysis indicates that all gsh - mutants belong to one complementation group. (Auth.)

  12. Salt-induced changes in cardiac phosphoproteome in a rat model of chronic renal failure.

    Directory of Open Access Journals (Sweden)

    Zhengxiu Su

    Full Text Available Heart damage is widely present in patients with chronic kidney disease. Salt diet is the most important environmental factor affecting development of chronic renal failure and cardiovascular diseases. The proteins involved in chronic kidney disease -induced heart damage, especially their posttranslational modifications, remain largely unknown to date. Sprague-Dawley rats underwent 5/6 nephrectomy (chronic renal failure model or sham operation were treated for 2 weeks with a normal-(0.4% NaCl, or high-salt (4% NaCl diet. We employed TiO2 enrichment, iTRAQ labeling and liquid-chromatography tandem mass spectrometry strategy for phosphoproteomic profiling of left ventricular free walls in these animals. A total of 1724 unique phosphopeptides representing 2551 non-redundant phosphorylation sites corresponding to 763 phosphoproteins were identified. During normal salt feeding, 89 (54% phosphopeptides upregulated and 76 (46% phosphopeptides downregulated in chronic renal failure rats relative to sham rats. In chronic renal failure rats, high salt intake induced upregulation of 84 (49% phosphopeptides and downregulation of 88 (51% phosphopeptides. Database searches revealed that most of the identified phospholproteins were important signaling molecules such as protein kinases, receptors and phosphatases. These phospholproteins were involved in energy metabolism, cell communication, cell differentiation, cell death and other biological processes. The Search Tool for the Retrieval of Interacting Genes analysis revealed functional links among 15 significantly regulated phosphoproteins in chronic renal failure rats compared to sham group, and 23 altered phosphoproteins induced by high salt intake. The altered phosphorylation levels of two proteins involved in heart damage, lamin A and phospholamban were validated. Expression of the downstream genes of these two proteins, desmin and SERCA2a, were also analyzed.

  13. Comparative study of the functional properties of three legume seed isolates: adzuki, pea and soy bean.

    Science.gov (United States)

    Barac, Miroljub B; Pesic, Mirjana B; Stanojevic, Sladjana P; Kostic, Aleksandar Z; Bivolarevic, Vanja

    2015-05-01

    The aim of this work was to compare functional properties including solubility, emulsifying and foaming properties of native and thermally treated adzuki, soy and pea protein isolates prepared under the same conditions. These functional properties were tested at four pH values: pH 3.0, pH 5.0, pH 7.0 and pH 8.0. The lowest solubility at all pH values were obtained for isolate of adzuki whereas isolates of soybean had the highest values at almost all pHs. Thermal treatment reduced solubility of soy and pea isolates at all pH values, whereas solubility of adzuki isolate was unchanged, except at pH 8. Native isolate of adzuki had the best emulsifying properties at pH 7.0 whereas at the other pH values some of native pea and soybean protein isolates were superior. After thermal treatment, depending on tested pH and selected variety all of three species could be a good emulsifier. Native soy protein isolates formed the most stable foams at all pHs. Thermal treatment significantly improved foaming properties of adzuki isolate, whereas reduced foaming capacity of soy and pea isolates, but could improve foam stability of these isolates at specific pH. Appropriate selection of legume seed as well as variety could have great importance in achievement of desirable functional properties of final products. All three tested species could find specific application in wide range of food products.

  14. Performance of culture media for the isolation and identification of Staphylococcus aureus from bovine mastitis.

    Science.gov (United States)

    Bautista-Trujillo, G U; Solorio-Rivera, J L; Rentería-Solórzano, I; Carranza-Germán, S I; Bustos-Martínez, J A; Arteaga-Garibay, R I; Baizabal-Aguirre, V M; Cajero-Juárez, M; Bravo-Patiño, A; Valdez-Alarcón, J J

    2013-03-01

    Rapid isolation and identification of pathogens is a major goal of diagnostic microbiology. In order to isolate and identify Staphylococcus aureus, a number of authors have used a variety of selective and/or differential culture media. However, to date, there are no reports comparing the efficacy of selective and differential culture media for S. aureus isolation from bovine mastitis cases using the 16S rRNA (rrs) gene sequence as a gold standard test. In the present study, we evaluated the efficacy of four selective and/or differential culture media for the isolation of S. aureus from milk samples collected from cows suffering from bovine mastitis. Four hundred and forty isolates were obtained using salt-mannitol agar (SMA, Bioxon), Staphylococcus-110 agar (S110, Bioxon), CHROMAgar Staph aureus (CSA, BD-BBL) and sheep's blood agar (SBA, BD-BBL). All bacterial isolates were identified by their typical colony morphology in the respective media, by secondary tests (for coagulase and β-haemolysis) and by partial 16S rRNA (rrs) gene sequencing as a gold standard test. Sensitivity, positive predictive and negative predictive values were higher for SMA (86.96, 52.63 and 95.95%, respectively) compared with S110 (70.00, 23.73 and 90.91%, respectively), CSA (69.23, 28.13 and 95.74%, respectively) and SBA (68.75, 37.93 and 89.58%, respectively) while specificity values were similar for all media. Data indicated that the use of culture media for S. aureus isolation combined with determination of coagulase activity and haemolysis as secondary tests improved accuracy of the identification and was in accordance with rrs gene sequence-analysis compared with the use of the culture media alone.

  15. Quantitative cardiac phosphoproteomics profiling during ischemia-reperfusion in an immature swine model

    Energy Technology Data Exchange (ETDEWEB)

    Ledee, Dolena R.; Kang, Min A.; Kajimoto, Masaki; Purvine, Samuel O.; Brewer, Heather M.; Pasa Tolic, Ljiljana; Portman, Michael A.

    2017-07-01

    Ischemia-reperfusion (I/R) results in altered metabolic and molecular responses, and phosphorylation is one of the most noted regulatory mechanisms mediating signaling mechanisms during physiological stresses. To expand our knowledge of the potential phosphoproteomic changes in the myocardium during I/R, we used Isobaric Tags for Relative and Absolute Quantitation-based analyses in left ventricular samples obtained from porcine hearts under control or I/R conditions. The data are available via ProteomeXchange with identifier PXD006066. We identified 1,896 phosphopeptides within left ventricular control and I/R porcine samples. Significant differential phosphorylation between control and I/R groups was discovered in 111 phosphopeptides from 86 proteins. Analysis of the phosphopeptides using Motif-x identified five motifs: (..R..S..), (..SP..), (..S.S..), (..S…S..), and (..S.T..). Semiquantitative immunoblots confirmed site location and directional changes in phosphorylation for phospholamban and pyruvate dehydrogenase E1, two proteins known to be altered by I/R and identified by this study. Novel phosphorylation sites associated with I/R were also identified. Functional characterization of the phosphopeptides identified by our methodology could expand our understanding of the signaling mechanisms involved during I/R damage in the heart as well as identify new areas to target therapeutic strategies.

  16. Characterization of Multilayer Piezoelectric Actuators for Use in Active Isolation Mounts

    Science.gov (United States)

    Wise, Stephanie A.; Hooker, Matthew W.

    1997-01-01

    Active mounts are desirable for isolating spacecraft science instruments from on-board vibrational sources such as motors and release mechanisms. Such active isolation mounts typically employ multilayer piezoelectric actuators to cancel these vibrational disturbances. The actuators selected for spacecraft systems must consume minimal power while exhibiting displacements of 5 to 10 micron under load. This report describes a study that compares the power consumption, displacement, and load characteristics of four commercially available multilayer piezoelectric actuators. The results of this study indicate that commercially available actuators exist that meet or exceed the design requirements used in spacecraft isolation mounts.

  17. Characterization of Staphylococcus simulans strains isolated from cases of bovine mastitis

    DEFF Research Database (Denmark)

    Aarestrup, Frank Møller; Larsen, H. D.; Jensen, N. E.

    1999-01-01

    This study was conducted to characterize Staphylococcus simulans isolated from cases of bovine mastitis. A total of 134 isolates of S. simulans selected from 80 quarters from 61 cows or heifers in 37 different herds were characterized by EcoRI ribotyping. From 22 quarters two to seven consecutive......, where 76 paired or multiple isolates were at disposal, the same ribotype was constantly found in the same quarter. This study showed that S. simulans causing bovine mastitis could be divided into relatively large number of different types, but that two types predominated. More than one type could...

  18. Site identification presentation: Basalt Waste Isolation Project

    International Nuclear Information System (INIS)

    1979-11-01

    The final step in the site identification process for the Basalt Waste Isolation Project is described. The candidate sites are identified. The site identification methodology is presented. The general objectives which must be met in selecting the final site are listed. Considerations used in the screening process are also listed. Summary tables of the guidelines used are included

  19. Novel psychrotolerant picocyanobacteria isolated from Chesapeake Bay in the winter.

    Science.gov (United States)

    Xu, Yongle; Jiao, Nianzhi; Chen, Feng

    2015-08-01

    Picocyanobacteria are major primary producers in the ocean, especially in the tropical or subtropical oceans or during warm seasons. Many "warm" picocyanobacterial species have been isolated and characterized. However, picocyanobacteria in cold environments or cold seasons are much less studied. In general, little is known about the taxonomy and ecophysiology of picocyanobacteria living in the winter. In this study, 17 strains of picocyanobacteria were isolated from Chesapeake Bay, a temperate estuarine ecosystem, during the winter months. These winter isolates belong to five distinct phylogenetic lineages, and are distinct from the picocyanobacteria previously isolated from the warm seasons. The vast majority of the winter isolates were closely related to picocyanobacteria isolated from other cold environments like Arctic or subalpine waters. The winter picocyanobacterial isolates were able to maintain slow growth or prolonged dormancy at 4°C. Interestingly, the phycoerythrin-rich strains outperformed the phycocyanin-rich strains at cold temperature. In addition, winter picocyanobacteria changed their morphology when cultivated at 4°C. The close phylogenetic relationship between the winter picocyanobacteria and the picocyanobacteria living in high latitude cold regions indicates that low temperature locations select specific ecotypes of picocyanobacteria. © 2015 Phycological Society of America.

  20. Seismic isolation structure for pool-type LMFBR - isolation building with vertically isolated floor for NSSS

    International Nuclear Information System (INIS)

    Sakurai, A.; Shiojiri, H.; Aoyagi, S.; Matsuda, T.; Fujimoto, S.; Sasaki, Y.; Hirayama, H.

    1987-01-01

    The NSSS isolation floor vibration characteristics were made clear. Especially, the side support bearing (rubber bearing) is effective for horizontal floor motion restraint and rocking motion control. Seismic isolation effects for responses of the reactor components can be sufficiently expected, using the vertical seismic isolation floor. From the analytical and experimental studies, the following has been concluded: (1) Seismic isolation structure, which is suitable for large pool-type LMFBR, were proposed. (2) Seismic response characteristics of the seismic isolation structure were investigated. It was made clear that the proposed seismic isolation (Combination of the isolated building and the isolated NSSS floor) was effective. (orig./HP)

  1. Antibiotic resistance of microbial contaminations isolated from husbandry animals and foodstuffs

    Directory of Open Access Journals (Sweden)

    Lukáš Hleba

    2014-05-01

    Full Text Available In this paper the antibiotic resistance of microbial contaminations isolated from husbandry animals and foodstuffs were investigated. Microorganisms isolated from animals and foodstuffs were contaminations of selective media as MacConkey agar for Enterobacteriaceae genera and MRS agar for lactobacilli strains. Microorganisms were isolated and puryfied by agar four ways streak plate method. Identification of isolated microorganisms was done by mass-spectrometry method in MALDI-TOF MS Biotyper. For investigation of antibiotic resistance disc diffusion method by EUCAST was used. In this study Gram-negative and Gram-positive bacteria were identified. The most resistant or multi-resistant bacteria as Pseudomonas aeruginosa, Acinetobacter lwoffi, Lysinibacillus sphaericus, Staphylococcus aureus and Staphylococcus epidermis were determined. Other identified microorganisms were resistant to one antibiotic or not at all.

  2. Bioremediation Of Heavy Metals By Pseudomonas Putida Isolated From Groundwater In Egypt.

    Directory of Open Access Journals (Sweden)

    Fawazy

    2015-08-01

    Full Text Available In this present study total four bacterial isolates were obtained from 34 collected groundwater samples in 10th of Ramadan Sharkia governorate Egypt. These isolate were grown on nutrient agar supplemented with 1mgl of iron manganese and combination between them VV. Further testing of the bacterial isolates were grown on nutrient agar supplemented with different concentrations 2 4 5 6 7 8 and 9 mgl of iron and manganese. Out of four isolates one bacterial isolate no.83 has shown the resistance to heavy metals at maximum concentration of 8mgl. Selected isolate no.S83 was identified as Pseudomonas putida S83 according to Bergeys manual depending on morphological and biochemical characteristics. Transmission electron microscopy study of P. putida isolate no. S83 showed accumulation of heavy metal salts within and external to bacterial cells. P. putida S83 have higher removal efficiency of Fe2 94.5 and Mn2 94 at concentration 2 mgl and 96 hours.

  3. Substitution rate and natural selection in parvovirus B19

    Science.gov (United States)

    Stamenković, Gorana G.; Ćirković, Valentina S.; Šiljić, Marina M.; Blagojević, Jelena V.; Knežević, Aleksandra M.; Joksić, Ivana D.; Stanojević, Maja P.

    2016-01-01

    The aim of this study was to estimate substitution rate and imprints of natural selection on parvovirus B19 genotype 1. Studied datasets included 137 near complete coding B19 genomes (positions 665 to 4851) for phylogenetic and substitution rate analysis and 146 and 214 partial genomes for selection analyses in open reading frames ORF1 and ORF2, respectively, collected 1973–2012 and including 9 newly sequenced isolates from Serbia. Phylogenetic clustering assigned majority of studied isolates to G1A. Nucleotide substitution rate for total coding DNA was 1.03 (0.6–1.27) x 10−4 substitutions/site/year, with higher values for analyzed genome partitions. In spite of the highest evolutionary rate, VP2 codons were found to be under purifying selection with rare episodic positive selection, whereas codons under diversifying selection were found in the unique part of VP1, known to contain B19 immune epitopes important in persistent infection. Analyses of overlapping gene regions identified nucleotide positions under opposite selective pressure in different ORFs, suggesting complex evolutionary mechanisms of nucleotide changes in B19 viral genomes. PMID:27775080

  4. Isolation, Identification, and Characterization of Cadmium Resistant Pseudomonas sp. M3 from Industrial Wastewater

    OpenAIRE

    Syed Zaghum Abbas; Mohd Rafatullah; Norli Ismail; Japareng Lalung

    2014-01-01

    The present study deals with the isolation, identification, and characterization of the cadmium resistant bacteria from wastewater collected from industrial area of Penang, Malaysia. The isolate was selected based on high level of the cadmium and antibiotic resistances. On the basis of morphological, biochemical characteristics, 16S rDNA gene sequencing and phylogeny analysis revealed that the strain RZCd1 was authentically identified as Pseudomonas sp. M3. The industrial isolate showed more ...

  5. Isolation and Characterization of Lactic Acid Bacteria from Inasua

    Directory of Open Access Journals (Sweden)

    Ferymon Mahulette

    2017-04-01

    Full Text Available Inasua is a traditionally product of wet salt fish fermentation produced by Teon, Nila and Serua (TNS Communities in Central Maluku, Indonesia. The community made this fermented fish to anticipate the lean time when fisherman could not go to sea.  The  fish that used as inasua raw material is demersal fishes that live around coral reefs, such as Samandar fish (Siganatus guttatus, Gala-gala fish (Lutjanus sp. and Sikuda fish (Lethrinus ornatus. The objective of the research was to isolate and characterize of bacterial indigenous in  Inasua from three producers in Seram Island. The measurement of pH from inasua samples were 5.9, 5.0 and 5.8, respectively. The highest number of lactic acid bacteria was found from  Gala – gala inasua was 2,5x107 cfu/g sample. Isolation of all isolates bacteria from inasua showed that a total of 7 isolates of bacteria was obtained  from Samadar inasua, 9 isolates from  Gala-gala inasua, and 7 isolates from  Sikuda inasua.  From a total of 23 isolates, only 6 isolates had characteristic as lactic acid bacteria that were Gram  positive, negative catalase, and cocci shape. The microscopic characteristics  of the isolates are coccid in pairs or uniforms which combine to form tetrads. Carbohydrate utilization test  of selected isolate by using API 50 CHB kit indicated that 13 carbohydrates are fermented by these isolates  after incubation for 48 hours. The research  was concluded that the dominant bacteria in inasua sample  is  cocci-lactic acid bacteria. Keywords : fermented fish, inasua, lactic acid bacteria, MRSA medium

  6. Development of selective media for the isolation of yeasts and filamentous fungi from the sputum of adult patients with cystic fibrosis (CF).

    Science.gov (United States)

    Nagano, Yuriko; Millar, B Cherie; Goldsmith, Colin E; Walker, James M; Elborn, J Stuart; Rendall, Jackie; Moore, John E

    2008-11-01

    Yeasts and filamentous fungi are beginning to emerge as significant microbial pathogens in patients with cystic fibrosis (CF), particularly in relation to allergic-type responses, as seen in patients with allergic bronchopulmonary aspergillosis (ABPA), Aspergillus bronchitis and in invasive fungal disease in lung transplant patients. Four fungal media were compared in this study, including Sabouraud Dextrose Agar (SDA) and Medium B, with and without the addition of selective antibiotics, where antibiotic-supplemented media were designated with (+). These media were compared for their ability to suppress contaminating, mainly Gram-ve pathogens, in CF sputa (Pseudomonas aeruginosa, Burkholderia cepacia complex [BCC] organisms) and to enhance the growth of fungi present in CF sputum. Medium B consisted of glucose (16.7 g/l), agar (20 g/l), yeast extract (30 g/l) and peptone (6.8 g/l) at pH 6.3 and both SDA(+) and Medium B(+) were supplemented with cotrimethoxazole, 128 mg/l; chloramphenicol, 50 mg/l; ceftazidime, 32 mg/l; colistin, 24 mg/l). Employment of SDA(+) or Medium B(+) allowed an increase in specificity in the detection of yeasts and moulds, by 42.8% and 39.3%, respectively, over SDA when used solely. SDA(+) had a greater ability than Medium B(+) to suppress bacterial growth from predominantly Gram-ve co-colonisers. This is a significant benefit when attempting to detect and isolate fungi from the sputum of CF patients, as it largely suppressed any bacterial growth, with the exception of the BCC organisms, thus allowing for an increased opportunity to detect target fungal organisms in sputum and represented a significant improvement over the commercial medium (SDA), which is currently used. Overall, both novel selective media were superior in their ability to suppress bacteria in comparison with the commercially available SDA medium, which is routinely employed in most clinical microbiology diagnostic laboratories presently. Alternatively, Medium B(+) had a

  7. Use of a primary isolation medium for recovery of methicillin-resistant Staphylococcus aureus.

    OpenAIRE

    Van Enk, R A; Thompson, K D

    1992-01-01

    Clinical specimens frequently contain methicillin-resistant Staphylococcus aureus (MRSA) isolates in low numbers or mixed with methicillin-susceptible staphylococci, which can obscure MRSA on nonselective media. By using an oxacillin-containing mannitol-salt-based selective and differential medium on 936 respiratory specimens, we recovered 45% more MRSA isolates (29 versus 20) than on nonselective media alone.

  8. Whole-Genome Scans Provide Evidence of Adaptive Evolution in Malawian Plasmodium falciparum Isolates

    DEFF Research Database (Denmark)

    Ocholla, Harold; Preston, Mark D; Mipando, Mwapatsa

    2014-01-01

    BACKGROUND:  Selection by host immunity and antimalarial drugs has driven extensive adaptive evolution in Plasmodium falciparum and continues to produce ever-changing landscapes of genetic variation. METHODS:  We performed whole-genome sequencing of 69 P. falciparum isolates from Malawi and used......, an area of high malaria transmission. Allele frequency-based tests provided evidence of recent population growth in Malawi and detected potential targets of host immunity and candidate vaccine antigens. Comparison of the sequence variation between isolates from Malawi and those from 5 geographically...... dispersed countries (Kenya, Burkina Faso, Mali, Cambodia, and Thailand) detected population genetic differences between Africa and Asia, within Southeast Asia, and within Africa. Haplotype-based tests of selection to sequence data from all 6 populations identified signals of directional selection at known...

  9. Biocontrol properties of indigenous Trichoderma isolates from North ...

    African Journals Online (AJOL)

    IBSD

    2012-04-26

    Apr 26, 2012 ... Based on their antifungal activity in dual plate assay, 25 isolates were selected for further analysis. The interaction between the Trichoderma and fungal pathogens were ... effective biocontrol agents based on their ability to overgrow and ... bean seeds were planted in the plastic pot, which was filled with 3.

  10. Antibiotic resistant pattern of environmental isolates of Listeria ...

    African Journals Online (AJOL)

    Incidence of Listeria monocytogenes in cow manure, agricultural soil, and common vegetables sold in major markets in Ado-Ekiti, Nigeria was determined. Antibiotic resistant pattern of the isolates was examined by paper disk assay. A total of 196 environmental samples were cultured on a selective medium out of which ...

  11. Screening and selection of most potent diazotrophic cyanobacterial isolate exhibiting natural tolerance to rice field herbicides for exploitation as biofertilizer.

    Science.gov (United States)

    Singh, Surendra; Datta, Pallavi

    2006-01-01

    Periodic applications of heavy dosages of herbicides in modern rice-agriculture are a necessary evil for obtaining high crop productivity. Such herbicides are not only detrimental to weeds but biofertilizer strains of diazotrophic cyanobacteria also. It is therefore, essential to screen and select such biofertilizer strains of diazotrophic cyanobacteria exhibiting natural tolerance to common rice-field herbicides that can be further improved by mutational techniques to make biofertilizer technology a viable one. Therefore, efforts have been made to screen five dominant diazotrophic cyanobacterial forms e.g. filamentous heterocystous Nostoc punctiforme , Nostoc calcicola , Anabaena variabilis and unicellular Gloeocapsa sp. and Aphanocapsa sp. along with standard laboratory strain Nostoc muscorum ISU against increasing concentrations (0-100 mg l(-1) of four commercial grade common rice-field herbicides i.e. Arozin, Butachlor, Alachlor and 2,4-D under diazotrophic growth conditions. The lethal and IGC(50) concentrations for all four herbicides tested were found highest for A. variabilis as compared to other test cyanobacteria. The lowest reduction in chlorophyll a content, photosynthetic oxygen evolution, and N(2)-fixation was found in A. variabilis as compared to other rice field isolates and standard laboratory strain N. muscorum ISU. On the basis of prolong survival potential and lowest reductions in vital metabolic activities tested at IGC(50) concentration of four herbicides, it is concluded that A. variabilis is the most potent and promising cyanobacterial isolate as compared with other forms. This could be further improved by mutational techniques for exploitation as most potential and viable biofertilizer strain.

  12. An altered Pseudomonas diversity is recovered from soil by using nutrient-poor Pseudomonas-selective soil extract media

    DEFF Research Database (Denmark)

    Aagot, N.; Nybroe, O.; Nielsen, P.

    2001-01-01

    We designed five Pseudomonas-selective soil extract NAA media containing the selective properties of trimethoprim and sodium lauroyl sarcosine and 0 to 100% of the amount of Casamino Acids used in the classical Pseudomonas-selective Gould's S1 medium. All of the isolates were confirmed to be Pseu......We designed five Pseudomonas-selective soil extract NAA media containing the selective properties of trimethoprim and sodium lauroyl sarcosine and 0 to 100% of the amount of Casamino Acids used in the classical Pseudomonas-selective Gould's S1 medium. All of the isolates were confirmed....... Several of these analyses showed that the amount of Casamino Acids significantly influenced the diversity of the recovered Pseudomonas isolates. Furthermore, the data suggested that specific Pseudomonas subpopulations were represented on the nutrient-poor media. The NAA 1:100 medium, containing ca. 15 mg...... of organic carbon per liter, consistently gave significantly higher Pseudomonas CFU counts than Gould's S1 when tested on four Danish soils. NAA 1:100 may, therefore, be a better medium than Gould's S1 for enumeration and isolation of Pseudomonas from the low-nutrient soil environment....

  13. Differences in virulence and sporulation of Phytophthora kernoviae isolates originating from two distinct geographical regions

    Science.gov (United States)

    Phytophthora kernoviae has only been isolated from the United Kingdom (U.K.) and New Zealand. To understand what differences may exist between isolates from these two distinct geographical regions, virulence studies on three host plants and sporulation on host leaves were conducted on select isolat...

  14. Integration trumps selection in object recognition

    Science.gov (United States)

    Saarela, Toni P.; Landy, Michael S.

    2015-01-01

    Summary Finding and recognizing objects is a fundamental task of vision. Objects can be defined by several “cues” (color, luminance, texture etc.), and humans can integrate sensory cues to improve detection and recognition [1–3]. Cortical mechanisms fuse information from multiple cues [4], and shape-selective neural mechanisms can display cue-invariance by responding to a given shape independent of the visual cue defining it [5–8]. Selective attention, in contrast, improves recognition by isolating a subset of the visual information [9]. Humans can select single features (red or vertical) within a perceptual dimension (color or orientation), giving faster and more accurate responses to items having the attended feature [10,11]. Attention elevates neural responses and sharpens neural tuning to the attended feature, as shown by studies in psychophysics and modeling [11,12], imaging [13–16], and single-cell and neural population recordings [17,18]. Besides single features, attention can select whole objects [19–21]. Objects are among the suggested “units” of attention because attention to a single feature of an object causes the selection of all of its features [19–21]. Here, we pit integration against attentional selection in object recognition. We find, first, that humans can integrate information near-optimally from several perceptual dimensions (color, texture, luminance) to improve recognition. They cannot, however, isolate a single dimension even when the other dimensions provide task-irrelevant, potentially conflicting information. For object recognition, it appears that there is mandatory integration of information from multiple dimensions of visual experience. The advantage afforded by this integration, however, comes at the expense of attentional selection. PMID:25802154

  15. Integration trumps selection in object recognition.

    Science.gov (United States)

    Saarela, Toni P; Landy, Michael S

    2015-03-30

    Finding and recognizing objects is a fundamental task of vision. Objects can be defined by several "cues" (color, luminance, texture, etc.), and humans can integrate sensory cues to improve detection and recognition [1-3]. Cortical mechanisms fuse information from multiple cues [4], and shape-selective neural mechanisms can display cue invariance by responding to a given shape independent of the visual cue defining it [5-8]. Selective attention, in contrast, improves recognition by isolating a subset of the visual information [9]. Humans can select single features (red or vertical) within a perceptual dimension (color or orientation), giving faster and more accurate responses to items having the attended feature [10, 11]. Attention elevates neural responses and sharpens neural tuning to the attended feature, as shown by studies in psychophysics and modeling [11, 12], imaging [13-16], and single-cell and neural population recordings [17, 18]. Besides single features, attention can select whole objects [19-21]. Objects are among the suggested "units" of attention because attention to a single feature of an object causes the selection of all of its features [19-21]. Here, we pit integration against attentional selection in object recognition. We find, first, that humans can integrate information near optimally from several perceptual dimensions (color, texture, luminance) to improve recognition. They cannot, however, isolate a single dimension even when the other dimensions provide task-irrelevant, potentially conflicting information. For object recognition, it appears that there is mandatory integration of information from multiple dimensions of visual experience. The advantage afforded by this integration, however, comes at the expense of attentional selection. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Technological and Probiotic Traits of the Lactobacilli Isolated From Vaginal Tract of the Healthy Women for Probiotic Use

    OpenAIRE

    Bouridane, Hamida; Sifour, Mohamed; Idoui, Tayeb; Annick, Lejeune; Thonard, Philip

    2016-01-01

    Background For biotechnological application, selected lactic acid bacteria strains belonging to the genera Lactobacillus (Lb) are proposed as an alternative to the antibiotics for the prevention and treatment of urogenital tract infections. Objectives Isolating and selecting vaginal lactobacilli strains for probiotic use based on their technological and probiotic aptitudes. Materials and Methods The vaginal isolates were examined for their essential characteristics as the potential probiotic ...

  17. Identifying patients at risk of intraoperative and postoperative transfusion in isolated CABG: toward selective conservation strategies.

    Science.gov (United States)

    Arora, Rakesh C; Légaré, Jean-Francois; Buth, Karen J; Sullivan, John A; Hirsch, Gregory M

    2004-11-01

    Allogeneic blood product use during cardiac operation is often reported to exceed 40% despite published guidelines and costly blood conservation strategies. We developed a predictive model, based on eight preoperative risk factors, of allogeneic blood product transfusion rates in patients undergoing a cardiac procedure. All 3,046 consecutive, isolated coronary artery bypass graft (CABG) procedures at a university hospital from 1995 to 1998 were included. A logistic regression model was created to identify independent predictors of allogeneic blood product transfusion. This model was validated using a prospective patient sample. Overall use of allogeneic blood products was 23% with a crude operative mortality of 2.1%. In isolated, elective, first-time CABG cases, 16.9% received allogeneic blood products. Independent predictors of blood product usage in CABG patients were preoperative hemoglobin 12.0 or less, emergent operation, renal failure, female sex, age 70 years or older, left ventricular ejection fraction 0.40 or less, redo procedure, and low body surface area. Prospective validation of this model on 2,117 consecutive isolated CABG patients demonstrated an observed-to-expected allogeneic blood product transfusion rate ratio of 1.06. This internally validated logistic regression risk model is a sensitive and specific predictor of allogeneic blood product use in patients undergoing isolated CABG. Utilization of this model allows for preoperative risk stratification and may allow for more rational resource allocation of costly blood conservation strategies and blood bank resources.

  18. Isolation of keratinophilic fungi from selected soils of Sanjay Gandhi National Park, Mumbai (India).

    Science.gov (United States)

    Deshmukh, S K; Verekar, S A

    2014-12-01

    One hundred and twenty-five samples were collected from eight different sites in the vicinity of Sanjay Gandhi National Park (SGNP) and screened for the presence of keratinophilic fungi using hair baiting technique for isolation. Seventy-three isolates were recovered and identified. The cultures were identified using macro- and micro-morphological features. Their identification was also confirmed by the BLAST search of sequences of the ITS1-5.8S-ITS2 rDNA region against the NCBI/Genbank data and compared with deposited sequences for identification purpose. Thirteen species of nine genera were isolated viz. Aphanoascus durus (2.4%), Arthroderma corniculatum (1.6%), Auxarthron umbrinum (0.8%), Chrysosporium evolceanui (1.6%), Chrysosporium indicum (16.0%), Chrysosporium tropicum (2.4%), Chrysosporium zonatum (4.0%), Chrysosporium states of Arthroderma tuberculatum (0.8%), Chrysosporium state of Ctenomyces serratus (11.2%), Gymnascella dankaliensis (3.2%), Microsporum gypseum (12.0%), Myriodontium keratinophilum (0.8%) and Trichophyton mentagrophytes (1.6%). Representative of all thirteen species can release the protein in the range of 152.2-322.4 μg/mL in liquid media when grown on human hair in shake flask culture and also decompose 18.4-40.2% of human hair after four weeks of incubation. This study indicates that the soils of SGNP, Mumbai may be significant reservoirs of certain keratinophilic fungi. The keratinolytic activity of these fungi may be playing significant role in superficial infections to man and animals and recycling of keratinic material of this environment. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  19. Association of bioassays and molecular characterization to select new Bacillus thuringiensis isolates effective against Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae)

    International Nuclear Information System (INIS)

    Fatoretto, Julio C.; Sena, Janete A.D.; Lemos, Manoel V.F.; Junior Boica, Arlindo L. , Jaboticabal, SP . Faculdade de Ciencias Agrarias e Veterinarias. Dept. de Fitossanidade; Brazil)

    2007-01-01

    The fall armyworm, Spodoptera frugiperda (J. E. Smith), is one of the main corn pests and Bacillus thuringiensis is important in its control because of its entomopathogenic property. The objective of this study was the molecular characterization of B. thuringiensis isolates for cry1 locus presence and the assessment of the efficiency of these isolates in controlling S. frugiperda caterpillars. Gral-cry1 was used in the PCR analyses to confirm the presence of the cry1 locus in 15 isolates. A 3 x 108 spore/ml suspension bathed the diet used to feed 30 caterpillars per isolate, with three replications. The cry1 locus type genes of the different isolates were identified for five gene subclasses; linear regression analyses were carried out to ascertain possible associations between the presence of an individual cry1 locus gene and high levels of toxicity. All the DNAs amplified with Gral-cry1 presented an amplification product with the expected size. Regarding the levels of insecticide efficiency against the cob worm, 41 isolates presented 100% mortality and 16 presented an index between 70% and 90%. The cry1Ab gene was present in 80 isolates, cryb in 69 isolates, cry1Ac in all the isolates and cryv and cry1E in 93 and 27 isolates, respectively. The values regarding the individual effect of each gene on caterpillar mortality were significant at 1% probability for the cry1Ac and cry1E genes. (author)

  20. Isolation of Indigenous Bacteria of Phosphate Solubilizing from Green Bean Rhizospheres

    Directory of Open Access Journals (Sweden)

    N Arfarita

    2017-04-01

    Full Text Available The use of phosphate-solubilizing bacteria (PSB as a biological fertilizer of Agricultural land is one solution to overcome problem of phosphate availability for plants. However, often application of a biological fertilizer is ineffective for certain places. The purpose of this study was to obtain indigenous phosphate solubilizing bacterial isolates that can be effective in the area of Malang. Samples were collected from rhizosphereof green bean plants at three locations in Malang, East Java. The study was conducted to determine the total bacterial population of soil samples, to select the best three bacterial isolates in phosphate solubilizing ability, which is not antagonistic and nonpathogenic for plants,along with observing its potential as a bacterial consortium. The highest total population was found in FHR samples of 1.5x1011 CFU / mL. We have selected three bacterial isolates namely SPP1, SPP2 and SPP3. They were not antagonistic to each other and nonpathogenic on mungbean sprouts. They had possibility of producing growth hormone which characterized by an increasing in length of plant and total root length, be compared to controls. Strain SPP2 has shown the highest activity of phosphate solubilization then was selected for 16S rRNA identification. Similarity test of genome sequence of strain SPP2 had 99% similarity with Pseudomonas plecoglossicida strain PR19

  1. Antimicrobial susceptibility profile of selected bacteraemic ...

    African Journals Online (AJOL)

    Extended-spectrum 13-lactamase (ESBL) production was determined in selected species of Enterobacteriaceae irrespective of source. Results. The overall prevalence of ampicillin resistance in blood culture isolates of E. coli (N = 471) was 84%, and 20% were resistant to the fluoroquinolones. Considerable geographical ...

  2. UCLA1 aptamer inhibition of human immunodeficiency virus type 1 subtype C primary isolates in macrophages and selection of resistance

    CSIR Research Space (South Africa)

    Mufhandu, Hazel T

    2016-09-01

    Full Text Available isolates in monocyte-derived macrophages (MDMs). Of 4 macrophage-tropic isolates tested, 3 were inhibited by UCLA1 in the low nanomolar range (IC80 <29 nM). One isolate that showed reduced susceptibility (<50 nM) to UCLA1 contained mutations in the a5 helix...

  3. Isolation and characterization of Bacillus thuringiensis strains active against Elasmopalpus lignosellus (Zeller, 1848 (Lepidoptera, Pyralidae

    Directory of Open Access Journals (Sweden)

    Janaina Zorzetti

    2017-08-01

    Full Text Available Elasmopalpus lignosellus (Zeller, 1848 (Lepidoptera, Pyralidae is an insect pest of 60 economically important crops, including sugarcane, wheat, soybean, rice, beans, sorghum, peanuts, and cotton. The aim of this work was to select and characterize Bacillus thuringiensis isolates with insecticidal activity against E. Lignosellus that could be used as an alternative method of control. Selective bioassays were done to evaluate the toxicity of 47 isolates against first instar larvae of E. lignosellus. For the most toxic bacterial strains, the lethal concentration (LC50 was estimated and morphological, biochemical and molecular methods were used to characterize the isolates. Among the 47 isolates tested, 12 caused mortality above 85% and showed LC50 values from 0.038E+8 to 0.855E+8 spores mL-1. Isolates BR83, BR145, BR09, BR78, S1534, and S1302 had the lowest LC50 values and did not differ from the standard HD-1 strain; the exception was BR83.The protein profiles produced bands with molecular masses of 60-130 kDa. The genes cry1, cry2, cry3, and cry11 were identified in the molecular characterization. The morphological analysis identified three different crystal inclusions: bipyramidal, spherical and cuboidal. Among the tested isolates, 12 isolates have potential for biotechnological control of E. Lignosellus by development of new biopesticides or genetically modified plants.

  4. Isolation and Characterization of Actinomycete Antagonists of a Fungal Root Pathogen †

    Science.gov (United States)

    Crawford, Don L.; Lynch, James M.; Whipps, John M.; Ousley, Margaret A.

    1993-01-01

    By use of selective media, 267 actinomycete strains were isolated from four rhizosphere-associated and four non-rhizosphere-associated British soils. Organic media with low nutrient concentrations were found to be best for isolating diverse actinomycetes while avoiding contamination and overgrowth of isolation media by eubacteria and fungi. While all isolates grew well at pHs 6.5 to 8.0, a few were unable to grow at pH 6.0 and a significant number failed to grow at pH 5.5. Eighty-two selected isolates were screened for in vitro antagonism towards Pythium ultimum by use of a Difco cornmeal agar assay procedure. Five isolates were very strong antagonists of the fungus, four were strong antagonists, and ten others were weakly antagonistic. The remaining isolates showed no antagonism by this assay. Additional studies showed that several of the P. ultimum antagonists also strongly inhibited growth of other root-pathogenic fungi. Twelve isolates showing antifungal activity in the in vitro assay were also tested for their effects on the germination and short-term growth of lettuce plants in glasshouse pot studies in the absence of pathogens. None of the actinomycetes prevented seed germination, although half of the isolates retarded seed germination and outgrowth of the plants by 1 to 3 days. During 18-day growth experiments, biomass yields of some actinomycete-inoculated plants were reduced in comparison with untreated control plants, although all plants appeared healthy and well rooted. None of the actinomycetes significantly enhanced plant growth over these short-term experiments. For some, but not all, actinomycetes, some correlations between delayed seed germination and reduced 18-day plant biomass yields were seen. For others, plant biomass yields were not reduced despite an actinomycete-associated delay in seed germination and plant outgrowth. Preliminary glasshouse experiments indicated that some of the actinomycetes protect germinating lettuce seeds against

  5. Nucleotide and amino acid sequences of a coat protein of an Ukrainian isolate of Potato virus Y: comparison with homologous sequences of other isolates and phylogenetic analysis

    Directory of Open Access Journals (Sweden)

    Budzanivska I. G.

    2014-03-01

    Full Text Available Aim. Identification of the widespread Ukrainian isolate(s of PVY (Potato virus Y in different potato cultivars and subsequent phylogenetic analysis of detected PVY isolates based on NA and AA sequences of coat protein. Methods. ELISA, RT-PCR, DNA sequencing and phylogenetic analysis. Results. PVY has been identified serologically in potato cultivars of Ukrainian selection. In this work we have optimized a method for total RNA extraction from potato samples and offered a sensitive and specific PCR-based test system of own design for diagnostics of the Ukrainian PVY isolates. Part of the CP gene of the Ukrainian PVY isolate has been sequenced and analyzed phylogenetically. It is demonstrated that the Ukrainian isolate of Potato virus Y (CP gene has a higher percentage of homology with the recombinant isolates (strains of this pathogen (approx. 98.8– 99.8 % of homology for both nucleotide and translated amino acid sequences of the CP gene. The Ukrainian isolate of PVY is positioned in the separate cluster together with the isolates found in Syria, Japan and Iran; these isolates possibly have common origin. The Ukrainian PVY isolate is confirmed to be recombinant. Conclusions. This work underlines the need and provides the means for accurate monitoring of Potato virus Y in the agroecosystems of Ukraine. Most importantly, the phylogenetic analysis demonstrated the recombinant nature of this PVY isolate which has been attributed to the strain group O, subclade N:O.

  6. Bacteriocin and cellulose production by lactic acid bacteria isolated ...

    African Journals Online (AJOL)

    Sixteen colonies of lactic acid bacteria (LAB) were selected and screened for their ability to produce bacteriocin by agar well diffusion method using the supernatant of centrifuged test cultures. Four isolates inhibited the growth of Listeria monocytogenes and Escherichia coli. Lactobacillus plantarum (6) and Lactobacillus ...

  7. Rapid isolation of gene homologs across taxa: Efficient identification and isolation of gene orthologs from non-model organism genomes, a technical report

    Directory of Open Access Journals (Sweden)

    Heffer Alison

    2011-03-01

    Full Text Available Abstract Background Tremendous progress has been made in the field of evo-devo through comparisons of related genes from diverse taxa. While the vast number of species in nature precludes a complete analysis of the molecular evolution of even one single gene family, this would not be necessary to understand fundamental mechanisms underlying gene evolution if experiments could be designed to systematically sample representative points along the path of established phylogenies to trace changes in regulatory and coding gene sequence. This isolation of homologous genes from phylogenetically diverse, representative species can be challenging, especially if the gene is under weak selective pressure and evolving rapidly. Results Here we present an approach - Rapid Isolation of Gene Homologs across Taxa (RIGHT - to efficiently isolate specific members of gene families. RIGHT is based upon modification and a combination of degenerate polymerase chain reaction (PCR and gene-specific amplified fragment length polymorphism (AFLP. It allows targeted isolation of specific gene family members from any organism, only requiring genomic DNA. We describe this approach and how we used it to isolate members of several different gene families from diverse arthropods spanning millions of years of evolution. Conclusions RIGHT facilitates systematic isolation of one gene from large gene families. It allows for efficient gene isolation without whole genome sequencing, RNA extraction, or culturing of non-model organisms. RIGHT will be a generally useful method for isolation of orthologs from both distant and closely related species, increasing sample size and facilitating the tracking of molecular evolution of gene families and regulatory networks across the tree of life.

  8. Isolation and identification of Pseudomonas azotoformans for induced calcite precipitation.

    Science.gov (United States)

    Heidari Nonakaran, Siamak; Pazhouhandeh, Maghsoud; Keyvani, Abdullah; Abdollahipour, Fatemeh Zahra; Shirzad, Akbar

    2015-12-01

    Biomineralization is a process by which living organisms produce minerals. The extracellular production of these biominerals by microbes has potential for various bioengineering applications. For example, crack remediation and improvement of durability of concrete is an important goal for engineers and biomineral-producing microbes could be a useful tool in achieving this goal. Here we report the isolation, biochemical characterization and molecular identification of Pseudomonas azotoformans, a microbe that produces calcite and which potentially be used to repair cracks in concrete structures. Initially, 38 bacterial isolates were isolated from soil and cements. As a first test, the isolates were screened using a urease assay followed by biochemical tests for the rate of urea hydrolysis, calcite production and the insolubility of calcite. Molecular amplification and sequencing of a 16S rRNA fragment of selected isolates permitted us to identify P. azotoformans as a good candidate for preparation of biotechnological concrete. This species was isolated from soil and the results show that among the tested isolates it had the highest rate of urea hydrolysis, produced the highest amount of calcite, which, furthermore was the most adhesive and insoluble. This species is thus of interest as an agent with the potential ability to repair cracks in concrete.

  9. Characterization of biocide-tolerant bacteria isolated from cheese and dairy small-medium enterprises.

    Science.gov (United States)

    Fernández Márquez, Ma Luisa; Grande Burgos, Ma José; López Aguayo, Ma Carmen; Pérez Pulido, Rubén; Gálvez, Antonio; Lucas, Rosario

    2017-04-01

    A collection of 120 bacterial isolates from small medium enterprises involved in the production of cow milk and the manufacture of goat cheese were screened for sensitivity to biocides benzalkonium chloride (BC), cetrimide (CT), hexadecylpyridinium chloride (HDP), triclosan (TC), hexachlorophene (CF) and poly-(hexamethylen guanidinium) hydrochloride (PHMG). Nineteen isolates were selected according to biocide tolerance and identified by 16S rDNA sequencing as Lactococcus sp. (6) Enterococcus sp. (1), Lactobacillus sp. (4), Bacillus sp. (1) Escherichia sp. (5), Enterobacter sp. (1) and Helicobacter sp. (1). These were further characterised regarding antimicrobial resistance phenotype and genotype. Several isolates were multiply (3 or more) tolerant to biocides or resistant to antibiotics, but only two Escherichia sp. isolates and Enterobacter sp. were multiply resistant to biocides and antibiotics. Statistical analysis of biocide tolerance and antibiotic resistance revealed significant positive correlations between different biocides and between biocides and antibiotics. The biocide tolerance genes most frequently found were qacEΔ1 and qacA/B. The sulfonamide resistance gene sul1 was found in two Escherichia sp. isolates and in Enterobacter sp., all of which also carried qacEΔ1. Beta-lactam (bla CTX-M , bla PSE ) and tetracycline resistance genes [tet(A), tet(C) and tet(D)] were detected. Efflux pump genes acrB and mdfA were found in most Gram-negative isolates. Results from the study suggest that exposure to biocides can indirectly select for antibiotic resistance. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Efficient and rapid C. elegans transgenesis by bombardment and hygromycin B selection.

    Directory of Open Access Journals (Sweden)

    Inja Radman

    Full Text Available We report a simple, cost-effective, scalable and efficient method for creating transgenic Caenorhabditis elegans that requires minimal hands-on time. The method combines biolistic bombardment with selection for transgenics that bear a hygromycin B resistance gene on agar plates supplemented with hygromycin B, taking advantage of our observation that hygromycin B is sufficient to kill wild-type C. elegans at very low concentrations. Crucially, the method provides substantial improvements in the success of bombardments for isolating transmitting strains, the isolation of multiple independent strains, and the isolation of integrated strains: 100% of bombardments in a large data set yielded transgenics; 10 or more independent strains were isolated from 84% of bombardments, and up to 28 independent strains were isolated from a single bombardment; 82% of bombardments yielded stably transmitting integrated lines with most yielding multiple integrated lines. We anticipate that the selection will be widely adopted for C. elegans transgenesis via bombardment, and that hygromycin B resistance will be adopted as a marker in other approaches for manipulating, introducing or deleting DNA in C. elegans.

  11. Antimicrobial resistance and typing of Salmonella isolated from street vended foods and associated environment.

    Science.gov (United States)

    Anukampa; Shagufta, Bi; Sivakumar, M; Kumar, Surender; Agarwal, Rajesh Kumar; Bhilegaonkar, Kiran Narayan; Kumar, Ashok; Dubal, Zunjar Baburao

    2017-07-01

    The present study was carried out to find out the occurrence and types of Salmonella present in street vended foods and associated environment, and their resistance pattern against various antibiotics. About 1075 street vended food and associated environment samples were processed for isolation and confirmation of different Salmonella spp. by targeting gene specific inv A gene and serotype specific Sdf I, Via B and Spy genes by PCR. Selected Salmonella isolates were screened for antibiotic resistance by using Baeur-Kirby disk diffusion test. Out of 1075 samples, only 31 (2.88%) isolates could be amplified the inv A gene of which 19 could be recovered from meat vendors; 8 from egg vendors while remaining 4 from milk vendors. Though, majority of Salmonella recovered from raw foods the ready-to-eat food like chicken gravy and rasmalai also showed its presence which pose a serious public health threat. Overall, 19, 6 and 1 isolates of S. Typhimurium, S. Enteritidis and S. Typhi could be detected by PCR while remaining 5 isolates could not be amplified suggesting other type of Salmonella. Selected Salmonella isolates were completely resistance to Oxacillin (100%) followed by Cefoxitin (30.43%) and Ampicillin (26.10%). Thus, it is observed that the street vended foods of animal origin and associated environment play an important role in transmission of food borne pathogens including Salmonella .

  12. Biological, serological and molecular typing of potato virus Y (PVY) isolates from Tunisia.

    Science.gov (United States)

    Tayahi, M; Gharsallah, C; Khamassy, N; Fakhfakh, H; Djilani-Khouadja, F

    2016-10-17

    In Tunisia, potato virus Y (PVY) currently presents a significant threat to potato production, reducing tuber yield and quality. Three hundred and eighty-five potato samples (six different cultivars) collected in autumn 2007 from nine regions in Tunisia were tested for PVY infection by DAS-ELISA. The virus was detected in all regions surveyed, with an average incidence of 80.26%. Subsequently, a panel of 82 Tunisian PVY isolates (PVY-TN) was subjected to systematic biological, serological and molecular typing using immunocapture reverse-transcription polymerase chain reaction and a series of PVY OC - and PVY N -specific monoclonal antibodies. Combined analyses revealed ~67% of PVY NTN variants of which 17 were sequenced in the 5'NTR-P1 region to assess the genetic diversity and phylogenetic relationship of PVY-TN against other worldwide PVY isolates. To investigate whether selective constraints could act on viral genomic RNA, synonymous and non-synonymous substitution rates and their ratio were analyzed. Averages of all pairwise comparisons obtained in the 5'NTR-P1 region allowed more synonymous changes, suggesting selective constraint acting in this region. Selective neutrality test was significantly negative, suggesting a rapid expansion of PVY isolates. Pairwise mismatch distribution gave a bimodal pattern and pointed to an eventually early evolution characterizing these sequences. Genetic haplotype network topology provided evidence of the existence of a distinct geographical structure. This is the first report of such genetic analyses conducted on PVY isolates from Tunisia.

  13. [Proposal of a five MIRU-VNTR panel to screen clinical isolates of Mycobacterium tuberculosis in Mexico].

    Science.gov (United States)

    Bolado-Martínez, Enrique; Candia-Plata, Maria Del Carmen; Zenteno-Cuevas, Roberto; Mendoza Damián, Fabiola; Avilés-Acosta, Magali; Álvarez-Hernández, Gerardo

    2015-11-01

    Tuberculosis is a public health problem across Mexico. This paper aims to select a panel, with a minimum number of repetitive elements (MIRU-VNTR) for genotypic characterization of Mycobacterium tuberculosis (M. tuberculosis) clinical isolates. In this study, a full panel of 24 MIRU-VNTR loci was used to discriminate 65 clinical isolates of M. tuberculosis from three different geographical regions of Mexico. Those loci with the highest discriminatory power were subsequently selected. The panel, including five loci, was obtained by selecting the highest values of allelic diversity among the genotypes obtained. The dendrogram, generated by the panel MIRU-VNTR 5, showed a high discriminatory power with 65 unique genotype profiles and formed clusters according to the geographical region of origin. The panel MIRU-VNTR 5 can be useful for characterizing clinical isolates of M. tuberculosis in Mexico. Copyright © 2014 Elsevier España, S.L.U. y Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  14. Social isolation and risk for malnutrition among older people.

    Science.gov (United States)

    Boulos, Christa; Salameh, Pascale; Barberger-Gateau, Pascale

    2017-02-01

    Social isolation and loneliness are emerging issues among the geriatric population. The relationships between both, and their impact on health and nutritional status in older people are complex. The purpose of the present study was to evaluate the association between three components of social isolation: social network, feeling of loneliness, commensality and nutritional status. A total of 1200 randomly selected elderly individuals aged ≥65 years and living in rural areas of Lebanon participated in the present study. Data were collected during a face-to-face interview including nutritional status (Mini-Nutritional Assessment), measures of social isolation (Lubben Social Network Scale), subjective loneliness (Jong-Gierveld Loneliness Scale), sociodemographic conditions, and health and functional status. Both social isolation and loneliness were independently associated with a higher risk of malnutrition (OR 1.58, P = 0.011; OR 1.15, P = 0.020, respectively). However no association was found between the frequency of sharing meals and the risk of malnutrition. The present study showed that social isolation and subjective loneliness are two independent risk factors for malnutrition among older people. Geriatr Gerontol Int 2017; 17: 286-294. © 2016 Japan Geriatrics Society.

  15. Alternative methodology for isolation of biosurfactant-producing bacteria.

    Science.gov (United States)

    Krepsky, N; Da Silva, F S; Fontana, L F; Crapez, M A C

    2007-02-01

    Wide biosurfactant application on biorremediation is limited by its high production cost. The search for cheaper biossurfactant production alternatives has guided our study. The use of selective media containing sucrose (10 g x L(-1)) and Arabian Light oil (2 g x L(-1)) as carbon sources showed to be effective to screen and maintain biosurfactant-producing consortia isolated from mangrove hydrocarbon-contaminated sediment. The biosurfactant production was assayed by kerosene, gasoline and Arabian Light Emulsification activity and the bacterial growth curve was determined by bacterial quantification. The parameters analyzed for biosurfactant production were the growth curve, salinity concentration, flask shape and oxygenation. All bacteria consortia screened were able to emulsify the petroleum derivatives tested. Biosurfactant production increased according to the incubation time; however the type of emulsification (non-aqueous phase or aqueous phase) did not change with time but with the compound tested. The methodology was able to isolate biosurfactant-producing consortia from superficial mangrove sediment contaminated by petroleum hydrocarbons and was recommended for selection of biosurfactant producing bacteria in tropical countries with low financial resources.

  16. Alternative methodology for isolation of biosurfactant-producing bacteria

    Directory of Open Access Journals (Sweden)

    N. Krepsky

    Full Text Available Wide biosurfactant application on biorremediation is limited by its high production cost. The search for cheaper biossurfactant production alternatives has guided our study. The use of selective media containing sucrose (10 g.L-1 and Arabian Light oil (2 g.L-1 as carbon sources showed to be effective to screen and maintain biosurfactant-producing consortia isolated from mangrove hydrocarbon-contaminated sediment. The biosurfactant production was assayed by kerosene, gasoline and Arabian Light Emulsification activity and the bacterial growth curve was determined by bacterial quantification. The parameters analyzed for biosurfactant production were the growth curve, salinity concentration, flask shape and oxygenation. All bacteria consortia screened were able to emulsify the petroleum derivatives tested. Biosurfactant production increased according to the incubation time; however the type of emulsification (non-aqueous phase or aqueous phase did not change with time but with the compound tested. The methodology was able to isolate biosurfactant-producing consortia from superficial mangrove sediment contaminated by petroleum hydrocarbons and was recommended for selection of biosurfactant producing bacteria in tropical countries with low financial resources.

  17. Selection against recombinant hybrids maintains reproductive isolation in hybridizing Populus species despite F1 fertility and recurrent gene flow.

    Science.gov (United States)

    Christe, Camille; Stölting, Kai N; Bresadola, Luisa; Fussi, Barbara; Heinze, Berthold; Wegmann, Daniel; Lexer, Christian

    2016-06-01

    Natural hybrid zones have proven to be precious tools for understanding the origin and maintenance of reproductive isolation (RI) and therefore species. Most available genomic studies of hybrid zones using whole- or partial-genome resequencing approaches have focused on comparisons of the parental source populations involved in genome admixture, rather than exploring fine-scale patterns of chromosomal ancestry across the full admixture gradient present between hybridizing species. We have studied three well-known European 'replicate' hybrid zones of Populus alba and P. tremula, two widespread, ecologically divergent forest trees, using up to 432 505 single-nucleotide polymorphisms (SNPs) from restriction site-associated DNA (RAD) sequencing. Estimates of fine-scale chromosomal ancestry, genomic divergence and differentiation across all 19 poplar chromosomes revealed strikingly contrasting results, including an unexpected preponderance of F1 hybrids in the centre of genomic clines on the one hand, and genomically localized, spatially variable shared variants consistent with ancient introgression between the parental species on the other. Genetic ancestry had a significant effect on survivorship of hybrid seedlings in a common garden trial, pointing to selection against early-generation recombinants. Our results indicate a role for selection against recombinant genotypes in maintaining RI in the face of apparent F1 fertility, consistent with the intragenomic 'coadaptation' model of barriers to introgression upon secondary contact. Whole-genome resequencing of hybridizing populations will clarify the roles of specific genetic pathways in RI between these model forest trees and may reveal which loci are affected most strongly by its cyclic breakdown. © 2016 John Wiley & Sons Ltd.

  18. Molecular detection of salmonella species from selected vegetables ...

    African Journals Online (AJOL)

    Molecular detection of salmonella species from selected vegetables sold in a north-central ... African Journal of Clinical and Experimental Microbiology ... of the pure isolates testing positive as being pathogenic after biochemical analysis.

  19. Molecular characterization of Salmonella isolates by REP-PCR and RAPD analysis.

    Science.gov (United States)

    Albufera, U; Bhugaloo-Vial, P; Issack, M I; Jaufeerally-Fakim, Y

    2009-05-01

    Eighteen Salmonella isolates from both human and food (non-human) sources (fish, meat, and poultry) were characterized using conventional culture methods, biochemical, serological, and molecular analyses. REP-PCR and RAPD produced DNA profiles for differentiation purposes. Enterobacterial repetitive intergenic consensus (ERIC), repetitive extragenic palindronic (REP) and BOXAIR primers were selected for REP-PCR and two arbitrary primers, namely OPP-16 and OPS-11 were used for RAPD to generate DNA fingerprints from the Salmonella isolates. REP-PCR method showed greater discriminatory power in differentiating closely related strains of the related strains of Salmonella and produced more complex banding patterns as compared with RAPD. A dendogram was constructed with both sets of profiles using SPSS Version 13.0 computer software and showed that most human isolates were separately clustered from the non-human isolates. Two of the human isolates were closely related to some of the non-human isolates. A good correlation was also observed between the serogrouping of the O antigen and the molecular profiles obtained from REP-PCR and RAPD data of the Salmonella isolates. The results of a principal coordinate analysis (PCA) corresponded to the clustering in the dendrogram.

  20. Radiation-resistant micro-organisms isolated from textiles

    International Nuclear Information System (INIS)

    Kristensen, H.; Christensen, E.A.

    1981-01-01

    Towels from private homes and public offices and underwear contaminated by being used by employees at a public health laboratory were examined for occurrence of radiation-resistant bacteria and fungi. Three different methods were used for isolation of the most resistant organisms, one with multiplication of the microbial population prior to an irradiation used for selection, and two witout this multiplication and with the organisms placed on membrane filters or in situ on the textiles, respectively. A total of 44 different strains were isolated. Differences in the three methods used for selection of the most radiation-resistant microorganisms were not reflected in the results. 16 pigmentproducing Gram-positive cocci, tentatively classified as Micrococcus radiodurans, were the most radiation-resistant and were isolated in about half of the examinations. Other Gram-positive cocci, nonspore forming rods, some Nocardia and Candida parapsilosis strains and two Bacillus strains constituted the rest of the collection. With few exceptions dose-response curves for the strains were upward convex. D-6 values determined to be between 1.5 megarad for the most radiation sensitive, a Candida, and 5.7 megarad for the most resistant, tentatively classified as M. radiodurans. The D-6 values for the Bacillus strains were in both cases 1.8 megarad, consistent with a D-value of 0.3 megarad. The same resistance is reported to be the maximum resistance for B. pumilus, strain E601, commonly used as reference strain in the literature on radiation sterilization of medical devices and supplies. (author)

  1. Radiation-resistant micro-organisms isolated from textiles

    Energy Technology Data Exchange (ETDEWEB)

    Kristensen, H; Christensen, E A [Statens Seruminstitut, Copenhagen (Denmark)

    1981-01-01

    Towels from private homes and public offices and underwear contaminated by being used by employees at a public health laboratory were examined for occurrence of radiation-resistant bacteria and fungi. Three different methods were used for isolation of the most resistant organisms, one with multiplication of the microbial population prior to an irradiation used for selection, and two witout this multiplication and with the organisms placed on membrane filters or in situ on the textiles, respectively. A total of 44 different strains were isolated. Differences in the three methods used for selection of the most radiation-resistant microorganisms were not reflected in the results. 16 pigmentproducing Gram-positive cocci, tentatively classified as Micrococcus radiodurans, were the most radiation-resistant and were isolated in about half of the examinations. Other Gram-positive cocci, nonspore forming rods, some Nocardia and Candida parapsilosis strains and two Bacillus strains constituted the rest of the collection. With few exceptions dose-response curves for the strains were upward convex. D-6 values determined to be between 1.5 megarad for the most radiation sensitive, a Candida, and 5.7 megarad for the most resistant, tentatively classified as M. radiodurans. The D-6 values for the Bacillus strains were in both cases 1.8 megarad, consistent with a D-value of 0.3 megarad. The same resistance is reported to be the maximum resistance for B. pumilus, strain E601, commonly used as reference strain in the literature on radiation sterilization of medical devices and supplies.

  2. Maladaptive learning and memory in hybrids as a reproductive isolating barrier.

    Science.gov (United States)

    Rice, Amber M; McQuillan, Michael A

    2018-05-30

    Selection against hybrid offspring, or postzygotic reproductive isolation, maintains species boundaries in the face of gene flow from hybridization. In this review, we propose that maladaptive learning and memory in hybrids is an important, but overlooked form of postzygotic reproductive isolation. Although a role for learning in premating isolation has been supported, whether learning deficiencies can contribute to postzygotic isolation has rarely been tested. We argue that the novel genetic combinations created by hybridization have the potential to impact learning and memory abilities through multiple possible mechanisms, and that any displacement from optima in these traits is likely to have fitness consequences. We review evidence supporting the potential for hybridization to affect learning and memory, and evidence of links between learning abilities and fitness. Finally, we suggest several avenues for future research. Given the importance of learning for fitness, especially in novel and unpredictable environments, maladaptive learning and memory in hybrids may be an increasingly important source of postzygotic reproductive isolation. © 2018 The Author(s).

  3. Novel, non-symbiotic isolates of Neorhizobium from a dryland agricultural soil.

    Science.gov (United States)

    Soenens, Amalia; Imperial, Juan

    2018-01-01

    Semi-selective enrichment, followed by PCR screening, resulted in the successful direct isolation of fast-growing Rhizobia from a dryland agricultural soil. Over 50% of these isolates belong to the genus Neorhizobium , as concluded from partial rpoB and near-complete 16S rDNA sequence analysis. Further genotypic and genomic analysis of five representative isolates confirmed that they form a coherent group within Neorhizobium , closer to N. galegae than to the remaining Neorhizobium species, but clearly differentiated from the former, and constituting at least one new genomospecies within Neorhizobium. All the isolates lacked nod and nif symbiotic genes but contained a repABC replication/maintenance region, characteristic of rhizobial plasmids, within large contigs from their draft genome sequences. These repABC sequences were related, but not identical, to repABC sequences found in symbiotic plasmids from N. galegae , suggesting that the non-symbiotic isolates have the potential to harbor symbiotic plasmids. This is the first report of non-symbiotic members of Neorhizobium from soil.

  4. Identification of novel anti-inflammatory probiotic strains isolated from pulque.

    Science.gov (United States)

    Torres-Maravilla, Edgar; Lenoir, Marion; Mayorga-Reyes, Lino; Allain, Thibault; Sokol, Harry; Langella, Philippe; Sánchez-Pardo, María E; Bermúdez-Humarán, Luis G

    2016-01-01

    Probiotics are live microorganisms which when administered in adequate amounts, confer health benefits on the host. Their use is more and more widespread for both prevention and treatment of diseases, including traveler’s diarrhea and inflammatory bowel diseases (IBDs). In this work, we isolated and characterized novel candidate probiotic strains from pulque (xaxtle), a traditional Mexican alcoholic fermented beverage. A total of 14 strains were obtained from xaxtle samples isolated from three different Mexican regions. Species identification was performed by biochemical methods and 16S rRNA gene targeted PCR. The isolates belonged to the Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus brevis, and Lactobacillus composti phylogenetic groups, with L. brevis being the most dominant group. Bacteria were tested for lysozyme, low pH, and bile acid resistance. Moreover, the strains were tested for adherence to human intestinal epithelial cells and screened for their immunomodulatory properties using a cellular model. Selected bacterial strains with anti-inflammatory properties were then tested in vivo in a dinitro-benzene sulfonic acid (DNBS)-induced chronic colitis mouse model, and weight loss, gut permeability, and cytokine profiles were measured as readouts of inflammation. One of the selected strains, Lactobacillus sanfranciscensis LBH1068, improved mice health as observed by a reduction of weight loss, significant decreases in gut permeability, and cytokine modulation. Altogether, our results highlighted the potential of lactobacilli isolated from pulque and in particular the strain L. sanfranciscensis LBH1068 as a novel probiotic to treat IBD.

  5. Genome-wide differentiation in closely related populations: the roles of selection and geographic isolation

    Czech Academy of Sciences Publication Activity Database

    Safran, R. J.; Scordato, E. S. C.; Wilkins, M. R.; Hubbard, J. K.; Jenkins, B. R.; Albrecht, Tomáš; Flaxman, S. M.; Karaardic, H.; Vortman, Y.; Lotem, A.; Nosil, P.; Pap, P.; Shen, S.; Chan, S.-F.; Parchman, T. L.; Kane, N. C.

    2016-01-01

    Roč. 25, č. 16 (2016), s. 3865-3883 ISSN 0962-1083 R&D Projects: GA MŠk(CZ) LH14045 Institutional support: RVO:68081766 Keywords : climate variability * climate variability * genotyping by sequencing * population genetics * reproductive isolation * speciation Subject RIV: EG - Zoology Impact factor: 6.086, year: 2016

  6. Probiotic features of two oral Lactobacillus isolates

    Directory of Open Access Journals (Sweden)

    Gordana Zavisic

    2012-03-01

    Full Text Available In this study, we checked lactobacilli strains of human origin for their potential as probiotic. Samples were collected from oral mucosa of 16 healthy individuals, out of which twenty isolates were obtained and two of them were selected and identified as Lactobacillus plantarum (G1 and L. casei (G3. Both isolates exhibited antagonistic action towards pathogenic microorganisms such as Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Salmonella abony, and Clostridium sporogenes, but not on the growth of Candida albicans. The bacteriocin activity against Staphylococcus aureus ATCC 6358-P was shown only by L. plantarum G1. Moreover, the isolates G1 and G3 showed good viability in the acid gastric environment and in the gut environment containing bovine bile salts. The viability of G1 and G3 isolates in the gastrointestinal tract, and the adhesion to the intestinal mucosa were also confirmed in vivo. The biochemical tests of blood samples revealed lower levels of serum triglycerides and cholesterol, as well as reduced activity of alkaline phosphatase in all lactobacilli-treated Wistar rats, compared to control ones. No toxicity for NMRI Ham mice was observed. According to our experimental results, these findings imply that L. plantarum G1 and L. casei G3 could be characterized as potential probiotics.

  7. Probiotic features of two oral Lactobacillus isolates.

    Science.gov (United States)

    Zavisic, Gordana; Petricevic, Sasa; Radulovic, Zeljka; Begovic, Jelena; Golic, Natasa; Topisirovic, Ljubisa; Strahinic, Ivana

    2012-01-01

    In this study, we checked lactobacilli strains of human origin for their potential as probiotic. Samples were collected from oral mucosa of 16 healthy individuals, out of which twenty isolates were obtained and two of them were selected and identified as Lactobacillus plantarum (G1) and L. casei (G3). Both isolates exhibited antagonistic action towards pathogenic microorganisms such as Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Salmonella abony, and Clostridium sporogenes, but not on the growth of Candida albicans. The bacteriocin activity against Staphylococcus aureus ATCC 6358-P was shown only by L. plantarum G1. Moreover, the isolates G1 and G3 showed good viability in the acid gastric environment and in the gut environment containing bovine bile salts. The viability of G1 and G3 isolates in the gastrointestinal tract, and the adhesion to the intestinal mucosa were also confirmed in vivo. The biochemical tests of blood samples revealed lower levels of serum triglycerides and cholesterol, as well as reduced activity of alkaline phosphatase in all lactobacilli-treated Wistar rats, compared to control ones. No toxicity for NMRI Ham mice was observed. According to our experimental results, these findings imply that L. plantarum G1 and L. casei G3 could be characterized as potential probiotics.

  8. Isolation of radiation-resistant bacteria without exposure to irradiation

    International Nuclear Information System (INIS)

    Sanders, S.W.; Maxcy, R.B.

    1979-01-01

    Resistance to desiccation was utilized in the selection of highly radiation-resistant asporogenous bacteria from nonirradiated sources. A bacterial suspension in phosphate buffer was dried in a thin film at 25 0 C and 33% relative humidity. Storage under these conditions for 15 days or more reduced the number of radiation-sensitive bacteria. Further selection for radiation-resistant bacteria was obtained by irradiation of bacteria on velveteen in the replication process, therby avoiding the toxic effect of irradiated media. The similarity of radiation resistance and identifying characteristics in irradiated and non-irradiated isolates should allay some concerns that highly radiation-resistance bacteria have been permanently altered by radiation selection

  9. Antibiotics susceptibility patterns of urine bacterial isolates in Zaria ...

    African Journals Online (AJOL)

    Purpose: The prevalence of E. coli, Ps. aeruginosa and Staph aureus isolates from urine of selected residents in Zaria was investigated. This was an attempt to elucidate the antibiotic susceptibility profiles of these bacteria commonly implicated in urinary tact infection. Methods: Urine samples collected from students of ...

  10. From retrospective assessment to prospective decisions in natural product isolation

    DEFF Research Database (Denmark)

    Johansen, Kenneth T; Wubshet, Sileshi G; Nyberg, Nils T

    2011-01-01

    An extract of Carthamus oxyacantha (wild safflower) was investigated using two approaches: a traditional, nontarget fractionation by VLC and HPLC, and the hyphenated technique HPLC-PDA-HRMS-SPE-NMR followed by targeted isolation of selected constituents for inclusion in a screening library of pure......, HPLC-HRMS-guided, targeted isolation scheme aiming exclusively at a series of identified spiro compounds. Thus, HPLC-PDA-HRMS-SPE-NMR is a dereplication technique of choice, allowing economical acquisition of comprehensive data about compounds in crude extracts, which can be used for rational...

  11. Isolation and Characterization of Seed-Borne Pathogenic Bacteria

    African Journals Online (AJOL)

    76 L.K. Ashura et aI. Table 4: Selected morphological and biochemical characteristics of bacteria isolates from rice seed as de- tected by the Liquid Assay on mXOS. Ace. No.1. Fluorescentl. Kovac's. OIF. NR. GL. SH. LS. HR. Pstho. Diolog Identification lsolale(s). Colour of·. Oxidase. TIP g on. (Similarity). I. Pigment. Rice.

  12. Diversity in UV sensitivity and recovery potential among bacterioneuston and bacterioplankton isolates.

    Science.gov (United States)

    Santos, A L; Lopes, S; Baptista, I; Henriques, I; Gomes, N C M; Almeida, A; Correia, A; Cunha, A

    2011-04-01

    To assess the variability in UV-B (280-320 nm) sensitivity of selected bacterial isolates from the surface microlayer and underlying water of the Ria de Aveiro (Portugal) estuary and their ability to recover from previous UV-induced stress. Bacterial suspensions were exposed to UV-B radiation (3·3 W m⁻²). Effects on culturability and activity were assessed from colony counts and (3) H-leucine incorporation rates, respectively. Among the tested isolates, wide variability in UV-B-induced inhibition of culturability (37·4-99·3%) and activity (36·0-98·0%) was observed. Incubation of UV-B-irradiated suspensions under reactivating regimes (UV-A, 3·65 W m⁻²; photosynthetic active radiation, 40 W m⁻²; dark) also revealed diversity in the extent of recovery from UV-B stress. Trends of enhanced resistance of culturability (up to 15·0%) and enhanced recovery in activity (up to 52·0%) were observed in bacterioneuston isolates. Bacterioneuston isolates were less sensitive and recovered more rapidly from UV-B stress than bacterioplankton isolates, showing enhanced reduction in their metabolism during the irradiation period and decreased culturability during the recovery process compared to bacterioplankton. UV exposure can affect the diversity and activity of microbial communities by selecting UV-resistant strains and alter their metabolic activity towards protective strategies. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

  13. Selecting Sample Preparation Workflows for Mass Spectrometry-Based Proteomic and Phosphoproteomic Analysis of Patient Samples with Acute Myeloid Leukemia.

    Science.gov (United States)

    Hernandez-Valladares, Maria; Aasebø, Elise; Selheim, Frode; Berven, Frode S; Bruserud, Øystein

    2016-08-22

    Global mass spectrometry (MS)-based proteomic and phosphoproteomic studies of acute myeloid leukemia (AML) biomarkers represent a powerful strategy to identify and confirm proteins and their phosphorylated modifications that could be applied in diagnosis and prognosis, as a support for individual treatment regimens and selection of patients for bone marrow transplant. MS-based studies require optimal and reproducible workflows that allow a satisfactory coverage of the proteome and its modifications. Preparation of samples for global MS analysis is a crucial step and it usually requires method testing, tuning and optimization. Different proteomic workflows that have been used to prepare AML patient samples for global MS analysis usually include a standard protein in-solution digestion procedure with a urea-based lysis buffer. The enrichment of phosphopeptides from AML patient samples has previously been carried out either with immobilized metal affinity chromatography (IMAC) or metal oxide affinity chromatography (MOAC). We have recently tested several methods of sample preparation for MS analysis of the AML proteome and phosphoproteome and introduced filter-aided sample preparation (FASP) as a superior methodology for the sensitive and reproducible generation of peptides from patient samples. FASP-prepared peptides can be further fractionated or IMAC-enriched for proteome or phosphoproteome analyses. Herein, we will review both in-solution and FASP-based sample preparation workflows and encourage the use of the latter for the highest protein and phosphorylation coverage and reproducibility.

  14. Natural and sexual selection against hybrid flycatchers

    NARCIS (Netherlands)

    Svedin, Nina; Wiley, Chris; Veen, Thor; Gustafsson, Lars; Qvarnstrom, Anna

    2008-01-01

    While sexual selection is generally assumed to quickly cause or strengthen prezygotic barriers between sister species, its role in causing postzygotic isolation, through the unattractiveness of intermediate hybrids, is less often examined. Combining 24 years of pedigree data and recently developed

  15. Restriction analysis of genetic variability of Polish isolates of Tomato black ring virus.

    Science.gov (United States)

    Jończyk, Magdalena; Borodynko, Natasza; Pospieszny, Henryk

    2004-01-01

    Several different isolates of Tomato black ring virus (TBRV) have been collected in Poland from cucumber, tomato, potato and black locust plants. Biological tests showed some differences in the range of infected plants and the type of symptoms, which was the basis for selection of seven the most biologically different TBRV isolates. According to the sequence of TBRV-MJ, several primer pairs were designed and almost the entire sequence of both genomic RNAs was amplified. The RT-PCR products derived from all tested TBRV isolates were digested by restriction enzymes. On the basis of the restriction patterns, the variable and the conserved regions of the TBRV genome were defined and the relationships between the Polish TBRV isolates established.

  16. Susceptibility testing of fish cell lines for virus isolation

    DEFF Research Database (Denmark)

    Ariel, Ellen; Skall, Helle Frank; Olesen, Niels Jørgen

    2009-01-01

    and laboratories, but also between lineages of the same cell line. To minimise the occurrence of false negatives in a cell culture based surveillance system, we have investigated methods, to select cell lineages that are relatively superior in their susceptibility to a panel of virus isolates. The procedures...... cell lineages, we increased the number of isolates of each virus, propagated stocks in a given cell line and tested all lineages of that line in use in the laboratory. Testing of relative cell line susceptibility between laboratories is carried out annually via the Inter-laboratory Proficiency Test...... sensitivity for surveillance purposes within a cell line and between laboratories.In terms of economic and practical considerations as well as attempting to approach a realistic test system, we suggest the optimal procedure for susceptibility testing of fish cell lines for virus isolation to be a combination...

  17. X-ray Diffraction from Isolated and Strongly Aligned Gas-Phase Molecules with a Free-Electron Laser

    DEFF Research Database (Denmark)

    Küpper, Jochen; Stern, Stephan; Holmegaard, Lotte

    2014-01-01

    We report experimental results on x-ray diffraction of quantum-state-selected and strongly aligned ensembles of the prototypical asymmetric rotor molecule 2,5-diiodobenzonitrile using the Linac Coherent Light Source. The experiments demonstrate first steps toward a new approach to diffractive...... imaging of distinct structures of individual, isolated gas-phase molecules. We confirm several key ingredients of single molecule diffraction experiments: the abilities to detect and count individual scattered x-ray photons in single shot diffraction data, to deliver state-selected, e. g., structural......-isomer-selected, ensembles of molecules to the x-ray interaction volume, and to strongly align the scattering molecules. Our approach, using ultrashort x-ray pulses, is suitable to study ultrafast dynamics of isolated molecules....

  18. Selection of the Chrome Reduction Bacteria in the Waste of Tanning Leather Industries by Ozonization Method

    International Nuclear Information System (INIS)

    Yazid, M.; Aris Bastianudin; Widdi Usada

    2007-01-01

    Selection of the chrome reduction bacteria in the waste of tanning leather industries by ozonization method has been done. The objectives of this research was to obtain isolate bacteria from the waste with chrome contain, so that expected can be used for chrome bioremediation agent for arrange to improved the waste treatment for tanning leather industries. Selection of bacteria in the waste was carried out by ozonization method with time variation 0 to 210 minutes by time interval 15 minutes. Isolation bacteria was carried out was grown on the BHI media for 24 hours at 37°C temperature. So be inoculated by streak plate method on the TBX, MC, EA, CTM and BP media. Characterization of bacteria was done by saw the colonies morphology, sel morphology and biochemical characterization. So, identification of isolate bacteria by matching profile method. The result of this research can be obtained 5 isolate bacteria BCR1, BCR2, BCR3, BCR4 and BCR5 with the different phenotypic character. From the five isolate can be selected resistance ozon isolate until 180 minutes time ozonization were BCR 2, were identified belong to the genus of Bacillus. The examination results showed that the isolate bacteria be able to reduction of the chrome concentration in the waste of tanning leather industries by 71.03 %. Efficiency. (author)

  19. Preliminary screening of bacterial isolates from mining wastes

    Directory of Open Access Journals (Sweden)

    Rodino S.,

    2016-05-01

    Full Text Available Developing innovative biotechnology for obtaining new resources of high tech critical metals is strongly influenced by the need to reduce the potential risk of shortages, to support the development of industry at European level. To set up these new technologies is essential to isolate strains with high potential in bioleaching of ore, tailings and mine wastes and bioaccumulation of high tech critical metals. Microorganisms are capable of mediating metal and mineral bioprecipitation. In this paper are presented preliminary studies performed for the isolation of strains existing in mining residues containing high tech critical metals. Were used samples collected from various depths in an area of mining wastes containing high tech critical metals. The samples were fine grounded and the powder was washed with sterile saline water. Exact quantities of samples were dispersed in sterile saline water, shaken for a period of 60 minutes, diluted and plated in triplicate on selective agar. After several steps were isolated 3 strains of gram negative bacteria.

  20. Screening of β-Glucosidase and β-Xylosidase Activities in Four Non-Saccharomyces Yeast Isolates.

    Science.gov (United States)

    López, María Consuelo; Mateo, José Juan; Maicas, Sergi

    2015-08-01

    The finding of new isolates of non-Saccharomyces yeasts, showing beneficial enzymes (such as β-glucosidase and β-xylosidase), can contribute to the production of quality wines. In a selection and characterization program, we have studied 114 isolates of non-Saccharomyces yeasts. Four isolates were selected because of their both high β-glucosidase and β-xylosidase activities. The ribosomal D1/D2 regions were sequenced to identify them as Pichia membranifaciens Pm7, Hanseniaspora vineae Hv3, H. uvarum Hu8, and Wickerhamomyces anomalus Wa1. The induction process was optimized to be carried on YNB-medium supplemented with 4% xylan, inoculated with 106 cfu/mL and incubated 48 h at 28 °C without agitation. Most of the strains had a pH optimum of 5.0 to 6.0 for both the β-glucosidase and β-xylosidase activities. The effect of sugars was different for each isolate and activity. Each isolate showed a characteristic set of inhibition, enhancement or null effect for β-glucosidase and β-xylosidase. The volatile compounds liberated from wine incubated with each of the 4 yeasts were also studied, showing an overall terpene increase (1.1 to 1.3-folds) when wines were treated with non-Saccharomyces isolates. In detail, terpineol, 4-vinyl-phenol and 2-methoxy-4-vinylphenol increased after the addition of Hanseniaspora isolates. Wines treated with Hanseniaspora, Wickerhamomyces, or Pichia produced more 2-phenyl ethanol than those inoculated with other yeasts. © 2015 Institute of Food Technologists®

  1. Molecular screening and isolation of Newcastle disease virus from ...

    African Journals Online (AJOL)

    Molecular screening and isolation of Newcastle disease virus from live poultry markets and chickens from commercial poultry farms in Zaria, Kaduna state, Nigeria. ... The PDF file you selected should load here if your Web browser has a PDF reader plug-in installed (for example, a recent version of Adobe Acrobat Reader).

  2. Isolation, characterization and development of bacteria in the Mine Gafsa for applications in bioremediation

    International Nuclear Information System (INIS)

    Heni, Sana

    2010-01-01

    Today pollution represents an important environmental problem. Bacterial ability to bioremediate many types of pollutants in different matrixes (soil, water, and air) have been widely acknowledged. The goal of the present work is to isolate from contaminated soil of Gafsa, in Tunisia, bacterial strains to evaluate their potential for bioremediation. Soil from the mining area of Gafsa was collected. Initially, many bacterial strains were isolated in TGY agar (Tryptone/Glucose/Yeast extract agar) based on the presence of pigments. The primary bacterial selection was performed using heavy metals and the minimal inhibitory concentrations (MICs) of a metal-resistant bacterium, Cupriavidus metallidurans CH34. Isolated metal-resistant bacterium was checked for its potential to resistant to gamma radiation. Selected strain, Micrococcus luteus S7, was assessed for its bioremediation potential of matrixes artificially contaminated under laboratory conditions for its future use in developing a bio product for contaminated soil inoculation.

  3. Selection and identification of oleaginous yeast isolated from soil, animal feed and ruminal fluid for use as feed supplement in dairy cattle.

    Science.gov (United States)

    Paserakung, A; Pattarajinda, V; Vichitphan, K; Froetschel, M A

    2015-10-01

    The purpose of this study was to select oleaginous yeast for microbial lipid production. Sixty-four yeast isolates were obtained from soil (GSY1-12), animal feeds (FDY1-21), and ruminal fluid (RMY1-31) using yeast extract peptone dextrose (YPD) agar. The cultivation of these isolates on nitrogen limited-medium revealed that GSY2 to GSY6, GSY10, FDY2, FDY12 and FDY14 accumulated lipid over 20% of dry biomass. Therefore, they were preliminarily classified as oleaginous yeast. In subsequent experiment, an 8 × 3 factorial in completely randomized design was conducted to examine the effect of eight oleaginous yeast strains and three nitrogen sources (peptone, (NH4 )2 SO4 , urea) on lipid accumulation when using molasses as substrate. The result illustrated that only GSY3 and GSY10 accumulated lipid over 20% of biomass when using peptone or (NH4 )2 SO4 but urea did not. However, GSY10 gave higher biomass and lipid yield than GSY3 (P yeast for microbial lipid production from molasses. This study illustrated the ability of T. asahii GSY10 to utilize molasses and (NH4 )2 SO4 for synthesizing and accumulating cellular lipid of which oleic acid (C18:1 ) was predominant. This yeast would be used for microbial lipid production used as feed supplement in dairy cattle. © 2015 The Society for Applied Microbiology.

  4. Functionality of selected strains of moulds and yeasts from Vietnamese rice wine starters

    NARCIS (Netherlands)

    Dung, N.T.P.; Rombouts, F.M.; Nout, M.J.R.

    2006-01-01

    The role of starch-degrading mycelial fungi, and the alcohol production and ethanol tolerance of the yeasts isolated from selected Vietnamese traditional rice wine starters were examined, and optimum conditions for these essential steps in rice wine fermentation were determined. Of pure isolates

  5. Reduced susceptibility to praziquantel among naturally occurring Kenyan isolates of Schistosoma mansoni.

    Directory of Open Access Journals (Sweden)

    Sandra D Melman

    2009-08-01

    Full Text Available The near exclusive use of praziquantel (PZQ for treatment of human schistosomiasis has raised concerns about the possible emergence of drug-resistant schistosomes.We measured susceptibility to PZQ of isolates of Schistosoma mansoni obtained from patients from Kisumu, Kenya continuously exposed to infection as a consequence of their occupations as car washers or sand harvesters. We used a an in vitro assay with miracidia, b an in vivo assay targeting adult worms in mice and c an in vitro assay targeting adult schistosomes perfused from mice. In the miracidia assay, in which miracidia from human patients were exposed to PZQ in vitro, reduced susceptibility was associated with previous treatment of the patient with PZQ. One isolate ("KCW" that was less susceptible to PZQ and had been derived from a patient who had never fully cured despite multiple treatments was studied further. In an in vivo assay of adult worms, the KCW isolate was significantly less susceptible to PZQ than two other isolates from natural infections in Kenya and two lab-reared strains of S. mansoni. The in vitro adult assay, based on measuring length changes of adults following exposure to and recovery from PZQ, confirmed that the KCW isolate was less susceptible to PZQ than the other isolates tested. A sub-isolate of KCW maintained separately and tested after three years was susceptible to PZQ, indicative that the trait of reduced sensitivity could be lost if selection was not maintained.Isolates of S. mansoni from some patients in Kisumu have lower susceptibility to PZQ, including one from a patient who was never fully cured after repeated rounds of treatment administered over several years. As use of PZQ continues, continued selection for worms with diminished susceptibility is possible, and the probability of emergence of resistance will increase as large reservoirs of untreated worms diminish. The potential for rapid emergence of resistance should be an important

  6. Studies on fibrolytic bacterium Butyrivibrio fibrisolvens isolated from sheep rumen

    Directory of Open Access Journals (Sweden)

    Sawanon, S.

    2007-03-01

    Full Text Available Fibrolytic Butyrivibrio fibrisolvens was an attractive target for genetic engineering in rumen bacteria. The experiment was initiated in making culture collection of this species, some of which may be useful ascandidate strain in the future. Hay suspended in sheep rumen was used as the source of isolates. The source was enriched with filter paper degradation, diluted with an anaerobic solution and used for pure culturing bya roll tube technique. After colony forming, Gram-negative curved rods bacteria were selected and screened for further identification with volatile fatty acid (VFA profiling and 16S rDNA sequencing. Fibrolyticstrains were selected to find fibrolytic enzymes and attachment to and digestion of various fibers. Fortyseven strains of Gram-negative curved rods were isolated. After determining cellulase, xylanase activities and VFA profile, 2 strains were chosen and employed for 16S rDNA sequencing. Both strains producingbutyrate were B. fibrisolvens. Of these 2 strains, most fibrolytic S-28 was selected. The strain S-28 could degrade natural fibers but not cellulose and showed strong attachment to them. A strong xylanase activitywas detected and presence of cellulase, β-glucosidase, β-xylosidase, α-L-arabinofuranosidase and β- cellobiosidase were also demonstrated.

  7. Environmental and Irradiation Effect on the Biosynthesis of Bio surfactant by Some Local Bacterial Isolates

    International Nuclear Information System (INIS)

    Ghoneimy, E.; Ahmed, S.; Desouki, S.; Rasmy, W.; El-Shahawy, M.

    2010-01-01

    Twenty eight bacterial isolates were isolated from The Suez Gulf sea water from the coast of the El-Nasr Petroleum Company on Suez Canal and formation water from overhead of an oil well in western desert of Egypt named (M68). Sixteen bacterial isolates were obtained from The Suez Gulf sea water. Twelve bacterial isolates were obtained from formation water (M68). The bacterial isolates were screened for bio surfactant production by using emulsification activity and haemolytic activity. The most potent two isolates N8 and S8 were selected according to three parameters; the ability of isolate to grow and produce surfactant on wide range of ph, temperature and salinity. The most promising bacterial isolates were subjected to different doses of gamma irradiation in a trial to improve their abilities for bio surfactant production which resulted in a passive effect on bio surfactant production

  8. Status of technology for isolating high-level radioactive wastes in geologic repositories

    International Nuclear Information System (INIS)

    Klingsberg, C.; Duguid, J.

    1980-10-01

    This report attempts to summarize the status of scientific and technological knowledge relevant to long-term isolation of high-level and transuranic wastes in a mined geologic repository. It also identifies and evaluates needed information and identifies topics in which work is under way or needed to reduce uncertainties. The major findings and conclusions on the following topics are presented: importance of the systems approach; prospects for successful isolation of wastes; need for site-specific investigations; human activities in the future; importance of modelling; disposal of transuranic wastes; status of technology of isolation barriers, performance assessment, site selection and characterization, and potential host rocks

  9. Isolation of microbial native Stumps with cellulolytic activity of a compost process

    International Nuclear Information System (INIS)

    Jaramillo G, Marisol; Ruiz V Orlando Simon; Yepes P, Maria del Socorro; Montoya C, Olga Ines

    2003-01-01

    The isolation, selection adaptation and handling of native microorganisms coming from organic waste are an alternative to avoid the accumulation and the lack of the proper use of these undesirable materials. This organic waste is a source for obtaining microbial strains, which are potentially producers of Industrial enzymes and, at the same time, it works as substrate so that these organisms can transform it into compost or organic manure. In this work, 39 native strains of microorganisms with potential cellulolytic activity coming from the organic waste of the urban and rural sector, from the Compost Plant of Marinilla Antioquia) municipality, were isolated, evaluated and purified. The waste was previously selected and then submitted to an aerobic degradation or compost. The microbial strains were isolated in a selective medium with carboxymethyl cellulose (CMC), of the phases mesophile, termophile, cooling and maturation of the compost process. Eighty-two percent (82%)of the obtained colonies were identified, in principle as Bacillus, because of their morphology and their reaction to the Gram coloration. The fungi population was seen only during the cooling phase. Then, the potential cellulolytic activity was evaluated qualitatively in a solid medium with the Congo Red coloration, with which the Beta-endoglucanase activity was evaluated through the formation of clarified zones. Such staining was applied in two mediums with CMC with and without glucose It was observed that 33.3% of the isolated organisms produced the enzyme In both mediums; however, 25.6% of microorganisms did not show the production of this enzyme, and only 15.8% did not require the inducers to produce it

  10. Identification of novel PAMP-triggered phosphorylation and dephosphorylation events in arabidopsis thaliana by quantitative phosphoproteomic analysis

    KAUST Repository

    Rayapuram, Naganand; Bonhomme, Ludovic; Bigeard, Jean; Haddadou, Kahina; Przybylski, Cé dric; Hirt, Heribert; Pflieger, Delphine

    2014-01-01

    Signaling cascades rely strongly on protein kinase-mediated substrate phosphorylation. Currently a major challenge in signal transduction research is to obtain high confidence substrate phosphorylation sites and assign them to specific kinases. In response to bacterial flagellin, a pathogen-associated molecular pattern (PAMP), we searched for rapidly phosphorylated proteins in Arabidopsis thaliana by combining multistage activation (MSA) and electron transfer dissociation (ETD) fragmentation modes, which generate complementary spectra and identify phosphopeptide sites with increased reliability. Of a total of 825 phosphopeptides, we identified 58 to be differentially phosphorylated. These peptides harbor kinase motifs of mitogen-activated protein kinases (MAPKs) and calcium-dependent protein kinases (CDPKs), as well as yet unknown protein kinases. Importantly, 12 of the phosphopeptides show reduced phosphorylation upon flagellin treatment. Since protein abundance levels did not change, these results indicate that flagellin induces not only various protein kinases but also protein phosphatases, even though a scenario of inhibited kinase activity may also be possible. © 2014 American Chemical Society.

  11. Phosphoproteomic investigation of a solvent producing bacterium Clostridium acetobutylicum.

    Science.gov (United States)

    Bai, Xue; Ji, Zhihong

    2012-07-01

    In this study, we employed TiO₂ enrichment and high accuracy liquid chromatography-mass spectrometry-mass spectrometry to identify the phosphoproteome of Clostridium acetobutyicum ATCC824 in acidogenesis and solventogenesis. As many as 82 phosphopeptides in 61 proteins, with 107 phosphorylated sites on serine, threonine, or tyrosine, were identified with high confidence. We detected 52 phosphopeptides from 44 proteins in acidogenesis and 70 phosphopeptides from 51 proteins in solventogenesis, respectively. Bioinformatic analysis revealed most of the phosphoproteins located in cytoplasm and participated in carbon metabolism. Based on comparison between the two stages, we found 27 stage-specific phosphorylated proteins (10 in acidogenesis and 17 in solventogenesis), some of which were solvent production-related enzymes and metabolic regulators, showed significantly different phosphorylated status. Further analysis indicated that protein phosphorylation could be involved in the shift of stages or in solvent production pathway directly. Comparison against several other organisms revealed the evolutionary diversity among them on phosphorylation level in spite of their high homology on protein sequence level.

  12. Identification of novel PAMP-triggered phosphorylation and dephosphorylation events in arabidopsis thaliana by quantitative phosphoproteomic analysis

    KAUST Repository

    Rayapuram, Naganand

    2014-04-04

    Signaling cascades rely strongly on protein kinase-mediated substrate phosphorylation. Currently a major challenge in signal transduction research is to obtain high confidence substrate phosphorylation sites and assign them to specific kinases. In response to bacterial flagellin, a pathogen-associated molecular pattern (PAMP), we searched for rapidly phosphorylated proteins in Arabidopsis thaliana by combining multistage activation (MSA) and electron transfer dissociation (ETD) fragmentation modes, which generate complementary spectra and identify phosphopeptide sites with increased reliability. Of a total of 825 phosphopeptides, we identified 58 to be differentially phosphorylated. These peptides harbor kinase motifs of mitogen-activated protein kinases (MAPKs) and calcium-dependent protein kinases (CDPKs), as well as yet unknown protein kinases. Importantly, 12 of the phosphopeptides show reduced phosphorylation upon flagellin treatment. Since protein abundance levels did not change, these results indicate that flagellin induces not only various protein kinases but also protein phosphatases, even though a scenario of inhibited kinase activity may also be possible. © 2014 American Chemical Society.

  13. Investigating quantitation of phosphorylation using MALDI-TOF mass spectrometry.

    Science.gov (United States)

    Parker, Laurie; Engel-Hall, Aaron; Drew, Kevin; Steinhardt, George; Helseth, Donald L; Jabon, David; McMurry, Timothy; Angulo, David S; Kron, Stephen J

    2008-04-01

    Despite advances in methods and instrumentation for analysis of phosphopeptides using mass spectrometry, it is still difficult to quantify the extent of phosphorylation of a substrate because of physiochemical differences between unphosphorylated and phosphorylated peptides. Here we report experiments to investigate those differences using MALDI-TOF mass spectrometry for a set of synthetic peptides by creating calibration curves of known input ratios of peptides/phosphopeptides and analyzing their resulting signal intensity ratios. These calibration curves reveal subtleties in sequence-dependent differences for relative desorption/ionization efficiencies that cannot be seen from single-point calibrations. We found that the behaviors were reproducible with a variability of 5-10% for observed phosphopeptide signal. Although these data allow us to begin addressing the issues related to modeling these properties and predicting relative signal strengths for other peptide sequences, it is clear that this behavior is highly complex and needs to be further explored. John Wiley & Sons, Ltd

  14. Isolation, screening and characterization of bio surfactant producing bacteria

    International Nuclear Information System (INIS)

    Kokub, D.; Shafeeq, M.; Khalid, Z.M.; Malik, K.A.

    1991-01-01

    Bio surfactant producing bacteria were enriched from oil, oil contaminated soil and formation water collected from some local oil wells; Balkassar, Joyamair, Dullian, Meyal and Khore, and oil-riched soils from Karachi coastal area and Petroleum Refinery Limited (PRL) Karachi, by rowing them on different growth media with various carbon sources. These enriched cultures were analysed qualitatively and quantitatively for various types of bacteria. Morphologically different colonies present in these enriched cultures were quantified and different bacterial strains were isolated by single colony isolation method. Sixty two isolates were screened out by growing them individually on Khaskheli crude oil and comparing for the above parameters. Two bacterial strains which did not fulfill this criteria were also used for comparison in further studies. The selected strains were grown on n-hexadecane/glucose and the spent culture broth were tested for reduction in surface tension (ST) and interfacial tension (IFT). The surface tension was checked after every 24 hours and the minimum time required for the reduction in surface tension 33 mN/m was noted. On the basis of these observation, six groups of bacteria were made. These cultures were also grown on blood agar plates to test for hemolysis. Sixty six percent of these selected cultures were found to reduce surface tension lesser than 33 mN/m and IFT lesser than 3 mN/m whereas 85% of them showed hemolytic activity. IFT of these culture broths was found to be positively correlated to surface tension. Among the isolates from different localities Pseudomonas spp. was found to be most prevalent while some Micrococcus and Acinetobacter were also found. (author)

  15. Comparative analysis of the genomes of two field isolates of the rice blast fungus Magnaporthe oryzae.

    Directory of Open Access Journals (Sweden)

    Minfeng Xue

    Full Text Available Rice blast caused by Magnaporthe oryzae is one of the most destructive diseases of rice worldwide. The fungal pathogen is notorious for its ability to overcome host resistance. To better understand its genetic variation in nature, we sequenced the genomes of two field isolates, Y34 and P131. In comparison with the previously sequenced laboratory strain 70-15, both field isolates had a similar genome size but slightly more genes. Sequences from the field isolates were used to improve genome assembly and gene prediction of 70-15. Although the overall genome structure is similar, a number of gene families that are likely involved in plant-fungal interactions are expanded in the field isolates. Genome-wide analysis on asynonymous to synonymous nucleotide substitution rates revealed that many infection-related genes underwent diversifying selection. The field isolates also have hundreds of isolate-specific genes and a number of isolate-specific gene duplication events. Functional characterization of randomly selected isolate-specific genes revealed that they play diverse roles, some of which affect virulence. Furthermore, each genome contains thousands of loci of transposon-like elements, but less than 30% of them are conserved among different isolates, suggesting active transposition events in M. oryzae. A total of approximately 200 genes were disrupted in these three strains by transposable elements. Interestingly, transposon-like elements tend to be associated with isolate-specific or duplicated sequences. Overall, our results indicate that gain or loss of unique genes, DNA duplication, gene family expansion, and frequent translocation of transposon-like elements are important factors in genome variation of the rice blast fungus.

  16. Social Isolation and Aging in Zambia: Examining the Possible Predictors

    Directory of Open Access Journals (Sweden)

    Christopher Chabila Mapoma

    2012-01-01

    Full Text Available This research paper examined social isolation and aging in Zambia by examining possible predictors. The paper produces evidence on risk factors likely to engender social isolation among the elderly population of Zambia. Snowball sampling was undertaken to select 690 adults aged 60 and over in communities as well as those living in homes for the aged. A structured questionnaire was used to solicit information from respondents. Results show that old people in Zambia experience forms of social isolation which exhibit themselves (but not limited to through such factors as loss of appetite, stress, moody, hopeless, useless, unhappy, and lonely. On balance, however, the direction of association and the number of statistically significant findings suggest that associations between variables examined and risk factors associated with social isolation amongst older people in this analysis could explain the overall situation occuring currently in Zambia and probably other developing countries. In view of this, this study recommends that further work is needed to identify and explain details of factors of social isolation using techniques such as focus group discussions as well as in-depth interviews with key informants. Such approaches may even help to explain why, for example, sex seems not to be significant in determining indicators of social isolation.

  17. Improvement of the Uranium Sequestration Ability of a Chlamydomonas sp. (ChlSP Strain) Isolated From Extreme Uranium Mine Tailings Through Selection for Potential Bioremediation Application.

    Science.gov (United States)

    Baselga-Cervera, Beatriz; Romero-López, Julia; García-Balboa, Camino; Costas, Eduardo; López-Rodas, Victoria

    2018-01-01

    The extraction and processing of uranium (U) have polluted large areas worldwide, rendering anthropogenic extreme environments inhospitable to most species. Noticeably, these sites are of great interest for taxonomical and applied bioprospection of extremotolerant species successfully adapted to U tailings contamination. As an example, in this work we have studied a microalgae species that inhabits extreme U tailings ponds at the Saelices mining site (Salamanca, Spain), characterized as acidic (pH between 3 and 4), radioactive (around 4 μSv h -1 ) and contaminated with metals, mainly U (from 25 to 48 mg L -1 ) and zinc (from 17 to 87 mg L -1 ). After isolation of the extremotolerant ChlSP strain, morphological characterization and internal transcribed spacer (ITS)-5.8S gene sequences placed it in the Chlamydomonadaceae , but BLAST analyses identity values, against the nucleotide datasets at the NCBI database, were very low (tailings waters based on newly evolved extremotolerants and outline the potential of artificial selection in the improvement of desired features in microalgae by experimental adaptation and selection.

  18. Production of extracellular laccase from the newly isolated Bacillus ...

    African Journals Online (AJOL)

    This study was carried out with aim of screening for extracellular thermostable laccase producing bacteria. Twenty-two (22) laccase positive strains were isolated from the selected environmental samples while extracellular laccase activity was detected only in six strains namely TM1, TMT1, PK4, PS1, TMS1 and ASP3.

  19. Isolation, Screening and Development of Local Bacterial Consortia With Azo Dyes Decolourising Capability

    Directory of Open Access Journals (Sweden)

    Khadijah, O.

    2009-01-01

    Full Text Available A total of 1540 bacterial isolates were isolated and screened for their ability to degrade selected azo dyes. Of these, nine isolates were chosen for further studies based on their ability to degrade a wide spectrum of dyes efficiently and rapidly. Several microbial consortia were developed and tested for their effectiveness. Overall the consortia were able to degrade 70 - 100% colour within 72 hours compared to 60 – 97% colour removed by individual isolates. A microbial consortium labelled C15 showed good growth in agitation culture but the colour removal was best in static culture with 80 - 100% colour removed in less than 72 hours. Based on the 16S rRNA sequencing, two of the bacterial isolates in C15 belong to the Chryseobacterium genus while the other one belongs to Flavobacterium genus.

  20. Peroxidase Activity in Poplar Inoculated with Compatible and Incompetent Isolates of Paxillus involutus

    Directory of Open Access Journals (Sweden)

    ABDUL GAFUR

    2007-06-01

    Full Text Available Peroxidase activity of the hybrid poplar Populus×canescens (Ait. Sm. (= P. tremula L. × P. alba L. inoculated with compatible and incompetent isolates of Paxillus involutus (Batsch Fr. was investigated. Screening of the ectomycorrhizal fungal isolates was initiated with exploration of mycelial growth characteristics and mycorrhizal ability in vitro with poplar. Both traits varied within the fungus although they did not seem to be genetically correlated. While isolates SCO1, NAU, and 031 grew faster than others, only isolates MAJ, SCO1, and 031 were able to form ectomycorrhiza with poplar. Isolates MAJ (compatible and NAU (incompetent were subsequently selected for further experiments. Activity of peroxidase, one of the defense-related enzymes, was examined in pure culture and short root components of compatible and incompetent interactions between poplar and P. involutus. Peroxidase activities increased significantly in poplar inoculated with incompetent isolate of the fungus compared to control, while induction of the same enzyme was not detected in compatible associations.

  1. Selection and Characterization of Potential Baker's Yeast from Indigenous Resources of Nepal.

    Science.gov (United States)

    Karki, Tika B; Timilsina, Parash Mani; Yadav, Archana; Pandey, Gyanu Raj; Joshi, Yogesh; Bhujel, Sahansila; Adhikari, Rojina; Neupane, Katyayanee

    2017-01-01

    The study aims to isolate the yeast strains that could be used effectively as baker's yeast and compare them with the commercial baker's yeast available in the market of Nepal. A total of 10 samples including locally available sources like fruits, Murcha, and a local tree "Dar" were collected from different localities of Bhaktapur, Kavre, and Syangja districts of Nepal, respectively. Following enrichment and fermentation of the samples, 26 yeast strains were isolated using selective medium Wallerstein Laboratory Nutrient Agar. From the differential tests which included morphological and microscopic observation and physiological and biochemical characterization such as nitrate reduction and lactose utilization tests, 8 strains were selected as possible Saccharomyces strain. The selected strains were further assessed for their efficient leavening ability by tests such as ethanol tolerance, osmotolerance, invertase test, and stress exclusion test. The three most potent strains ENG, MUR3B, and SUG1 isolated from grape, Murcha, and sugarcane, respectively, were used in the fermentation and baking of dough. These strains also carried a possibility of being used as industrial baker's yeast.

  2. Monocytes isolated by positive and negative magnetic sorting techniques show different molecular characteristics and immunophenotypic behaviour [version 3; referees: 2 approved

    Directory of Open Access Journals (Sweden)

    Jashdeep Bhattacharjee

    2018-03-01

    Full Text Available Background: Magnetic sorting of cells, based on  microbead conjugated antibodies (Abs, employs positive as well as negative immunomagnetic separation methods, for isolation of a specific cell population. These microbeads are suggested to be nontoxic, biodegradable carriers conjugated to various antibodies. Isolation of cells through positive selection involves the attachment of antibody conjugated microbeads to the cells of interest, followed by their isolation in the presence of a strong magnetic field to obtain higher purity. Negative selection involves attachment of microbead conjugated antibodies to all other cell populations except the cells of interest, which remain untagged. In the present study, we compared the two methods for their effect on functional and immunophenotypic behavior of isolated CD14+ monocytes. Methods: Peripheral blood mononuclear cells (PBMCs were isolated from blood collected from healthy volunteers by density gradient centrifugation. Human blood derived monocytes were isolated through positive selection and negative selection, making use of the appropriate monocyte isolation kit. Monocytes were then stimulated with lipopolysaccharide (LPS and their activation and proliferation capacity were examined. The degradation or dissociation of cell-bound microbeads was also investigated. Results: We observed an impaired LPS sensitivity as well as poor activation and proliferation capacity upon stimulation by LPS in positively sorted CD14+ monocytes as compared to negatively sorted CD14+ monocytes. The attached microbeads did not degrade and remained attached to the cells even after 6 days of culture. Conclusions: Our results suggest that positively sorted CD14+ cells exhibit hampered functionality and may result in inaccurate analysis and observations in downstream applications. However, these cells can be used for immediate analytical procedures.

  3. K. OXYTOCA BACTERIOPHAGES ISOLATION METHODS IMPROVEMENT

    Directory of Open Access Journals (Sweden)

    G. R. Sadrtdinova

    2017-01-01

    Full Text Available The article presents the results of a study related to increasing the efficiency of phage isolation of bacteria of the species K. oxytoca, by developing the optimal composition of the medium used in the work. In scientific research, in almost all methods associated with the isolation of bacteriophages, meat-peptone broth and meat-peptone agar are used as the nutrient basis. The peculiarities of growth and cultivation of microorganisms create certain difficulties for the isolation of phages active against bacteria of the species K. oxytoca. The selection of components and the creation of an environment that would ensure the optimal growth of both the bacterial culture and the reproduction of the virus makes it possible to facilitate the isolation of bacteriophages. The number of bacterial strains used in the work was 7. All strains of cultures were obtained from the Museum of the Department of Microbiology, Virology, Epizootology and Veterinary and Sanitary Expertise of the Federal State Budget Educational Institution of Higher Education “Ulyanovsk State Agrarian University named after P.A. Stolypin”. The studies included 2 main stages. The first stage consisted in isolation of bacteriophages by the method of isolation from the external environment by the method of Adelson L.I., Lyashenko E.A. The material for the studies were samples: soil, sewage sample, fecal samples (2. Only 4 samples. According to the chosen method, the sowing of the putative phagolysate was carried out on meat-peptone agar (1.5% and the agar for isolating bacteriophages (Aph (1.5%. A positive result was the presence on the environment of negative colonies, clearly visible on the matt background of deep growth of bacteria. A negative result is a continuous growth (“lawn” of bacterial culture. As a control, the culture of the microorganism studied was used for the media. In the course of the conducted studies for the first stage, 2 bacteriophages were isolated, active

  4. Occurrence and characterization of Staphylococcus bacteria isolated from poultry in Western Poland.

    Science.gov (United States)

    Marek, Agnieszka; Stepień-Pyśniak, Dagmara; Pyzik, Ewelina; Adaszek, Łukasz; Wilczyński, Jarosław; Winiarczyk, Stanisław

    2016-01-01

    In the pathology of poultry, infections caused by Staphylococcus spp. are taking on increasing significance. Although the Staphylococcus species most frequently isolated from these animals is Staphylococcus aureus, the literature data indicate that other species, both coagulase-positive and coagulase-negative, can also cause infections in birds. The aim of the study was to assess the frequency of occurrence of Staphylococcus infections in various poultry species in Western Poland and to test the susceptibility of isolated strains to selected antibiotics. The results obtained showed a relatively high rate of Staphylococcus infection in the poultry. From 2805 samples tested 302 strains (10.8%) of Staphylococcus were isolated. As many as 25 Staphylococcus species were distinguished among the strains isolated. S. cohnii (23.50%), S. aureus (15.89%) and S. lentus (13.90%) accounted for the highest percentages. Over half of the isolated staphylococci exhibited resistance to five of the antibiotics applied, with the highest percentage of resistant strains, 65%, noted for enrofloxacin.

  5. Molecular characterization of partial fusion gene and C-terminus extension length of haemagglutinin-neuraminidase gene of recently isolated Newcastle disease virus isolates in Malaysia

    Directory of Open Access Journals (Sweden)

    Berhanu Ayalew

    2010-08-01

    Full Text Available Abstract Background Newcastle disease (ND, caused by Newcastle disease virus (NDV, is a highly contagious disease of birds and has been one of the major causes of economic losses in the poultry industry. Despite routine vaccination programs, sporadic cases have occasionally occurred in the country and remain a constant threat to commercial poultry. Hence, the present study was aimed to characterize NDV isolates obtained from clinical cases in various locations of Malaysia between 2004 and 2007 based on sequence and phylogenetic analysis of partial F gene and C-terminus extension length of HN gene. Results The coding region of eleven NDV isolates fusion (F gene and carboxyl terminal region of haemagglutinin-neuraminidase (HN gene including extensions were amplified by reverse transcriptase PCR and directly sequenced. All the isolates have shown to have non-synonymous to synonymous base substitution rate ranging between 0.081 - 0.264 demonstrating presence of negative selection. Analysis based on F gene showed the characterized isolates possess three different types of protease cleavage site motifs; namely 112RRQKRF117, 112RRRKRF117 and 112GRQGRL117 and appear to show maximum identities with isolates in the region such as cockatoo/14698/90 (Indonesia, Ch/2000 (China, local isolate AF2240 indicating the high similarity of isolates circulating in the South East Asian countries. Meanwhile, one of the isolates resembles commonly used lentogenic vaccine strains. On further characterization of the HN gene, Malaysian isolates had C-terminus extensions of 0, 6 and 11 amino acids. Analysis of the phylogenetic tree revealed that the existence of three genetic groups; namely, genotype II, VII and VIII. Conclusions The study concluded that the occurrence of three types of NDV genotypes and presence of varied carboxyl terminus extension lengths among Malaysian isolates incriminated for sporadic cases.

  6. Molecular characterization of partial fusion gene and C-terminus extension length of haemagglutinin-neuraminidase gene of recently isolated Newcastle disease virus isolates in Malaysia.

    Science.gov (United States)

    Berhanu, Ayalew; Ideris, Aini; Omar, Abdul R; Bejo, Mohd Hair

    2010-08-08

    Newcastle disease (ND), caused by Newcastle disease virus (NDV), is a highly contagious disease of birds and has been one of the major causes of economic losses in the poultry industry. Despite routine vaccination programs, sporadic cases have occasionally occurred in the country and remain a constant threat to commercial poultry. Hence, the present study was aimed to characterize NDV isolates obtained from clinical cases in various locations of Malaysia between 2004 and 2007 based on sequence and phylogenetic analysis of partial F gene and C-terminus extension length of HN gene. The coding region of eleven NDV isolates fusion (F) gene and carboxyl terminal region of haemagglutinin-neuraminidase (HN) gene including extensions were amplified by reverse transcriptase PCR and directly sequenced. All the isolates have shown to have non-synonymous to synonymous base substitution rate ranging between 0.081 - 0.264 demonstrating presence of negative selection. Analysis based on F gene showed the characterized isolates possess three different types of protease cleavage site motifs; namely 112RRQKRF117, 112RRRKRF117 and 112GRQGRL117 and appear to show maximum identities with isolates in the region such as cockatoo/14698/90 (Indonesia), Ch/2000 (China), local isolate AF2240 indicating the high similarity of isolates circulating in the South East Asian countries. Meanwhile, one of the isolates resembles commonly used lentogenic vaccine strains. On further characterization of the HN gene, Malaysian isolates had C-terminus extensions of 0, 6 and 11 amino acids. Analysis of the phylogenetic tree revealed that the existence of three genetic groups; namely, genotype II, VII and VIII. The study concluded that the occurrence of three types of NDV genotypes and presence of varied carboxyl terminus extension lengths among Malaysian isolates incriminated for sporadic cases.

  7. In vitro antimicrobial activity and characterization of mangrove isolates of streptomycetes effective against bacteria and fungi of nosocomial origin

    Directory of Open Access Journals (Sweden)

    Arijit Das

    2014-06-01

    Full Text Available The study aimed at determining the in vitro antimicrobial activity of alkaliphilic and halotolerant actinomycetes isolated from a mangrove ecosystem and identification of a potent strain. Twenty five isolates of actinomycetes were isolated from the sediment samples of Valapattanam mangrove swamp in Kerala, India. Antimicrobial activity of four selected actinomycete isolates was determined against bacterial and fungal pathogens of nosocomial origin by agar well diffusion method. Molecular characterization of the potent isolate was performed by 16S rDNA sequencing. Isolate no I-1 significantly inhibited Staphylococcus aureus ATCC 25923 (12 mm, S. aureus (15±0.05 mm, S. citreus (20±0.5 mm, Bacillus cereus (17±0.2 mm and Serratia marcescens (12 mm. It also demonstrated effective antifungal action against Penicillium sp. (12±0.2 mm, Candida albicans (20±0.5 mm, C. parapsilosis (12 mm and Cryptococcus neoformans (12 mm. Morphological study revealed that all the isolated actinomycetes belonged to the genus Streptomyces. Based on 16S rDNA sequence data, the selected isolate I-1 was shown to be closely related to Streptomyces xiamenensis. The results revealed that the mangrove ecosystem of Valapattanam harboured a rich consortium of many potent actinomycetes, which could synthesize novel bioactive compounds of pharmacological significance.

  8. Isolation of Lactic Acid Bacteria with High Biological Activity from Local Fermented Dairy Products

    Directory of Open Access Journals (Sweden)

    B. Munkhtsetseg

    2009-12-01

    Full Text Available The thirty-two strains of lactic acid bacteria were isolated from the Mongolian traditional fermented dairy products, among them 25 strains show antimicrobial activity against test microorganisms including Escherichia coli , Staphylococcus aureus , Enterococcus faecalis , Pseudom о nas aeruginosa . Protease sensitivity assay demonstrated that the antimicrobial substances produced by isolates А 23, Т 2 are bacteriocins as their antibacterial activities were eliminated completely after treatment with protease. Identi fi cation of bacteria is being carried out. Among the isolates 22 strains show protease enzyme producing activity. The selected strains isolated from mare’s fermented milk (airag or kumis and yoghurt (tarag show the speci fi c protease activity from 7.9 μ g/ml to 11.9 μ g/ml. The strain T2, isolated from yoghurt exhibited the highest proteolytic activity.

  9. Novel, non-symbiotic isolates of Neorhizobium from a dryland agricultural soil

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    Amalia Soenens

    2018-05-01

    Full Text Available Semi-selective enrichment, followed by PCR screening, resulted in the successful direct isolation of fast-growing Rhizobia from a dryland agricultural soil. Over 50% of these isolates belong to the genus Neorhizobium, as concluded from partial rpoB and near-complete 16S rDNA sequence analysis. Further genotypic and genomic analysis of five representative isolates confirmed that they form a coherent group within Neorhizobium, closer to N. galegae than to the remaining Neorhizobium species, but clearly differentiated from the former, and constituting at least one new genomospecies within Neorhizobium. All the isolates lacked nod and nif symbiotic genes but contained a repABC replication/maintenance region, characteristic of rhizobial plasmids, within large contigs from their draft genome sequences. These repABC sequences were related, but not identical, to repABC sequences found in symbiotic plasmids from N. galegae, suggesting that the non-symbiotic isolates have the potential to harbor symbiotic plasmids. This is the first report of non-symbiotic members of Neorhizobium from soil.

  10. Screening and characterization of bacteriocins produced by some Strains of Lactobacillus spp isolated from Iranian Dairy products

    OpenAIRE

    S Mirdamadi; M Tangestani

    2011-01-01

    In this study, the inhibitory effects of bacteriocins of lactobacilli which were isolated from Iranian traditional dairy products was determined against known gram positive, gram negative and yeast by well diffusion technique. Among 8 isolates with higher capability of bacteriocin production, 2 isolates were selected for further investigations. The bacteriocins were purified by iso-propanol and ammonium sulfate precipitation following by dialysis and chromatography technique. The molecular we...

  11. Characteristics of primary infection of a European human immunodeficiency virus type 1 clade B isolate in chimpanzees

    NARCIS (Netherlands)

    Bogers, W. M.; Koornstra, W. H.; Dubbes, R. H.; ten Haaft, P. J.; Verstrepen, B. E.; Jhagjhoorsingh, S. S.; Haaksma, A. G.; Niphuis, H.; Laman, J. D.; Norley, S.; Schuitemaker, H.; Goudsmit, J.; Hunsmann, G.; Heeney, J. L.; Wigzell, H.

    1998-01-01

    The aim of the study was to select, from a panel of candidate European human immunodeficiency virus type 1 (HIV-1) clade B primary virus isolates, one isolate based on replication properties in chimpanzee peripheral blood mononuclear cells (PBMC). Secondly, to evaluate the in vivo kinetics of

  12. Inhibition of Clostridium activities in silage and cheese using anticlostridial Lactobacillus Isolated from Danish semi-hard cheese

    DEFF Research Database (Denmark)

    Christiansen, Pia

    Growth of Clostridium, originating mainly from silage, may cause serious late blowing defects in semi-hard cheeses during ripening. In the present project, the possibilities were investigated to use anticlostridial non-starter Lactobacillus (mainly Lb. paracasei), isolated from Danish semi......-hard cheeses of high quality, as protective adjunct cultures against clostridia activities in silage and cheese. Screening for anticlostridial activity among non-starter Lactobacillus isolates against selected Clostridium strains showed that almost half (44%) of the naturally occurring non......-starter Lactobacillus in Danish semi-hard cheeses possessed anticlostridial activities and 10% possessed a broad anticlostridial activity, and these were selected for further investigations. Antagonistic antimicrobial interactions between some of the selected anticlostridial Lactobacillus strains were demonstrated...

  13. Isolation of Panels of Llama Single-Domain Antibody Fragments Binding All Nine Neuraminidase Subtypes of Influenza A Virus

    Directory of Open Access Journals (Sweden)

    Guus Koch

    2013-04-01

    Full Text Available Avian influenza A virus comprises sixteen hemagglutinin (HA and nine neuraminidase (NA subtypes (N1–N9. To isolate llama single-domain antibody fragments (VHHs against all N subtypes, four llamas were immunized with mixtures of influenza viruses. Selections using influenza virus yielded predominantly VHHs binding to the highly immunogenic HA and nucleoprotein. However, selection using enzymatically active recombinant NA (rNA protein enabled us to isolate NA binding VHHs. Some isolated VHHs cross-reacted to other N subtypes. These were subsequently used for the capture of N subtypes that could not be produced as recombinant protein (rN6 or were enzymatically inactive (rN1, rN5 in phage display selection, yielding novel VHHs. In total we isolated 188 NA binding VHHs, 64 of which were expressed in yeast. Most VHHs specifically recognize a single N subtype, but some VHHs cross-react with other N-subtypes. At least one VHH bound to all N subtypes, except N4, identifying a conserved antigenic site. Thus, this work (1 describes methods for isolating NA binding VHHs, (2 illustrates the suitability of llama immunization with multiple antigens for retrieving many binders against different antigens and (3 describes 64 novel NA binding VHHs, including a broadly reactive VHH, which can be used in various assays for influenza virus subtyping, detection or serology.

  14. Isolation, diversity, and antimicrobial activity of rare actinobacteria from medicinal plants of tropical rain forests in Xishuangbanna, China.

    Science.gov (United States)

    Qin, Sheng; Li, Jie; Chen, Hua-Hong; Zhao, Guo-Zhen; Zhu, Wen-Yong; Jiang, Cheng-Lin; Xu, Li-Hua; Li, Wen-Jun

    2009-10-01

    Endophytic actinobacteria are relatively unexplored as potential sources of novel species and novel natural products for medical and commercial exploitation. Xishuangbanna is recognized throughout the world for its diverse flora, especially the rain forest plants, many of which have indigenous pharmaceutical histories. However, little is known about the endophytic actinobacteria of this tropical area. In this work, we studied the diversity of actinobacteria isolated from medicinal plants collected from tropical rain forests in Xishuangbanna. By the use of different selective isolation media and methods, a total of 2,174 actinobacteria were isolated. Forty-six isolates were selected on the basis of their morphologies on different media and were further characterized by 16S rRNA gene sequencing. The results showed an unexpected level of diversity, with 32 different genera. To our knowledge, this is the first report describing the isolation of Saccharopolyspora, Dietzia, Blastococcus, Dactylosporangium, Promicromonospora, Oerskovia, Actinocorallia, and Jiangella species from endophytic environments. At least 19 isolates are considered novel taxa by our current research. In addition, all 46 isolates were tested for antimicrobial activity and were screened for the presence of genes encoding polyketide synthetases and nonribosomal peptide synthetases. The results confirm that the medicinal plants of Xishuangbanna represent an extremely rich reservoir for the isolation of a significant diversity of actinobacteria, including novel species, that are potential sources for the discovery of biologically active compounds.

  15. Geo-scientific considerations on evaluation of possible sites for radioactive waste isolation in Japan

    Energy Technology Data Exchange (ETDEWEB)

    Doi, K [Radioactive Waste Management Center, Tokyo (Japan)

    1980-08-01

    Japan is located on the Circum-Pacific Arc, which is a geoscientifically difficult area for selecting sites suitable as repositories for isolating radioactive waste. The writer has analyzed the problems relevant to radioactive waste isolation in this aqueous and active structural territory, with a view to examining the possibility of finding geological formations suitable for such repositories. As a result, certain parts in Neogene sedimentary rocks and Palaeozoic calcarious rocks were found to present geological characteristics that appeared favorable for radioactive waste isolation, while, on the other hand, the study indicated that much difficulty would be foreseen in crystalline rocks such as are currently suitable in the US and in Europe for high level radioactive waste isolation.

  16. New Zealand; Selected Issues

    OpenAIRE

    International Monetary Fund

    2009-01-01

    This Selected Issues paper conducts a comparative analysis of the main determinants of GDP per capita growth in New Zealand and in other OECD countries to assess the relative importance of macroeconomic factors, institutional settings, and geographical location in New Zealand’s growth performance during the last 30 years. The estimation results find strong support for the view that geographical isolation has significantly hampered growth in New Zealand. The paper also reviews the internationa...

  17. Pattern of Bacterial Pathogens and Their Susceptibility Isolated from Surgical Site Infections at Selected Referral Hospitals, Addis Ababa, Ethiopia

    Directory of Open Access Journals (Sweden)

    Walelign Dessie

    2016-01-01

    Full Text Available Background. The emergence of multidrug resistant bacterial pathogens in hospitals is becoming a challenge for surgeons to treat hospital acquired infections. Objective. To determine bacterial pathogens and drug susceptibility isolated from surgical site infections at St. Paul Specialized Hospital Millennium Medical College and Yekatit 12 Referral Hospital Medical College, Addis Ababa, Ethiopia. Methods. A cross-sectional study was conducted between October 2013 and March 2014 on 107 surgical site infected patients. Wound specimens were collected using sterile cotton swab and processed as per standard operative procedures in appropriate culture media; and susceptibility testing was done using Kirby-Bauer disc diffusion technique. The data were analyzed by using SPSS version 20. Result. From a total of 107 swabs collected, 90 (84.1% were culture positive and 104 organisms were isolated. E. coli (24 (23.1% was the most common organism isolated followed by multidrug resistant Acinetobacter species (23 (22.1%. More than 58 (75% of the Gram negative isolates showed multiple antibiotic resistance (resistance ≥ 5 drugs. Pan-antibiotic resistance was noted among 8 (34.8% Acinetobacter species and 3 (12.5% E. coli. This calls for abstinence from antibiotic abuse. Conclusion. Gram negative bacteria were the most important isolates accounting for 76 (73.1%. Ampicillin, amoxicillin, penicillin, cephazoline, and tetracycline showed resistance while gentamicin and ciprofloxacin were relatively effective antimicrobials.

  18. Genetic relatedness of selected clinical Vibrio cholerae O139 isolates from the southern coastal area of China over a 20-year period.

    Science.gov (United States)

    Li, B S; Xiao, Y; Wang, D C; Tan, H L; Ke, B X; He, D M; Ke, C W; Zhang, Y H

    2016-09-01

    Vibrio cholerae O139 emerged as a causative agent of epidemic cholera in 1992 in India and Bangladesh, and was subsequently reported in China in 1993. The genetic relatedness and molecular characteristics of V. cholerae O139 in Guangdong Province, located in the southern coastal area of China, remains undetermined. In this study, we investigated 136 clinical V. cholerae O139 isolates from 1993 to 2013 in Guangdong. By conventional PCR, 123 (90·4%) isolates were positive for ctxB, ace and zot. Sequencing of the positive amplicons indicated 113 (91·7%) isolates possessed the El Tor allele of ctxB (genotype 3); seven carried the classical ctxB type (genotype 1) and three harboured a novel ctxB type (genotype 5). With respect to tcpA, 123 (90·4%) isolates were positive for the El Tor allele. In addition, pulsed-field gel electrophoresis (with NotI digestion) differentiated the isolates into clusters A and B. Cluster A contained seven of the non-toxigenic isolates from 1998 to 2000; another six non-toxigenic isolates (from 1998 and 2007) and all of the toxigenic isolates formed cluster B. Our results suggest that over a 20-year period, the predominant O139 clinical isolates have maintained a relatively tight clonal structure, although some genetic variance and shift has occurred. Our data highlight the persistence of toxigenic V. cholerae O139 in clinical settings in the southern coastal area of China.

  19. Seleção de isolados de estreptomicetos para controle de Ralstonia solanacearum em tomateiro Selection of streptomycetes isolates for the control of Ralstonia solanacearum in tomato plants

    Directory of Open Access Journals (Sweden)

    Carlos Alberto Tuão Gava

    2002-10-01

    Full Text Available Em condições tropicais, a murcha-bacteriana, causada por Ralstonia solanacearum, tem provocado danos severos à cultura do tomate (Lycopersicon esculentum Mill., principalmente em condições de temperaturas acima de 25°C, com umidade relativa elevada. Neste contexto, o controle biológico pode representar uma alternativa viável. Este trabalho teve como objetivo selecionar isolados de estreptomicetos para o controle de R. solanacearum em tomateiro. Entre os isolados avaliados verificaram-se diferenças na inibição do patógeno in vitro, tanto em relação à faixa de pH avaliada, quanto ao tempo de observação. Constataram-se, também, variações no estabelecimento dos isolados de estreptomicetos na rizosfera das plântulas e nas mudas preparadas para transplante, e os isolados SP164, SF232, SAc326 e SG384 apresentaram as densidades populacionais mais elevadas. O experimento conduzido em canteiros com solo infestado com R. solanacearum mostrou que as plantas tratadas com o isolado SG384 apresentaram melhor nível de controle, 48 dias após a semeadura, período no qual todas as testemunhas haviam morrido.The bacterial wilt caused by Ralstonia solanacearum causes high damages in tomato (Lycopersicon esculentum Mill. culture, mainly under temperature above 25ºC and high relative humidity. Because conventional methods of control have shown low efficiency, biological control is a promising alternative. This work aimed to select isolates of streptomycetes for R. solanacearum control in tomato. Differences were found among the isolates in the in vitro inhibition of the pathogen either in time or pH range. The streptomycetes isolates grew differently in the rhizosphere of seedlings and plantlets of tomato. The isolates SP164, SF232, SAc326 and SG384 showed the highest population densities. In the experiment conducted in soil infested with R. solanacearum, plants treated with the isolate SG384 presented best control level 48 days after sowing

  20. Antimicrobial Susceptibility and Clonality of Clinical Ureaplasma Isolates in the United States.

    Science.gov (United States)

    Fernández, Javier; Karau, Melissa J; Cunningham, Scott A; Greenwood-Quaintance, Kerryl E; Patel, Robin

    2016-08-01

    Ureaplasma urealyticum and Ureaplasma parvum are pathogens involved in urogenital tract and intrauterine infections and also in systemic diseases in newborns and immunosuppressed patients. There is limited information on the antimicrobial susceptibility and clonality of these species. In this study, we report the susceptibility of 250 contemporary isolates of Ureaplasma (202 U. parvum and 48 U. urealyticum isolates) recovered at Mayo Clinic, Rochester, MN. MICs of doxycycline, azithromycin, ciprofloxacin, tetracycline, erythromycin, and levofloxacin were determined by broth microdilution, with MICS of the last three interpreted according to CLSI guidelines. Levofloxacin resistance was found in 6.4% and 5.2% of U. parvum and U. urealyticum isolates, respectively, while 27.2% and 68.8% of isolates, respectively, showed ciprofloxacin MICs of ≥4 μg/ml. The resistance mechanism of levofloxacin-resistant isolates was due to mutations in parC, with the Ser83Leu substitution being most frequent, followed by Glu87Lys. No macrolide resistance was found among the 250 isolates studied; a single U. parvum isolate was tetracycline resistant. tet(M) was found in 10 U. parvum isolates, including the single tetracycline-resistant isolate, as well as in 9 isolates which had low tetracycline and doxycycline MICs. Multilocus sequence typing (MLST) performed on a selection of 46 isolates showed high diversity within the clinical Ureaplasma isolates studied, regardless of antimicrobial susceptibility. The present work extends previous knowledge regarding susceptibility to antimicrobial agents, resistance mechanisms, and clonality of Ureaplasma species in the United States. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  1. A Tunable Cyclic Container: Guest-Induced Conformational Switching, Efficient Guest Exchange, and Selective Isolation of C70 from a Fullerene Mixture.

    Science.gov (United States)

    Mondal, Pritam; Rath, Sankar Prasad

    2017-07-18

    An adaptable cyclic porphyrin dimer with highly flexible linkers has been used as an artificial molecular container that can efficiently encapsulate various aromatic guests (TCNQ/C 60 /C 70 ) through strong π-π interactions by adjusting its cavity size and conformation. The planar aromatic guest (TCNQ) can be easily and selectively exchanged with larger aromatic guests (C 60 /C 70 ). During the guest-exchange process, the two porphyrin rings switch their relative orientation according to the size and shape of the guests. This behavior of the cyclic container has been thoroughly investigated by using UV/Vis spectroscopy, NMR spectroscopy, and X-ray crystal structure determination of the host-guest assemblies. The electrochemical and photophysical studies demonstrated the occurrence of photoinduced electron transfer from bisporphyrin to TCNQ/C 60 /C 70 in the respective host-guest assemblies. The cyclic host can form complexes with C 60 and C 70 with association constants of (2.8±0.2)×10 5 and (1.9±0.3)×10 8  m -1 , respectively; the latter value represents the highest binding affinity for C 70 reported so far for zinc(II) bisporphyrinic receptors. This high selectivity for the binding of C 70 versus C 60 allows the easy extraction and efficient isolation of C 70 from a C 60 /C 70 fullerene mixture. Experimental evidence was substantiated by DFT calculations. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Biopathologic Characterization of Three Mixed Poultry Eimeria spp. Isolates

    Directory of Open Access Journals (Sweden)

    B Mansoori

    2011-09-01

    Full Text Available Background: Coccidiosis of domestic fowl, caused by species of the Genus Eimeria, is responsi­ble for important economic losses in poultry production. Because different species and/or strains can vary in pathogenicity and other biological parameters, their precise characteriza­tion is important for epizootiological studies.Methods: Fifty samples from litter, whole intestinal tract and feces were collected from poul­try houses located in different provinces of Iran. One hundred twenty male day-old broi­ler chicks were challenged with three selected isolates. Data on weight gain, Food Conversion Ratio (FCR, food intake, lesion scoring and shedding of oocysts per gram of feces were rec­orded and analyzed by the Duncan's test.Results: In all treatments, the challenged groups had statistically significant lower weight gain than that of unchallenged control group. Isolate three caused the lowest weight gain and food intake and the worst lesion score as well as FCR. Despite originating from close geographi­cal regions for isolates 1 and 2, the difference in biopathologic factors may be either due to different proportion of identified species or the different pathogenicity of the species present in the isolates.Conclusion: The results highlight the importance of considering various species of Eimeria in designing the preventive, control and treatment strategies to prevent coccidiosis in different regions of Iran. Further characterization of each isolate would be the next step to provide a basis for coccidiosis research with well-characterized local isolates.

  3. Exogenous selection rather than cytonuclear incompatibilities shapes asymmetrical fitness of reciprocal Arabidopsis hybrids.

    Science.gov (United States)

    Muir, Graham; Ruiz-Duarte, Paola; Hohmann, Nora; Mable, Barbara K; Novikova, Polina; Schmickl, Roswitha; Guggisberg, Alessia; Koch, Marcus A

    2015-04-01

    Reciprocal crosses between species often display an asymmetry in the fitness of F1 hybrids. This pattern, referred to as isolation asymmetry or Darwin's corollary to Haldane's rule, is a general feature of reproductive isolation in plants, yet factors determining its magnitude and direction remain unclear. We evaluated reciprocal species crosses between two naturally hybridizing diploid species of Arabidopsis to assess the degree of isolation asymmetry at different postmating life stages. We found that pollen from Arabidopsis arenosa will usually fertilize ovules from Arabidopsis lyrata; the reverse receptivity being less complete. Maternal A. lyrata parents set more F1 hybrid seed, but germinate at lower frequency, reversing the asymmetry. As predicted by theory, A. lyrata (the maternal parent with lower seed viability in crosses) exhibited accelerated chloroplast evolution, indicating that cytonuclear incompatibilities may play a role in reproductive isolation. However, this direction of asymmetrical reproductive isolation is not replicated in natural suture zones, where delayed hybrid breakdown of fertility at later developmental stages, or later-acting selection against A. arenosa maternal hybrids (unrelated to hybrid fertility, e.g., substrate adaptation) may be responsible for an excess of A. lyrata maternal hybrids. Exogenous selection rather than cytonuclear incompatibilities thus shapes the asymmetrical postmating isolation in nature.

  4. Probiotic attributes of indigenous Lactobacillus spp. isolated from traditional fermented foods and beverages of north-western Himalayas using in vitro screening and principal component analysis.

    Science.gov (United States)

    Kumari, Anila; Angmo, Kunzes; Monika; Bhalla, Tek Chand

    2016-05-01

    The present research was designed to explore indigenous probiotic Lactic acid bacteria from traditional fermented foods and beverages of North-western Himalayas for their probiotic potential. It was achieved through a step-by step approach focused on the technological characterization, evaluation of the probiotic traits and adherence ability. Fifty one LAB isolates from traditional fermented foods and beverages were initially screened for their technological properties and among them twenty isolates were selected. These isolates were further characterized and identified using 16S rRNA gene sequencing as Lactobacillus brevis (7 isolates), Lactobacillus casei (5), Lactobacillus paracasei (2), Lactobacillus buchneri (1), Lactobacillus plantarum (1) and Lactobacillus sp. (3). Identified isolates were evaluated by in vitro methods including survival in gastrointestinal tract, antibiotic susceptibility, antimicrobial activity, cell surface characteristics, exopolysacharride production and haemolytic activity. The results of these experiments were used as input data for Principal Component Analysis; thus, to select the most promising probiotic isolates. Three isolates (L. brevis PLA2, L. paracasei PLA8 and L. brevis PLA16) were found to be most technological relevant and promising probiotic candidates in comparison to commercial probiotic strains. L. brevis PLA2 was selected as best isolate with probiotic potential by in vitro adherence to the human intestinal HT-29 cell line.

  5. Zinc bioaccumulation by microbial consortium isolated from nickel smelter sludge disposal site

    Directory of Open Access Journals (Sweden)

    Kvasnová Simona

    2017-06-01

    Full Text Available Heavy metal pollution is one of the most important environmental issues of today. Bioremediation by microorganisms is one of technologies extensively used for pollution treatment. In this study, we investigated the heavy metal resistance and zinc bioaccumulation by microbial consortium isolated from nickel sludge disposal site near Sereď (Slovakia. The composition of consortium was analyzed based on MALDI-TOF MS of cultivable bacteria and we have shown that the consortium was dominated by bacteria of genus Arthrobacter. While consortium showed very good growth in the zinc presence, it was able to remove only 15 % of zinc from liquid media. Selected members of consortia have shown lower growth rates in the zinc presence but selected isolates have shown much higher bioaccumulation abilities compared to whole consortium (up to 90 % of zinc removal for NH1 strain. Bioremediation is frequently accelerated through injection of native microbiota into a contaminated area. Based on data obtained in this study, we can conclude that careful selection of native microbiota could lead to the identification of bacteria with increased bioaccumulation abilities.

  6. Genetic and phenotypic diversity of naturally isolated wild strains of Aspergillus niger with hyper glucose oxidase production

    Directory of Open Access Journals (Sweden)

    MAHMOUD EL-HARIRI

    2015-12-01

    Full Text Available Glucose oxidase (GOx is the basic stone for many of biological industry worldwide. The improvement of GOx production basically depends on selection of hyper producer strain of Aspergillus niger. Selective isolation and screening for natural hyper producer strains of A. niger and sequence analysis of the GOD gene, which is responsible for production of the enzyme, are very effective approaches to investigate the naturally modified strains of A. niger with hyper productive capacity of GOx enzyme. The aims of the current study were selective isolation of naturally hyper GOx producing strains of A. niger and evaluation of their GOx activities under optimized parameters in the laboratory. Five wild Egyptian isolates of A. niger were screened for GOx and catalase activity using two types of modified basal liquid media. The GOx activity was evaluated by high throughout liquid phase system. The isolates showed a variable activity for GOx production ranged from 0 to 28.7 U.ml-1. One isolate coded Strain 7 was negative GOx producer on Vogel's broth medium in comparison to other isolates, while its GOx activity on Cazpek Dox was considered as positive (7.28 U.ml-1. It was concluded that GOx production is affected by three controllable factors – the basal media components, time of incubation, and the strain with its adaption to the media components‎. Also, the catalase activity was tested and it was produced with a different degree of variability, which may be reflected on GOx stability. GOD genes of these wild variant of A. niger were cloned and sequenced to determine intraspecies diversity of GOD between the wild variants. The comparison of isolated wild variants to other reference hyper GOx producer strains of A. niger was performed to determine if the GOD sequence analysis of these strains can be distinguished based on their GOx activity. This is the first report for isolation and detection of naturally A. niger hyper GOx-producer strains with

  7. Isolation and characterization of putative endophytic bacteria antagonistic to Phoma tracheiphila and Verticillium albo-atrum.

    Science.gov (United States)

    Kalai-Grami, Leila; Saidi, Sabrine; Bachkouel, Sarra; Ben Slimene, Imen; Mnari-Hattab, Monia; Hajlaoui, Mohamed Rebah; Limam, Ferid

    2014-09-01

    A collection of 200 bacterial isolates recovered from citrus plants (Citrus limon, Citrus sinensis, and Citrus reticulata), Medicago truncatula and Laurus nobilis, was established. In vitro screening indicated that 28 isolates exhibited an inhibitory activity against the vascular pathogens Phoma tracheiphila and Verticillium albo-atrum. Isolates were screened according to their hydrolytic activities, plant growth-promoting bacteria (PGPB) abilities, as well as for the presence of nonribosomal peptide synthetase (NRPS) genes responsible of the lipopeptide biosynthesis. The results were positive for 16 isolates which exhibited at least two PGPB activities and a single NRPS gene. Genetic diversity of the selected isolates was studied using random amplified polymorphic DNA (RAPD) and repetitive element PCR (REP) tools that showed clustering of strains into three major groups (I, II, and III) (i, ii, and iii), respectively. Clustering was further confirmed by the 16S rDNA sequencing that assigned nine isolates to Bacillus velezensis, four isolates to Bacillus methyltrophicus, one isolate to Bacillus amyloliquefaciens, and two isolates to Bacillus mojavensis. Organ-bacterial genotype interaction as well as positive correlation with NRPS genes are discussed.

  8. Antioxidant activity directed isolations form punica granatum

    International Nuclear Information System (INIS)

    Siddiqi, R.; Saeed, M.G.; Sayeed, S.A.

    2012-01-01

    The extracts derived from pomegranate juice following antioxidant activity directed isolation were screened for their antioxidant activity through their ability to scavenge 2,2- diphenyl-l-picrylhydrazyl (DPPH) radicals. Only fractions which exhibited >50 / 0 DPPH scavenging effect at each step of isolation were selected for further purification and their ability to reduce peroxide formation (peroxide value) in heated com oil. Phytochemical analysis of the pure compounds finally obtained, revealed the presence of pelargonidin-3- galactose (Pg-3-galactose), cyanidin-3-glucose (Cy-3-Glucose), gallic acid, quercetin and myricetin in the fractions exhibiting >50% DPPH scavenging potential. The order of antioxidant activity of these pure compounds by DPPH method was found to be gallic acid> quercetin> myricetin> Cy-3-galactose> Pg-3-Glucose while order with respect to reduction in peroxide value (PV) was the reverse of DPPH. (author)

  9. Isolated galaxies

    International Nuclear Information System (INIS)

    Einasto, Maret

    1990-01-01

    To test for the possible presence of really isolated galaxies, which form a randomly distributed population in voids, we compare the distribution of most isolated galaxies in an observed sample with distributions of the same number of random points using the nearest neighbour test. The results show that the random population of really isolated galaxies does not exist - even the most isolated galaxies are connected with systems of galaxies, forming their outlying parts. (author)

  10. Thermodynamic tables for nuclear waste isolation

    International Nuclear Information System (INIS)

    Phillips, S.L.; Hale, F.V.; Silvester, L.F.; Siegel, M.D.

    1988-05-01

    Tables of consistent thermodynamic property values for nuclear waste isolation are given. The tables include critically assessed values for Gibbs energy of formation, enthalpy of formation, entropy and heat capacity for minerals; solids; aqueous ions; ion pairs and complex ions of selected actinide and fission decay products at 25 degree C and zero ionic strength. These intrinsic data are used to calculate equilibrium constants and standard potentials which are compared with typical experimental measurements and other work. Recommendations for additional research are given. 13 figs., 23 tabs

  11. Antimicrobial resistance and prevalence of resistance genes of obligate anaerobes isolated from periodontal abscesses.

    Science.gov (United States)

    Xie, Yi; Chen, Jiazhen; He, Junlin; Miao, Xinyu; Xu, Meng; Wu, Xingwen; Xu, Beiyun; Yu, Liying; Zhang, Wenhong

    2014-02-01

    This study attempts to determine the antimicrobial resistance profiles of obligate anaerobic bacteria that were isolated from a periodontal abscess and to evaluate the prevalence of resistance genes in these bacteria. Forty-one periodontal abscess samples were cultivated on selective and non-selective culture media to isolate the oral anaerobes. Their antibiotic susceptibilities to clindamycin, doxycycline, amoxicillin, imipenem, cefradine, cefixime, roxithromycin, and metronidazole were determined using the agar dilution method, and polymerase chain reaction assays were performed to detect the presence of the ermF, tetQ, nim, and cfxA drug resistance genes. A total of 60 different bacterial colonies was isolated and identified. All of the isolates were sensitive to imipenem. Of the strains, 6.7%, 13.3%, 16.7%, and 25% were resistant to doxycycline, metronidazole, cefixime, and amoxicillin, respectively. The resistance rate for both clindamycin and roxithromycin was 31.7%. Approximately 60.7% of the strains had the ermF gene, and 53.3% of the amoxicillin-resistant strains were found to have the cfxA gene. Two nim genes that were found in eight metronidazole-resistant strains were identified as nimB. In the present study, the Prevotella species are the most frequently isolated obligate anaerobes from periodontal abscesses. The current results show their alarmingly high resistance rate against clindamycin and roxithromycin; thus, the use of these antibiotics is unacceptable for the empirical therapy of periodontal abscesses. A brief prevalence of four resistance genes in the anaerobic bacteria that were isolated was also demonstrated.

  12. Adaptive evolution of simian immunodeficiency viruses isolated from two conventional progressor macaques with neuroaids

    Energy Technology Data Exchange (ETDEWEB)

    Foley, Brian T [Los Alamos National Laboratory; Korber, Bette T [Los Alamos National Laboratory

    2008-01-01

    Simian immunodeficiency virus infection of macaques may result in neuroAIDS, a feature more commonly observed in macaques with rapid progressive disease than in those with conventional disease. This is the first report of two conventional progressors (H631 and H636) with encephalitis in rhesus macaques inoculated with a derivative of SIVsmES43-3. Phylogenetic analyses of viruses isolated from the cerebral spinal fluid (CSF) and plasma from both animals demonstrated tissue compartmentalization. Additionally, virus from the central nervous system (CNS) was able to infect primary macaque monocyte-derived macrophages more efficiently than virus from plasma. Conversely, virus isolated from plasma was able to replicate better in peripheral blood mononuclear cells than virus from CNS. We speculate that these viruses were under different selective pressures in their separate compartments. Furthermore, these viruses appear to have undergone adaptive evolution to preferentially replicate in their respective cell targets. Analysis of the number of potential N-linked glycosylation sites (PNGS) in gp160 showed that there was a statistically significant loss of PNGS in viruses isolated from CNS in both macaques compared to SIVsmE543-3. Moreover, virus isolated from the brain in H631, had statistically significant loss of PNGS compared to virus isolated from CSF and plasma of the same animal. It is possible that the brain isolate may have adapted to decrease the number of PNGS given that humoral immune selection pressure is less likely to be encountered in the brain. These viruses provide a relevant model to study the adaptations required for SIV to induce encephalitis.

  13. Basalt waste isolation project overview

    International Nuclear Information System (INIS)

    Dahlem, D.H.

    1987-01-01

    The proposed candidate site for a high-level nuclear waste repository is located beneath the Hanford Nuclear Reservation in southeastern Washington State. At this point, the Hanford Reservation has been selected as one of three preferred candidates in the draft environmental assessment. Project activities have concentrated on understanding the site location with respect to the 10CFR60, 40CFR191, and 10CFR960, identifying critical parameters for design of water package and repository seals, and identifying parameters for repository design. This paper describes the program to evaluate the site and to identify the natural processes that would effect isolation

  14. Antibiotic susceptibility of Brachyspira hyodysenteriae isolates from Czech swine farms: a 10-year follow-up study

    Directory of Open Access Journals (Sweden)

    Josef Prášek

    2014-01-01

    Full Text Available Brachyspira hyodysenteriae is the causative agent of swine dysentery. Loss of clinical efficacy of some antimicrobial agents authorized for treating swine dysentery was observed on certain Czech pig farms. The aim of the present study was to evaluate the antimicrobial sensitivity of six antibiotics using a set of 202 randomly selected B. hyodysenteriae isolates obtained from farms in the Czech Republic between years 1997 and 2006. Minimum inhibitory concentration of antibiotics tylosin, lincomycin, tylvalosin, chlortetracyclin, tiamulin and valnemulin were tested, using an agar dilution method. All antibiotics tested showed an increase in minimal inhibitory concentrations. Continual decrease in susceptibility of B. hyodysenteriae isolates to tiamulin and valnemulin was observed. Multiresistant B. hyodysenteriae were isolated more frequently in the past years. Only a careful use of antibiotics can ensure their efficacy, especially in case of pleuromutilins, in the strategic therapy of swine dysentery. This rare study demonstrates the minimal inhibitory concentration changes of selected antidysenterics among Czech isolates of Brachyspira hyodysenteriae during a ten-year period.

  15. Isolation and characterization of novel chitinolytic bacteria

    Science.gov (United States)

    Gürkök, Sümeyra; Görmez, Arzu

    2016-04-01

    Chitin, a linear polymer of β-1,4-N-acetylglucosamine units, is one of the most abundant biopolymers widely distributed in the marine and terrestrial environments. It is found as a structural component of insects, crustaceans and the cell walls of fungi. Chitinases, the enzymes degrading chitin by cleaving the β-(1-4) bond, have gained increased attention due to their wide range of biotechnological applications, especially for biocontrol of harmful insects and phytopathogenic fungi in agriculture. In the present study, 200 bacterial isolates from Western Anatolia Region of Turkey were screened for chitinolytic activity on agar media amended with colloidal chitin. Based on the chitin hydrolysis zone, 13 isolates were selected for further study. Bacterial isolates with the highest chitinase activity were identified as Acinetobacter calcoaceticus, Arthrobacter oxydans, Bacillus cereus, Bacillus megaterium, Brevibacillus reuszeri, Kocuria erythromyxa, Kocuria rosea, Novosphingobium capsulatum, Rhodococcus bratislaviensis, Rhodococcus fascians and Staphylococcus cohnii by MIS and BIOLOG systems. The next aims of the study are to compare the productivity of these bacteria quantitatively, to purify the enzyme from the most potent producer and to apply the pure enzyme for the fight against the phytopathogenic fungi and harmful insects.

  16. Commonalities in Development of Pure Breeds and Population Isolates Revealed in the Genome of the Sardinian Fonni's Dog

    Science.gov (United States)

    Dreger, Dayna L.; Davis, Brian W.; Cocco, Raffaella; Sechi, Sara; Di Cerbo, Alessandro; Parker, Heidi G.; Polli, Michele; Marelli, Stefano P.; Crepaldi, Paola; Ostrander, Elaine A.

    2016-01-01

    The island inhabitants of Sardinia have long been a focus for studies of complex human traits due to their unique ancestral background and population isolation reflecting geographic and cultural restriction. Population isolates share decreased genomic diversity, increased linkage disequilibrium, and increased inbreeding coefficients. In many regions, dogs and humans have been exposed to the same natural and artificial forces of environment, growth, and migration. Distinct dog breeds have arisen through human-driven selection of characteristics to meet an ideal standard of appearance and function. The Fonni’s Dog, an endemic dog population on Sardinia, has not been subjected to an intensive system of artificial selection, but rather has developed alongside the human population of Sardinia, influenced by geographic isolation and unregulated selection based on its environmental adaptation and aptitude for owner-desired behaviors. Through analysis of 28 dog breeds, represented with whole-genome sequences from 13 dogs and ∼170,000 genome-wide single nucleotide variants from 155 dogs, we have produced a genomic illustration of the Fonni’s Dog. Genomic patterns confirm within-breed similarity, while population and demographic analyses provide spatial identity of Fonni’s Dog to other Mediterranean breeds. Investigation of admixture and fixation indices reveals insights into the involvement of Fonni’s Dogs in breed development throughout the Mediterranean. We describe how characteristics of population isolates are reflected in dog breeds that have undergone artificial selection, and are mirrored in the Fonni’s Dog through traditional isolating factors that affect human populations. Lastly, we show that the genetic history of Fonni’s Dog parallels demographic events in local human populations. PMID:27519604

  17. A comparative study of coastal and clinical isolates of Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Anusree V. Nair

    2015-09-01

    Full Text Available Pseudomonas aeruginosa is a ubiquitous Gram-negative bacterium having a versatile metabolic potential and great ecological and clinical significance. The geographical distribution of P. aeruginosahas revealed the existence of an unbiased genetic arrangement in terrestrial isolates. In contrast, there are very few reports about P. aeruginosa strains from marine environments. The present work was aimed at studying the distribution of P. aeruginosa in coastal waters along the Indian Peninsula and understanding the environmental influence on genotypic, metabolic and phenotypic characteristics by comparing marine and clinical isolates. Of the 785 marine isolates obtained on selective media, only 32 (~4.1% were identified as P. aeruginosa, based on their fatty acid methyl ester profiles. A low Euclidian distance value (P. aeruginosa. While biogeographical separation was not evident based solely on phenotypic and metabolic typing, genomic and metatranscriptomic studies are more likely to show differences between these isolates. Thus, newer and more insightful methods are required to understand the ecological distribution of this complex group of bacteria.

  18. Isolation of Primary Murine Retinal Ganglion Cells (RGCs) by Flow Cytometry.

    Science.gov (United States)

    Chintalapudi, Sumana R; Patel, Need N; Goldsmith, Zachary K; Djenderedjian, Levon; Wang, Xiang Di; Marion, Tony N; Jablonski, Monica M; Morales-Tirado, Vanessa M

    2017-07-05

    Neurodegenerative diseases often have a devastating impact on those affected. Retinal ganglion cell (RGC) loss is implicated in an array of diseases, including diabetic retinopathy and glaucoma, in addition to normal aging. Despite their importance, RGCs have been extremely difficult to study until now due in part to the fact that they comprise only a small percentage of the wide variety of cells in the retina. In addition, current isolation methods use intracellular markers to identify RGCs, which produce non-viable cells. These techniques also involve lengthy isolation protocols, so there is a lack of practical, standardized, and dependable methods to obtain and isolate RGCs. This work describes an efficient, comprehensive, and reliable method to isolate primary RGCs from mice retinae using a protocol based on both positive and negative selection criteria. The presented methods allow for the future study of RGCs, with the goal of better understanding the major decline in visual acuity that results from the loss of functional RGCs in neurodegenerative diseases.

  19. Characterization of Microorganisms Isolated from Petroleum Hydrocarbon Polluted Soil

    Directory of Open Access Journals (Sweden)

    Adriana Criste

    2016-02-01

    Full Text Available Bioremediation has received a great deal of attention, and bacteria isolated from polluted soil can be usedin that process. In this study, we performed an evaluation of the physiological groups of microorganisms fromsoil contaminated with petroleum. Bacterial strains were isolated from contaminated soil using the selectiveenrichment technique. Minimal Salt Media was used for serial dilutions to determine viable cell count. Thenumber of total viable cells and different types of microorganisms in the original sample was determined by serialdilution, agar plating procedure using selective media. The plates were incubated at 300C for 24-72 hours. Distinctcolonies growing on each plate were selected, and stored at freezing temperatures. The bacterial colonies werethen identified by Gram staining and biochemical tests. Following our research, it was observed that although thetotal microbial load of soil is relatively close in value, there are differences regarding the physiological group ofmicroorganisms. In the oil contaminated soil sample the largest group of microorganisms was the nitrous nitrifyingbacteria followed by nitrate bacteria. All bacterial strains that were isolated from soil samples contaminated withhydrocarbons but also the Pseudomonas putida and Bacillus subtillis strains can use diesel fuel as a food source.With the increase of diesel fuel concentration from culture medium, the majority of the bacterial strains that wereused in our experiments showed an increased value of absorbance. This fact suggests that these strains can be usedin bioremediation processes.

  20. Isolation and Selection of Microalgal Strains from Natural Water Sources in Viet Nam with Potential for Edible Oil Production.

    Science.gov (United States)

    Thao, Tran Yen; Linh, Dinh Thi Nhat; Si, Vo Chi; Carter, Taylor W; Hill, Russell T

    2017-06-23

    Industrial vegetable oil production in Viet Nam depends on oil seeds and crude plant oils that are currently more than 90% imported. As the first step in investigating the feasibility of using microalgae to provide Viet Nam with a domestic source of oil for food and edible oil industries, fifty lipid-producing microalgae were isolated and characterized. The microalgae were isolated from water sources ranging from freshwater to brackish and marine waters from a wide geographic distribution in Viet Nam. Initial analyses showed that 20 of the 50 strains had good growth rates, produced high biomass and had high lipid content, ranging up to 50% of dry weight biomass. 18S rRNA gene sequence analyses of the 50 strains showed a great diversity in this assemblage of microalgae, comprising at least 38 species and representatives of 25 genera : Chlamydomonas , Poterioochromonas , Scenedesmus , Desmodesmus , Chlorella , Bracteacoccus , Monoraphidium , Selenastrum , Acutodesmus , Mychonastes , Ankistrodesmus , Kirchneriella , Raphidocelis , Dictyosphaerium , Coelastrella , Schizochlamydella , Oocystidium , Nannochloris , Auxenochlorella , Chlorosarcinopsis , Stichococcus , Picochlorum , Prasinoderma , Chlorococcum , and Marvania. Some of the species are closely related to well-known lipid producers such as Chlorella sorokiniana , but some other strains are not closely related to the strains found in public sequence databases and likely represent new species. Analysis of oil quality showed that fatty acid profiles of the microalgal strains were very diverse and strain-dependent. Fatty acids in the microalgal oils comprised saturated fatty acids (SFAs), poly-unsaturated fatty acids (PUFAs), and mono-unsaturated fatty acids (MUFAs). The main SFA was palmitic acid. MUFAs and PUFAs were dominated by oleic acid, and linoleic and linolenic acids, respectively. Some strains were especially rich in the essential fatty acid α-linolenic acid (ALA), which comprised more than 20% of the