Molecular biological enhancement of coal biodesulfurization. [Rhodococcus rhodochrous

Energy Technology Data Exchange (ETDEWEB)

Kilbane, J.J.; Bielaga, B.A.

1990-07-01

The overall objective of this project is to sue molecular genetics to develop strains of bacteria with enhanced ability to remove sulfur from coal and to obtain data that will allow the performance and economics of a coal biodesulfurization process to be predicted. The work planned for the current quarter (May 1990 to July 1990) includes the following activities: (1) Construct a cloning vector that can be used in Rhodococcus rhodochrous IGTS8 from the small cryptic plasmid found in Rhodococcus rhodochrous ATCC 190607; (2) Develop techniques for the genetic analysis of IGTS8; (3) Continue biochemical experiments, particularly those that may allow the identification of desulfurization-related enzymes; (4) Continue experiments with coal to determine the kinetics of organic sulfur removal.

Plant exudates promote PCB degradation by a rhodococcal rhizobacteria

Energy Technology Data Exchange (ETDEWEB)

Toussaint, Jean-Patrick; Pham, Thi Thanh My; Barriault, Diane; Sylvestre, Michel [Instiut National de la Recherche Scientifique INRS, Laval, QC (Canada). Inst. Armand-Frappier

2012-09-15

Rhodococcus erythropolis U23A is a polychlorinated biphenyl (PCB)-degrading bacterium isolated from the rhizosphere of plants grown on a PCB-contaminated soil. Strain U23A bphA exhibited 99% identity with bphA1 of Rhodococcus globerulus P6. We grew Arabidopsis thaliana in a hydroponic axenic system, collected, and concentrated the plant secondary metabolite-containing root exudates. Strain U23A exhibited a chemotactic response toward these root exudates. In a root colonizing assay, the number of cells of strain U23A associated to the plant roots (5.7 x 105 CFU g{sup -1}) was greater than the number remaining in the surrounding sand (4.5 x 104 CFU g{sup -1}). Furthermore, the exudates could support the growth of strain U23A. In a resting cell suspension assay, cells grown in a minimal medium containing Arabidopsis root exudates as sole growth substrate were able to metabolize 2,3,4'- and 2,3',4-trichlorobiphenyl. However, no significant degradation of any of congeners was observed for control cells grown on Luria-Bertani medium. Although strain U23A was unable to grow on any of the flavonoids identified in root exudates, biphenyl-induced cells metabolized flavanone, one of the major root exudate components. In addition, when used as co-substrate with sodium acetate, flavanone was as efficient as biphenyl to induce the biphenyl catabolic pathway of strain U23A. Together, these data provide supporting evidence that some rhodococci can live in soil in close association with plant roots and that root exudates can support their growth and trigger their PCB-degrading ability. This suggests that, like the flagellated Gram-negative bacteria, non-flagellated rhodococci may also play a key role in the degradation of persistent pollutants. (orig.)

Structural and biochemical characterization of 3-hydroxybenzoate 6-hydroxylase

NARCIS (Netherlands)

Montersino, S.

2012-01-01

<p> The thesis deals with the characterization of a new flavoprotein hydroxylase 3 hydroxybenzoate 6-hydroxylase (3HB6H) from Rhodococcus jostii RHA1. 3HB6H is able to insert exclusively oxygen in para-position and the enzyme has been chosen to study the structural basis of such regioselectivity. As

Successful treatment of Rhodococcus equi pulmonary infection in a renal transplant recipient.

OpenAIRE

Marsh, H. P.; Bowler, I. C.; Watson, C. J.

2000-01-01

The rhodococcus is a mycobacterium-like organism which is normally a pathogen in foals. It usually spreads by direct contact or by aerosol from horse faeces and causes pyogranulomatous pulmonary infections. Occasionally, it acts opportunistically to infect immuno-compromised human hosts, most commonly those with the acquired immune deficiency syndrome (AIDS). Here we report a pulmonary infection by Rhodococcus equi in a renal transplant recipient who was successfully treated. The literature o...

Isolation and characterization of Rhodococcus ruber CGMCC3090 ...

African Journals Online (AJOL)

STORAGESEVER

2009-10-19

Oct 19, 2009 ... Available online at http://www.academicjournals.org/AJB. ISSN 1684–5315 .... the soil samples by an enrichment culture method in basal medium containing 0.5 ..... The phylogeny of the genus Rhodococcus currently remains ...

Complete Genome Sequence of a Rhodococcus Species Isolated from the Winter Skate Leucoraja ocellata.

Science.gov (United States)

Wiens, Julia; Ho, Ryan; Fernando, Dinesh; Kumar, Ayush; Loewen, Peter C; Brassinga, Ann Karen C; Anderson, W Gary

2016-09-01

We report here a genome sequence for Rhodococcus sp. isolate UM008 isolated from the renal/interrenal tissue of the winter skate Leucoraja ocellata Genome sequence analysis suggests that Rhodococcus bacteria may act in a novel mutualistic relationship with their elasmobranch host, serving as biocatalysts in the steroidogenic pathway of 1α-hydroxycorticosterone. Copyright © 2016 Wiens et al.

The detection and phylogenetic analysis of the alkane 1-monooxygenase gene of members of the genus Rhodococcus.

Science.gov (United States)

Táncsics, András; Benedek, Tibor; Szoboszlay, Sándor; Veres, Péter G; Farkas, Milán; Máthé, István; Márialigeti, Károly; Kukolya, József; Lányi, Szabolcs; Kriszt, Balázs

2015-02-01

Naturally occurring and anthropogenic petroleum hydrocarbons are potential carbon sources for many bacteria. The AlkB-related alkane hydroxylases, which are integral membrane non-heme iron enzymes, play a key role in the microbial degradation of many of these hydrocarbons. Several members of the genus Rhodococcus are well-known alkane degraders and are known to harbor multiple alkB genes encoding for different alkane 1-monooxygenases. In the present study, 48 Rhodococcus strains, representing 35 species of the genus, were investigated to find out whether there was a dominant type of alkB gene widespread among species of the genus that could be used as a phylogenetic marker. Phylogenetic analysis of rhodococcal alkB gene sequences indicated that a certain type of alkB gene was present in almost every member of the genus Rhodococcus. These alkB genes were common in a unique nucleotide sequence stretch absent from other types of rhodococcal alkB genes that encoded a conserved amino acid motif: WLG(I/V/L)D(G/D)GL. The sequence identity of the targeted alkB gene in Rhodococcus ranged from 78.5 to 99.2% and showed higher nucleotide sequence variation at the inter-species level compared to the 16S rRNA gene (93.9-99.8%). The results indicated that the alkB gene type investigated might be applicable for: (i) differentiating closely related Rhodococcus species, (ii) properly assigning environmental isolates to existing Rhodococcus species, and finally (iii) assessing whether a new Rhodococcus isolate represents a novel species of the genus. Copyright © 2014 Elsevier GmbH. All rights reserved.

Genetically engineered micro-organisms: Aromatic hydrocarbon biodegradation genes from Rhodococcus

International Nuclear Information System (INIS)

Kendall, K.

1993-01-01

DNA known to encode toluene biodegradation genes in Pseudomonas putida was used in Southern Blots to identify homologous DNA in the unrelated toluene degrading Actinomycete, Rhodococcus sp. ATCC 19070. Two strongly hybridizing EcoRI fragments of 2.3 kb and 2.7 kb respectively were cloned into E. coli. Sequence analysis of a 400 bp section of the 2.3 kb fragment demonstrated that it encodes proteins with similar amino acid sequences to the xylX and xylY genes of P. putida. These proteins are components of toluate oxygenase, the enzyme catalyzing the first step in the metabolism of benzoic acid

Vanillin Catabolism in Rhodococcus jostii RHA1

Science.gov (United States)

Chen, Hao-Ping; Chow, Mindy; Liu, Chi-Chun; Lau, Alice; Liu, Jie

2012-01-01

Genes encoding vanillin dehydrogenase (vdh) and vanillate O-demethylase (vanAB) were identified in Rhodococcus jostii RHA1 using gene disruption and enzyme activities. During growth on vanillin or vanillate, vanA was highly upregulated while vdh was not. This study contributes to our understanding of lignin degradation by RHA1 and other actinomycetes. PMID:22057861

Expression control of nitrile hydratase and amidase genes in Rhodococcus erythropolis and substrate specificities of the enzymes

Czech Academy of Sciences Publication Activity Database

Rucká, Lenka; Volkova, Olga; Pavlík, Adam; Kaplan, Ondřej; Kracík, M.; Nešvera, Jan; Martínková, Ludmila; Pátek, Miroslav

2014-01-01

Roč. 105, č. 6 (2014), s. 1179-1190 ISSN 0003-6072 R&D Projects: GA MŠk(CZ) LC06010; GA ČR(CZ) GAP504/11/0394 Institutional support: RVO:61388971 Keywords : Rhodococcus erythropolis * Amidase * Nitrile hydratase Subject RIV: EE - Microbiology, Virology Impact factor: 1.806, year: 2014

Fluorene transformation by bacteria of the genus Rhodococcus

NARCIS (Netherlands)

Finkelstein, Z.I.; Baskunov, B.P.; Golovlev, E.L.; Vervoort, J.J.M.; Rietjens, I.M.C.M.; Baboshin, M.A.; Golovleva, L.A.

2003-01-01

Of the four investigated Rhodococcus strains (R. rhodochrous172, R. opacus 4a and 557, and R. rhodnii 135), the first three strains were found to be able to completely transform fluorene when it was present in the medium as the sole source of carbon at a concentration of 12¿25 mg/l. At a fluorene

Rhodococcus equi Parte 2: imunologia e profilaxia Rhodococcus equi Part 2: immunology and profilaxy

Directory of Open Access Journals (Sweden)

Ana Carolina Rusca Correa Porto

2011-12-01

Full Text Available Rhodococcus equi é um patógeno ubíquo e habitante da flora intestinal dos equinos de importância na neonatologia equina. Todos os potros são expostos à doença ao nascimento, porém alguns a desenvolvem e outros não. Este artigo revisa características da resposta imune, tanto em adultos competentes quanto em potros suscetíveis ao patógeno. A resposta imune humoral é abordada, incluindo uma discussão sobre o uso do plasma hiperimune como ferramenta profilática. O papel dos mecanismos de imunidade inata na suscetibilidade de alguns potros ao R. equi é também abordado. Da mesma maneira, os componentes envolvidos na resposta cito-mediada são revisados, com atenção particular às pesquisas direcionadas ao desenvolvimento de uma vacina efetiva para ser utilizada em potros.Rhodococcus equi is an important pathogen in equine neonatology that is ubiquitous and a normal intestinal inhabitant of equines. All foals are exposed at birth, however, some foals develop disease and others do not. This article reviews concepts of the equine immune response, both in the immune adult and susceptible neonate, with respect to this pathogen. Humoral immune responses are addressed, with a discussion on the use of hyperimmune plasma as a prophylactic tool. The role that innate immune mechanisms play in the susceptibility of some foals to R. equi infection is also highlighted. Likewise, cell-mediated immune components are reviewed, with particular attention directed towards research to develop an effective vaccine for foals.

Growth kinetics and biodeterioration of polypropylene microplastics by Bacillus sp. and Rhodococcus sp. isolated from mangrove sediment.

Science.gov (United States)

Auta, H S; Emenike, C U; Jayanthi, B; Fauziah, S H

2018-02-01

Interest in the biodegradation of microplastics is due to their ubiquitous distribution, availability, high persistence in the environment and deleterious impact on marine biota. The present study evaluates the growth response and mechanism of polypropylene (PP) degradation by Bacillus sp. strain 27 and Rhodococcus sp. strain 36 isolated from mangrove sediments upon exposure to PP microplastics. Both bacteria strains were able to utilise PP microplastic for growth as confirmed by the reduction of the polymer mass. The weight loss was 6.4% by Rhodococcus sp. strain 36 and 4.0% by Bacillus sp. strain 27 after 40days of incubation. PP biodegradation was further confirmed using Fourier-transform infrared spectroscopy and scanning electron microscopy analyses, which revealed structural and morphological changes in the PP microplastics with microbial treatment. These analyses showed that the isolates can colonise, modify and utilise PP microplastics as carbon source. Copyright © 2017 Elsevier Ltd. All rights reserved.

Microbial biodiesel production from oil palm biomass hydrolysate using marine Rhodococcus sp. YHY01.

Science.gov (United States)

Bhatia, Shashi Kant; Kim, Junyoung; Song, Hun-Seok; Kim, Hyun Joong; Jeon, Jong-Min; Sathiyanarayanan, Ganesan; Yoon, Jeong-Jun; Park, Kyungmoon; Kim, Yun-Gon; Yang, Yung-Hun

2017-06-01

The effect of various biomass derived inhibitors (i.e. furfural, hydroxymethylfurfural (HMF), vanillin, 4-hydroxy benzaldehyde (4-HB) and acetate) was investigated for fatty acid accumulation in Rhodococcus sp. YHY 01. Rhodococcus sp. YHY01 was able to utilize acetate, vanillin, and 4-HB for biomass production and fatty acid accumulation. The IC 50 value for furfural (3.1mM), HMF (3.2mM), vanillin (2.0mM), 4-HB (2.7mM) and acetate (3.7mM) was calculated. HMF and vanillin affect fatty acid composition and increase saturated fatty acid content. Rhodococcus sp. YHY 01 cultured with empty fruit bunch hydrolysate (EFBH) as the main carbon source resulted in enhanced biomass (20%) and fatty acid productivity (37%), in compression to glucose as a carbon source. Overall, this study showed the beneficial effects of inhibitory molecules on growth and fatty acid production, and support the idea of biomass hydrolysate utilization for biodiesel production by avoiding complex efforts to remove inhibitory compounds. Copyright © 2017 Elsevier Ltd. All rights reserved.

Biodegradation of phenol using recombinant plasmid-carrying Rhodococcus erythropolis strains

Czech Academy of Sciences Publication Activity Database

Zídková, L.; Szököl, Juraj; Rucká, Lenka; Pátek, Miroslav; Nešvera, Jan

2013-01-01

Roč. 84, OCT 2013 (2013), s. 179-184 ISSN 0964-8305 R&D Projects: GA MŠk 2B08062 Institutional research plan: CEZ:AV0Z50200510 Keywords : Rhodococcus erythropolis * Phenol degradation * Wastewater Subject RIV: EE - Microbiology, Virology Impact factor: 2.235, year: 2013

Rhodococcus equi infection after alemtuzumab therapy for T-cell prolymphocytic leukemia

NARCIS (Netherlands)

Meeuse, Jan J.; Sprenger, Herman G.; Van Assen, Sander; Leduc, Dominique; Daenen, Simon M.G.J.; Arends, Jan P.; van der Werf, Tjipke

2007-01-01

Rhodococcus equi, mainly known from veterinary medicine as a pathogen in domestic animals, can also cause infections in immunocompromised humans, especially in those with defects in cellular immunity. Alemtuzumab, an anti-CD52 monoclonal antibody, causes lymphocytopenia by eliminating CD52-positive

Serological survey of Rhodococcus equi infection in horses in Hokkaido.

Science.gov (United States)

Sanada, Y; Noda, H; Nagahata, H

1992-08-01

Serological survey of Rhodococcus equi infection in horses in Hokkaido was performed using ELISA. Of 2,879 horse sera, 318 (11.0%) gave antibody-positive (OD greater than or equal to 0.3) reactions. The antibody-positive rate of female was significantly higher (p less than 0.01) than that of male, and no statistical difference between Anglo-Arab and thoroughbred was detected in the antibody-positive rate. The maximum antibody-positive rate (27.1%) was shown at 14 years of age. The antibody-positive rates on the 160 farms were found to vary widely from 0 to 78.9%. A significant difference (p less than 0.01) in the antibody-positive rate was detected among horse farms. It was elucidated that 100 (62.5%) out of 160 horse farms had an antibody-positive horse. These results indicate that R. equi was widespread on horse farms, and the level of environmental contamination with R. equi differed among horse farms.

Severe Rhodococcus equi pneumonia: case report and literature review

DEFF Research Database (Denmark)

Vestbo, Jørgen; Lundgren, Jens Dilling; Gaub, J

1991-01-01

or sputum on standard media, but is frequently regarded as a contaminant. Mortality from Rhodococcus equi pneumonia is high (25%) and early surgical intervention has been recommended. Based on this review, the benefit of surgery seems dubious, whereas good results have been obtained using long......-term antibiotic treatment with erythromycin plus rifampicin, or vancomycin in combination with either of these antibiotics....

Mathematic Modeling for Optimum Conditions on Aflatoxin B1 Degradation by the Aerobic Bacterium Rhodococcus erythropolis

Directory of Open Access Journals (Sweden)

Jiujiang Yu

2012-11-01

Full Text Available Response surface methodology was employed to optimize the degradation conditions of AFB1 by Rhodococcus erythropolis in liquid culture. The most important factors that influence the degradation, as identified by a two-level Plackett-Burman design with six variables, were temperature, pH, liquid volume, inoculum size, agitation speed and incubation time. Central composite design (CCD and response surface analysis were used to further investigate the interactions between these variables and to optimize the degradation efficiency of R. erythropolis based on a second-order model. The results demonstrated that the optimal parameters were: temperature, 23.2 °C; pH, 7.17; liquid volume, 24.6 mL in 100-mL flask; inoculum size, 10%; agitation speed, 180 rpm; and incubation time, 81.9 h. Under these conditions, the degradation efficiency of R. erythropolis could reach 95.8% in liquid culture, which was increased by about three times as compared to non-optimized conditions. The result by mathematic modeling has great potential for aflatoxin removal in industrial fermentation such as in food processing and ethanol production.

[Respiratory infections caused by slow-growing bacteria: Nocardia, Actinomyces, Rhodococcus].

Science.gov (United States)

Eschapasse, E; Hussenet, C; Bergeron, A; Lebeaux, D

2017-06-01

Pneumonia caused by slow-growing bacteria is rare but sometimes severe. These infections share many similarities such as several differential diagnoses, difficulties to identify the pathogen, the importance of involving the microbiologist in the diagnostic investigation and the need for prolonged antibiotic treatment. However, major differences distinguish them: Nocardia and Rhodococcus infect mainly immunocompromised patients while actinomycosis also concerns immunocompetent patients; the severity of nocardioses is related to their hematogenous spread while locoregional extension by contiguity makes the gravity of actinomycosis. For these diseases, molecular diagnostic tools are essential, either to obtain a species identification and guide treatment in the case of nocardiosis or to confirm the diagnosis from a biological sample. Treatment of these infections is complex due to: (1) the limited data in the literature; (2) the need for prolonged treatment of several months; (3) the management of toxicities and drug interactions for the treatment of Nocardia and Rhodococcus. Close cooperation between pneumonologists, infectious disease specialists and microbiologists is essential for the management of these patients. Copyright © 2017 SPLF. Published by Elsevier Masson SAS. All rights reserved.

Effect of Tween 80 on 9a-steroid hydroxylating activity and ultrastructural characteristics of Rhodococcus sp. cells

OpenAIRE

Avramova , Tatyana; Spassova , Dimitrina; Mutafov , Sava; Momchilova , Svetlana; Boyadjieva , Lilyana; Damyanova , Boryana; Angelova , Blaga

2010-01-01

International audience; Studied is the effect of the non-ionic surfactant Tween 80 on the microbial transformation of 4-androstene- 3,17-dione into its 9a-hydroxy-derivative by resting Rhodococcus sp. cells. The surfactant was applied in the cultivation medium as an additional source of carbon, in the transformation reaction medium as a mediator of the steroid substrate solubility or was used for permeabilization of the glucose grown Rhodococcus sp. cells. Special attention is paid to the fac...

Potential of Rhodococcus strains for biotechnological vanillin production from ferulic acid and eugenol.

Science.gov (United States)

Plaggenborg, Rainer; Overhage, Jörg; Loos, Andrea; Archer, John A C; Lessard, Philip; Sinskey, Anthony J; Steinbüchel, Alexander; Priefert, Horst

2006-10-01

The potential of two Rhodococcus strains for biotechnological vanillin production from ferulic acid and eugenol was investigated. Genome sequence data of Rhodococcus sp. I24 suggested a coenzyme A-dependent, non-beta-oxidative pathway for ferulic acid bioconversion, which involves feruloyl-CoA synthetase (Fcs), enoyl-CoA hydratase/aldolase (Ech), and vanillin dehydrogenase (Vdh). This pathway was proven for Rhodococcus opacus PD630 by physiological characterization of knockout mutants. However, expression and functional characterization of corresponding structural genes from I24 suggested that degradation of ferulic acid in this strain proceeds via a beta-oxidative pathway. The vanillin precursor eugenol facilitated growth of I24 but not of PD630. Coniferyl aldehyde was an intermediate of eugenol degradation by I24. Since the genome sequence of I24 is devoid of eugenol hydroxylase homologous genes (ehyAB), eugenol bioconversion is most probably initiated by a new step in this bacterium. To establish eugenol bioconversion in PD630, the vanillyl alcohol oxidase gene (vaoA) from Penicillium simplicissimum CBS 170.90 was expressed in PD630 together with coniferyl alcohol dehydrogenase (calA) and coniferyl aldehyde dehydrogenase (calB) genes from Pseudomonas sp. HR199. The recombinant strain converted eugenol to ferulic acid. The obtained data suggest that genetically engineered strains of I24 and PD630 are suitable candidates for vanillin production from eugenol.

Comparative Genomics of Rhodococcus equi Virulence Plasmids Indicates Host-Driven Evolution of the vap Pathogenicity Island.

Science.gov (United States)

MacArthur, Iain; Anastasi, Elisa; Alvarez, Sonsiray; Scortti, Mariela; Vázquez-Boland, José A

2017-05-01

The conjugative virulence plasmid is a key component of the Rhodococcus equi accessory genome essential for pathogenesis. Three host-associated virulence plasmid types have been identified the equine pVAPA and porcine pVAPB circular variants, and the linear pVAPN found in bovine (ruminant) isolates. We recently characterized the R. equi pangenome (Anastasi E, et al. 2016. Pangenome and phylogenomic analysis of the pathogenic actinobacterium Rhodococcus equi. Genome Biol Evol. 8:3140-3148.) and we report here the comparative analysis of the virulence plasmid genomes. Plasmids within each host-associated type were highly similar despite their diverse origins. Variation was accounted for by scattered single nucleotide polymorphisms and short nucleotide indels, while larger indels-mostly in the plasticity region near the vap pathogencity island (PAI)-defined plasmid genomic subtypes. Only one of the plasmids analyzed, of pVAPN type, was exceptionally divergent due to accumulation of indels in the housekeeping backbone. Each host-associated plasmid type carried a unique PAI differing in vap gene complement, suggesting animal host-specific evolution of the vap multigene family. Complete conservation of the vap PAI was observed within each host-associated plasmid type. Both diversity of host-associated plasmid types and clonality of specific chromosomal-plasmid genomic type combinations were observed within the same R. equi phylogenomic subclade. Our data indicate that the overall strong conservation of the R. equi host-associated virulence plasmids is the combined result of host-driven selection, lateral transfer between strains, and geographical spread due to international livestock exchanges. © The Author(s) 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

Bio desulfurization of a system containing synthetic fuel by rhodococcus erythropolis ATCC 4277; Remocao de compostos sulfurosos de sitema bifasico contendo combustivel sintetico por Rhodococcus erythropolis ATCC 4277

Energy Technology Data Exchange (ETDEWEB)

Maass, Danielle; Souza, Antonio Augusto Ulson de; Souza, Selene Maria de Arruda Guelli Ulson de [Universidade Federal de Santa Catarina (UFSC), SC (Brazil)

2012-07-01

For decades the burning of fossil fuels released a lot of pollutants in the atmosphere. Among the most harmful is sulfur dioxide (SO{sub 2}), which reacts with the moisture in the air and turns into sulfuric acid, being the main cause of acid rain. Acid rain is very harmful to animal and plant kingdoms; accelerates the corrosion's processes of buildings and monuments, and causes serious health problems for humans. As a result, many countries have reformed their legislation to require the sale of fuels with very low sulfur content. The existing processes of desulfurization are not capable of removing sulfur so low. Therefore, there has developed a new process called bio desulfurization. In this process, the degradation of sulfur occurs through the action of microorganisms that act as catalysts. The bacterium Rhodococcus erythropolis has emerged as one of the most promising for bio desulfurization because it removes the sulfur without breaking the benzene rings, thereby maintaining the potential energy of the same. Using dibenzothiophene as a model of sulfur compounds, the products of the bio desulfurization process are 2- hydroxybiphenyl and sulfate. In this study we sought to examine the desulfurizing capacity of national Rhodococcus erythropolis strain ATCC4277 in a batch reactor using concentrations of organic phase (n-dodecane) of 20 and 80% (v/v). Rhodococcus erythropolis ATCC4277 was capable of degrading DBT in 93.3 and 98.0% in the presence of 20 and 80% (v/v) of synthetic fuel, respectively. (author)

Pneumonia por Rhodococcus equi em doente VIH (+: A propósito de uma associação rara

Directory of Open Access Journals (Sweden)

Paula Esteves

2007-09-01

Full Text Available Resumo: A infecção humana por Rhodococcus equi, mesmo no contexto da infecção pelo VIH, é relativamente rara. Os autores apresentam o caso clínico de um indivíduo, infectado pelo VIH, que apresentava um quadro clínico e imagiológico compatível com pneumonia, cujos exames iniciais revelaram a presença de Pneumocystis jiroveci. Após realização de terapêutica específica, perante a manutenção do quadro clínico-imagiológico, procedeu-se à realização de novos exames complementares, tendo sido identificado a presença de Rhodococcus equi no lavado broncoalveolar. Com instituição de terapêutica dirigida a este agente, verificou--se progressiva melhoria clínica e imagiológica. Os autores efectuam uma revisão bibliográfica da pneumonia por Rhodococcus equi, salientando a raridade desta associação e a elevada sobrevida observada. Abstract: The human infection by Rhodococcus equi, even in the presence of HIV infection, remains a rare disease. The authors present a case report of pneumonia, occurring in a HIV (+ man. After identifying Pneumocystis jiroveci in the BAL, despite proper medication, the patient didn’t improve. Another BAL was performed and a Rhodoccus equi isolated. The therapeutic regimen was changed according to this finding and the patient improved. The authors make a review of the literature, focusing on the rarity of this association and the high survival observed. Palavras-chave: Rhodococcus equi, Pneumocystis jiroveci, VIH, Key-words: Rhodococcus equi, Pneumocystis jiroveci, HIV

Genome-based exploration of the specialized metabolic capacities of the genus Rhodococcus

NARCIS (Netherlands)

Ceniceros, Ana; Dijkhuizen, Lubbert; Petrusma, Mirjan; Medema, M.H.

2017-01-01

Background Bacteria of the genus Rhodococcus are well known for their ability to degrade a large range of organic compounds. Some rhodococci are free-living, saprophytic bacteria; others are animal and plant pathogens. Recently, several studies have shown that their genomes encode putative pathways

Genome-based exploration of the specialized metabolic capacities of the genus Rhodococcus

NARCIS (Netherlands)

Ceniceros, Ana; Dijkhuizen, Lubbert; Petrusma, Mirjan; Medema, Marnix H.

2017-01-01

Background: Bacteria of the genus Rhodococcus are well known for their ability to degrade a large range of organic compounds. Some rhodococci are free-living, saprophytic bacteria; others are animal and plant pathogens. Recently, several studies have shown that their genomes encode putative pathways

Genome-based exploration of the specialized metabolic capacities of the genus Rhodococcus

NARCIS (Netherlands)

Ceniceros, Ana; Dijkhuizen, Lubbert; Petrusma, Mirjan; Medema, Marnix H

2017-01-01

BACKGROUND: Bacteria of the genus Rhodococcus are well known for their ability to degrade a large range of organic compounds. Some rhodococci are free-living, saprophytic bacteria; others are animal and plant pathogens. Recently, several studies have shown that their genomes encode putative pathways

Rhodococcus erythropolis MTHt3 biotransforms ergopeptines to lysergic acid

OpenAIRE

Thamhesl, Michaela; Apfelthaler, Elisabeth; Schwartz-Zimmermann, Heidi Elisabeth; Kunz-Vekiru, Elisavet; Krska, Rudolf; Kneifel, Wolfgang; Schatzmayr, Gerd; Moll, Wulf-Dieter

2015-01-01

Background Ergopeptines are a predominant class of ergot alkaloids produced by tall fescue grass endophyte Neotyphodium coenophialum or cereal pathogen Claviceps purpurea. The vasoconstrictive activity of ergopeptines makes them toxic for mammals, and they can be a problem in animal husbandry. Results We isolated an ergopeptine degrading bacterial strain, MTHt3, and classified it, based on its 16S rDNA sequence, as a strain of Rhodococcus erythropolis (Nocardiaceae, Actinobacteria). For strai...

Virulence of Rhodococcus equi Isolated from Cats and Dogs

OpenAIRE

Takai, Shinji; Martens, Ronald J.; Julian, Alan; Garcia Ribeiro, Márcio; Rodrigues de Farias, Marconi; Sasaki, Yukako; Inuzuka, Kazuho; Kakuda, Tsutomu; Tsubaki, Shiro; Prescott, John F.

2003-01-01

Nine cat isolates and nine dog isolates of Rhodococcus equi from clinical material were investigated for the presence of the virulence-associated antigens (VapA and VapB) and virulence plasmids. Five of the cat isolates and one dog isolate were VapA positive and contained an 85-kb type I or an 87-kb type I plasmid. The remaining 12 isolates were avirulent R. equi strains and contained no virulence plasmids.

[Cloning and expression of Micrococcus luteus IAM 14879 Rpf and its role in the recovery of the VBNC state in Rhodococcus sp. DS471].

Science.gov (United States)

Ding, Linxian; Zhang, Pinghua; Hong, Huachang; Lin, Hongjun; Yokota, Akira

2012-01-01

The purpose of the present study was to produce the Rpf (resuscitation promoting factor) protein by cloning and expressing the rpf gene, secreted by Micrococcus luteus IAM 14879, in Escherichia coli and to evaluate its role in the recovery of the VBNC (viable but non-culturable) state in high-GC Gram-positive bacteria. Genomic DNA was extracted from Micrococcus luteus IAM 14879 and the rpf gene was amplified by PCR using specific primers. The PCR products was purified, cloned into a pET15b expression vector, and transformed into Escherichia coli BL21 (DE3). Then the pET15b plasmid expression vector was used to confirm the purification of the recombinant proteins via SDS-PAGE. The VBNC state cells from the high-GC Gram-positive bacteria, Rhodococcus sp. DS471, were used to confirm the promotion and recovery of growth capacity. Rhodococcus sp. DS471 were isolated from soil and closely related to Micrococcus luteus IAM 14879. The gene sequences confirmed that the rpf gene from Micrococcus luteus IAM 14879 that was expressed in Escherichia coli, was 672 bp. SDS-PAGE analysis showed that the recombinant Rpf protein was obtained successfully, and further studies showed it capable of promoting the recovery of the VBNC state by about 100-fold relative to the control. Rpf of Micrococus luteus IAM 14879 can be successfully cloned and expressed in Escherichia coli and shows a strong ability to promote the recovery of the VBNC state of cells of Rhodococcus sp. DS471.

Biofouling inhibition in MBR by Rhodococcus sp. BH4 isolated from real MBR plant.

Science.gov (United States)

Oh, Hyun-Suk; Kim, Sang-Ryoung; Cheong, Won-Suk; Lee, Chung-Hak; Lee, Jung-Kee

2013-12-01

It has been reported that an indigenous quorum quenching bacterium, Rhodococcus sp. BH4, which was isolated from a real plant of membrane bioreactor (MBR) has promising potential to control biofouling in MBR. However, little is known about quorum quenching mechanisms by the strain BH4. In this study, various characteristics of strain BH4 were investigated to elucidate its behavior in more detail in the mixed liquor of MBR. The N-acyl homoserine lactone hydrolase (AHL-lactonase) gene of strain BH4 showed a high degree of identity to qsdA in Rhodococcus erythropolis W2. The LC-ESI-MS analysis of the degradation product by strain BH4 confirmed that it inactivated AHL activity by hydrolyzing the lactone bond of AHL. It degraded a wide range of N-acyl homoserine lactones (AHLs), but there was a large difference in the degradation rate of each AHL compared to other reported AHL-lactonase-producing strains belonging to Rhodococcus genus. Its quorum quenching activity was confirmed not only in the Luria-Bertani medium, but also in the synthetic wastewater. Furthermore, the amount of strain BH4 encapsulated in the vessel as well as the material of the vessel substantially affected the quorum quenching activity of strain BH4, which provides useful information, particularly for the biofouling control in a real MBR plant from an engineering point of view.

Bacteremia due to Rhodococcus equi in an immunocompetent infant

OpenAIRE

P Devi; S Malhotra; A Chadha

2011-01-01

Rhodococcus equi , previously known as Corynebacterium equi, is one of the most important causes of zoonotic infection in grazing animals. Increased cases of human infection with R. equi have been reported especially in immunocompromised patients. Infection in immunocompetent patients is extremely rare. We report a case of R. equi bacteremia in a 26-day-old immunocompetent infant with recurrent swellings on different parts of the body. To the best of our knowledge, this is the first ever repo...

Disseminated Rhodococcus equi infection in a kidney transplant patient without initial pulmonary involvement

NARCIS (Netherlands)

Rahamat-Langendoen, Janette C.; van Meurs, Matijs; Zijlstra, Jan G.; Lo-Ten-Foe, Jerome R.

2009-01-01

Rhodococcus equi is increasingly recognized as an opportunistic pathogen in solid organ transplant recipients. Primary pulmonary involvement is the most common finding. We report a case of a 42-year-old female kidney transplant recipient who developed multiple disseminated abscesses caused by R.

Genome Sequence of Rhodococcus erythropolis Strain CCM2595, a Phenol Derivative-Degrading Bacterium

Czech Academy of Sciences Publication Activity Database

Strnad, Hynek; Pátek, Miroslav; Fousek, Jan; Szököl, Juraj; Ulbrich, P.; Nešvera, Jan; Pačes, Václav; Vlček, Čestmír

2014-01-01

Roč. 2, č. 2 (2014) ISSN 2169-8287 R&D Projects: GA ČR GA13-28283S; GA MŠk 2B08062 Institutional support: RVO:68378050 ; RVO:61388971 Keywords : Rhodococcus erythropolis * genome sequence Subject RIV: EB - Genetics ; Molecular Biology

Biodegradation of cyanide by acetonitrile-induced cells of Rhodococcus sp. UKMP-5M.

Science.gov (United States)

Nallapan Maniyam, Maegala; Sjahrir, Fridelina; Ibrahim, Abdul Latif; Cass, Anthony E G

2013-01-01

A Rhodococcus sp. UKMP-5M isolate was shown to detoxify cyanide successfully, suggesting the presence of an intrinsic property in the bacterium which required no prior cyanide exposure for induction of this property. However, in order to promote growth, Rhodococcus sp. UKMP-5M was fully acclimatized to cyanide after 7 successive subcultures in 0.1 mM KCN for 30 days. To further shorten the lag phase and simultaneously increase the tolerance towards higher cyanide concentrations, the bacterium was induced with various nitrile compounds sharing a similar degradatory pathway to cyanide. Acetonitrile emerged as the most favored inducer and the induced cells were able to degrade 0.1 mM KCN almost completely within 18 h. With the addition of subsequent aliquots of 0.1 mM KCN a shorter period for complete removal of cyanide was required, which proved to be advantageous economically. Both resting cells and crude enzyme of Rhodococcus sp. UKMP-5M were able to biodegrade cyanide to ammonia and formate without the formation of formamide, implying the identification of a simple hydrolytic cyanide degradation pathway involving the enzyme cyanidase. Further verification with SDS-PAGE revealed that the molecular weight of the active enzyme was estimated to be 38 kDa, which is consistent with previously reported cyanidases. Since the recent advancement in the application of biological methods in treating cyanide-bearing wastewater has been promising, the discovery of this new bacterium will add value by diversifying the existing microbial populations capable of cyanide detoxification.

Infecção por Rhodococcus em doente imunocompetente - a propósito de um caso

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Susana Barbosa

2015-12-01

Full Text Available Resumo: O Rhodococcus equi raramente causa infecção no humano, sendo que a maioria dos casos descritos estão associados a doentes imunodeprimidos. Os autores apresentam o caso de um homem de 65 anos que foi admitido no serviço de urgência por agravamento da dispneia habitual, tosse e febre. Foi internado com o diagnóstico de sépsis com ponto de partida respiratório em pneumonia de focos múltiplos e peritonite. Medicado empiricamente com ceftriaxone, com melhoria clínica. Isolamento no líquido ascítico de Rhodococcus spp. Cumpriu 2 meses de antibioterapia com doxiciclina, com boa evolução clínica, analítica e radiológica. Os autores apresentam este caso dada a raridade da situação e pretendem realçar a necessidade de um elevado índice de suspeição para o diagnóstico da infecção por este agente em doentes imunocompetentes. Abstract: Rhodococcus equi rarely causes human infection, and most of the described cases are associated with immunosuppressed patients. The authors present a 65-year-old man who was admitted to the emergency department with worsening of dyspnoea, cough and fever. He was admitted with a diagnosis of sepsis with respiratory starting point in multiple foci pneumonia and peritonitis. He was medicated empirically with ceftriaxone, with clinical improvement. Isolation in the ascites fluid of Rhodococcus spp. He completed two months of antibiotic treatment with doxycycline, with clinical, analytical and radiological improvement. The authors present this case due to the rarity of the situation and highlight the need for a high index of suspicion for the infection diagnosis by this agent in immunocompetent patients.

Analysis of catRABC operon for catechol degradation from phenol-degrading Rhodococcus erythropolis

Czech Academy of Sciences Publication Activity Database

Veselý, Martin; Knoppová, Monika; Nešvera, Jan; Pátek, Miroslav

2007-01-01

Roč. 76, - (2007), s. 159-168 ISSN 0175-7598 R&D Projects: GA ČR GA526/04/0542 Institutional research plan: CEZ:AV0Z50200510 Keywords : rhodococcus erythropolis * catrabc operon * catechol degradation Subject RIV: EE - Microbiology, Virology Impact factor: 2.475, year: 2007

Biosurfactant production by halotolerant Rhodococcus fascians from Casey Station, Wilkes Land, Antarctica.

Science.gov (United States)

Gesheva, Victoria; Stackebrandt, Erko; Vasileva-Tonkova, Evgenia

2010-08-01

Isolate A-3 from Antarctic soil in Casey Station, Wilkes Land, was characterized for growth on hydrocarbons. Use of glucose or kerosene as a sole carbon source in the culture medium favoured biosynthesis of surfactant which, by thin-layer chromatography, indicated the formation of a rhamnose-containing glycolipid. This compound lowered the surface tension at the air/water interface to 27 mN/m as well as inhibited the growth of B. subtilis ATCC 6633 and exhibited hemolytic activity. A highly hydrophobic surface of the cells suggests that uptake occurs via a direct cell-hydrocarbon substrate contact. Strain A-3 is Gram-positive, halotolerant, catalase positive, urease negative and has rod-coccus shape. Its cell walls contained meso-diaminopimelic acid. Phylogenetic analysis based on comparative analysis of 16S rRNA gene sequences revealed that strain A-3 is closely related to Rhodococcus fascians with which it shares 100% sequence similarity. This is the first report on rhamnose-containing biosurfactant production by Rhodococcus fascians isolated from Antarctic soil.

MICs of Oxazolidinones for Rhodococcus equi Strains Isolated from Humans and Animals

OpenAIRE

Bowersock, Terry L.; Salmon, Sarah A.; Portis, Ellen S.; Prescott, John F.; Robison, Denise A.; Ford, Charles W.; Watts, Jeffrey L.

2000-01-01

Eperezolid and linezolid are representatives of a new class of orally active, synthetic antimicrobial agents. The in vitro activity values (MICs) of linezolid, eperezolid, and comparator antibiotics against 102 strains of Rhodococcus equi isolated from humans and animals were determined. Linezolid was more active than eperezolid against the strains tested; premafloxacin was the most active comparator antibiotic.

A new zearalenone biodegradation strategy using non-pathogenic Rhodococcus pyridinivorans K408 strain.

Science.gov (United States)

Kriszt, Rókus; Krifaton, Csilla; Szoboszlay, Sándor; Cserháti, Mátyás; Kriszt, Balázs; Kukolya, József; Czéh, Arpád; Fehér-Tóth, Szilvia; Török, Lívia; Szőke, Zsuzsanna; Kovács, Krisztina J; Barna, Teréz; Ferenczi, Szilamér

2012-01-01

Zearalenone (hereafter referred to as ZEA) is a nonsteroidal estrogenic mycotoxin produced by several Fusarium spp. on cereal grains. ZEA is one of the most hazardous natural endocrine disrupting chemicals (EDC) which induces hyper estrogenic responses in mammals. This can result in reproductive disorders in farm animals as well as in humans. Consequently, detoxification strategies for contaminated crops are crucial for food safety. In this study we have developed a bacterial based detoxification system using a non-pathogen Rhodococcus pyridinivorans K408 strain. Following 5 days treatment of ZEA with R. pyridinivorans K408 strain HPLC analyses showed an 87.21% ZEA-degradation efficiency of the bacterial enzyme systems. In another approach, the strain biotransformation ability has also been confirmed by a bioluminescent version of the yeast estrogen screening system (BLYES), which detected an 81.75% of biodegradability of ZEA, in a good agreement with the chemical analyses. Furthermore, the capacity of R. pyridinivorans to eliminate the estrogenic effects of ZEA was tested by using an immature uterotrophic assay. Prepubertal female rats were treated with vehicle (olive oil), 17β-estradiol, ZEA (0.1-1-5-10 mg/kg body weight) and LB broth containing 500 mg/l ZEA that has already been incubated with or without Rhodococcus pyridinivorans K408 strain. Uterine weights were measured and the mRNA level changes relating to apelin, aquaporin 5, complement component 2, and calbindin-3 genes were measured by qRT-PCR. These genes represent the major pathways that are affected by estromimetic compounds. Zearalenone feeding significantly increased the uterus weight in a dose dependent manner and at the same time upregulated complement component 2 and calbindin-3 expression as well as decreased apelin and aquaporin 5 mRNA levels comparable to that seen in 17β-estradiol exposed rats. In contrast, LB broth in which ZEA was incubated with Rhodococcus pyridinivorans K408 prior to

A new zearalenone biodegradation strategy using non-pathogenic Rhodococcus pyridinivorans K408 strain.

Directory of Open Access Journals (Sweden)

Rókus Kriszt

Full Text Available Zearalenone (hereafter referred to as ZEA is a nonsteroidal estrogenic mycotoxin produced by several Fusarium spp. on cereal grains. ZEA is one of the most hazardous natural endocrine disrupting chemicals (EDC which induces hyper estrogenic responses in mammals. This can result in reproductive disorders in farm animals as well as in humans. Consequently, detoxification strategies for contaminated crops are crucial for food safety. In this study we have developed a bacterial based detoxification system using a non-pathogen Rhodococcus pyridinivorans K408 strain. Following 5 days treatment of ZEA with R. pyridinivorans K408 strain HPLC analyses showed an 87.21% ZEA-degradation efficiency of the bacterial enzyme systems. In another approach, the strain biotransformation ability has also been confirmed by a bioluminescent version of the yeast estrogen screening system (BLYES, which detected an 81.75% of biodegradability of ZEA, in a good agreement with the chemical analyses. Furthermore, the capacity of R. pyridinivorans to eliminate the estrogenic effects of ZEA was tested by using an immature uterotrophic assay. Prepubertal female rats were treated with vehicle (olive oil, 17β-estradiol, ZEA (0.1-1-5-10 mg/kg body weight and LB broth containing 500 mg/l ZEA that has already been incubated with or without Rhodococcus pyridinivorans K408 strain. Uterine weights were measured and the mRNA level changes relating to apelin, aquaporin 5, complement component 2, and calbindin-3 genes were measured by qRT-PCR. These genes represent the major pathways that are affected by estromimetic compounds. Zearalenone feeding significantly increased the uterus weight in a dose dependent manner and at the same time upregulated complement component 2 and calbindin-3 expression as well as decreased apelin and aquaporin 5 mRNA levels comparable to that seen in 17β-estradiol exposed rats. In contrast, LB broth in which ZEA was incubated with Rhodococcus pyridinivorans K

Bacteremia due to Rhodococcus equi in an immunocompetent infant

Directory of Open Access Journals (Sweden)

P Devi

2011-01-01

Full Text Available Rhodococcus equi , previously known as Corynebacterium equi, is one of the most important causes of zoonotic infection in grazing animals. Increased cases of human infection with R. equi have been reported especially in immunocompromised patients. Infection in immunocompetent patients is extremely rare. We report a case of R. equi bacteremia in a 26-day-old immunocompetent infant with recurrent swellings on different parts of the body. To the best of our knowledge, this is the first ever report of R. equi bacteremia from an immunocompetent patient from Northern India.

Complete Genome Sequence of Rhodococcus sp. Strain WMMA185, a Marine Sponge-Associated Bacterium

OpenAIRE

Adnani, Navid; Braun, Doug R.; McDonald, Bradon R.; Chevrette, Marc G.; Currie, Cameron R.; Bugni, Tim S.

2016-01-01

The Rhodococcus strain WMMA185 was isolated from the marine sponge Chondrilla nucula as part of ongoing drug discovery efforts. Analysis of the 4.44-Mb genome provides information regarding interspecies interactions as pertains to regulation of secondary metabolism and natural product biosynthetic potentials.

Analysis of genome sequences from plant pathogenic Rhodococcus reveals genetic novelties in virulence loci

Science.gov (United States)

Members of Gram-positive Actinobacteria cause economically important diseases to plants. Within the Rhodococcus genus, some members can cause growth deformities and persist as pathogens on a wide range of host plants. The current model predicts that phytopathogenic isolates require a cluster of thre...

Cold adaptive traits revealed by comparative genomic analysis of the eurypsychrophile Rhodococcus sp. JG3 isolated from high elevation McMurdo Dry Valley permafrost, Antarctica.

Science.gov (United States)

Goordial, Jacqueline; Raymond-Bouchard, Isabelle; Zolotarov, Yevgen; de Bethencourt, Luis; Ronholm, Jennifer; Shapiro, Nicole; Woyke, Tanja; Stromvik, Martina; Greer, Charles W; Bakermans, Corien; Whyte, Lyle

2016-02-01

The permafrost soils of the high elevation McMurdo Dry Valleys are the most cold, desiccating and oligotrophic on Earth. Rhodococcus sp. JG3 is one of very few bacterial isolates from Antarctic Dry Valley permafrost, and displays subzero growth down to -5°C. To understand how Rhodococcus sp. JG3 is able to survive extreme permafrost conditions and be metabolically active at subzero temperatures, we sequenced its genome and compared it to the genomes of 14 mesophilic rhodococci. Rhodococcus sp. JG3 possessed a higher copy number of genes for general stress response, UV protection and protection from cold shock, osmotic stress and oxidative stress. We characterized genome wide molecular adaptations to cold, and identified genes that had amino acid compositions favourable for increased flexibility and functionality at low temperatures. Rhodococcus sp. JG3 possesses multiple complimentary strategies which may enable its survival in some of the harshest permafrost on Earth. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Infecção pulmonar por "Rhodococcus equi": relato dos dois primeiros casos brasileiros Pulmonary Rhodococcus equi infection: report of the first two Brazilian cases

Directory of Open Access Journals (Sweden)

LUIZ CARLOS SEVERO

2001-05-01

Full Text Available O Rhodococcus equi, principal agente da rodococose, é um cocobacilo pleomórfico, gram-positivo e aeróbio, que infecta humanos por via inalatória ou transcutânea e se manifesta clinicamente como abscesso pulmonar. Relatam-se os dois primeiros casos brasileiros da doença. Ambos os pacientes eram imunocomprometidos e apresentavam quadro infeccioso pulmonar. O primeiro tinha AIDS e apresentava pneumonia cavitada em lobo superior esquerdo, que teve evolução fatal. O segundo tinha doença de Goodpasture, insuficiência renal crônica e fazia uso de corticosteróides. Apresentava uma lesão pulmonar escavada no lobo superior direito, que foi tratada com sulfametoxazol-trimetoprim, com resolução do processo.Rhodococcus equi, the principal agent of rhodococcosis, is a pleomorphic, gram-positive, aerobic coccus bacillus that infects humans by inhalation or through a transcutaneous route. It is clinically manifested as a pulmonary abscess. The first two Brazilian cases of rhodococcosis are reported on. Both patients were immunocompromised and showed pulmonary infection. The first patient had AIDS and cavitating pneumonia in the left upper lobe, that was fatal. The second case presented Goodpasture syndrome and was under chronic corticotherapy. He displayed a cavitating nodular lesion in the right upper lobe, that was successfully treated with sulfametoxazol-trimethoprim.

Biosurfactant Production by Rhodococcus Erythropolis and its Application to Oil Removal

OpenAIRE

Pacheco, Graziela Jardim; Ciapina, Elisa Mara Prioli; Gomes, Edelvio de Barros; Junior, Nei Pereira

2010-01-01

The influence of different nutrients on biosurfactant production by Rhodococcus erythropolis was investigated. Increasing the concentration of phosphate buffer from 30 up through 150 mmol/L stimulated an increase in biosurfactant production, which reached a maximum concentration of 285 mg/L in shaken flasks. Statistical analysis showed that glycerol, NaNO3, MgSO4 and yeast extract had significant effects on production. The results were confirmed in a batchwise bioreactor, and semi-growth-asso...

Rhodococcus erythropolis DCL14 Contains a Novel Degradation Pathway for Limonene

OpenAIRE

van der Werf, Mariët J.; Swarts, Henk J.; de Bont, Jan A. M.

1999-01-01

Strain DCL14, which is able to grow on limonene as a sole source of carbon and energy, was isolated from a freshwater sediment sample. This organism was identified as a strain of Rhodococcus erythropolis by chemotaxonomic and genetic studies. R. erythropolis DCL14 also assimilated the terpenes limonene-1,2-epoxide, limonene-1,2-diol, carveol, carvone, and (−)-menthol, while perillyl alcohol was not utilized as a carbon and energy source. Induction tests with cells grown on limonene revealed t...

Analysis and optimization of triacylglycerol synthesis in novel oleaginous Rhodococcus and Streptomyces strains isolated from desert soil.

Science.gov (United States)

Röttig, Annika; Hauschild, Philippa; Madkour, Mohamed H; Al-Ansari, Ahmed M; Almakishah, Naief H; Steinbüchel, Alexander

2016-05-10

As oleaginous microorganisms represent an upcoming novel feedstock for the biotechnological production of lipids or lipid-derived biofuels, we searched for novel, lipid-producing strains in desert soil. This was encouraged by the hypothesis that neutral lipids represent an ideal storage compound, especially under arid conditions, as several animals are known to outlast long periods in absence of drinking water by metabolizing their body fat. Ten lipid-accumulating bacterial strains, affiliated to the genera Bacillus, Cupriavidus, Nocardia, Rhodococcus and Streptomyces, were isolated from arid desert soil due to their ability to synthesize poly(β-hydroxybutyrate), triacylglycerols or wax esters. Particularly two Streptomyces sp. strains and one Rhodococcus sp. strain accumulate significant amounts of TAG under storage conditions under optimized cultivation conditions. Rhodococcus sp. A27 and Streptomyces sp. G49 synthesized approx. 30% (w/w) fatty acids from fructose or cellobiose, respectively, while Streptomyces isolate G25 reached a cellular fatty acid content of nearly 50% (w/w) when cultivated with cellobiose. The stored triacylglycerols were composed of 30-40% branched fatty acids, such as anteiso-pentadecanoic or iso-hexadecanoic acid. To date, this represents by far the highest lipid content described for streptomycetes. A biotechnological production of such lipids using (hemi)cellulose-derived raw material could be used to obtain sustainable biodiesel with a high proportion of branched-chain fatty acids to improve its cold-flow properties and oxidative stability. Copyright © 2016 Elsevier B.V. All rights reserved.

Steady-state and pre-steady-state kinetic analysis of halopropane conversion by a Rhodococcus haloalkane dehalogenase

NARCIS (Netherlands)

Bosma, T; Pikkemaat, MG; Kingma, Jacob; Dijk, J; Janssen, DB

2003-01-01

Haloalkane dehalogenase from Rhodococcus rhodochrous NCIMB 13064 (DhaA) catalyzes the hydrolysis of carbon-halogen bonds in a wide range of haloalkanes. We examined the steady-state and pre-steady-state kinetics of halopropane conversion by DhaA to illuminate mechanistic details of the

An Adenoviral Vector Based Vaccine for Rhodococcus equi.

Directory of Open Access Journals (Sweden)

Carla Giles

Full Text Available Rhodococcus equi is a respiratory pathogen which primarily infects foals and is endemic on farms around the world with 50% mortality and 80% morbidity in affected foals. Unless detected early and treated appropriately the disease can be fatal. Currently, there is no vaccine available to prevent this disease. For decades researchers have endeavoured to develop an effective vaccine to no avail. In this study a novel human adenoviral vector vaccine for R. equi was developed and tested in the mouse model. This vaccine generated a strong antibody and cytokine response and clearance of R. equi was demonstrated following challenge. These results show that this vaccine could potentially be developed further for use as a vaccine to prevent R. equi disease in foals.

Pulmonary Rhodococcus equi infection: report of the first two Brazilian cases

OpenAIRE

SEVERO, LUIZ CARLOS; RITTER, PATRICIA; PETRILLO, VICTOR FLÁVIO; DIAS, CÍCERO ARMÍDIO GOMES; PORTO, NELSON DA SILVA

2001-01-01

O Rhodococcus equi, principal agente da rodococose, é um cocobacilo pleomórfico, gram-positivo e aeróbio, que infecta humanos por via inalatória ou transcutânea e se manifesta clinicamente como abscesso pulmonar. Relatam-se os dois primeiros casos brasileiros da doença. Ambos os pacientes eram imunocomprometidos e apresentavam quadro infeccioso pulmonar. O primeiro tinha AIDS e apresentava pneumonia cavitada em lobo superior esquerdo, que teve evolução fatal. O segundo tinha doença de Goodpas...

Genetic and biochemical characterization of a novel monoterpene epsilon-lactone hydrolase from Rhodococcus erythropolis DCL14

NARCIS (Netherlands)

Vlugt-Bergmans, van der C.J.B.; Werf, van der M.J.

2001-01-01

A monoterpene ε-lactone hydrolase (MLH) from Rhodococcus erythropolis DCL14, catalyzing the ring opening of lactones which are formed during degradation of several monocyclic monoterpenes, including carvone and menthol, was purified to apparent homogeneity. It is a monomeric enzyme of 31 kDa that is

Genetic and biochemical characterization of a novel monoterpene e-lactone hydrolase from Rhodococcus erythropolis DCL14

NARCIS (Netherlands)

Vlugt-Bergmans, C.J.B. van der; Werf, M.J. van der

2001-01-01

A monoterpene ε-lactone hydrolase (MLH) from Rhodococcus erythropolis DCL14, catalyzing the ring opening of lactones which are formed during degradation of several monocyclic monoterpenes, including carvone and menthol, was purified to apparent homogeneity. It is a monomeric enzyme of 31 kDa that is

Molecular characterization of three 3-ketosteroid-Delta(1)-dehydrogenase isoenzynnes of Rhodococcus ruber strain Chol-4

NARCIS (Netherlands)

Fernandez de las Heras, Laura; van der Geize, Robert; Drzyzga, Oliver; Perera, Julian; Navarro Llorens, Juana Maria

2012-01-01

Rhodococcus ruber strain Chol-4 isolated from a sewage sludge sample is able to grow on minimal medium supplemented with steroids, showing a broad catabolic capacity. This paper reports the characterization of three different 3-ketosteroid-Delta(1)-dehydrogenases (KstDs) in the genome of R. ruber

Metabolomics of the Bio-Degradation Process of Aflatoxin B1 by Actinomycetes at an Initial pH of 6.0

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Manal Eshelli

2015-02-01

Full Text Available Contamination of food and feed by Aflatoxin B1 (AFB1 is a cause of serious economic and health problems. Different processes have been used to degrade AFB1. In this study, biological degradation of AFB1 was carried out using three Actinomycete species, Rhodococcus erythropolis ATCC 4277, Streptomyces lividans TK 24, and S. aureofaciens ATCC 10762, in liquid cultures. Biodegradation of AFB1 was optimised under a range of temperatures from 25 to 40 °C and pH values of 4.0 to 8.0. An initial concentration of 20 µg/mL of AFB1 was used in this study. The amount of AFB1 remaining was measured against time by thin layer chromatography (TLC and high-performance liquid chromatography (HPLC, coupled with UV and mass spectrometry (LC-MS. All species were able to degrade the AFB1, and no significant difference was found between them. AFB1 remained in the liquid culture for R. erythropolis, S. lividans and S. aureofaciens were 0.81 µg/mL, 2.41 µg/mL and 2.78 µg/mL respectively, at the end of the first 24 h. Degradation occurred at all incubation temperatures and the pH with the optimal conditions for R. erythropolis was achieved at 30 °C and pH 6, whereas for S. lividans and S. aureofaciens the optimum conditions for degradation were 30 °C and pH 5. Analysis of the degradative route indicated that each microorganism has a different way of degrading AFB1. The metabolites produced by R. erythropolis were significantly different from the other two microorganisms. Products of degradation were identified through metabolomic studies by utilizing high-resolution mass spectral data. Mass spectrometric analysis indicated that the degradation of AFB1 was associated with the appearance of a range of lower molecular weight compounds. The pathway of degradation or chemical alteration of AFB1 was followed by means of high resolution Fourier transform mass spectrometry (HR-FTMS analysis as well as through the MS2 fragmentation to unravel the degradative pathway for

Calcium carbonate formation on mica supported extracellular polymeric substance produced by Rhodococcus opacus

Energy Technology Data Exchange (ETDEWEB)

Szcześ, Aleksandra, E-mail: aszczes@poczta.umcs.lublin.pl [Department of Physical Chemistry – Interfacial Phenomena, Faculty of Chemistry, Maria Curie-Skłodowska University, Lublin 20-031 (Poland); Czemierska, Magdalena; Jarosz-Wilkołazka, Anna [Department of Biochemistry, Maria Curie-Skłodowska University, Lublin 20-031 (Poland)

2016-10-15

Extracellular polymeric substance (EPS) extracted from Rhodococcus opacus bacterial strain was used as a matrix for calcium carbonate precipitation using the vapour diffusion method. The total exopolymer and water-soluble exopolymer fraction of different concentrations were spread on the mica surface by the spin-coating method. The obtained layers were characterized using the atomic force microscopy measurement and XPS analysis. The effects of polymer concentration, initial pH of calcium chloride solution and precipitation time on the obtained crystals properties were investigated. Raman spectroscopy and scanning electron microscopy were used to characterize the precipitated minerals. It was found that the type of precipitated CaCO{sub 3} polymorph and the crystal size depend on the kind of EPS fraction. The obtained results indicates that the water soluble fraction favours vaterite dissolution and calcite growth, whereas the total EPS stabilizes vaterite and this effect is stronger at basic pH. It seems to be due to different contents of the functional group of EPS fractions. - Highlights: • CaCO{sub 3} crystal size and polymorph can be controlled by EPS substance obtained from R. opacus. • The water soluble fraction favours vaterite dissolution and calcite growth. • The total EPS stabilizes vaterite. • This effect is stronger at basic pH.

Calcium carbonate formation on mica supported extracellular polymeric substance produced by Rhodococcus opacus

International Nuclear Information System (INIS)

Szcześ, Aleksandra; Czemierska, Magdalena; Jarosz-Wilkołazka, Anna

2016-01-01

Extracellular polymeric substance (EPS) extracted from Rhodococcus opacus bacterial strain was used as a matrix for calcium carbonate precipitation using the vapour diffusion method. The total exopolymer and water-soluble exopolymer fraction of different concentrations were spread on the mica surface by the spin-coating method. The obtained layers were characterized using the atomic force microscopy measurement and XPS analysis. The effects of polymer concentration, initial pH of calcium chloride solution and precipitation time on the obtained crystals properties were investigated. Raman spectroscopy and scanning electron microscopy were used to characterize the precipitated minerals. It was found that the type of precipitated CaCO 3 polymorph and the crystal size depend on the kind of EPS fraction. The obtained results indicates that the water soluble fraction favours vaterite dissolution and calcite growth, whereas the total EPS stabilizes vaterite and this effect is stronger at basic pH. It seems to be due to different contents of the functional group of EPS fractions. - Highlights: • CaCO 3 crystal size and polymorph can be controlled by EPS substance obtained from R. opacus. • The water soluble fraction favours vaterite dissolution and calcite growth. • The total EPS stabilizes vaterite. • This effect is stronger at basic pH.

Infection by rhodococcus fascians maintains cotyledons as a sink tissue for the pathogen

Czech Academy of Sciences Publication Activity Database

Dhandapani, P.; Song, J.; Novák, Ondřej; Jameson, P. E.

2017-01-01

Roč. 119, č. 5 (2017), s. 841-852 ISSN 0305-7364 R&D Projects: GA MŠk(CZ) LO1204; GA MŠk LK21306 Institutional support: RVO:61389030 Keywords : Amino acid transporter * Apical dominance * Cell wall invertase * Cytokinin * Cytokinin oxidase/dehydro-genase * Pea * Pisum sativum L. * Rhodococcus fascians * Seed * Sink and source * Sucrose transporter * sweet Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Plant sciences, botany Impact factor: 4.041, year: 2016

Evidence for an Inducible Nucleotide-Dependent Acetone Carboxylase in Rhodococcus rhodochrous B276

OpenAIRE

Clark, Daniel D.; Ensign, Scott A.

1999-01-01

The metabolism of acetone was investigated in the actinomycete Rhodococcus rhodochrous (formerly Nocardia corallina) B276. Suspensions of acetone- and isopropanol-grown R. rhodochrous readily metabolized acetone. In contrast, R. rhodochrous cells cultured with glucose as the carbon source lacked the ability to metabolize acetone at the onset of the assay but gained the ability to do so in a time-dependent fashion. Chloramphenicol and rifampin prevented the time-dependent increase in this acti...

[Effects of nitriles and amides on the growth and the nitrile hydratase activity of the Rhodococcus sp. strain gt1].

Science.gov (United States)

Maksimov, A Iu; Kuznetsova, M V; Ovechkina, G V; Kozlov, S V; Maksimova, Iu G; Demakov, V A

2003-01-01

Effects of some nitriles and amides, as well as glucose and ammonium, on the growth and the nitrile hydratase (EC 4.2.1.84) activity of the Rhodococcus sp. strain gt1 isolated from soil were studied. The activity of nitrile hydratase mainly depended on carbon and nitrogen supply to cells. The activity of nitrile hydratase was high in the presence of glucose and ammonium at medium concentrations and decreased at concentrations of glucose more than 0.3%. Saturated unsubstituted aliphatic nitriles and amides were found to be a good source of nitrogen and carbon. However, the presence of nitriles and amides in the medium was not absolutely necessary for the expression of the activity of nitrile hydratase isolated from the Rhodococcus sp. strain gt1.

Biocatalytic properties and structural analysis of eugenol oxidase from Rhodococcus jostii RHA1 : a versatile oxidative biocatalyst

NARCIS (Netherlands)

Nguyen, Quoc-Thai; De Gonzalo, Gonzalo; Binda, Claudia; Rioz, Ana; Mattevi, Andrea; Fraaije, Marco

2016-01-01

Eugenol oxidase (EUGO) from Rhodococcus jostii RHA1 was previously shown to convert only a limited set of phenolic compounds. In this study, we have explored the biocatalytic potential of this flavoprotein oxidase resulting in a broadened substrate scope and a deeper insight into its structural

Identification of microbial carotenoids and isoprenoid quinones from Rhodococcus sp. B7740 and its stability in the presence of iron in model gastric conditions.

Science.gov (United States)

Chen, Yashu; Xie, Bijun; Yang, Jifang; Chen, Jigang; Sun, Zhida

2018-02-01

Rhodococcus sp. B7740 is a newfound bacterium which was isolated from 25m deep seawater in the arctic. In this paper, Rhodococcus sp. B7740 was firstly discovered to produce abundant natural isoprenoids, including ubiquinone-4(UQ-4), 13 kinds of menaquinones, three rare aromatic carotenoids and more than one common carotenoid. These compounds were identified by UV-Visible, HPLC-APCI-MS/MS and HRMS spectra. Results demonstrated that Rhodococcus sp. B7740 might be a worthy source of natural isoprenoids especially for scarce aromatic carotenoids. Among them, isorenieratene with 528.3762Da (calculated for 528.3756Da, error: 1.1ppm), a carotenoid with aromatic ring, was purified by HSCCC. The stability of isorenieratene under the mimical gastric conditions was measured compared with common dietary carotenoids, β-carotene and lutein. Unlike β-carotene and lutein, isorenieratene exhibited rather stable in the presence of free iron or heme iron. Its high retention rate in gastrointestinal tract after ingestion indicates the benefits for health. Copyright © 2017. Published by Elsevier Ltd.

El género Rhodococcus. Una revisión didáctica

OpenAIRE

Sánchez, N; Sandoval, AH; Díaz-Corrales, F; Serrano, JA

2004-01-01

Los autores presentan una extensa revisión sobre el género Rhodococcus, grupo de microorganismos pertenecientes a la familia de los Actinomicetales. Se revisan aspectos relativos a la microbiología, taxonomía, citoquímica, patología, clínica y terapéutica de las afecciones que, como agentes patógenos, producen estos microorganismos, en especial en su papel de agentes productores de infecciones nosocomiales y oportunistas. Así mismo se revisan aspectos relativos a la importancia de estos micro...

Aerobic degradation of buprofezin via novel degradation intermediates by Rhodococcus sp. strain RX-3

OpenAIRE

Ruixue Li; Chun Dai; Guangli Wang; Shaoxian Wu; Yubao Gong; Yuanyuan Jiang; Zhijia Wang; Naiyue Sun

2016-01-01

Buprofezin is a commonly used chemical with satisfactory efficacy against sucking insect pests, but its disposal causes serious environmental problems. In this study, a bacterial strain RX-3 isolated by continuous enrichment from buprofezin-treated soil was tested for biodegradation of buprofezin. The bacteria were most similar to Rhodococcus sp. based on their morphological, physiological and biochemical characteristics, as well as phylogenetic placement inferred from 16S rRNA gene sequence....

RP-HPLC/MS-APCI Analysis of Branched Chain TAG Prepared by Precursor-Directed Biosynthesis with Rhodococcus erythropolis

Czech Academy of Sciences Publication Activity Database

Schreiberová, O.; Krulikovská, T.; Sigler, Karel; Čejková, A.; Řezanka, Tomáš

2010-01-01

Roč. 45, č. 8 (2010), s. 743-756 ISSN 0024-4201 R&D Projects: GA MŠk 2B08062 Institutional research plan: CEZ:AV0Z50200510 Keywords : Rhodococcus erythropolis * RP-HPLC/MS-APCI * Branched chain triacylglycerols Subject RIV: EE - Microbiology, Virology Impact factor: 2.151, year: 2010

Lag phase and biomass determination of Rhodococcus pyridinivorans GM3 for degradation of phenol

Science.gov (United States)

Al-Defiery, M. E. J.; Reddy, G.

2018-05-01

Among various techniques available for removal of phenol, biodegradation is an eco-friendly and cost effective method. Thus, it is required to understand the process of biodegradation of phenol, such as investigate on lag phase and biomass concentration. Phenol degrading bacteria were isolated from soil samples of industrial sites in enriched mineral salts medium (MSM) with phenol as a sole source of energy and carbon. One isolate of potential phenol degradation from consortium for phenol degrading studies was identified as Rhodococcus pyridinivorans GM3. Lag phase and biomass determination of R. pyridinivorans GM3 was studied with different phenol concentrations under pH 8.5 at temperature 32 Co and 200 rpm. Microbial biomass was directly proportional to increasing phenol concentration between 1.0 to 2.0 g/L with a maximum dry biomass of 1.745 g/L was noted after complete degradation of 2.0 g/L phenol in 48 hours.

Optimization of biosurfactant production in soybean oil by rhodococcus rhodochrous and its utilization in remediation of cadmium-contaminated solution

Science.gov (United States)

Suryanti, Venty; Hastuti, Sri; Andriani, Dewi

2016-02-01

Biosurfactant production by Rhodococcus rhodochrous in soybean oil was developed, where the effect of medium composition and fermentation time were evaluated. The optimum condition for biosurfactant production was achieved when a medium containing 30 g/L TSB (tryptic soy broth) and 20% v/v soybean oil was used as media with 7 days of fermentation. Biosurfactant was identified as glycolipids type biosurfactant which had critical micelle concentration (CMC) value of 896 mg/L. The biosurfactant had oil in water emulsion type and was able to reduce the surface tension of palm oil about 52% which could stabilize the emulsion up to 12 days. The batch removal of cadmium metal ion by crude and partially purified biosurfactants have been examined from synthetic aqueous solution at pH 6. The results exhibited that the crude biosurfactant had a much better adsorption ability of Cd(II) than that of partially purified biosurfactant. However, it was found that there was no significant difference in the adsorption of Cd(II) with 5 and 10 minutes of contact time. The results indicated that the biosurfactant could be used in remediation of heavy metals from contaminated aqueous solution.

Haloalkane-utilizing Rhodococcus strains isolated from geographically distinct locations possess a highly conserved gene cluster encoding haloalkane catabolism

NARCIS (Netherlands)

Poelarends, GJ; Bosma, T; Kulakov, LA; Larkin, MJ; Marchesi, [No Value; Weightman, AJ; Janssen, DB; Kulakov, Leonid A.; Larkin, Michael J.; Marchesi, Julian R.; Weightman, Andrew J.

The sequences of the 16S rRNA and haloalkane dehalogenase (dhaA) genes of five gram-positive haloalkane-utilizing bacteria isolated from contaminated sites in Europe, Japan, and the United States and of the archetypal haloalkane-degrading bacterium Rhodococcus sp. strain NCIMB13064 were compared.

Failure of pulmonary clearance of Rhodococcus equi infection in CD4+ T-lymphocyte-deficient transgenic mice.

OpenAIRE

Kanaly, S T; Hines, S A; Palmer, G H

1993-01-01

Pulmonary clearance of Rhodococcus equi requires functional T lymphocytes. In this study, CD8+ T-lymphocyte-deficient transgenic mice cleared virulent R. equi from the lungs while infection in CD4+ T-lymphocyte-deficient transgenic mice persisted. Although both CD4+ and CD8+ T cells function early in pulmonary defense against R. equi, clearance is dependent on CD4+ T lymphocytes.

Improved Method for the Isolation of Biosurfactant Glycolipids from Rhodococcus sp. Strain H13A

OpenAIRE

Bryant, Frank O.

1990-01-01

An improved method for the isolation of the biosurfactant glycolipids from Rhodococcus sp. strain H13A by using XM 50 diafiltration and isopropanol precipitation was devised. This procedure was advantageous since it removes protein coisolated when the glycolipids are obtained by organic extraction and silicic acid chromatography. The protein apparently does not contribute any biosurfactant characteristics to the glycolipids. The deacylated glycolipid backbone included only a disaccharide.

Benzoate Catabolite Repression of the Phthalate Degradation Pathway in Rhodococcus sp. Strain DK17▿

OpenAIRE

Choi, Ki Young; Zylstra, Gerben J.; Kim, Eungbin

2006-01-01

Rhodococcus sp. strain DK17 exhibits a catabolite repression-like response when provided simultaneously with benzoate and phthalate as carbon and energy sources. Benzoate in the medium is depleted to detection limits before the utilization of phthalate begins. The transcription of the genes encoding benzoate and phthalate dioxygenase paralleled the substrate utilization profile. Two mutant strains with defective benzoate dioxygenases were unable to utilize phthalate in the presence of benzoat...

Cholesterol oxidase (ChoE) is not important in the virulence of Rhodococcus equi.

Science.gov (United States)

Pei, Yanlong; Dupont, Chris; Sydor, Tobias; Haas, Albert; Prescott, John F

2006-12-20

To analyze further the role in virulence of the prominent cholesterol oxidase (ChoE) of Rhodococcus equi, an allelic exchange choE mutant from strain 103+ was constructed and assessed for virulence in macrophages, in mice, and in foals. There was no difference between the mutant and parent strain in cytotoxic activity for macrophages or in intra-macrophage multiplication. No evidence of attenuation was obtained in macrophages and in mice, but there was slight attenuation apparent in four intra-bronchially infected foals compared to infection of four foals with the virulent parent strain, based on a delayed rise in temperature of the choE-mutant infected foals. However, bacterial colony counts in the lung 2 weeks after infection were not significantly different, although there was a slight but non-significant (P=0.12) difference in lung:body weight ratio of the choE mutant versus virulent parent infected foals (mean 2.67+/-0.25% compared to 4.58+/-0.96%). We conclude that the cholesterol oxidase is not important for the virulence of R. equi.

Differences in Rhodococcus equi Infections Based on Immune Status and Antibiotic Susceptibility of Clinical Isolates in a Case Series of 12 Patients and Cases in the Literature

Science.gov (United States)

Suzuki, Yasuhiro; Ribes, Julie A.; Thornton, Alice

2016-01-01

Rhodococcus equi is an unusual zoonotic pathogen that can cause life-threatening diseases in susceptible hosts. Twelve patients with R. equi infection in Kentucky were compared to 137 cases reported in the literature. Although lungs were the primary sites of infection in immunocompromised patients, extrapulmonary involvement only was more common in immunocompetent patients (P antibiotics, preferably selected from vancomycin, imipenem, clarithromycin/azithromycin, ciprofloxacin, rifampin, or cotrimoxazole. Local antibiograms should be checked prior to using cotrimoxazole due to developing resistance. PMID:27631004

Production of carotenoids and lipids by Rhodococcus opacus PD630 in batch and fed-batch culture.

Science.gov (United States)

Thanapimmetha, Anusith; Suwaleerat, Tharatron; Saisriyoot, Maythee; Chisti, Yusuf; Srinophakun, Penjit

2017-01-01

Production of carotenoids by Rhodococcus opacus PD630 is reported. A modified mineral salt medium formulated with glycerol as an inexpensive carbon source was used for the fermentation. Ammonium acetate was the nitrogen source. A dry cell mass concentration of nearly 5.4 g/L could be produced in shake flasks with a carotenoid concentration of 0.54 mg/L. In batch culture in a 5 L bioreactor, without pH control, the maximum dry biomass concentration was ~30 % lower than in shake flasks and the carotenoids concentration was 0.09 mg/L. Both the biomass concentration and the carotenoids concentration could be raised using a fed-batch operation with a feed mixture of ammonium acetate and acetic acid. With this strategy, the final biomass concentration was 8.2 g/L and the carotenoids concentration was 0.20 mg/L in a 10-day fermentation. A control of pH proved to be unnecessary for maximizing the production of carotenoids in this fermentation.

Genome-based exploration of the specialized metabolic capacities of the genus Rhodococcus.

Science.gov (United States)

Ceniceros, Ana; Dijkhuizen, Lubbert; Petrusma, Mirjan; Medema, Marnix H

2017-08-09

Bacteria of the genus Rhodococcus are well known for their ability to degrade a large range of organic compounds. Some rhodococci are free-living, saprophytic bacteria; others are animal and plant pathogens. Recently, several studies have shown that their genomes encode putative pathways for the synthesis of a large number of specialized metabolites that are likely to be involved in microbe-microbe and host-microbe interactions. To systematically explore the specialized metabolic potential of this genus, we here performed a comprehensive analysis of the biosynthetic coding capacity across publicly available rhododoccal genomes, and compared these with those of several Mycobacterium strains as well as that of their mutual close relative Amycolicicoccus subflavus. Comparative genomic analysis shows that most predicted biosynthetic gene cluster families in these strains are clade-specific and lack any homology with gene clusters encoding the production of known natural products. Interestingly, many of these clusters appear to encode the biosynthesis of lipopeptides, which may play key roles in the diverse environments were rhodococci thrive, by acting as biosurfactants, pathogenicity factors or antimicrobials. We also identified several gene cluster families that are universally shared among all three genera, which therefore may have a more 'primary' role in their physiology. Inactivation of these clusters by mutagenesis might help to generate weaker strains that can be used as live vaccines. The genus Rhodococcus thus provides an interesting target for natural product discovery, in view of its large and mostly uncharacterized biosynthetic repertoire, its relatively fast growth and the availability of effective genetic tools for its genomic modification.

Pneumonia due to Rhodococcus equi in a non-Hodgkin's lymphoma patient: case report

Directory of Open Access Journals (Sweden)

Iuri de França Bonilha

Full Text Available The authors reported a lung infection by Rhodococcus equi in a 25 years-old male patient admitted to hospital with cough, dyspnea, fever, and previous diagnosis of pleural effusion. R. equi was isolated from pleural fluid and the patient acquired nosocomial infection by Acinetobacter baumannii, isolated from chest drain. The patient was treated with antibiotics. During hospitalization, he was diagnosed with non-Hodgkin lymphoma of precursor T-cell lymphoblastic lymphoma subtype in biopsy of pleura. After undergoing surgery for pulmonary decortication for drain empyema, the patient died due to septicemia.

Enzymatic cyanide degradation by cell-free extract of Rhodococcus UKMP-5M.

Science.gov (United States)

Nallapan Maniyam, Maegala; Sjahrir, Fridelina; Latif Ibrahim, Abdul; Cass, Anthony E G

2015-01-01

The cell-free extract of locally isolated Rhodococcus UKMP-5M strain was used as an alternative to develop greener and cost effective cyanide removal technology. The present study aims to assess the viability of the cell-free extract to detoxify high concentrations of cyanide which is measured through the monitoring of protein concentration and specific cyanide-degrading activity. When cyanide-grown cells were subjected to grinding in liquid nitrogen which is relatively an inexpressive and fast cell disruption method, highest cyanide-degrading activity of 0.63 mM min(-1) mg(-1) protein was obtained in comparison to enzymatic lysis and agitation with fine glass beads. The cell-free extracts managed to degrade 80% of 20 mM KCN within 80 min and the rate of cyanide consumption increased linearly as the concentration of protein was raised. In both cases, the addition of co-factor was not required which proved to be advantageous economically. The successful formation of ammonia and formate as endproducts indicated that the degradation of cyanide by Rhodococcus UKMP-5M proceeded via the activity of cyanidase and the resulting non-toxic products are safe for disposal into the environment. Further verification with SDS-PAGE revealed that the molecular weight of the active enzyme was estimated to be 38 kDa, which is consistent with previously reported cyanidases. Thus, the utilization of cell-free extracts as an alternative to live microbial in cyanide degradation offers numerous advantageous such as the potential to tolerate and degrade higher concentration of cyanide and total reduction in the overall cost of operation since the requirement for nutrient support is irrelevant.

Variación de la composición de ácidos grasos de membrana celular de Rhodococcus rodochrous GNP-OHP-38r en respuesta a la temperatura y salinidad Variation in the composition of Rhodococcus rodochrous GNP-OHP-38r cell membrane fatty acids in response to temperature and salinity

Directory of Open Access Journals (Sweden)

G.N. Pucci

2004-06-01

Full Text Available Los integrantes del género Rhodococcus son habitantes frecuentes y abundantes de áreas contaminadas con hidrocarburos y resisten la creciente salinidad que se presenta en la Patagonia central. Este género tiene buena capacidad de eliminar contaminantes hidrocarburos que constituyen el mayor contaminante de la región. En el presente trabajo se estudió la respuesta en la composición de sus ácidos grasos de una cepa aislada de un sistema de landfarming, ante la acción combinada de diferentes temperaturas y concentraciones salinas. La estrategia de la cepa Rhodococcus rodochrous GNP-OHP-38r frente al incremento de temperatura, es el aumento del porcentaje de los ácidos grasos saturados totales (n:0; ácidos grasos ramificados en el carbono terminal con grupos oxidrilo en posición 2 (n:0 iso 2 OH y saturados con grupo metilo en carbono 10 (n:0 10 metil, a expensas de la disminución del porcentaje de los n:1 cis.The members of the genus Rhodococcus are frequent and abundant inhabitants of polluted areas with hydrocarbons and they resist the salinity present in the central Patagonia. This genus has good capacity to eliminate pollution produced by hydrocarbons that constitutes the biggest pollutant agent in the region. The present work studies the answer in the composition of its fatty acids under the combined action of the temperature and saline concentration of an isolated stump of a landfarming system. The strategy of Rhodococcus rodochrous strain GNP-OHP-38r in front of the thermal-osmotic stress is the increase of the percentage of the total saturated fatty acids (n:0; fatty acids branched in the terminal carbon with hidroxyl group in position 2 (n:0 iso 2 OH and saturated with group methyl in carbon 10 (n:0 10 metil when the temperature is increased. These acids increase while the percentage of n:1 cis decrease.

Rhodococcus rhodochrous DSM 43269 3-Ketosteroid 9 alpha-Hydroxylase, a Two-Component Iron-Sulfur-Containing Monooxygenase with Subtle Steroid Substrate Specificity

NARCIS (Netherlands)

Petrusma, M.; Dijkhuizen, L.; van der Geize, R.

2009-01-01

This paper reports the biochemical characterization of a purified and reconstituted two-component 3-ketosteroid 9 alpha-hydroxylase (KSH). KSH of Rhodococcus rhodochrous DSM 43269, consisting of a ferredoxin reductase (KshB) and a terminal oxygenase (KshA), was heterologously expressed in

Bruker Biotyper Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry System for Identification of Nocardia, Rhodococcus, Kocuria, Gordonia, Tsukamurella, and Listeria Species

Science.gov (United States)

Lee, Tai-Fen; Du, Shin-Hei; Teng, Shih-Hua; Liao, Chun-Hsing; Sheng, Wang-Hui; Teng, Lee-Jene

2014-01-01

We evaluated whether the Bruker Biotyper matrix-associated laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) system provides accurate species-level identifications of 147 isolates of aerobically growing Gram-positive rods (GPRs). The bacterial isolates included Nocardia (n = 74), Listeria (n = 39), Kocuria (n = 15), Rhodococcus (n = 10), Gordonia (n = 7), and Tsukamurella (n = 2) species, which had all been identified by conventional methods, molecular methods, or both. In total, 89.7% of Listeria monocytogenes, 80% of Rhodococcus species, 26.7% of Kocuria species, and 14.9% of Nocardia species (n = 11, all N. nova and N. otitidiscaviarum) were correctly identified to the species level (score values, ≥2.0). A clustering analysis of spectra generated by the Bruker Biotyper identified six clusters of Nocardia species, i.e., cluster 1 (N. cyriacigeorgica), cluster 2 (N. brasiliensis), cluster 3 (N. farcinica), cluster 4 (N. puris), cluster 5 (N. asiatica), and cluster 6 (N. beijingensis), based on the six peaks generated by ClinProTools with the genetic algorithm, i.e., m/z 2,774.477 (cluster 1), m/z 5,389.792 (cluster 2), m/z 6,505.720 (cluster 3), m/z 5,428.795 (cluster 4), m/z 6,525.326 (cluster 5), and m/z 16,085.216 (cluster 6). Two clusters of L. monocytogenes spectra were also found according to the five peaks, i.e., m/z 5,594.85, m/z 6,184.39, and m/z 11,187.31, for cluster 1 (serotype 1/2a) and m/z 5,601.21 and m/z 11,199.33 for cluster 2 (serotypes 1/2b and 4b). The Bruker Biotyper system was unable to accurately identify Nocardia (except for N. nova and N. otitidiscaviarum), Tsukamurella, or Gordonia species. Continuous expansion of the MALDI-TOF MS databases to include more GPRs is necessary. PMID:24759706

Members of the genera Paenibacillus and Rhodococcus harbor genes homologous to enterococcal glycopeptide resistance genes vanA and vanB

DEFF Research Database (Denmark)

Guardabassi, L.; Christensen, H.; Hasman, Henrik

2004-01-01

Genes homologous to enterococcal glycopeptide resistance genes vanA and vanB were found in glycopeptide-resistant Paenibacillus and Rhodococcus strains from soil. The putative D-Ala:D-Lac ligase genes in Paenibacillus thiaminolyticus PT-2B1 and Paenibacillus apiarius PA-B2B were closely related...

Rhodococcus erythropolis MTHt3 biotransforms ergopeptines to lysergic acid.

Science.gov (United States)

Thamhesl, Michaela; Apfelthaler, Elisabeth; Schwartz-Zimmermann, Heidi Elisabeth; Kunz-Vekiru, Elisavet; Krska, Rudolf; Kneifel, Wolfgang; Schatzmayr, Gerd; Moll, Wulf-Dieter

2015-03-28

Ergopeptines are a predominant class of ergot alkaloids produced by tall fescue grass endophyte Neotyphodium coenophialum or cereal pathogen Claviceps purpurea. The vasoconstrictive activity of ergopeptines makes them toxic for mammals, and they can be a problem in animal husbandry. We isolated an ergopeptine degrading bacterial strain, MTHt3, and classified it, based on its 16S rDNA sequence, as a strain of Rhodococcus erythropolis (Nocardiaceae, Actinobacteria). For strain isolation, mixed microbial cultures were obtained from artificially ergot alkaloid-enriched soil, and provided with the ergopeptine ergotamine in mineral medium for enrichment. Individual colonies derived from such mixed cultures were screened for ergotamine degradation by high performance liquid chromatography and fluorescence detection. R. erythropolis MTHt3 converted ergotamine to ergine (lysergic acid amide) and further to lysergic acid, which accumulated as an end product. No other tested R. erythropolis strain degraded ergotamine. R. erythropolis MTHt3 degraded all ergopeptines found in an ergot extract, namely ergotamine, ergovaline, ergocristine, ergocryptine, ergocornine, and ergosine, but the simpler lysergic acid derivatives agroclavine, chanoclavine, and ergometrine were not degraded. Temperature and pH dependence of ergotamine and ergine bioconversion activity was different for the two reactions. Degradation of ergopeptines to ergine is a previously unknown microbial reaction. The reaction end product, lysergic acid, has no or much lower vasoconstrictive activity than ergopeptines. If the genes encoding enzymes for ergopeptine catabolism can be cloned and expressed in recombinant hosts, application of ergopeptine and ergine degrading enzymes for reduction of toxicity of ergot alkaloid-contaminated animal feed may be feasible.

Cloning, overexpression, purification, crystallization and preliminary X-ray analysis of 3-ketosteroid Delta(4)-(5 alpha)-dehydrogenase from Rhodococcus jostii RHA1

NARCIS (Netherlands)

van Oosterwijk, Niels; Knol, Jan; Dijkhuizen, Lubbert; van der Geize, Robert; Dijkstra, Bauke

2011-01-01

3-Ketosteroid dehydrogenases are flavoproteins which play key roles in steroid ring degradation. The enzymes are abundantly present in actinobacteria, including the catabolic powerhouse Rhodococcus jostii and the pathogenic species R. equi and Mycobacterium tuberculosis. The gene for 3-ketosteroid

Rhodococcus equi: the many facets of a pathogenic actinomycete.

Science.gov (United States)

Vázquez-Boland, José A; Giguère, Steeve; Hapeshi, Alexia; MacArthur, Iain; Anastasi, Elisa; Valero-Rello, Ana

2013-11-29

Rhodococcus equi is a soil-dwelling pathogenic actinomycete that causes pulmonary and extrapulmonary pyogranulomatous infections in a variety of animal species and people. Young foals are particularly susceptible and develop a life-threatening pneumonic disease that is endemic at many horse-breeding farms worldwide. R. equi is a facultative intracellular parasite of macrophages that replicates within a modified phagocytic vacuole. Its pathogenicity depends on a virulence plasmid that promotes intracellular survival by preventing phagosome-lysosome fusion. Species-specific tropism of R. equi for horses, pigs and cattle appears to be determined by host-adapted virulence plasmid types. Molecular epidemiological studies of these plasmids suggest that human R. equi infection is zoonotic. Analysis of the recently determined R. equi genome sequence has identified additional virulence determinants on the bacterial chromosome. This review summarizes our current understanding of the clinical aspects, biology, pathogenesis and immunity of this fascinating microbe with plasmid-governed infectivity. Copyright © 2013 Elsevier B.V. All rights reserved.

Biodesulfurization of Naphthothiophene and Benzothiophene through Selective Cleavage of Carbon-Sulfur Bonds by Rhodococcus sp. Strain WU-K2R

Science.gov (United States)

Kirimura, Kohtaro; Furuya, Toshiki; Sato, Rika; Ishii, Yoshitaka; Kino, Kuniki; Usami, Shoji

2002-01-01

Naphtho[2,1-b]thiophene (NTH) is an asymmetric structural isomer of dibenzothiophene (DBT), and in addition to DBT derivatives, NTH derivatives can also be detected in diesel oil following hydrodesulfurization treatment. Rhodococcus sp. strain WU-K2R was newly isolated from soil for its ability to grow in a medium with NTH as the sole source of sulfur, and growing cells of WU-K2R degraded 0.27 mM NTH within 7 days. WU-K2R could also grow in the medium with NTH sulfone, benzothiophene (BTH), 3-methyl-BTH, or 5-methyl-BTH as the sole source of sulfur but could not utilize DBT, DBT sulfone, or 4,6-dimethyl-DBT. On the other hand, WU-K2R did not utilize NTH or BTH as the sole source of carbon. By gas chromatography-mass spectrometry analysis, desulfurized NTH metabolites were identified as NTH sulfone, 2′-hydroxynaphthylethene, and naphtho[2,1-b]furan. Moreover, since desulfurized BTH metabolites were identified as BTH sulfone, benzo[c][1,2]oxathiin S-oxide, benzo[c][1,2]oxathiin S,S-dioxide, o-hydroxystyrene, 2-(2′-hydroxyphenyl)ethan-1-al, and benzofuran, it was concluded that WU-K2R desulfurized NTH and BTH through the sulfur-specific degradation pathways with the selective cleavage of carbon-sulfur bonds. Therefore, Rhodococcus sp. strain WU-K2R, which could preferentially desulfurize asymmetric heterocyclic sulfur compounds such as NTH and BTH through the sulfur-specific degradation pathways, is a unique desulfurizing biocatalyst showing properties different from those of DBT-desulfurizing bacteria. PMID:12147483

Restriction Fragment Length Polymorphisms of Virulence Plasmids in Rhodococcus equi

Science.gov (United States)

Takai, Shinji; Shoda, Masato; Sasaki, Yukako; Tsubaki, Shiro; Fortier, Guillaume; Pronost, Stephane; Rahal, Karim; Becu, Teotimo; Begg, Angela; Browning, Glenn; Nicholson, Vivian M.; Prescott, John F.

1999-01-01

Virulent Rhodococcus equi, which is a well-known cause of pyogranulomatous pneumonia in foals, possesses a large plasmid encoding virulence-associated 15- to 17-kDa antigens. Foal and soil isolates from five countries—Argentina, Australia, Canada, France, and Japan—were investigated for the presence of 15- to 17-kDa antigens by colony blotting, using the monoclonal antibody 10G5, and the gene coding for 15- to 17-kDa antigens by PCR. Plasmid DNAs extracted from positive isolates were digested with restriction endonucleases BamHI, EcoRI, EcoT22I, and HindIII, and the digestion patterns that resulted divided the plasmids of virulent isolates into five closely related types. Three of the five types had already been reported in Canadian and Japanese isolates, and the two new types had been found in French and Japanese isolates. Therefore, we tentatively designated these five types 85-kb type I (pREAT701), 85-kb type II (a new type), 87-kb type I (EcoRI and BamHI type 2 [V. M. Nicholson and J. F. Prescott, J. Clin. Microbiol. 35:738–740, 1997]), 87-kb type II (a new type), and 90-kb (pREL1) plasmids. The 85-kb type I plasmid was found in isolates from Argentina, Australia, Canada, and France. Plasmid 87-kb type I was isolated in specimens from Argentina, Canada, and France. The 85-kb type II plasmid appeared in isolates from France. On the other hand, plasmids 87-kb type II and 90-kb were found only in isolates from Japan. These results revealed geographic differences in the distribution of the virulence plasmids found in the five countries and suggested that the restriction fragment length polymorphism of virulence plasmids might be useful to elucidate the molecular epidemiology of virulent R. equi in the world. PMID:10488224

Individual or synchronous biodegradation of di-n-butyl phthalate and phenol by Rhodococcus ruber strain DP-2

Energy Technology Data Exchange (ETDEWEB)

He, Zhixing; Niu, Chengzhen; Lu, Zhenmei, E-mail: lzhenmei@zju.edu.cn

2014-05-01

Highlights: • A Rhodococcus ruber strain degraded DBP and phenol. • Degradation kinetics of DBP or phenol fit modified first-order models. • Degradation interaction between DBP and phenol was studied by strain DP-2. • The degradation genes transcriptional were quantified by RT-qPCR. - Abstract: The bacterial strain DP-2, identified as Rhodococcus ruber, is able to effectively degrade di-n-butyl phthalate (DBP) and phenol. Degradation kinetics of DBP and phenol at different initial concentrations revealed DBP and phenol degradation to fit modified first-order models. The half-life of DBP degradation ranged from 15.81 to 27.75 h and phenol degradation from 14.52 to 45.52 h under the initial concentrations of 600–1200 mg/L. When strain DP-2 was cultured with a mixture of DBP (800 mg/L) and phenol (700 mg/L), DBP degradation rate was found to be only slightly influenced; however, phthalic acid (PA) accumulated, and phenol degradation was clearly inhibited during synchronous degradation. Transcriptional levels of degradation genes, phenol hydroxylase (pheu) and phthalate 3,4-dioxygenase (pht), decreased significantly more during synchronous degradation than during individual degradation. Quantitative estimation of individual or synchronous degradation kinetics is essential to manage mixed hazardous compounds through biodegradation in industrial waste disposal.

New reactions and products resulting from alternative interactions between the P450 enzyme and redox partners.

Science.gov (United States)

Zhang, Wei; Liu, Yi; Yan, Jinyong; Cao, Shaona; Bai, Fali; Yang, Ying; Huang, Shaohua; Yao, Lishan; Anzai, Yojiro; Kato, Fumio; Podust, Larissa M; Sherman, David H; Li, Shengying

2014-03-05

Cytochrome P450 enzymes are capable of catalyzing a great variety of synthetically useful reactions such as selective C-H functionalization. Surrogate redox partners are widely used for reconstitution of P450 activity based on the assumption that the choice of these auxiliary proteins or their mode of action does not affect the type and selectivity of reactions catalyzed by P450s. Herein, we present an exceptional example to challenge this postulate. MycG, a multifunctional biosynthetic P450 monooxygenase responsible for hydroxylation and epoxidation of 16-membered ring macrolide mycinamicins, is shown to catalyze the unnatural N-demethylation(s) of a range of mycinamicin substrates when partnered with the free Rhodococcus reductase domain RhFRED or the engineered Rhodococcus-spinach hybrid reductase RhFRED-Fdx. By contrast, MycG fused with the RhFRED or RhFRED-Fdx reductase domain mediates only physiological oxidations. This finding highlights the larger potential role of variant redox partner protein-protein interactions in modulating the catalytic activity of P450 enzymes.

Biocatalytic Properties and Structural Analysis of Eugenol Oxidase from Rhodococcus jostii RHA1: A Versatile Oxidative Biocatalyst

OpenAIRE

Nguyen, Quoc-Thai; de Gonzalo, Gonzalo; Binda, Claudia; Rioz-Martínez, Ana; Mattevi, Andrea; Fraaije, Marco W.

2016-01-01

Abstract Eugenol oxidase (EUGO) from Rhodococcus jostii RHA1 had previously been shown to convert only a limited set of phenolic compounds. In this study, we have explored the biocatalytic potential of this flavoprotein oxidase, resulting in a broadened substrate scope and a deeper insight into its structural properties. In addition to the oxidation of vanillyl alcohol and the hydroxylation of eugenol, EUGO can efficiently catalyze the dehydrogenation of various phenolic ketones and the selec...

Bio-surfactants production from low cost substrate and degradation of diesel oil by a Rhodococcus strain; Production de biosurfactants sur un substrat economique et degradation du gasoil par une souche du genre Rhodococcus

Energy Technology Data Exchange (ETDEWEB)

Sadouk, Z.; Tazerouti, A. [Universite des Sciences et de la Technologie H. Boumediene (USTHB), Lab. de Synthese Organique, Faculte de Chimie, Algiers (Algeria); Sadouk, Z.; Hacene, H. [Universite des Sciences et de la Technologie H. Boumediene (USTHB), Lab. de Microbiologie, Faculte des Sciences Biologiques, Algiers (Algeria)

2008-07-01

The ability of a Rhodococcus strain to produce surface-active agents from residual sunflower frying oil (RSFO) has been screened in batch cultures. During cultivation with RSFO at the concentration 3% (vol/vol), the strain has synthesized extra-cellular compounds which increase the E{sub 24} emulsion index of the culture medium up to 63%. In their crude form, these substances lower the surface tension of water until 31.9 mN m{sup -1}. The exponential growth with RSFO as the sole carbon source has developed at a specific growth rate {mu} = 0.55 d{sup -1}. The critical micelle concentration of the crude product reached the value 287 mg L{sup -1} ({gamma}CMC = 31.9 mN m{sup -1}). After methyl-esterification, the lipid fraction of bio-surfactants has been analyzed by GC-MS in EI, which reveals the presence of fatty acid methyl esters. The microorganism was also cultivated with the diesel oil as the sole carbon source at the concentration 1% (vol/vol): the active growth phase has developed at rate = 0.02 d{sup -1}, without production of emulsifying substance: the microorganism seems to develop different modes of substrate uptake, according to the nature of the carbon source. The potential use of surface-active agents synthesized on RSFO by Rhodococcus erythropolis 16 LM.USTHB is in the oil industry with minimum purity specification, so that crude preparation could be used, at low cost, in clean-up of hydrocarbons contaminated sites and for enhanced oil recovery. (authors)

vanO, a new glycopeptide resistance operon in environmental Rhodococcus equi isolates

DEFF Research Database (Denmark)

Gudeta, Dereje Dadi; Moodley, Arshnee; Bortolaia, Valeria

2014-01-01

We describe sequence and gene organization of a new glycopeptide resistance operon (vanO) in Rhodococcus equi from soil. The vanO operon has low homology to enterococccal van operons and harbors a vanHOX cluster transcribed in opposite direction to the vanS-vanR regulatory system and comprised be...... between three open reading frames with unknown function. This finding has clinical interest since glycopeptides are used to treat R. equi infections and resistance has been reported in clinical isolates....

Biotransformation of nitriles to amides using soluble and immobilized nitrile hydratase from Rhodococcus erythropolis A4

Czech Academy of Sciences Publication Activity Database

Kubáč, David; Kaplan, Ondřej; Elišáková, Veronika; Pátek, Miroslav; Vejvoda, Vojtěch; Slámová, Kristýna; Tóthová, A.; Lemaire, M.; Gallienne, E.; Lutz-Wahl, S.; Fischer, L.; Kuzma, Marek; Pelantová, Helena; van Pelt, S.; Bolte, J.; Křen, Vladimír; Martínková, Ludmila

2008-01-01

Roč. 50, 2-4 (2008), s. 107-113 ISSN 1381-1177 R&D Projects: GA ČR GA203/05/2267; GA MŠk(CZ) LC06010; GA MŠk OC 171 Grant - others:XE(XE) ESF COST D25/0002/02; CZ(CZ) D10-CZ25/06-07; CZ(CZ) D-25 Institutional research plan: CEZ:AV0Z50200510 Keywords : rhodococcus erythropolis * nitrile hydratase * amidase Subject RIV: EE - Microbiology, Virology Impact factor: 2.015, year: 2008

Crystals of DhaA mutants from Rhodococcus rhodochrous NCIMB 13064 diffracted to ultrahigh resolution: crystallization and preliminary diffraction analysis.

Science.gov (United States)

Stsiapanava, Alena; Koudelakova, Tana; Lapkouski, Mikalai; Pavlova, Martina; Damborsky, Jiri; Smatanova, Ivana Kuta

2008-02-01

The enzyme DhaA from Rhodococcus rhodochrous NCIMB 13064 belongs to the haloalkane dehalogenases, which catalyze the hydrolysis of haloalkanes to the corresponding alcohols. The haloalkane dehalogenase DhaA and its variants can be used to detoxify the industrial pollutant 1,2,3-trichloropropane (TCP). Three mutants named DhaA04, DhaA14 and DhaA15 were constructed in order to study the importance of tunnels connecting the buried active site with the surrounding solvent to the enzymatic activity. All protein mutants were crystallized using the sitting-drop vapour-diffusion method. The crystals of DhaA04 belonged to the orthorhombic space group P2(1)2(1)2(1), while the crystals of the other two mutants DhaA14 and DhaA15 belonged to the triclinic space group P1. Native data sets were collected for the DhaA04, DhaA14 and DhaA15 mutants at beamline X11 of EMBL, DESY, Hamburg to the high resolutions of 1.30, 0.95 and 1.15 A, respectively.

Evaluation of biosurfactants grown in corn oil by Rhodococcus rhodochrous on removing of heavy metal ion from aqueous solution

International Nuclear Information System (INIS)

Suryanti, Venty; Hastuti, Sri; Pujiastuti, Dwi

2016-01-01

The potential application of biosurfactants to remove heavy metal ion from aqueous solution by batch technique was examined. The glycolipids type biosurfactants were grown in a media containing of 20% v/v corn oil with 7 days of fermentation by Rhodococcus rhodochrous. The biosurfactants reduced the surface tension of water of about 51% from 62 mN/m to 30 mN/m. The biosurfactant increased the E24 of water-palm oil emulsion of about 55% from 43% to 97% and could maintain this E24 value of above 50% for up to 9 days. Heavy metal ion removal, in this case cadmium ion, by crude and patially purified biosurfactants has been investigated from aqueous solution at pH 6. Adsorption capacity of Cd(II) ion by crude biosurfactant with 5 and 10 minutes of contact times were 1.74 and 1.82 mg/g, respectively. Additionally, the adsorption capacity of Cd(II) ion by partially purified biosurfactant with 5 and 10 minutes of contact times were 0.79 and 1.34 mg/g, respectively. The results demonstrated that the adsorption capacity of Cd(II) ion by crude biosurfactant was higher than that of by partially purified biosurfactant. The results suggested that the biosurfactant could be used in the removal of heavy metal ions from aqueous solution

Evaluation of biosurfactants grown in corn oil by Rhodococcus rhodochrous on removing of heavy metal ion from aqueous solution

Science.gov (United States)

Suryanti, Venty; Hastuti, Sri; Pujiastuti, Dwi

2016-02-01

The potential application of biosurfactants to remove heavy metal ion from aqueous solution by batch technique was examined. The glycolipids type biosurfactants were grown in a media containing of 20% v/v corn oil with 7 days of fermentation by Rhodococcus rhodochrous. The biosurfactants reduced the surface tension of water of about 51% from 62 mN/m to 30 mN/m. The biosurfactant increased the E24 of water-palm oil emulsion of about 55% from 43% to 97% and could maintain this E24 value of above 50% for up to 9 days. Heavy metal ion removal, in this case cadmium ion, by crude and patially purified biosurfactants has been investigated from aqueous solution at pH 6. Adsorption capacity of Cd(II) ion by crude biosurfactant with 5 and 10 minutes of contact times were 1.74 and 1.82 mg/g, respectively. Additionally, the adsorption capacity of Cd(II) ion by partially purified biosurfactant with 5 and 10 minutes of contact times were 0.79 and 1.34 mg/g, respectively. The results demonstrated that the adsorption capacity of Cd(II) ion by crude biosurfactant was higher than that of by partially purified biosurfactant. The results suggested that the biosurfactant could be used in the removal of heavy metal ions from aqueous solution.

Evaluation of biosurfactants grown in corn oil by Rhodococcus rhodochrous on removing of heavy metal ion from aqueous solution

Energy Technology Data Exchange (ETDEWEB)

Suryanti, Venty, E-mail: venty@mipa.uns.ac.id; Hastuti, Sri; Pujiastuti, Dwi [Department of Chemistry, Faculty of Mathematics and Natural Sciences, Sebelas Maret University Jl. Ir. Sutami 36A, Surakarta, Central Java 57126 (Indonesia)

2016-02-08

The potential application of biosurfactants to remove heavy metal ion from aqueous solution by batch technique was examined. The glycolipids type biosurfactants were grown in a media containing of 20% v/v corn oil with 7 days of fermentation by Rhodococcus rhodochrous. The biosurfactants reduced the surface tension of water of about 51% from 62 mN/m to 30 mN/m. The biosurfactant increased the E24 of water-palm oil emulsion of about 55% from 43% to 97% and could maintain this E24 value of above 50% for up to 9 days. Heavy metal ion removal, in this case cadmium ion, by crude and patially purified biosurfactants has been investigated from aqueous solution at pH 6. Adsorption capacity of Cd(II) ion by crude biosurfactant with 5 and 10 minutes of contact times were 1.74 and 1.82 mg/g, respectively. Additionally, the adsorption capacity of Cd(II) ion by partially purified biosurfactant with 5 and 10 minutes of contact times were 0.79 and 1.34 mg/g, respectively. The results demonstrated that the adsorption capacity of Cd(II) ion by crude biosurfactant was higher than that of by partially purified biosurfactant. The results suggested that the biosurfactant could be used in the removal of heavy metal ions from aqueous solution.

Structure of the virulence-associated protein VapD from the intracellular pathogen Rhodococcus equi

International Nuclear Information System (INIS)

Whittingham, Jean L.; Blagova, Elena V.; Finn, Ciaran E.; Luo, Haixia; Miranda-CasoLuengo, Raúl; Turkenburg, Johan P.; Leech, Andrew P.; Walton, Paul H.; Murzin, Alexey G.; Meijer, Wim G.; Wilkinson, Anthony J.

2014-01-01

VapD is one of a set of highly homologous virulence-associated proteins from the multi-host pathogen Rhodococcus equi. The crystal structure reveals an eight-stranded β-barrel with a novel fold and a glycine rich ‘bald’ surface. Rhodococcus equi is a multi-host pathogen that infects a range of animals as well as immune-compromised humans. Equine and porcine isolates harbour a virulence plasmid encoding a homologous family of virulence-associated proteins associated with the capacity of R. equi to divert the normal processes of endosomal maturation, enabling bacterial survival and proliferation in alveolar macrophages. To provide a basis for probing the function of the Vap proteins in virulence, the crystal structure of VapD was determined. VapD is a monomer as determined by multi-angle laser light scattering. The structure reveals an elliptical, compact eight-stranded β-barrel with a novel strand topology and pseudo-twofold symmetry, suggesting evolution from an ancestral dimer. Surface-associated octyl-β-d-glucoside molecules may provide clues to function. Circular-dichroism spectroscopic analysis suggests that the β-barrel structure is preceded by a natively disordered region at the N-terminus. Sequence comparisons indicate that the core folds of the other plasmid-encoded virulence-associated proteins from R. equi strains are similar to that of VapD. It is further shown that sequences encoding putative R. equi Vap-like proteins occur in diverse bacterial species. Finally, the functional implications of the structure are discussed in the light of the unique structural features of VapD and its partial structural similarity to other β-barrel proteins

Structure of the virulence-associated protein VapD from the intracellular pathogen Rhodococcus equi

Energy Technology Data Exchange (ETDEWEB)

Whittingham, Jean L.; Blagova, Elena V. [University of York, Heslington, York YO10 5DD (United Kingdom); Finn, Ciaran E.; Luo, Haixia; Miranda-CasoLuengo, Raúl [University College Dublin, Dublin (Ireland); Turkenburg, Johan P.; Leech, Andrew P.; Walton, Paul H. [University of York, Heslington, York YO10 5DD (United Kingdom); Murzin, Alexey G. [MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH (United Kingdom); Meijer, Wim G. [University College Dublin, Dublin (Ireland); Wilkinson, Anthony J., E-mail: tony.wilkinson@york.ac.uk [University of York, Heslington, York YO10 5DD (United Kingdom)

2014-08-01

VapD is one of a set of highly homologous virulence-associated proteins from the multi-host pathogen Rhodococcus equi. The crystal structure reveals an eight-stranded β-barrel with a novel fold and a glycine rich ‘bald’ surface. Rhodococcus equi is a multi-host pathogen that infects a range of animals as well as immune-compromised humans. Equine and porcine isolates harbour a virulence plasmid encoding a homologous family of virulence-associated proteins associated with the capacity of R. equi to divert the normal processes of endosomal maturation, enabling bacterial survival and proliferation in alveolar macrophages. To provide a basis for probing the function of the Vap proteins in virulence, the crystal structure of VapD was determined. VapD is a monomer as determined by multi-angle laser light scattering. The structure reveals an elliptical, compact eight-stranded β-barrel with a novel strand topology and pseudo-twofold symmetry, suggesting evolution from an ancestral dimer. Surface-associated octyl-β-d-glucoside molecules may provide clues to function. Circular-dichroism spectroscopic analysis suggests that the β-barrel structure is preceded by a natively disordered region at the N-terminus. Sequence comparisons indicate that the core folds of the other plasmid-encoded virulence-associated proteins from R. equi strains are similar to that of VapD. It is further shown that sequences encoding putative R. equi Vap-like proteins occur in diverse bacterial species. Finally, the functional implications of the structure are discussed in the light of the unique structural features of VapD and its partial structural similarity to other β-barrel proteins.

Both epiphytic and endophytic strains of Rhodococcus fascians influence transporter gene expression and cytokinins in infected Pisum sativum L. seedlings

Czech Academy of Sciences Publication Activity Database

Dhandapani, P.; Song, J.; Novák, Ondřej; Jameson, P. E.

2018-01-01

Roč. 85, č. 2 (2018), s. 231-242 ISSN 0167-6903 R&D Projects: GA ČR(CZ) GA17-06613S Institutional support: RVO:61389030 Keywords : Amino acid transporter * Cell wall invertase * Cytokinin * Pea * Rhodococcus fascians * Sucrose transporter * Sugar Will Eventually be Exported Transporter (SWEET) Subject RIV: EF - Botanics OBOR OECD: Plant sciences, botany Impact factor: 2.646, year: 2016

Bio-surfactants production from low cost substrate and degradation of diesel oil by a Rhodococcus strain

International Nuclear Information System (INIS)

Sadouk, Z.; Tazerouti, A.; Sadouk, Z.; Hacene, H.

2008-01-01

The ability of a Rhodococcus strain to produce surface-active agents from residual sunflower frying oil (RSFO) has been screened in batch cultures. During cultivation with RSFO at the concentration 3% (vol/vol), the strain has synthesized extra-cellular compounds which increase the E 24 emulsion index of the culture medium up to 63%. In their crude form, these substances lower the surface tension of water until 31.9 mN m -1 . The exponential growth with RSFO as the sole carbon source has developed at a specific growth rate μ = 0.55 d -1 . The critical micelle concentration of the crude product reached the value 287 mg L -1 (γCMC = 31.9 mN m -1 ). After methyl-esterification, the lipid fraction of bio-surfactants has been analyzed by GC-MS in EI, which reveals the presence of fatty acid methyl esters. The microorganism was also cultivated with the diesel oil as the sole carbon source at the concentration 1% (vol/vol): the active growth phase has developed at rate = 0.02 d -1 , without production of emulsifying substance: the microorganism seems to develop different modes of substrate uptake, according to the nature of the carbon source. The potential use of surface-active agents synthesized on RSFO by Rhodococcus erythropolis 16 LM.USTHB is in the oil industry with minimum purity specification, so that crude preparation could be used, at low cost, in clean-up of hydrocarbons contaminated sites and for enhanced oil recovery. (authors)

Biodegradation of Di-(2-ethylhexyl) Phthalate by Rhodococcus ruber YC-YT1 in Contaminated Water and Soil

OpenAIRE

Ting Yang; Lei Ren; Yang Jia; Shuanghu Fan; Junhuan Wang; Jiayi Wang; Ruth Nahurira; Haisheng Wang; Yanchun Yan

2018-01-01

Di-(2-ethylehxyl) phthalate (DEHP) is one of the most broadly representative phthalic acid esters (PAEs) used as a plasticizer in polyvinyl chloride (PVC) production, and is considered to be an endocrine-disrupting chemical. DEHP and its monoester metabolites are responsible for adverse effects on human health. An efficient DEHP-degrading bacterial strain Rhodococcus ruber YC-YT1, with super salt tolerance (0–12% NaCl), is the first DEHP-degrader isolated from marine plastic debris found in c...

The 4p-5d, 6d and 4p-6s, 7s transitions of Mo IX

International Nuclear Information System (INIS)

Khatoon, S.; Chaghtai, M.S.Z.; Rahimullah, K.

1979-01-01

The transitions 4p-5d, 6d and 4p-6s, 7s have been studied for the first time in Mo IX. The authors have identified 42 4p-5d, 36 4p-6d, 22 4p-6s and 22 4p-7s transitions, establishing 16 4p 3 5d, 14 4p 3 6d and all the ten 4p 3 6s, 7s levels of the spectrum concerned. The ionization energy is estimated to be (1 323 700 +- 700)cm -1 or (164.11 +- 0.09)eV. The spectrum was recorded in sliding and open spark discharges with a 5 m grazing incidence spectrograph of Lund University (Sweden) from about 40 A to 440 A. (Auth.)

Mechanisms of the p(He 6,He 5)d, p(He 6,α)t and p(He 6,t)α reactions

International Nuclear Information System (INIS)

Heiberg-Andersen, Henning

2002-07-01

This work was devoted to nucleon induced transfer reactions having the potential to probe the sub-cluster structures of the benchmark halo nucleus He 6, without the question marks the necessarily omitted exchange effects tend to put behind the CRC results when both collision partners are composite systems. Still, the exchange complications entered the analysis in an ironic way: The high Q-value of the p(He 6,α)t and p(He 6, t)α reactions caused sensitivity to the t - α optical potential at small radii, where the one-nucleon exchange effects are strongest. Since the attempt to throw them out of the extracted tau - α potential failed, it was necessary to extend the model space to avoid a too difficult modelling of the local equivalent t - α potential. By this step, all the complications originating from antisymmetrization within a larger model space entered the analysis. However, the persistent failures of the two-channel calculations of this and previous works can hardly be due to incorrect treatment of exchange effects only, so the loss of simplicity is probably illusory. Even at small angles, where the surface processes dominate, none of the two-channel calculations with various choices of t - α optical potentials managed to reproduce the p(He 6, α)t (p(He 6,t)α) data. This motivated inclusion of sequential transfers through the d + He 5 channel, where the sequential triton transfer process, included just for consistency in the coupling scheme of the four-channel calculation, turned out to be more influent than expected. The satisfactory reproduction of both the p(He 6, He 5)d and the p(He 6,α)t (p(He 6,t)α) data by the four-channel approach and the required re-normalization the real part of the p - He 6 optical potential are strong indications of substantial contributions from sequential transfer of the halo neutrons at this energy. The conclusions that can be drawn from this work are limited by the modest amount of available data. This limitation does

Mechanisms of the p(He 6,He 5)d, p(He 6,{alpha})t and p(He 6,t){alpha} reactions

Energy Technology Data Exchange (ETDEWEB)

Heiberg-Andersen, Henning

2002-07-01

This work was devoted to nucleon induced transfer reactions having the potential to probe the sub-cluster structures of the benchmark halo nucleus He 6, without the question marks the necessarily omitted exchange effects tend to put behind the CRC results when both collision partners are composite systems. Still, the exchange complications entered the analysis in an ironic way: The high Q-value of the p(He 6,{alpha})t and p(He 6, t){alpha} reactions caused sensitivity to the t - {alpha} optical potential at small radii, where the one-nucleon exchange effects are strongest. Since the attempt to throw them out of the extracted tau - {alpha} potential failed, it was necessary to extend the model space to avoid a too difficult modelling of the local equivalent t - {alpha} potential. By this step, all the complications originating from antisymmetrization within a larger model space entered the analysis. However, the persistent failures of the two-channel calculations of this and previous works can hardly be due to incorrect treatment of exchange effects only, so the loss of simplicity is probably illusory. Even at small angles, where the surface processes dominate, none of the two-channel calculations with various choices of t - {alpha} optical potentials managed to reproduce the p(He 6, {alpha})t (p(He 6,t){alpha}) data. This motivated inclusion of sequential transfers through the d + He 5 channel, where the sequential triton transfer process, included just for consistency in the coupling scheme of the four-channel calculation, turned out to be more influent than expected. The satisfactory reproduction of both the p(He 6, He 5)d and the p(He 6,{alpha})t (p(He 6,t){alpha}) data by the four-channel approach and the required re-normalization the real part of the p - He 6 optical potential are strong indications of substantial contributions from sequential transfer of the halo neutrons at this energy. The conclusions that can be drawn from this work are limited by the

Biosurfactant production by Rhodococcus erythropolis and its application to oil removal

Directory of Open Access Journals (Sweden)

Graziela Jardim Pacheco

2010-10-01

Full Text Available The influence of different nutrients on biosurfactant production by Rhodococcus erythropolis was investigated. Increasing the concentration of phosphate buffer from 30 up through 150 mmol/L stimulated an increase in biosurfactant production, which reached a maximum concentration of 285 mg/L in shaken flasks. Statistical analysis showed that glycerol, NaNO3,MgSO4 and yeast extract had significant effects on production. The results were confirmed in a batchwise bioreactor, and semi-growth-associated production was detected. Reduction in the surface tension, which indicates the presence of biosurfactant, reached a value of 38 mN/m at the end of 35 hours. Use of the produced biosurfactant for washing crude oil-contaminated soil showed that 2 and 4 times the critical micellar concentration (CMC were able to remove 97 and 99% of the oil, respectively, after 1 month of impregnation.

Identificação diferencial de Rhodococcus equi e Dietzia maris em bubalinos Differential identification of Rhodococcus equi and Dietzia maris in buffaloes

Directory of Open Access Journals (Sweden)

L.R. Viana

2009-08-01

Full Text Available Foram analisados 24 isolados bacterianos oriundos de leite e pele de búfalas (Bubalus bubalis, os quais foram previamente identificados como Rhodococcus equi com o auxílio de fenotipia concisa. Testes fenotípicos complementares e ferramentas moleculares foram utilizados com o objetivo de caracterizar esses isolados, bem como diferenciá-los de outros microrganismos intimamente relacionados. Observaram-se três fenótipos distintos, porém a identificação dos isolados foi inconclusiva. Apenas um dos isolados foi comprovado como sendo R. equi com a realização da PCR espécie-específica, sequenciamento e análise dos fragmentos de DNA. Os demais isolados só foram identificados pelo sequenciamento de fragmento do gene que codifica a região 16S do rRNA universal de bactérias, indicando tratar-se de Dietzia maris. O perfil de susceptibilidade aos antimicrobianos revelou maior resistência dos isolados de D. maris para oxacilina (96% e rifampicina (87%. O isolado de R. equi apresentou resistência à amicacina, oxacilina, penicilina, rifampicina e tetraciclina. Alerta-se para o risco da incorreta identificação dos isolados baseados em testes fenotípicos concisos e para a necessidade de utilização de testes complementares para diferenciação entre R. equi e D. maris.Twenty-four bacterial isolates from milk and skin of buffalo females (Bubalus bubalis, which previously had been identified as Rhodococcus equi by using a restricted number of phenotypical tests for bacterial characterization, were analyzed. The goal of this study was to perform the characterization of these isolates, as well as the differentiation of other microorganisms closely related by using additional phenotypical tests and molecular tools. Based on the phenotypical results, three different biotypes were obtained. However, the identification of the isolates was inconclusive. Only one of the isolates was confirmed as R. equi by the PCR specifically for this species, as

Induction of leafy galls in Acacia mearnsii De Wild seedlings infected by Rhodococcus fascians

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Marguerite Quoirin

2004-07-01

Full Text Available Plantlets of blackwattle (Acacia mearnsii De Wild were inoculated with the bacterium Rhodococcus fascians and cultured in vitro. Leafy galls appeared at the cotyledonary nodes in 75% of the infected plants. The galls were separated from the plants and cultured on a medium containing three-quarters-strength MS salts (Murashige and Skoog, 1962, MS vitamins, 2% sucrose and an antibiotic (cephalothin, supplemented with or without 0.2% activated charcoal. Histological studies conducted from the sixth to the twenty-second day after plant infection revealed the presence of newly formed meristematic centers, first in the axillary region, then on the petioles and lamina of the leaflets around the apical meristem. Approximately 37% of the galls developed one shoot with both concentrations of cephalothin.Plantas recém germinadas de acácia negra (Acacia mearnsii De Wild. foram inoculadas com a bactéria Rhodococcus fascians e cultivadas in vitro. Galhas cobertas por folhas apareceram na altura do nó cotiledonar em 75% das plantas infectadas. As galhas foram separadas das plantas e cultivadas num meio de cultura contendo os sais do meio MS (Murashige e Skoog, 1962 reduzidos a 3/4, as vitaminas do mesmo meio, 2% de sacarose e um antibiótico (cefalotina, adicionado ou não de 0,2% de carvão ativo. Estudos histológicos realizados entre o sexto e o vigésimo segundo dia depois da inoculação, revelaram a presença de centros meristemáticos novos, primeiro nas regiões axilares, em seguida nos pecíolos e limbos dos folíolos ao redor do meristema apical. Aproximadamente 37% das galhas desenvolveram um broto na presença de cefalotina.

Crystals of DhaA mutants from Rhodococcus rhodochrous NCIMB 13064 diffracted to ultrahigh resolution: crystallization and preliminary diffraction analysis

International Nuclear Information System (INIS)

Stsiapanava, Alena; Koudelakova, Tana; Lapkouski, Mikalai; Pavlova, Martina; Damborsky, Jiri; Kuta Smatanova, Ivana

2008-01-01

Three mutants of the haloalkane dehalogenase DhaA derived from R. rhodochrous NCIMB 13064 were crystallized and diffracted to ultrahigh resolution. The enzyme DhaA from Rhodococcus rhodochrous NCIMB 13064 belongs to the haloalkane dehalogenases, which catalyze the hydrolysis of haloalkanes to the corresponding alcohols. The haloalkane dehalogenase DhaA and its variants can be used to detoxify the industrial pollutant 1,2,3-trichloropropane (TCP). Three mutants named DhaA04, DhaA14 and DhaA15 were constructed in order to study the importance of tunnels connecting the buried active site with the surrounding solvent to the enzymatic activity. All protein mutants were crystallized using the sitting-drop vapour-diffusion method. The crystals of DhaA04 belonged to the orthorhombic space group P2 1 2 1 2 1 , while the crystals of the other two mutants DhaA14 and DhaA15 belonged to the triclinic space group P1. Native data sets were collected for the DhaA04, DhaA14 and DhaA15 mutants at beamline X11 of EMBL, DESY, Hamburg to the high resolutions of 1.30, 0.95 and 1.15 Å, respectively

APPLICATION OF PSEUDOMONAS PUTIDA AND RHODOCOCCUS SP. BY BIODEGRADATION OF PAH(S, PCB(S AND NEL SOIL SAMPLES FROM THE HAZARDOUS WASTE DUMP IN POZĎÁTKY (CZECH REPUBLIC

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Radmila Kucerova

2006-12-01

Full Text Available The objective of the project was a laboratory check of biodegradation of soil samples contaminated by PAH(s, PCB(s and NEL from the hazardous waste dump in the Pozďátky locality. For the laboratory check, pure bacterial cultures of Rhodococcus sp. and Pseudomonas putida have been used. It is apparent from the laboratory experiments results that after one-month bacterial leaching, applying the bacterium of Rhodococcus sp. there is a 83 % removal of NEL, a 79 % removal of PAH(s and a 14 % removal of PCB(s. Applying a pure culture of Pseudomonas putida there is a 87 % removal of NEL, a 81 % removal of PAH(s and a 14 % removal of PCB(s.

Successful bioaugmentation of an activated sludge reactor with Rhodococcus sp. YYL for efficient tetrahydrofuran degradation

International Nuclear Information System (INIS)

Yao, Yanlai; Lu, Zhenmei; Zhu, Fengxiang; Min, Hang; Bian, Caimiao

2013-01-01

Highlights: • Rhodococcus sp. YYL is an efficient tetrahydrofuran-degrading strain. • Strain YYL was used to augment an activated sludge system for THF degradation. • Successful bioaugmentation was achieved only by coinoculation of strain YYL and the two bacilli. • Successful bioaugmentation of the system resulted in efficient THF degradation. -- Abstract: The exchange of tetrahydrofuran (THF)-containing wastewater should significantly affect the performance of an activated sludge system. In this study, the feasibility of using THF-degrading Rhodococcus sp. strain YYL to bioaugment an activated sludge system treating THF wastewater was explored. As indicated by a DGGE analysis, strain YYL alone could not dominate the system, with the concentration of mixed liquor suspended solids (MLSS) decreasing to nearly half of the initial concentration after 45 d, and the microbial diversity was found to be significantly reduced. However, after the reactor was augmented with the mixed culture of strain YYL and two bacilli initially coexisting in the enriched isolation source, strain YYL quickly became dominant in the system and was incorporated into the activated sludge. The concentration of MLSS increased from 2.1 g/L to 7.3 g/L in 20 d, and the efficiency of THF removal from the system was remarkably improved. After the successful bioaugmentation, more than 95% of THF was completely removed from the wastewater when 20 mM THF was continuously loaded into the system. In conclusion, our research first demonstrates that bioaugmentation of activated sludge system for THF degradation is feasible but that successful bioaugmentation should utilize a THF-degrading mixed culture as the inoculum, in which the two bacilli might help strain YYL colonize in activated sludge by co-aggregation

Biotechnological methods for chalcone reduction using whole cells of Lactobacillus, Rhodococcus and Rhodotorula strains as a way to produce new derivatives.

Science.gov (United States)

Stompor, Monika; Kałużny, Mateusz; Żarowska, Barbara

2016-10-01

Microbial strains of the genera Dietzia, Micrococcus, Pseudomonas, Rhodococcus, Gordonia, Streptomyces, Pseudomonas, Bacillus, Penicillium, Rhodotorula and Lactobacillus were screened for the ability to convert chalcones. Synthesis of chalcones was performed by the Claisen-Schmidt reaction. There were three groups of chalcones obtained as the products, which included the derivatives containing 4-substituted chalcone, 2'-hydroxychalcone and 4'-methoxychalcone. The B ring of the chalcones was substituted in the para position with different groups, such as halide, hydroxyl, nitro, methyl, ethyl and ethoxy one. The structure-activity relationship of the tested chalcones in biotransformation processes was studied. It has been proven that Gram-positive bacterial strains Rhodococcus and Lactobacillus catalyzed reduction of C=C bond in the chalcones to give respective dihydrochalcones. The strain Rhodotorula rubra AM 82 transformed chalcones into dihydrochalcones and respective secondary alcohols. These results suggest that the probiotic strain of Lactobacillus can be used for biotransformations of chalcones, which has not been described before. The structure of new metabolites 14a and 15b were established as 4-ethoxy-4'-methoxydihydrochalcone and 3-(4-bromophenyl)-1-(4'-O-methylphenyl)-2-propan-1-ol, respectively, which was confirmed by (1)H NMR and (13)C NMR analysis.

Characterization of Virulence Plasmids and Serotyping of Rhodococcus equi Isolates from Submaxillary Lymph Nodes of Pigs in Hungary

OpenAIRE

Makrai, László; Takayama, Saki; Dénes, Béla; Hajtós, István; Sasaki, Yukako; Kakuda, Tsutomu; Tsubaki, Shiro; Major, Andrea; Fodor, László; Varga, János; Takai, Shinji

2005-01-01

The plasmid types and serotypes of 164 Rhodococcus equi strains obtained from submaxillary lymph nodes of swine from different piggeries in 28 villages and towns located throughout the country were examined. The strains were tested by PCR for the presence of 15- to 17-kDa virulence-associated protein antigen (VapA) and 20-kDa virulence-associated protein antigen (VapB) genes. Plasmid DNAs were isolated and analyzed by digestion with restriction endonucleases to estimate size and compare their...

Intracellular pH and 42.00 C heat response of CHO cells cultured at pH 6.6

International Nuclear Information System (INIS)

Cook, J.A.; Fox, M.H.

1987-01-01

The authors previously reported that cells under chronic low pH (6.6) conditions have altered thermotolerance. They further characterized both the doubling time (t/sub d/) and the internal pH (pH/sub 1/) of CHO cells continuously cultured at pH 6.6 for times greater than one year. The following differences were noted: 1) A t/sub d/ of 16 hr compared to a t/sub d/ of 12 hr for cells at normal pH (7.3) and a t/sub d/ of 25 hr for the acute low pH cells (pH = 6.6; incubation time = 4 hr). 2) A pH/sub i/ 0.1-0.15 pH units > normal cells and 0.3 pH units > acute low pH cells. 3) Survival at 42.0 0 C which differed from both normal and acute low pH cells. The chronic culture was still quite sensitive to 42.0 0 C treatments during the first 5 hr, but developed tolerance at a higher level than cells under acute low pH conditions. The pH/sub i/ of the chronic culture responded to 42.0 0 C heating in a manner similar to that for acute low pH cells. Whether this culture represents a normal response to long term low pH exposure, or was the response of a mutant population is at the present unknown

In vitro and intra-macrophage gene expression by Rhodococcus equi strain 103.

Science.gov (United States)

Rahman, Md Tanvir; Parreira, Valeria; Prescott, John F

2005-09-30

Rhodococcus equi is a facultative intracellular respiratory pathogen of foals that persists and multiplies within macrophages. In foals, virulence is associated with 80-90 kb plasmids, which include a pathogenicity island (PI) containing the virulence-associated protein (vap) gene family, but detailed understanding of the basis of virulence is still poor. A 60 spot-based DNA microarray was developed containing eight PI genes and 42 chromosomal putative virulence or virulence-associated genes selected from a recent partial genome sequence in order to study transcription of these genes by R. equi grown inside macrophages and under in vitro conditions thought to simulate those of macrophages. In addition to seven PI genes, nine chromosomal genes involved in fatty acid and lipid metabolism (choD, fadD13, fbpB), heme biosynthesis (hemE), iron utilization (mbtF), heat shock resistance and genes encoding chaperones (clpB, groEL), a sigma factor (sigK), and a transcriptional regulator (moxR) were significantly induced in R. equi growing inside macrophages. The pattern of R. equi chromosomal genes significantly transcribed inside macrophages largely differed from those transcribed under in vitro conditions (37 degrees C, pH 5.0 or 50mM H2O2 for 30 min). This study has identified genes, other than those of the virulence plasmid, the transcription of which is enhanced within equine macrophages. These genes should be investigated further to improve understanding of how this organism survives intracellularly.

Biodegradation waste of the stations service by Rhodococcus erythropolis ohp-al-gp

International Nuclear Information System (INIS)

Pucci, Oscar Hector; Acuna, Adrian Javier; Pucci, Graciela Natalia

2013-01-01

The strain Rhodococcus erythropolis ohp-al-gp was isolated from turbine oil contaminated soil from northern Santa Cruz province, Argentina. Because of its potential in bioremediation, the aim was to know the abilities for degradation of pure compounds and mixtures of hydrocarbons, as well as degradation in the presence and absence of diesel nitrogen measured by gas chromatography. The strain possesses the ability to use diesel, kerosene, lubricating oil, pristane, hexane, heptane, octane, pentadecane and hexadecane. R. erythropolis ohp-al-gp has excellent potential for bioremediation of hydrocarbons, which are conflictive as lubricating oils, their potential use in removing mud from washing engines or gas stations would be its most important application. The degradation rate in optimal culture conditions, gives it an additional advantage. It also has a low degradation in the absence of nitrogen, a frequent limiting factor in Patagonian soils.

Seroepidemiological survey of Rhodococcus equi infection in asymptomatic horses and donkeys from southeast Turkey.

Science.gov (United States)

Tel, O Y; Arserim, N B; Keskin, O

2011-12-01

In order to assess the level of Rhodococcus equi infection in southeast Turkey, 679 sera from healthy foals and adult horses and 78 sera from donkeys were tested by indirect ELISA using a R. equi reference strain (ATCC 33701) as antigen. Eighty (11.7%) sera from horses and 9 (11.5%) sera from donkeys with titres >0.85 were positive. The prevalence of seropositive horses in Sanliurfa Province was higher than in Diyarbakir Province; 56 (13.9%) horses in Sanliurfa Province and 24 (8.7%) horses in Diyarbakir Province were defined as seropositive. In Sanliurfa Province 14.5% of female (n=343) and 10.1% of male (n = 59) horses tested were defined as seropositive, while in Diyarbakir Province more males (11.4%, n=114) were seropositive than females (6.7%, n=163). Horses 1 to 5 years of age were found to have the highest seropositivity rate in both provinces. A total of 78 sera from donkeys were investigated in Sanliurfa Province, of which 9 (11.5%) were positive by ELISA. Among the 9 positive sera, 6 (12.8%) were from donkeys 1-5 years old and 3 (13.6%) were from donkeys >5 years of age. No positive sera were found in donkeys less than 1 year old. Five (12.5%) sera of females and 4 (10.5%) sera of males tested were positive. These results indicate the existence of R. equi in the horse populations in Sanliurfa and Diyarbakir Provinces. Similar infection rates were found for donkeys in Sanliurfa. This suggests the importance of serological surveys to diagnose R. equi infection in the region and to prevent the zoonotic risk.

Seroepidemiological survey of Rhodococcus equi infection in asymptomatic horses and donkeys from southeast Turkey

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O. Y. Tel

2011-05-01

Full Text Available n order to assess the level of Rhodococcus equi infection in southeast Turkey, 679 sera from healthy foals and adult horses and 78 sera from donkeys were tested by indirect ELISA using a R. equi reference strain (ATCC 33701 as antigen. Eighty (11.7 % sera from horses and 9 (11.5 % sera from donkeys with titres >0.85 were positive. The prevalence of seropositive horses in Sanliurfa Province was higher than in Diyarbakir Province; 56 (13.9 % horses in Sanliurfa Province and 24 (8.7 % horses in Diyarbakir Province were defined as seropositive. In Sanliurfa Province 14.5 % of female (n = 343 and 10.1 % of male (n = 59 horses tested were defined as seropositive, while in Diyarbakir Province more males (11.4 %, n = 114 were seropositive than females (6.7 %, n = 163. Horses 1 to 5 years of age were found to have the highest seropositivity rate in both provinces. A total of 78 sera from donkeys were investigated in Sanliurfa Province, of which 9 (11.5 % were positive by ELISA. Among the 9 positive sera, 6 (12.8 % were from donkeys 1-5 years old and 3 (13.6 % were from donkeys >5years of age. Nopositive sera were found in donkeys less than 1 year old. Five(12.5 % sera of females and 4(10.5 % sera of males tested were positive. These results indicate the existence of R. equi in the horse populations in Sanliurfa and Diyarbakir Provinces. Similar infection rates were found for donkeys in Sanliurfa. This suggests the importance of serological surveys to diagnose R. equi infection in the region and to prevent the zoonotic risk.

A preliminary report on the contact-independent antagonism of Pseudogymnoascus destructans by Rhodococcus rhodochrous strain DAP96253.

Science.gov (United States)

Cornelison, Christopher T; Keel, M Kevin; Gabriel, Kyle T; Barlament, Courtney K; Tucker, Trudy A; Pierce, George E; Crow, Sidney A

2014-09-26

The recently-identified causative agent of White-Nose Syndrome (WNS), Pseudogymnoascus destructans, has been responsible for the mortality of an estimated 5.5 million North American bats since its emergence in 2006. A primary focus of the National Response Plan, established by multiple state, federal and tribal agencies in 2011, was the identification of biological control options for WNS. In an effort to identify potential biological control options for WNS, multiply induced cells of Rhodococcus rhodochrous strain DAP96253 was screened for anti-P. destructans activity. Conidia and mycelial plugs of P. destructans were exposed to induced R. rhodochrous in a closed air-space at 15°C, 7°C and 4°C and were evaluated for contact-independent inhibition of conidia germination and mycelial extension with positive results. Additionally, in situ application methods for induced R. rhodochrous, such as fixed-cell catalyst and fermentation cell-paste in non-growth conditions, were screened with positive results. R. rhodochrous was assayed for ex vivo activity via exposure to bat tissue explants inoculated with P. destructans conidia. Induced R. rhodochrous completely inhibited growth from conidia at 15°C and had a strong fungistatic effect at 4°C. Induced R. rhodochrous inhibited P. destructans growth from conidia when cultured in a shared air-space with bat tissue explants inoculated with P. destructans conidia. The identification of inducible biological agents with contact-independent anti- P. destructans activity is a major milestone in the development of viable biological control options for in situ application and provides the first example of contact-independent antagonism of this devastating wildlife pathogen.

Engineering and improvement of the efficiency of a chimeric [P450cam-RhFRed reductase domain] enzyme.

Science.gov (United States)

Robin, Aélig; Roberts, Gareth A; Kisch, Johannes; Sabbadin, Federico; Grogan, Gideon; Bruce, Neil; Turner, Nicholas J; Flitsch, Sabine L

2009-05-14

A chimeric oxygenase, in which the P450cam domain was fused to the reductase host domains of a P450RhF from Rhodococcus sp. strain NCIMB 9784 was optimised to allow for a biotransformation at 30 mM substrate in 80% overall yield, with the linker region between P450 and FMN domain proving to be important for the effective biotransformation of (+)-camphor to 5-exo-hydroxycamphor.

Growth Kinetics of Rhodococcus Erythropolis IGTS8 on Thiophene and Dimethylsulphoxide

International Nuclear Information System (INIS)

El-Temtamy, S.A.; Farahat, L.A.; Al-Shatnawi, D.F.; AI-Sayed, S.

2004-01-01

Sulfur oxides emissions through fossil fuel combustion posse environmental problems because these oxides are major cause of acid rain, which dissolve buildings, kills forests and poisons lakes. That is why world environmental regulations are becoming harder with respect to sulfur content in fossil fuels. The demand for new low-cost desulfurization technologies has led to renewed interest in bio desulfurization. In this work the growth kinetics of Rhodococcus Erythopolis IGTS8 have been studied on two model sulfur compounds that are found among other sulfur compounds in petroleum fractions. These are namely thiophene and dimethyl sulfoxide, DEMSO. Batch reactor and different substrate concentrations ranging between 1 and 8 gl -1 were used to study the growth kinetics. Growth on thiophene was shown to follow Monode kinetics with μ m ax =0.064 h -1 and K s = 2.8 gl -1 , on the other hand, growth on DEMSO was found to be substrate inhibited

Development of Chemical and Metabolite Sensors for Rhodococcus opacus PD630.

Science.gov (United States)

DeLorenzo, Drew M; Henson, William R; Moon, Tae Seok

2017-10-20

Rhodococcus opacus PD630 is a nonmodel, Gram-positive bacterium that possesses desirable traits for biomass conversion, including consumption capabilities for lignocellulose-based sugars and toxic lignin-derived aromatic compounds, significant triacylglycerol accumulation, relatively rapid growth rate, and genetic tractability. However, few genetic elements have been directly characterized in R. opacus, limiting its application for lignocellulose bioconversion. Here, we report the characterization and development of genetic tools for tunable gene expression in R. opacus, including: (1) six fluorescent reporters for quantifying promoter output, (2) three chemically inducible promoters for variable gene expression, and (3) two classes of metabolite sensors derived from native R. opacus promoters that detect nitrogen levels or aromatic compounds. Using these tools, we also provide insights into native aromatic consumption pathways in R. opacus. Overall, this work expands the ability to control and characterize gene expression in R. opacus for future lignocellulose-based fuel and chemical production.

In vitro effect of josamycin in strains of Rhodococcus equi isolated from pulmonar infections in foals / Efeito in vitro da josamicina em cepas de rhodocaccus equi isoladas de afecções pulmonares em potros

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Jaime Galvão Dias Júnior

2000-12-01

Full Text Available Antmicrobial susceptibility test was performed in thirty-one samples of Rhodococcus equi isolated from iung infections in foals. Among the antimicrobial tosted, erythromycin (100,0 %, rifampin (96.3%. neomycin (93,6 %Josamycin (90,4% and gentamicin (90,4 %presented the highest sensitivity against R- equi- The most-common occurrence of resistance was observed from cephalexim (100,0 %}, lincomycin (100,0 %, cephalothin (96,3 %. oxacillin (96,8 %, penicillin G (96,8 %, amoxicillin (90,3 % and trimethoprim/sulfamethoxazole (83,8 %. The highest susceptibility of the R. equi to josamycin, suggest the drug as alternative for therapy of R. equi infections in foals.Procedeu-se o teste de sensibilidade microbiana in vitro pelo teste de difusão com discos em 31 cepas de Rhodococcus equi. isoladas de afecções pulmonares cm potros. frente a 20 antimicrobianos. Os maioires índices de sensibilidade de R. equi foram constatados para entromicina (100,0 %, níampicina (96.8 %}t ncomicina (93. 6 %. josamicina (90.4% e gentamicina (90A %. Os maiores índices de resistência do agente foram verificados para cefalexina (100,0%, lincomicina (100.0%, cefalotina (96.8 %, oxacilino (96,8 %}, penicilina G (96,3 %, amoxicilina (90.3 % e sulfametoxazol/trimetoprim (83,8 %. A alta sensibilidade das cepas de R. equi para a josamicina, sugere a possibilidade de utilização da droga como alternativa no tratamento da rodococose em potros.

Observation of the strongest 5s2 5p6 5d-(5s2 5p5 5d6s+5s25p6 7p) transitions in Au XI to Bi XV ions

International Nuclear Information System (INIS)

Churilov, S.S.; Joshi, Y.N.

2001-01-01

The spectra of gold till bismuth were studied in the 90-135 A region. Nine most intense lines belonging to the 5s 2 5p 6 5d-5s 2 5p 5 5d6s array were identified in Au XI to Bi XV ions. The 5s 2 5p 6 7p 2 P 3/2,1/2 levels in Au XI and the 5s 2 5p 6 7p 2 P 3/2 level in Hg XII were also identified. The observed wavelengths and intensities agree quite well with the Hartree-Fock calculations. (orig.)

Styrene Oxide Isomerase of Rhodococcus opacus 1CP, a Highly Stable and Considerably Active Enzyme

Science.gov (United States)

Gröning, Janosch A. D.; Tischler, Dirk; Kaschabek, Stefan R.; Schlömann, Michael

2012-01-01

Styrene oxide isomerase (SOI) is involved in peripheral styrene catabolism of bacteria and converts styrene oxide to phenylacetaldehyde. Here, we report on the identification, enrichment, and biochemical characterization of a novel representative from the actinobacterium Rhodococcus opacus 1CP. The enzyme, which is strongly induced during growth on styrene, was shown to be membrane integrated, and a convenient procedure was developed to highly enrich the protein in active form from the wild-type host. A specific activity of about 370 U mg−1 represents the highest activity reported for this enzyme class so far. This, in combination with a wide pH and temperature tolerance, the independence from cofactors, and the ability to convert a spectrum of substituted styrene oxides, makes a biocatalytic application imaginable. First, semipreparative conversions were performed from which up to 760 μmol of the pure phenylacetaldehyde could be obtained from 130 U of enriched SOI. Product concentrations of up to 76 mM were achieved. However, due to the high chemical reactivity of the aldehyde function, SOI was shown to be the subject of an irreversible product inhibition. A half-life of 15 min was determined at a phenylacetaldehyde concentration of about 55 mM, indicating substantial limitations of applicability and the need to modify the process. PMID:22504818

Spectral analysis of 5s25p2(6p+6d+7s) configurations of Ba VI

International Nuclear Information System (INIS)

Sharma, M.K.; Tauheed, A.; Rahimullah, K.

2014-01-01

The sixth spectrum of barium (Ba VI) has been investigated with the aid of experimental recordings made on a 3-m normal incidence vacuum spectrograph of Antigonish laboratory (Canada) in the wavelength region 300–2080 Å using triggered spark as an excitation source. The spectral analysis has been extended considerably to include new configuration the 5s 2 5p 2 6p in odd parity matrix and the 5s 2 5p 2 6d and 5s 2 5p 2 7s configurations in even parity matrix. Previously reported levels of the ground configuration (5s 2 5p 3 ) and three lowest excited configurations the 5s5p 4 , 5s 2 5p 2 5d and 5s 2 5p 2 6s have been confirmed and the two unknown levels of the 5s 2 5p 2 5d configuration with J=9/2, have now been established through the identification of transitions from the 5s 2 5p 2 6p levels. All twenty one levels of the 5s 2 5p 2 6p configuration and twenty nine levels out of thirty six of the 5s 2 5p 2 6d and 5s 2 5p 2 7s configurations have now been established. Hartree–Fock calculations involving configuration interactions support the analyses. The accuracy of our wavelength measurement is ±0.005 Å for sharp lines. - Highlights: • The spectrum of Ba was recorded on a 3-m spectrograph with triggered spark source. • Atomic transitions for Ba VI were identified to established new energy levels. • CI calculations with relativistic corrections were made for theoretical predictions. • Weighted oscillator strength (gf) and transition probabilities (gA) were calculated

Crystallization and crystallographic analysis of the Rhodococcus rhodochrous NCIMB 13064 DhaA mutant DhaA31 and its complex with 1,2,3-trichloropropane

Czech Academy of Sciences Publication Activity Database

Lahoda, M.; Chaloupková, R.; Stsiapanava, A.; Damborský, J.; Kutá-Smatanová, Ivana

2011-01-01

Roč. 67, Part 3 (2011), s. 397-400 ISSN 1744-3091 R&D Projects: GA MŠk(CZ) LC06010; GA MŠk(CZ) ED1.1.00/02.0073 Institutional research plan: CEZ:AV0Z60870520 Keywords : haloalkane dehalogenases * DhaA * Rhodococcus rhodochrous Subject RIV: CE - Biochemistry Impact factor: 0.506, year: 2011

Isolation and characterization of styrene metabolism genes from styrene-assimilating soil bacteria Rhodococcus sp. ST-5 and ST-10.

Science.gov (United States)

Toda, Hiroshi; Itoh, Nobuya

2012-01-01

Styrene metabolism genes were isolated from styrene-assimilating bacteria Rhodococcus sp. ST-5 and ST-10. Strain ST-5 had a gene cluster containing four open reading frames which encoded styrene degradation enzymes. The genes showed high similarity to styABCD of Pseudomonas sp. Y2. On the other hand, strain ST-10 had only two genes which encoded styrene monooxygenase and flavin oxidoreductase (styAB). Escherichia coli transformants possessing the sty genes of strains ST-5 and ST-10 produced (S)-styrene oxide from styrene, indicating that these genes function as styrene degradation enzymes. Metabolite analysis by resting-cell reaction with gas chromatography-mass spectrometry revealed that strain ST-5 converts styrene to phenylacetaldehyde via styrene oxide by styrene oxide isomerase (styC) reaction. On the other hand, strain ST-10 lacked this enzyme, and thus accumulated styrene oxide as an intermediate. HPLC analysis showed that styrene oxide was spontaneously isomerized to phenylacetaldehyde by chemical reaction. The produced phenylacetaldehyde was converted to phenylacetic acid (PAA) in strain ST-10 as well as in strain ST-5. Furthermore, phenylacetic acid was converted to phenylacetyl-CoA by the catalysis of phenylacetate-CoA ligase in strains ST-5 and ST-10. This study proposes possible styrene metabolism pathways in Rhodococcus sp. strains ST-5 and ST-10. Copyright © 2011 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Synthesis and biological activity of M6-P and M6-P analogs on fibroblast and keratinocyte proliferation.

Science.gov (United States)

Clavel, Caroline; Barragan-Montero, Véronique; Garric, Xavier; Molès, Jean-Pierre; Montero, Jean-Louis

2005-09-01

A new synthetic route to obtain the carboxylate analog of mannose 6-phosphate (M6-P) is presented. The effects of the M6-P, the carboxylate and two other analogs (the phosphonate and the alpha,beta ethylenic carboxylate) on the proliferation of human keratinocytes and dermal fibroblasts as well as on the proliferation of a murine fibroblast cell line, 3T3-J2 are tested. We observed that M6-P is a potent inhibitor of proliferation of both fibroblasts and keratinocytes. Among its analogs, the phosphonate showed a similar effect on human dermal fibroblasts but not on keratinocytes.

Aerobic degradation of 4-nitroaniline (4-NA) via novel degradation intermediates by Rhodococcus sp. strain FK48

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Khan, Fazlurrahman; Pandey, Janmejay; Vikram, Surendra; Pal, Deepika; Cameotra, Swaranjit Singh, E-mail: ssc@imtech.res.in

2013-06-15

Highlights: • This study reports isolation of a novel bacterium capable of mineralizing 4-nitroaniline (4-NA). • This bacterium has been identified as Rhodococcus sp. strain FK48. • Strain FK48 degrades 4-NA via a novel aerobic degradation pathway that involves 4-AP and 1,2,4-BT. • Subsequent degradation proceeds via ring fission and formation of maleylacetate. • This is the first report showing elucidation of catabolic pathway for microbial degradation 4-NA. -- Abstract: An aerobic strain, Rhodococcus sp. strain FK48, capable of growing on 4-nitroaniline (4-NA) as the sole source of carbon, nitrogen, and energy has been isolated from enrichment cultures originating from contaminated soil samples. During growth studies with non- induced cells of FK48 catalyzed sequential denitrification (release of NO{sub 2} substituent) and deamination (release of NH{sub 2} substituent) of 4-NA. However, none of the degradation intermediates could be identified with growth studies. During resting cell studies, 4-NA-induced cells of strain FK48 transformed 4-NA via a previously unknown pathway which involved oxidative hydroxylation leading to formation of 4-aminophenol (4-AP). Subsequent degradation involved oxidated deamination of 4-AP and formation of 1,2,4-benzenetriol (BT) as the major identified terminal aromatic intermediate. Identification of these intermediates was ascertained by HPLC, and GC–MS analyses of the culture supernatants. 4-NA-induced cells of strain FK48 showed positive activity for 1,2,4-benzenetriol dioxygenase in spectrophotometric assay. This is the first conclusive study on aerobic microbial degradation of 4-NA and elucidation of corresponding metabolic pathway.

Aerobic degradation of 4-nitroaniline (4-NA) via novel degradation intermediates by Rhodococcus sp. strain FK48

International Nuclear Information System (INIS)

Khan, Fazlurrahman; Pandey, Janmejay; Vikram, Surendra; Pal, Deepika; Cameotra, Swaranjit Singh

2013-01-01

Highlights: • This study reports isolation of a novel bacterium capable of mineralizing 4-nitroaniline (4-NA). • This bacterium has been identified as Rhodococcus sp. strain FK48. • Strain FK48 degrades 4-NA via a novel aerobic degradation pathway that involves 4-AP and 1,2,4-BT. • Subsequent degradation proceeds via ring fission and formation of maleylacetate. • This is the first report showing elucidation of catabolic pathway for microbial degradation 4-NA. -- Abstract: An aerobic strain, Rhodococcus sp. strain FK48, capable of growing on 4-nitroaniline (4-NA) as the sole source of carbon, nitrogen, and energy has been isolated from enrichment cultures originating from contaminated soil samples. During growth studies with non- induced cells of FK48 catalyzed sequential denitrification (release of NO 2 substituent) and deamination (release of NH 2 substituent) of 4-NA. However, none of the degradation intermediates could be identified with growth studies. During resting cell studies, 4-NA-induced cells of strain FK48 transformed 4-NA via a previously unknown pathway which involved oxidative hydroxylation leading to formation of 4-aminophenol (4-AP). Subsequent degradation involved oxidated deamination of 4-AP and formation of 1,2,4-benzenetriol (BT) as the major identified terminal aromatic intermediate. Identification of these intermediates was ascertained by HPLC, and GC–MS analyses of the culture supernatants. 4-NA-induced cells of strain FK48 showed positive activity for 1,2,4-benzenetriol dioxygenase in spectrophotometric assay. This is the first conclusive study on aerobic microbial degradation of 4-NA and elucidation of corresponding metabolic pathway

Multiplicity of 3-Ketosteroid-9 alpha-Hydroxylase Enzymes in Rhodococcus rhodochrous DSM43269 for Specific Degradation of Different Classes of Steroids

OpenAIRE

Petrusma, Mirjan; Hessels, Gerda; Dijkhuizen, Lubbert; van der Geize, Robert

2011-01-01

The well-known large catabolic potential of rhodococci is greatly facilitated by an impressive gene multiplicity. This study reports on the multiplicity of kshA, encoding the oxygenase component of 3-ketosteroid 9 alpha-hydroxylase, a key enzyme in steroid catabolism. Five kshA homologues (kshA1 to kshA5) were previously identified in Rhodococcus rhodochrous DSM43269. These KshA(DSM43269) homologues are distributed over several phylogenetic groups. The involvement of these KshA homologues in ...

Cloning, overexpression, purification, crystallization and preliminary X-ray analysis of 3-ketosteroid Δ4-(5α)-dehydrogenase from Rhodococcus jostii RHA1

International Nuclear Information System (INIS)

Oosterwijk, Niels van; Knol, Jan; Dijkhuizen, Lubbert; Geize, Robert van der; Dijkstra, Bauke W.

2011-01-01

The gene for 3-ketosteroid Δ 4 -(5α)-dehydrogenase from R. jostii RHA1 was cloned and overexpressed in E. coli and the protein product was purified and crystallized using the hanging-drop vapour-diffusion method. The crystals belonged to space group C222 1 and diffraction data were collected to a resolution of 1.6 Å. 3-Ketosteroid dehydrogenases are flavoproteins which play key roles in steroid ring degradation. The enzymes are abundantly present in actinobacteria, including the catabolic powerhouse Rhodococcus jostii and the pathogenic species R. equi and Mycobacterium tuberculosis. The gene for 3-ketosteroid Δ 4 -(5α)-dehydrogenase [Δ 4 -(5α)-KSTD] from R. jostii RHA1 was cloned and overexpressed in Escherichia coli. His-tagged Δ 4 -(5α)-KSTD enzyme was purified by Ni 2+ –NTA affinity chromatography, anion-exchange chromatography and size-exclusion chromatography and was crystallized using the hanging-drop vapour-diffusion method. Seeding greatly improved the number of crystals obtained. The crystals belonged to space group C222 1 , with unit-cell parameters a = 99.2, b = 114.3, c = 110.2 Å. Data were collected to a resolution of 1.6 Å

Enhanced biodegradation of diesel oil by a newly identified Rhodococcus baikonurensis EN3 in the presence of mycolic acid.

Science.gov (United States)

Lee, M; Kim, M K; Singleton, I; Goodfellow, M; Lee, S-T

2006-02-01

The aim of the present study was to isolate and characterize a bacterium, strain EN3, capable of using diesel oil as a major carbon and energy source, and to analyse the enhancement of diesel oil degradation by this organism using synthetic mycolic acid (2-hexyl-3-hydroxyldecanoic acid). An actinomycete with the ability to degrade diesel oil was isolated from oil contaminated soil and characterized. The strain had phenotypic properties consistent with its classification in the genus Rhodococcus showing a 16S rRNA gene similarity of 99.7% with Rhodococcus baikonurensis DSM 44587(T). The ability of the characterized strain to degrade diesel oil at various concentrations (1000, 5000, 10 000 and 20 000 mg l(-1)) was determined. The effect of synthetic mycolic acid on the biodegradation of diesel oil was investigated at the 20 000 mg l(-1) concentration; the surfactant was added to the flask cultures at three different concentrations (10, 50 and 100 mg l(-1)) and degradation followed over 7 days. Enhanced degradation was found at all three concentrations of the surfactant. In addition, the enhancement of diesel oil degradation by other surfactants was observed. The synthetic mycolic acid has potential for the remediation of petroleum-contaminated sites from both an economic and applied perspective as it can stimulate biodegradation at low concentrations. This study showed that the synthesized mycolic acid can be used for potential applications in the bioremediation industries, for example, in oil spill clean-up, diesel fuel remediation and biostimulation.

Functional characterization and stability improvement of a ‘thermophilic-like’ ene-reductase from Rhodococcus opacus 1CP

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Anika eRiedel

2015-10-01

Full Text Available Ene-reductases are widely applied for the asymmetric synthesis of relevant industrial chemicals. A novel ene-reductase OYERo2 was found within a set of 14 putative Old Yellow Enzymes (OYEs obtained by genome mining of the actinobacterium Rhodococcus opacus 1CP. Multiple sequence alignment suggested that the enzyme belongs to the group of ‘thermophilic-like’ OYEs. OYERo2 was produced in Escherichia coli and biochemically characterized. The enzyme is strongly NADPH dependent and uses non-covalently bound FMNH2 for the reduction of activated α,β-unsaturated alkenes. In the active form OYERo2 is a dimer. Optimal catalysis occurs at pH 7.3 and 37 °C. OYERo2 showed highest specific activities (4550 U mg-1 on maleimides, which are efficiently converted to the corresponding succinimides. The OYERo2-mediated reduction of prochiral alkenes afforded the (R-products with excellent optical purity (ee > 99%. OYERo2 is not as thermo-resistant as related OYEs. Introduction of a characteristic intermolecular salt bridge by site-specific mutagenesis raised the half-life of enzyme inactivation at 32 °C from 28 min to 87 min and improved the tolerance towards organic co-solvents. The suitability of OYERo2 for application in industrial biocatalysis is discussed.

EVALUACIÓN DE LA BIOTRANSFORMACIÓN DE GERANIOL Y (R-(+-α-PINENO EMPLEANDO CÉLULAS DE Rhodococcus opacus DSM 44313 EVALUATION OF THE BIOTRANSFORMATION OF GERANIOL AND (R-(+-α-PINENE USING CELL OF Rhodococcus opacus DSM 44313

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JENNIFER PILAR ROJAS

2009-12-01

Full Text Available La alta biodisponibilidad de los monoterpenos los hace precursores promisorios en los procesos de biotransformación, mediante los que se producen compuestos de valor agregado que pueden considerarse naturales, debido a que se obtienen por métodos enzimáticos a partir de precursores aislados de la naturaleza. En el presente estudio se evaluó la biotransformación de geraniol y (R-(+-α-pineno empleando la cepa bacteriana Rhodococcus opacus DSM 44313, para ello se determinó la influencia del tiempo de crecimiento de la bacteria, tomando suspensiones celulares en la mitad y finalizando la fase exponencial. También se evaluaron tres tiempos de reacción (12, 24 y 48 h y el efecto de un cosolvente, agregando los sustratos puros y disueltos en etanol al 10%. A partir del geraniol se produjeron geranial, ácido geránico y 6-metil-5-hepten-2-ona, sólo cuando el sustrato se adicionó puro, y se formaron en mayor concentración cuando la bacteria estaba finalizando su fase de crecimiento exponencial. Con el (R-(+-α-pineno se produjo como compuesto principal el (R-(+-cis-verbenol, siendo mayor su concentración agregando el pineno disuelto en etanol, y cuando la bacteria se encontraba en la mitad de su fase de crecimiento exponencial. Los resultados indican que el comportamiento de la bacteria cambia según el sustrato adicionado, debido a las propiedades de cada monoterpeno, pero los compuestos obtenidos con ambos sustratos tienen importantes aplicaciones en las industrias farmacéutica, alimenticia y de perfumeríaThe high bioavailability of monoterpenes make them promising precursors in biotransformation processes, through which produce value-added compounds that can be considered natural because they are obtained by enzymatic methods from precursors isolated from nature. In the present study we evaluated the biotransformation of geraniol and (R-(+-α-pinene using a bacterial strain of Rhodococcus opacus DSM 44313; the influence of growth time

Characterization of biosurfactants produced by the oil-degrading bacterium Rhodococcus erythropolis S67 at low temperature.

Science.gov (United States)

Luong, T M; Ponamoreva, O N; Nechaeva, I A; Petrikov, K V; Delegan, Ya A; Surin, A K; Linklater, D; Filonov, A E

2018-01-04

Production of trehalolipid biosurfactants by Rhodococcus erythropolis S67 depending on the growth temperature was studied. R. erythropolis S67 produced glycolipid biosurfactants such as 2,3,4-succinoyl-octanoyl-decanoyl-2'-decanoyl trehalose and 2,3,4-succinoyl-dioctanoyl-2'-decanoyl trehalose during the growth in n-hexadecane medium at 26 and 10 °C, despite the different aggregate state of the hydrophobic substrate at low temperature. The surface tension of culture medium was found being reduced from 72 to 27 and 45 mN m -1 , respectively. Production of trehalolipid biosurfactants by R. erythropolis S67 at low temperature could be useful for the biodegradation of petroleum hydrocarbons at low temperatures by enhancing the bioremediation performance in cold regions.

Rhodococcus erythropolis DCL14 Contains a Novel Degradation Pathway for Limonene

Science.gov (United States)

van der Werf, Mariët J.; Swarts, Henk J.; de Bont, Jan A. M.

1999-01-01

Strain DCL14, which is able to grow on limonene as a sole source of carbon and energy, was isolated from a freshwater sediment sample. This organism was identified as a strain of Rhodococcus erythropolis by chemotaxonomic and genetic studies. R. erythropolis DCL14 also assimilated the terpenes limonene-1,2-epoxide, limonene-1,2-diol, carveol, carvone, and (−)-menthol, while perillyl alcohol was not utilized as a carbon and energy source. Induction tests with cells grown on limonene revealed that the oxygen consumption rates with limonene-1,2-epoxide, limonene-1,2-diol, 1-hydroxy-2-oxolimonene, and carveol were high. Limonene-induced cells of R. erythropolis DCL14 contained the following four novel enzymatic activities involved in the limonene degradation pathway of this microorganism: a flavin adenine dinucleotide- and NADH-dependent limonene 1,2-monooxygenase activity, a cofactor-independent limonene-1,2-epoxide hydrolase activity, a dichlorophenolindophenol-dependent limonene-1,2-diol dehydrogenase activity, and an NADPH-dependent 1-hydroxy-2-oxolimonene 1,2-monooxygenase activity. Product accumulation studies showed that (1S,2S,4R)-limonene-1,2-diol, (1S,4R)-1-hydroxy-2-oxolimonene, and (3R)-3-isopropenyl-6-oxoheptanoate were intermediates in the (4R)-limonene degradation pathway. The opposite enantiomers [(1R,2R,4S)-limonene-1,2-diol, (1R,4S)-1-hydroxy-2-oxolimonene, and (3S)-3-isopropenyl-6-oxoheptanoate] were found in the (4S)-limonene degradation pathway, while accumulation of (1R,2S,4S)-limonene-1,2-diol from (4S)-limonene was also observed. These results show that R. erythropolis DCL14 metabolizes both enantiomers of limonene via a novel degradation pathway that starts with epoxidation at the 1,2 double bond forming limonene-1,2-epoxide. This epoxide is subsequently converted to limonene-1,2-diol, 1-hydroxy-2-oxolimonene, and 7-hydroxy-4-isopropenyl-7-methyl-2-oxo-oxepanone. This lactone spontaneously rearranges to form 3-isopropenyl-6-oxoheptanoate. In

Immunogenicity of an electron beam inactivated Rhodococcus equi vaccine in neonatal foals.

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Angela I Bordin

Full Text Available Rhodococcus equi is an important pathogen of foals that causes severe pneumonia. To date, there is no licensed vaccine effective against R. equi pneumonia of foals. The objectives of our study were to develop an electron beam (eBeam inactivated vaccine against R. equi and evaluate its immunogenicity. A dose of eBeam irradiation that inactivated replication of R. equi while maintaining outer cell wall integrity was identified. Enteral administration of eBeam inactivated R. equi increased interferon-γ production by peripheral blood mononuclear cells in response to stimulation with virulent R. equi and generated naso-pharyngeal R. equi-specific IgA in newborn foals. Our results indicate that eBeam irradiated R. equi administered enterally produce cell-mediated and upper respiratory mucosal immune responses, in the face of passively transferred maternal antibodies, similar to those produced in response to enteral administration of live organisms (a strategy which previously has been documented to protect foals against intrabronchial infection with virulent R. equi. No evidence of adverse effects was noted among vaccinated foals.

Molecular characterization of three 3-ketosteroid-Δ(1)-dehydrogenase isoenzymes of Rhodococcus ruber strain Chol-4.

Science.gov (United States)

Fernández de las Heras, Laura; van der Geize, Robert; Drzyzga, Oliver; Perera, Julián; María Navarro Llorens, Juana

2012-11-01

Rhodococcus ruber strain Chol-4 isolated from a sewage sludge sample is able to grow on minimal medium supplemented with steroids, showing a broad catabolic capacity. This paper reports the characterization of three different 3-ketosteroid-Δ(1)-dehydrogenases (KstDs) in the genome of R. ruber strain Chol-4. The genome of this strain does not contain any homologues of a 3-keto-5α-steroid-Δ(4)-dehydrogenase (Kst4d or TesI) that appears in the genomes of Rhodococcus erythropolis SQ1 or Comamonas testosteroni. Growth experiments with kstD2 mutants, either a kstD2 single mutant, kstD2 double mutants in combination with kstD1 or kstD3, or the triple kstD1,2,3 mutant, proved that KstD2 is involved in the transformation of 4-androstene-3,17-dione (AD) to 1,4-androstadiene-3,17-dione (ADD) and in the conversion of 9α-hydroxy-4-androstene-3,17-dione (9OHAD) to 9α-hydroxy-1,4-androstadiene-3,17-dione (9OHADD). kstD2,3 and kstD1,2,3 R. ruber mutants (both lacking KstD2 and KstD3) did not grow in minimal medium with cholesterol as the only carbon source, thus demonstrating the involvement of KstD2 and KstD3 in cholesterol degradation. In contrast, mutation of kstD1 does not alter the bacterial growth on the steroids tested in this study and therefore, the role of this protein still remains unclear. The absence of a functional KstD2 in R. ruber mutants provoked in all cases an accumulation of 9OHAD, as a branch product probably formed by the action of a 3-ketosteroid-9α-hydroxylase (KshAB) on the AD molecule. Therefore, KstD2 is a key enzyme in the AD catabolism pathway of R. ruber strain Chol-4 while KstD3 is involved in cholesterol catabolism. Copyright © 2012 Elsevier Ltd. All rights reserved.

Biodegradation of benzo[α]pyrene, toluene, and formaldehyde from the gas phase by a consortium of Rhodococcus erythropolis and Fusarium solani.

Science.gov (United States)

Morales, Paulina; Cáceres, Manuel; Scott, Felipe; Díaz-Robles, Luis; Aroca, Germán; Vergara-Fernández, Alberto

2017-09-01

Polycyclic aromatic hydrocarbons (PAHs) and volatile organic compounds (VOCs) are important indoor contaminants. Their hydrophobic nature hinders the possibility of biological abatement using biofiltration. Our aim was to establish whether the use of a consortium of Fusarium solani and Rhodococcus erythropolis shows an improved performance (in terms of mineralization rate and extent) towards the degradation of formaldehyde, as a slightly polar VOC; toluene, as hydrophobic VOC; and benzo[α]pyrene (BaP) as PAH at low concentrations compared to a single-species biofilm in serum bottles with vermiculite as solid support to mimic a biofilter and to relate the possible improvements with the surface hydrophobicity and partition coefficient of the biomass at three different temperatures. Results showed that the hydrophobicity of the surface of the biofilms was affected by the hydrophobicity of the carbon source in F. solani but it did not change in R. erythropolis. Similarly, the partition coefficients of toluene and BaP in F. solani biomass (both as pure culture and consortium) show a reduction of up to 38 times compared to its value in water, whereas this reduction was only 1.5 times in presence of R. erythropolis. Despite that increments in the accumulated CO 2 and its production rate were found when F. solani or the consortium was used, the mineralization extent of toluene was below 25%. Regarding BaP degradation, the higher CO 2 production rates and percent yields were obtained when a consortium of F. solani and R. erythropolis was used, despite a pure culture of R. erythropolis exhibits poor mineralization of BaP.

Diastereoselective reduction of 1-(4-fluorophenyl)-3(R)-[3-oxo-3-(4-fluorophenyl) -propyl]-4 (S)-(4-hydroxyphenyl)azetidin-2-one to Ezetimibe by the whole cell catalyst Rhodococcus fascians MO22

Czech Academy of Sciences Publication Activity Database

Kyslíková, Eva; Babiak, P.; Marešová, Helena; Palyzová, Andrea; Hajíček, J.; Kyslík, Pavel

2010-01-01

Roč. 67, 3-4 (2010), s. 266-270 ISSN 1381-1177 R&D Projects: GA MŠk 2B08064 Institutional research plan: CEZ:AV0Z50200510 Keywords : Biotransformation * Rhodococcus fascians MO22 * Ezetimibe Subject RIV: CE - Biochemistry Impact factor: 2.330, year: 2010

Emulsification of crude oil by an alkane-oxidizing Rhodococcus species isolated from seawater

Energy Technology Data Exchange (ETDEWEB)

Bredholt, H.; Bruheim, P.; Eimhjellen, K. [Norwegian Univ. of Scince and Technology, Trondheim (Norway); Josefsen, K.; Vatland, A. [SINTEF SI, Oslo (Norway). Industrial Chemistry Div.

1998-04-01

A Rhodococcus species, which has proven to be the best of 99 oil-emulsifying bacteria isolated from seawater, was characterized. This bacterium produced very stable oil-in-water emulsions from different crude oils with various content of aliphatic and aromatic compounds, by utilizing C{sub 1}1 and C{sub 3}3 n-alkanes as carbon and energy sources. Bacteria that produce stable emulsions are often able to adhere strongly to hydrocarbons or hydrophobic surfaces. It was at these surfaces that extensive emulsification of the residual oil and accumulation of acidic oxidation products occurred. The acidic products were consumed in a second step. This step was characterized by linear growth and an increasing number of cells growing in the water phase. The most extensive emulsification occurred at the end of the exponential phase. There was no evidence of surfactants at the end of the exponential phase, however, a polymeric compound with emulsifying activity, tightly bound to the oil droplets, was isolated, suggesting that the emulsification resulted from the release of the hydrophobic cell surface discarded during growth limitations. 38 refs., 7 figs.

Management of Rhodococcus equi pneumonia in foals

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Johns I

2013-11-01

Full Text Available Imogen Johns Department of Clinical Sciences and Services, Royal Veterinary College, North Mymms, UK Abstract: Rhodococcus equi, a gram-positive facultative intracellular bacterial pathogen, is the most important cause of pneumonia in foals aged 3 weeks to 5 months. The disease occurs worldwide, resulting in significant morbidity and mortality on endemically affected farms. Foals appear to become infected early in life, but clinical signs are typically delayed until 1–3 months of age because of the insidious nature of the disease. Although pneumonia is the most common clinical manifestation, up to 74% of foals may concurrently have extrapulmonary disorders, including both extrapulmonary infections (abdominal abscessation, colitis, osteomyelitis and immune-mediated disorders (nonseptic synovitis, uveitis. Diagnosis is based on the combination of clinical signs and abnormalities on hematologic screening and thoracic imaging in an appropriately aged foal and is confirmed by bacteriologic culture of the organism. Management of R. equi infections, in particular on farms with endemic disease, combines appropriate treatment of affected foals with preventative measures targeted at preventing infection and identifying foals before the development of severe disease. The combination of rifampin and a macrolide antimicrobial is recommended for treatment, as the combination is synergistic, reaches high intracellular concentrations, and should minimize the development of antimicrobial resistance. The prognosis for survival for foals with R. equi pneumonia is good, especially in foals mildly or subclinically affected, as is the prognosis for future athletic performance. Screening for early identification before the development of clinical signs has been advocated on endemically affected farms, although the most appropriate method, the timing of screening, and the selection of foals requiring treatment have yet to be determined. Recent evidence suggests that

Biodegradation of Di-(2-ethylhexyl Phthalate by Rhodococcus ruber YC-YT1 in Contaminated Water and Soil

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Ting Yang

2018-05-01

Full Text Available Di-(2-ethylehxyl phthalate (DEHP is one of the most broadly representative phthalic acid esters (PAEs used as a plasticizer in polyvinyl chloride (PVC production, and is considered to be an endocrine-disrupting chemical. DEHP and its monoester metabolites are responsible for adverse effects on human health. An efficient DEHP-degrading bacterial strain Rhodococcus ruber YC-YT1, with super salt tolerance (0–12% NaCl, is the first DEHP-degrader isolated from marine plastic debris found in coastal saline seawater. Strain YC-YT1 completely degraded 100 mg/L DEHP within three days (pH 7.0, 30 °C. According to high-performance liquid chromatography–mass spectrometry (HPLC-MS analysis, DEHP was transformed by strain YC-YT1 into phthalate (PA via mono (2-ethylehxyl phthalate (MEHP, then PA was used for cell growth. Furthermore, YC-YT1 metabolized initial concentrations of DEHP ranging from 0.5 to 1000 mg/L. Especially, YC-YT1 degraded up to 60% of the 0.5 mg/L initial DEHP concentration. Moreover, compared with previous reports, strain YC-YT1 had the largest substrate spectrum, degrading up to 13 kinds of PAEs as well as diphenyl, p-nitrophenol, PA, benzoic acid, phenol, protocatechuic acid, salicylic acid, catechol, and 1,2,3,3-tetrachlorobenzene. The excellent environmental adaptability of strain YC-YT1 contributed to its ability to adjust its cell surface hydrophobicity (CSH so that 79.7–95.9% of DEHP-contaminated agricultural soil, river water, coastal sediment, and coastal seawater were remedied. These results demonstrate that R. ruber YC-YT1 has vast potential to bioremediate various DEHP-contaminated environments, especially in saline environments.

Cloning, overexpression, purification, crystallization and preliminary X-ray analysis of 3-ketosteroid Δ{sup 4}-(5α)-dehydrogenase from Rhodococcus jostii RHA1

Energy Technology Data Exchange (ETDEWEB)

Oosterwijk, Niels van; Knol, Jan; Dijkhuizen, Lubbert; Geize, Robert van der; Dijkstra, Bauke W., E-mail: b.w.dijkstra@rug.nl [University of Groningen, Nijenborgh 7, 9747 AG Groningen (Netherlands)

2011-10-01

The gene for 3-ketosteroid Δ{sup 4}-(5α)-dehydrogenase from R. jostii RHA1 was cloned and overexpressed in E. coli and the protein product was purified and crystallized using the hanging-drop vapour-diffusion method. The crystals belonged to space group C222{sub 1} and diffraction data were collected to a resolution of 1.6 Å. 3-Ketosteroid dehydrogenases are flavoproteins which play key roles in steroid ring degradation. The enzymes are abundantly present in actinobacteria, including the catabolic powerhouse Rhodococcus jostii and the pathogenic species R. equi and Mycobacterium tuberculosis. The gene for 3-ketosteroid Δ{sup 4}-(5α)-dehydrogenase [Δ{sup 4}-(5α)-KSTD] from R. jostii RHA1 was cloned and overexpressed in Escherichia coli. His-tagged Δ{sup 4}-(5α)-KSTD enzyme was purified by Ni{sup 2+}–NTA affinity chromatography, anion-exchange chromatography and size-exclusion chromatography and was crystallized using the hanging-drop vapour-diffusion method. Seeding greatly improved the number of crystals obtained. The crystals belonged to space group C222{sub 1}, with unit-cell parameters a = 99.2, b = 114.3, c = 110.2 Å. Data were collected to a resolution of 1.6 Å.

Enzymatic degradation of aliphatic nitriles by Rhodococcus rhodochrous BX2, a versatile nitrile-degrading bacterium.

Science.gov (United States)

Fang, Shumei; An, Xuejiao; Liu, Hongyuan; Cheng, Yi; Hou, Ning; Feng, Lu; Huang, Xinning; Li, Chunyan

2015-06-01

Nitriles are common environmental pollutants, and their removal has attracted increasing attention. Microbial degradation is considered to be the most acceptable method for removal. In this work, we investigated the biodegradation of three aliphatic nitriles (acetonitrile, acrylonitrile and crotononitrile) by Rhodococcus rhodochrous BX2 and the expression of their corresponding metabolic enzymes. This organism can utilize all three aliphatic nitriles as sole carbon and nitrogen sources, resulting in the complete degradation of these compounds. The degradation kinetics were described using a first-order model. The degradation efficiency was ranked according to t1/2 as follows: acetonitrile>trans-crotononitrile>acrylonitrile>cis-crotononitrile. Only ammonia accumulated following the three nitriles degradation, while amides and carboxylic acids were transient and disappeared by the end of the assay. mRNA expression and enzyme activity indicated that the tested aliphatic nitriles were degraded via both the inducible NHase/amidase and the constitutive nitrilase pathways, with the former most likely preferred. Copyright © 2015 Elsevier Ltd. All rights reserved.

Isolation of endosulfan sulfate-degrading Rhodococcus koreensis strain S1-1 from endosulfan contaminated soil and identification of a novel metabolite, endosulfan diol monosulfate

International Nuclear Information System (INIS)

Ito, Koji; Kawashima, Fujimasa; Takagi, Kazuhiro; Kataoka, Ryota; Kotake, Masaaki; Kiyota, Hiromasa; Yamazaki, Kenichi; Sakakibara, Futa; Okada, Sanae

2016-01-01

An aerobic endosulfan sulfate-degrading bacterium, Rhodococcus koreensis strain S1-1, was isolated from soil to which endosulfan had been applied annually for more than 10 years until 2008. The strain isolated in this work reduced the concentration of endosulfan sulfate (2) from 12.25 μM to 2.11 μM during 14 d at 30 °C. Using ultra performance liquid chromatography-electrospray ionization-mass spectroscopy (UPLC-ESI-MS), a new highly water-soluble metabolite possessing six chlorine atoms was found to be endosulfan diol monosulfate (6), derived from 2 by hydrolysis of the cyclic sulfate ester ring. The structure of 6 was elucidated by chemical synthesis of the candidate derivatives and by HR-MS and UPLC-MS analyses. Therefore, it was suggested that the strain S1-1 has a new metabolic pathway of 2. In addition, 6 was expected to be less toxic among the metabolites of 1 because of its higher water-solubility. -- Highlights: •A novel endosulfan sulfate-degrading bacterium was isolated and named strain S1-1. •Strain S1-1 degraded endosulfan sulfate into a novel metabolite endosulfan diol monosulfate. •Endosulfan diol monosulfate showed higher polarity than other known metabolites of endosulfan. •We proposed the plausible metabolic pathway of endosulfan in terms of organic chemistry.

Isolation of endosulfan sulfate-degrading Rhodococcus koreensis strain S1-1 from endosulfan contaminated soil and identification of a novel metabolite, endosulfan diol monosulfate

Energy Technology Data Exchange (ETDEWEB)

Ito, Koji; Kawashima, Fujimasa [Department of Applied Biology and Chemistry, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo, 156-8502 (Japan); Organochemicals Division, National Institute for Agro-Environmental Sciences, 3-1-3 Kannondai, Tsukuba, Ibaraki, 305-8604 (Japan); Takagi, Kazuhiro, E-mail: ktakagi@affrc.go.jp [Department of Applied Biology and Chemistry, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo, 156-8502 (Japan); Organochemicals Division, National Institute for Agro-Environmental Sciences, 3-1-3 Kannondai, Tsukuba, Ibaraki, 305-8604 (Japan); Kataoka, Ryota [Department of Environmental Science, University of Yamanashi, 41-4-37 Takeda, Kofu, Yamanashi (Japan); Kotake, Masaaki [Graduate School of Agricultural Science, Tohoku University, Aoba-ku, Sendai 981-8555 (Japan); Kiyota, Hiromasa [Graduate School of Environmental & Life Science, Okayama University, 1-1-1 Tsushima-naka, Kita-ku, Okayama 700-8530 (Japan); Yamazaki, Kenichi [Organochemicals Division, National Institute for Agro-Environmental Sciences, 3-1-3 Kannondai, Tsukuba, Ibaraki, 305-8604 (Japan); Sakakibara, Futa [Organochemicals Division, National Institute for Agro-Environmental Sciences, 3-1-3 Kannondai, Tsukuba, Ibaraki, 305-8604 (Japan); The Japan Society for the Promotion of Science(JSPS), 1-8 Chiyoda-ku, Tokyo (Japan); Okada, Sanae [Department of Applied Biology and Chemistry, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo, 156-8502 (Japan)

2016-05-13

An aerobic endosulfan sulfate-degrading bacterium, Rhodococcus koreensis strain S1-1, was isolated from soil to which endosulfan had been applied annually for more than 10 years until 2008. The strain isolated in this work reduced the concentration of endosulfan sulfate (2) from 12.25 μM to 2.11 μM during 14 d at 30 °C. Using ultra performance liquid chromatography-electrospray ionization-mass spectroscopy (UPLC-ESI-MS), a new highly water-soluble metabolite possessing six chlorine atoms was found to be endosulfan diol monosulfate (6), derived from 2 by hydrolysis of the cyclic sulfate ester ring. The structure of 6 was elucidated by chemical synthesis of the candidate derivatives and by HR-MS and UPLC-MS analyses. Therefore, it was suggested that the strain S1-1 has a new metabolic pathway of 2. In addition, 6 was expected to be less toxic among the metabolites of 1 because of its higher water-solubility. -- Highlights: •A novel endosulfan sulfate-degrading bacterium was isolated and named strain S1-1. •Strain S1-1 degraded endosulfan sulfate into a novel metabolite endosulfan diol monosulfate. •Endosulfan diol monosulfate showed higher polarity than other known metabolites of endosulfan. •We proposed the plausible metabolic pathway of endosulfan in terms of organic chemistry.

Biodegradation of buprofezin by Rhodococcus sp. strain YL-1 isolated from rice field soil.

Science.gov (United States)

Li, Chao; Zhang, Ji; Wu, Zhi-Guo; Cao, Li; Yan, Xin; Li, Shun-Peng

2012-03-14

A buprofezin-degrading bacterium, YL-1, was isolated from rice field soil. YL-1 was identified as Rhodococcus sp. on the basis of the comparative analysis of 16S rDNA sequences. The strain could use buprofezin as the sole source of carbon and nitrogen for growth and was able to degrade 92.4% of 50 mg L(-1) buprofezin within 48 h in liquid culture. During the degradation of buprofezin, four possible metabolites, 2-tert-butylimino-3-isopropyl-1,3,5-thiadiazinan-4-one, N-tert-butyl-thioformimidic acid formylaminomethyl ester, 2-isothiocyanato-2-methyl-propane, and 2-isothiocyanato-propane, were identified using gas chromatography-mass spectrometry (GC-MS) analysis. The catechol 2,3-dioxygenase activity was strongly induced during the degradation of buprofezin. A novel microbial biodegradation pathway for buprofezin was proposed on the basis of these metabolites. The inoculation of soils treated with buprofezin with strain YL-1 resulted in a higher degradation rate than that observed in noninoculated soils, indicating that strain YL-1 has the potential to be used in the bioremediation of buprofezin-contaminated environments.

Effect of growth media on cell envelope composition and nitrile hydratase stability in Rhodococcus rhodochrous strain DAP 96253.

Science.gov (United States)

Tucker, Trudy-Ann; Crow, Sidney A; Pierce, George E

2012-11-01

Rhodococcus is an important industrial microorganism that possesses diverse metabolic capabilities; it also has a cell envelope, composed of an outer layer of mycolic acids and glycolipids. Selected Rhodococcus species when induced are capable of transforming nitriles to the corresponding amide by the enzyme nitrile hydratase (NHase), and subsequently to the corresponding acid via an amidase. This nitrile biochemistry has generated interest in using the rhodococci as biocatalysts. It was hypothesized that altering sugars in the growth medium might impact cell envelope components and have effects on NHase. When the primary carbon source in growth media was changed from glucose to fructose, maltose, or maltodextrin, the NHase activity increased. Cells grown in the presence of maltose and maltodextrin showed the highest activities against propionitrile, 197 and 202 units/mg cdw, respectively. Stability of NHase was also affected as cells grown in the presence of maltose and maltodextrin retained more NHase activity at 55 °C (45 and 23 %, respectively) than cells grown in the presence of glucose or fructose (19 and 10 %, respectively). Supplementation of trehalose in the growth media resulted in increased NHase stability at 55 °C, as cells grown in the presence of glucose retained 40 % NHase activity as opposed to 19 % without the presence of trehalose. Changes in cell envelope components, such mycolic acids and glycolipids, were evaluated by high-performance liquid chromatography (HPLC) and thin-layer chromatography (TLC), respectively. Changing sugars and the addition of inducing components for NHase, such as cobalt and urea in growth media, resulted in changes in mycolic acid profiles. Mycolic acid content increased 5 times when cobalt and urea were added to media with glucose. Glycolipids levels were also affected by the changes in sugars and addition of inducing components. This research demonstrates that carbohydrate selection impacts NHase activity and

Transcriptional Response of Rhodococcus aetherivorans I24 to Polychlorinated Biphenyl-Contaminated Sediments

KAUST Repository

Puglisi, Edoardo

2010-04-06

We used a microarray targeting 3,524 genes to assess the transcriptional response of the actinomycete Rhodococcus aetherivorans I24 in minimal medium supplemented with various substrates (e. g., PCBs) and in both PCB-contaminated and non-contaminated sediment slurries. Relative to the reference condition (minimal medium supplemented with glucose), 408 genes were upregulated in the various treatments. In medium and in sediment, PCBs elicited the upregulation of a common set of 100 genes, including gene-encoding chaperones (groEL), a superoxide dismutase (sodA), alkyl hydroperoxide reductase protein C (ahpC), and a catalase/peroxidase (katG). Analysis of the R. aetherivorans I24 genome sequence identified orthologs of many of the genes in the canonical biphenyl pathway, but very few of these genes were upregulated in response to PCBs or biphenyl. This study is one of the first to use microarrays to assess the transcriptional response of a soil bacterium to a pollutant under conditions that more closely resemble the natural environment. Our results indicate that the transcriptional response of R. aetherivorans I24 to PCBs, in both medium and sediment, is primarily directed towards reducing oxidative stress, rather than catabolism. © 2010 Springer Science+Business Media, LLC.

Aerobic Biodegradation of N-Nitrosodimethylamine by the Propanotroph Rhodococcus ruber ENV425▿

Science.gov (United States)

Fournier, Diane; Hawari, Jalal; Halasz, Annamaria; Streger, Sheryl H.; McClay, Kevin R.; Masuda, Hisako; Hatzinger, Paul B.

2009-01-01

The propanotroph Rhodococcus ruber ENV425 was observed to rapidly biodegrade N-nitrosodimethylamine (NDMA) after growth on propane, tryptic soy broth, or glucose. The key degradation intermediates were methylamine, nitric oxide, nitrite, nitrate, and formate. Small quantities of formaldehyde and dimethylamine were also detected. A denitrosation reaction, initiated by hydrogen atom abstraction from one of the two methyl groups, is hypothesized to result in the formation of n-methylformaldimine and nitric oxide, the former of which decomposes in water to methylamine and formaldehyde and the latter of which is then oxidized further to nitrite and then nitrate. Although the strain mineralized more than 60% of the carbon in [14C]NDMA to 14CO2, growth of strain ENV425 on NDMA as a sole carbon and energy source could not be confirmed. The bacterium was capable of utilizing NDMA, as well as the degradation intermediates methylamine and nitrate, as sources of nitrogen during growth on propane. In addition, ENV425 reduced environmentally relevant microgram/liter concentrations of NDMA to <2 ng/liter in batch cultures, suggesting that the bacterium may have applications for groundwater remediation. PMID:19542346

Humoral immune responses of pregnant Guinea pigs Immunized with live attenuated Rhodococcus equi

Directory of Open Access Journals (Sweden)

Mawlood Abass Ali Al- Graibawi

2018-02-01

Full Text Available The potential to increase passive transfer of specific Rhodococcus equi (R.equi humoral immunity to newborn by preparturient vaccination of their dams was investigated in Pregnant Guinea pigs as a pilot study. Attenuated autogenous vaccine was prepared from a Congo red negative (CR- R.equi local isolate mixed with adjuvant (potassium alum sulphate, tested for sterility, safety and potency prior to vaccination .Two groups of pregnant G. pigs were used, the first group was vaccinated twice subcutaneously (S.C with the prepared vaccine at five and three weeks prior parturition, the second group was inoculated with adjuvant plus phosphate buffer saline (PBS twice s.c and kept as control. Offspring from the vaccinated dams had revealed high titers of specific R. equi antibody as detected by tube agglutination (TA and passive haemagglutination (PH test and showed protection against challenge dose. The results revealed that vaccination of pregnant G. pigs with the prepared attenuated vaccine was safe and efficient method to protect their offspring against experimental challenge with virulent R.equi. Vaccination was associated with increased humoral immune response in vaccinated group.

Tumour suppressor protein p53 regulates the stress activated bilirubin oxidase cytochrome P450 2A6

Energy Technology Data Exchange (ETDEWEB)

Hu, Hao, E-mail: hao.hu1@uqconnect.edu.au [The University of Queensland, National Research Centre for Environmental Toxicology (Entox), 4072 Brisbane, Queensland (Australia); Yu, Ting, E-mail: t.yu2@uq.edu.au [The University of Queensland, National Research Centre for Environmental Toxicology (Entox), 4072 Brisbane, Queensland (Australia); Arpiainen, Satu, E-mail: Satu.Juhila@orion.fi [Institute of Biomedicine, Department of Pharmacology and Toxicology and Medical Research Center Oulu, Oulu University Hospital and University of Oulu, Oulu (Finland); Lang, Matti A., E-mail: m.lang@uq.edu.au [The University of Queensland, National Research Centre for Environmental Toxicology (Entox), 4072 Brisbane, Queensland (Australia); Hakkola, Jukka, E-mail: Jukka.hakkola@oulu.fi [Institute of Biomedicine, Department of Pharmacology and Toxicology and Medical Research Center Oulu, Oulu University Hospital and University of Oulu, Oulu (Finland); Abu-Bakar, A' edah, E-mail: a.abubakar@uq.edu.au [The University of Queensland, National Research Centre for Environmental Toxicology (Entox), 4072 Brisbane, Queensland (Australia)

2015-11-15

Human cytochrome P450 (CYP) 2A6 enzyme has been proposed to play a role in cellular defence against chemical-induced oxidative stress. The encoding gene is regulated by various stress activated transcription factors. This paper demonstrates that p53 is a novel transcriptional regulator of the gene. Sequence analysis of the CYP2A6 promoter revealed six putative p53 binding sites in a 3 kb proximate promoter region. The site closest to transcription start site (TSS) is highly homologous with the p53 consensus sequence. Transfection with various stepwise deletions of CYP2A6-5′-Luc constructs – down to − 160 bp from the TSS – showed p53 responsiveness in p53 overexpressed C3A cells. However, a further deletion from − 160 to − 74 bp, including the putative p53 binding site, totally abolished the p53 responsiveness. Electrophoretic mobility shift assay with a probe containing the putative binding site showed specific binding of p53. A point mutation at the binding site abolished both the binding and responsiveness of the recombinant gene to p53. Up-regulation of the endogenous p53 with benzo[α]pyrene – a well-known p53 activator – increased the expression of the p53 responsive positive control and the CYP2A6-5′-Luc construct containing the intact p53 binding site but not the mutated CYP2A6-5′-Luc construct. Finally, inducibility of the native CYP2A6 gene by benzo[α]pyrene was demonstrated by dose-dependent increases in CYP2A6 mRNA and protein levels along with increased p53 levels in the nucleus. Collectively, the results indicate that p53 protein is a regulator of the CYP2A6 gene in C3A cells and further support the putative cytoprotective role of CYP2A6. - Highlights: • CYP2A6 is an immediate target gene of p53. • Six putative p53REs located on 3 kb proximate CYP2A6 promoter region. • The region − 160 bp from TSS is highly homologous with the p53 consensus sequence. • P53 specifically bind to the p53RE on the − 160 bp region. • HNF4�

Dimetilsulfóxido - DMSO no teste de sensibilidade microbiana in vitro em cepas de Rhodococcus equi isoladas de afecções pulmonares em potros

Directory of Open Access Journals (Sweden)

Ribeiro Márcio Garcia

2001-01-01

Full Text Available Comparou-se a sensibilidade microbiana in vitro de isolados de Rhodococcus equi pelo teste padrão de difusão com discos, com o modificado, pela adição de 5% de dimetilsulfóxido-DMSO. Observou-se aumento da sensibilidade do R. equi no teste com DMSO, frente a aminoglicosídeos (canamicina, amicacina, estreptomicina e ao cloranfenicol, enquanto para a eritromicina e derivados ß-lactâmicos (penicilina G, cefalosporinas, amoxicilina, oxacilina, constatou-se redução da sensibilidade do agente.

Anisotropic thermal properties and ferroelectric phase transitions in layered CuInP2S6 and CuInP2Se6 crystals

Science.gov (United States)

Liubachko, V.; Shvalya, V.; Oleaga, A.; Salazar, A.; Kohutych, A.; Pogodin, A.; Vysochanskii, Yu. M.

2017-12-01

Thermal diffusivity and thermal conductivity have been studied for the layered crystals CuInP2S6, CuInP2Se6 from 30 K to 350 K, showing a relevant thermal anisotropy. Heat is much more efficiently transferred within the layers than perpendicular to them. The ferrielectric transition in CuInP2S6 is proven to be clearly first order while the ferroelectric one in CuInP2Se6 has a weak first order character. The behavior of the thermal conductivity as a function of temperature in the ferroelectric phases shows that heat conduction is phonon driven. Disorder in the paraelectric phases due to hopping motions of Cu ions significantly reduces the thermal conductivity to extremely low values.

Tungsten phosphanylarylthiolato complexes [W{PhP(2-SC6H4)2-kappa3S,S',P} 2] and [W{P(2-SC6H4)3-kappa4S,S',S",P}2]: synthesis, structures and redox chemistry.

Science.gov (United States)

Hildebrand, Alexandra; Lönnecke, Peter; Silaghi-Dumitrescu, Luminita; Hey-Hawkins, Evamarie

2008-09-14

PhP(2-SHC6H4)2 (PS2H2) reacts with WCl6 with reduction of tungsten to give the air-sensitive tungsten(IV) complex [W{PhP(2-SC6H4)2-kappa(3)S,S',P}2] (1). 1 is oxidised in air to [WO{PhPO(2-SC6H4)2-kappa(3)S,S',O}{PhP(2-SC6H4)2-kappa(3)S,S',P}] (2). The attempted synthesis of 2 by reaction of 1 with iodosobenzene as oxidising agent was unsuccessful. [W{P(2-SC6H4)3-kappa(4)S,S',S",P}2] (3) was formed in the reaction of P(2-SHC6H4)3 (PS3H3) with WCl6. The W(VI) complex 3 contains two PS3(3-) ligands, each coordinated in a tetradentate fashion resulting in a tungsten coordination number of eight. The reaction of 3 with AgBF4 yields the dinuclear tungsten complex [W2{P(2-SC6H4)3-kappa(4)S,S',S",P}3]BF4 (4). Complexes 1-4 were characterised by spectral methods and X-ray structure determination.

Production, Purification, and Identification of Cholest-4-en-3-one Produced by Cholesterol Oxidase from Rhodococcus sp. in Aqueous/Organic Biphasic System.

Science.gov (United States)

Wu, Ke; Li, Wei; Song, Jianrui; Li, Tao

2015-01-01

Cholest-4-en-3-one has positive uses against obesity, liver disease, and keratinization. It can be applied in the synthesis of steroid drugs as well. Most related studies are focused on preparation of cholest-4-en-3-one by using whole cells as catalysts, but production of high-quality cholest-4-en-3-one directly from cholesterol oxidase (COD) using an aqueous/organic two-phase system has been rarely explored. This study set up an enzymatic reaction system to produce cholest-4-en-3-one. We developed and optimized the enzymatic reaction system using COD from COX5-6 (a strain of Rhodococcus) instead of whole-cell biocatalyst. This not only simplifies and accelerates the production but also benefits the subsequent separation and purification process. Through extraction, washing, evaporation, column chromatography, and recrystallization, we got cholest-4-en-3-one with purity of 99.78%, which was identified by nuclear magnetic resonance, mass spectroscopy, and infrared spectroscopy. In addition, this optimized process of cholest-4-en-3-one production and purification can be easily scaled up for industrial production, which can largely decrease the cost and guarantee the purity of the product.

Restriction of human herpesvirus 6B replication by p53

DEFF Research Database (Denmark)

Øster, Bodil; Kofod-Olsen, Emil; Bundgaard, Bettina

2008-01-01

Human herpesvirus 6B (HHV-6B) induces significant accumulation of p53 in both the nucleus and cytoplasm during infection. Activation of p53 by DNA damage is known to induce either growth arrest or apoptosis; nevertheless, HHV-6B-infected cells are arrested in their cell cycle independently of p53...

Nucleus-associated phosphorylation of Ins(1,4,5)P3 to InsP6 in Dictyostelium

NARCIS (Netherlands)

Kaay, Jeroen van der; Wesseling, Jelle; Haastert, Peter J.M. van

1995-01-01

Although many cells contain large amounts of InsP(6), its metabolism and function is still largely unknown. In Dictyostelium lysates, the formation of InsP(6) by sequential phosphorylation of inositol via Ins(3,4,6)P-3 has been described [Stevens and Irvine (1990) Nature (London) 346, 580-583]; the

Bioremediation of soil contaminated by dichlorodiphenyltrichloroethane with the use of aerobic strain Rhodococcus wratislaviensis Ch628

Science.gov (United States)

Egorova, D. O.; Farafonova, V. V.; Shestakova, E. A.; Andreyev, D. N.; Maksimov, A. S.; Vasyanin, A. N.; Buzmakov, S. A.; Plotnikova, E. G.

2017-10-01

The concentration of dichlorodiphenyltrichloroethane (DDT) was determined in a sandy soil of specially Protected Natural Area Osinskaya Lesnaya Dacha (Perm region) 45 years after the last application of the insecticide in this area. The concentration of DDT in the soil exceeded the maximum permissible concentration by 250 times and reached 25.05 mg/kg of soil. Under the conditions of model experiment, efficient decontamination of the soil was recorded in the system with the introduced strain Rhodococcus wratislaviensis Ch628; the DDT concentration decreased by 99.7% and equaled 0.07 mg/kg. The process of DDT degradation proceeded slower in the model soil system with autochthonous microbial complex. In this case, 58.2% DDT degraded in 70 days, and the final concentration was 10.47 mg/kg. The soil lost its toxicity for animal and plant test objects by the end of the experiment only in the model system containing the R. wratislaviensis Ch628 strain.

GenBank blastx search result: AK062096 [KOME

Lifescience Database Archive (English)

Full Text Available AK062096 001-044-H12 U17130.1 Rhodococcus erythropolis ORF6' gene, partial cds, regulatory protein ThcR (thc...R), cytochrome P-450 (thcB), ORF4, rhodocoxin (thcC), rhodocoxin reductase (thcD) and ORF5 genes, complete cds.|BCT BCT 3e-18 +3 ...

Comparison between p6P thin films and nanofibers on transistor platforms

DEFF Research Database (Denmark)

Tavares, Luciana; Kjelstrup-Hansen, Jakob; Thilsing-Hansen, Kasper

2010-01-01

The organic semiconductor para-hexaphenylene (p-6P) can be used as the light-emitting material in organic light-emitting field-effect transistors (OLEFETs), has a low cost, and is easy to process which could allow, for example, the development of mechanically flexible optoelectronics. In this work...

Photoionization from the 6p 2P3/2 state of neutral cesium

International Nuclear Information System (INIS)

Haq, S. U.; Nadeem, Ali

2010-01-01

We report the photoionization studies of cesium from the 6p 2 P 3/2 excited state to measure the photoionization cross section at and above the first ionization threshold, oscillator strength of the highly excited transitions, and extension in the Rydberg series. The photoionization cross section at the first ionization threshold is measured as 25 (4) Mb and at excess energies 0.02, 0.04, 0.07, and 0.09 eV as 21, 19, 17, and 16 Mb, respectively. Oscillator strength of the 6p 2 P 3/2 → nd 2 D 5/2 (23 ≤ n ≤ 60) Rydberg transitions has been extracted utilizing the threshold value of photoionization cross section and the recorded nd 2 D 5/2 photoionization spectra.

The sensor kinase MprB is required for Rhodococcus equi virulence.

Science.gov (United States)

MacArthur, Iain; Parreira, Valeria R; Lepp, Dion; Mutharia, Lucy M; Vazquez-Boland, José A; Prescott, John F

2011-01-10

Rhodococcus equi is a soil bacterium and, like Mycobacterium tuberculosis, a member of the mycolata. Through possession of a virulence plasmid, it has the ability to infect the alveolar macrophages of foals, resulting in pyogranulomatous bronchopneumonia. The virulence plasmid has an orphan two-component system (TCS) regulatory gene, orf8, mutation of which completely attenuates virulence. This study attempted to find the cognate sensor kinase (SK) of orf8. Annotation of the R. equi strain 103 genome identified 23 TCSs encoded on the chromosome, which were used in a DNA microarray to compare TCS gene transcription in murine macrophage-like cells to growth in vitro. This identified six SKs as significantly up-regulated during growth in macrophages. Mutants of these SKs were constructed and their ability to persist in macrophages was determined with one SK, MprB, found to be required for intracellular survival. The attenuation of the mprB- mutant, and its complementation, was confirmed in a mouse virulence assay. In silico analysis of the R. equi genome sequence identified an MprA binding box motif homologous to that of M. tuberculosis, on mprA, pepD, sigB and sigE. The results of this study also show that R. equi responds to the macrophage environment differently from M. tuberculosis. MprB is the first SK identified as required for R. equi virulence and intracellular survival. Copyright © 2010 Elsevier B.V. All rights reserved.

Quasimolecular emission near the Xe(5p 56s 1,3 P 1 - 5p 6 1 S 0) and Kr (4p 55s 1,3 P 1 - 4p 6 1 S 0) resonance lines induced by collisions with He atoms

Science.gov (United States)

Alekseeva, O. S.; Devdariani, A. Z.; Grigorian, G. M.; Lednev, M. G.; Zagrebin, A. L.

2017-02-01

This study is devoted to the theoretical investigation of the quasimolecular emission of Xe*-He and Kr*-He collision pairs near the Xe (5p 56s 1,3 P 1 - 5p 6 1 S 0) and Kr (4p 55s 1,3 P 1 - 4p 6 1 S 0) resonance atomic lines. The potential curves of the quasimolecules Xe(5p 56s) + He and Kr(4p 55s) + He have been obtained with the use of the effective Hamiltonian and pseudopotential methods. Based on these potential curves the processes of quasimolecular emission of Xe*+He and Kr*+He mixtures have been considered and the spectral distributions I(ħΔω) of photons emitted have been obtained in the framework of quasistatic approximation.

Ge 3P 6Si 2O 25: A cage structure closely related to the intersecting tunnel structure KMo 3P 6Si 2O 25

Science.gov (United States)

Leclaire, A.; Raveau, B.

1988-08-01

A germanosilicophosphate Ge 3P 6Si 2O 25 has been isolated. Its structure was solved from a single-crystal study in the space group P overline31c . Its cell parameters are a = b = 7.994(1) Å, c = 16.513(2) Å, Z = 2. The refinement by full-matrix least-squares calculations leads to R = 0.043 with 686 independent reflections. The structure of this oxide is built up from corner-sharing PO 4 and SiO 4 tetrahedra and GeO 6 octahedra. One observes a feature common to several silicophosphates: the presence of the structural unit P 6Si 2O 25 built up from a disilicate group sharing its corners with six PO 4 tetrahedra. The structural relationships between this oxide and the silicophosphates AMo 3P 6Si 2O 25 and Si 3P 6Si 2O 25 (or Ge 3P 6 Ge 2O 25) are described.

Rhodococcus equi venous catheter infection: a case report and review of the literature

Directory of Open Access Journals (Sweden)

Nahleh Zeina

2011-08-01

Full Text Available Abstract Introduction Rhodococcus equi is an animal pathogen that was initially isolated from horses and is being increasingly reported as a cause of infection in humans with impaired cellular immunity. However, this pathogen is underestimated as a challenging antagonist and is frequently considered to be a mere contaminant despite the potential for life-threatening infections. Most case reports have occurred in immunocompromised patients who have received organ transplants (for example kidney, heart, bone marrow or those with human immunodeficiency virus infection. Infections often manifest as pulmonary involvement or soft tissue abscesses. Bacteremia related to R. equi infections of tunneled central venous catheters has rarely been described. Case presentation We report the case of a 63-year-old non-transplant recipient, non-HIV infected Caucasian woman with endometrial carcinoma who developed recurrent bloodstream infections and septic shock due to R. equi and ultimately required the removal of her port catheter, a subcutaneous implantable central venous catheter. We also review the medical literature related to human infections with R. equi. Conclusion R. equi should be considered a serious pathogen, not a contaminant, particularly in an immunocompromised patient who presents with a central venous catheter-related bloodstream infection. Counseling patients with central venous catheters who participate in activities involving exposure to domesticated animals is recommended.

Degradation Signals Recognized by the Ubc6p-Ubc7p Ubiquitin-Conjugating Enzyme Pair

Science.gov (United States)

Gilon, Tamar; Chomsky, Orna; Kulka, Richard G.

2000-01-01

Proteolysis by the ubiquitin-proteasome system is highly selective. Specificity is achieved by the cooperation of diverse ubiquitin-conjugating enzymes (Ubcs or E2s) with a variety of ubiquitin ligases (E3s) and other ancillary factors. These recognize degradation signals characteristic of their target proteins. In a previous investigation, we identified signals directing the degradation of β-galactosidase and Ura3p fusion proteins via a subsidiary pathway of the ubiquitin-proteasome system involving Ubc6p and Ubc7p. This pathway has recently been shown to be essential for the degradation of misfolded and regulated proteins in the endoplasmic reticulum (ER) lumen and membrane, which are transported to the cytoplasm via the Sec61p translocon. Mutant backgrounds which prevent retrograde transport of ER proteins (hrd1/der3Δ and sec61-2) did not inhibit the degradation of the β-galactosidase and Ura3p fusions carrying Ubc6p/Ubc7p pathway signals. We therefore conclude that the ubiquitination of these fusion proteins takes place on the cytosolic face of the ER without prior transfer to the ER lumen. The contributions of different sequence elements to a 16-amino-acid-residue Ubc6p-Ubc7p-specific signal were analyzed by mutation. A patch of bulky hydrophobic residues was an essential element. In addition, positively charged residues were found to be essential. Unexpectedly, certain substitutions of bulky hydrophobic or positively charged residues with alanine created novel degradation signals, channeling the degradation of fusion proteins to an unidentified proteasomal pathway not involving Ubc6p and Ubc7p. PMID:10982838

Dual Two-Component Regulatory Systems Are Involved in Aromatic Compound Degradation in a Polychlorinated-Biphenyl Degrader, Rhodococcus jostii RHA1 ▿ †

OpenAIRE

Takeda, Hisashi; Shimodaira, Jun; Yukawa, Kiyoshi; Hara, Naho; Kasai, Daisuke; Miyauchi, Keisuke; Masai, Eiji; Fukuda, Masao

2010-01-01

A Gram-positive polychlorinated-biphenyl (PCB) degrader, Rhodococcus jostii RHA1, degrades PCBs by cometabolism with biphenyl. A two-component BphS1T1 system encoded by bphS1 and bphT1 (formerly bphS and bphT) is responsible for the transcription induction of the five gene clusters, bphAaAbAcAdC1B1, etbAa1Ab1CbphD1, etbAa2Ab2AcD2, etbAdbphB2, and etbD1, which constitute multiple enzyme systems for biphenyl/PCB degradation. The bphS2 and bphT2 genes, which encode BphS2 and BphT2, virtually ide...

Nutrient starvation leading to triglyceride accumulation activates the Entner Doudoroff pathway in Rhodococcus jostii RHA1.

Science.gov (United States)

Juarez, Antonio; Villa, Juan A; Lanza, Val F; Lázaro, Beatriz; de la Cruz, Fernando; Alvarez, Héctor M; Moncalián, Gabriel

2017-02-27

Rhodococcus jostii RHA1 and other actinobacteria accumulate triglycerides (TAG) under nutrient starvation. This property has an important biotechnological potential in the production of sustainable oils. To gain insight into the metabolic pathways involved in TAG accumulation, we analysed the transcriptome of R jostii RHA1 under nutrient-limiting conditions. We correlate these physiological conditions with significant changes in cell physiology. The main consequence was a global switch from catabolic to anabolic pathways. Interestingly, the Entner-Doudoroff (ED) pathway was upregulated in detriment of the glycolysis or pentose phosphate pathways. ED induction was independent of the carbon source (either gluconate or glucose). Some of the diacylglycerol acyltransferase genes involved in the last step of the Kennedy pathway were also upregulated. A common feature of the promoter region of most upregulated genes was the presence of a consensus binding sequence for the cAMP-dependent CRP regulator. This is the first experimental observation of an ED shift under nutrient starvation conditions. Knowledge of this switch could help in the design of metabolomic approaches to optimize carbon derivation for single cell oil production.

Mathematical Modeling of E6-p53 interactions in Cervical Cancer

Science.gov (United States)

Khattak, Faryal; Haseeb, Muhammad; Fazal, Sahar; Bhatti, A I; Ullah, Mukhtar

2017-04-01

Background: Cervical cancer is the third most common cancer in women throughout the world. The human papillomavirus (HPV) E6 viral protein plays an essential role in proteasomal degradation of the cancer suppressant protein p53. As a result, p53 negative regulation and apoptosis relevant activities are abrogated, facilitating development of cervical cancer. Methods: A mathematical model of E6-p53 interactions was developed using mathematical laws. In-silico simulations were carried out on CellDesigner and as a test case the small molecule drug RITA was considered for its ability to rescue the functions of tumor suppressor p53 by inhibiting E6 mediated proteasomal degradation. Results: Using a computational model we scrutinized how p53 responds to RITA, and chemical reactions of this small molecule drug were incorporated to perceive the full effects. The evolved strategy allowed the p53 response and rescue of its tumor suppressor function to be delineated, RITA being found to block p53 interactions with E6 associated proteins. Conclusion: We could develop a model of E6-p53 interactions with incorporation of actions of the small molecule drug RITA. Suppression of E6 associated proteins by RITA induces accumulation of tumor suppressant p53. Using CellDesigner to encode the model ensured that it can be easily modified and extended as more data become available. This strategy should play an effective role in the development of therapies against cancer. Creative Commons Attribution License

Identification of 6H1 as a P2Y purinoceptor: P2Y5.

Science.gov (United States)

Webb, T E; Kaplan, M G; Barnard, E A

1996-02-06

We have determined the identity of the orphan G-protein coupled receptor cDNA, 6H1, present in activated chicken T cells, as a subtype of P2Y purinoceptor. This identification is based on first on the degree of sequence identity shared with recently cloned members of the P2Y receptor family and second on the pharmacological profile. Upon transient expression in COS-7 cells the 6H1 receptor bound the radiolabel [35S]dATP alpha S specifically and with high affinity (Kd, 10 nM). This specific binding could be competitively displaced by a range of ligands active at P2 purinoceptors, with ATP being the most active (K (i)), 116 nM). Such competition studies have established the following rank order of activity: ATP ADP 2-methylthioATP alpha, beta-methylene ATP, UTP, thus confirming 6H1 as a member of the growing family of P2Y purinoceptors. As the fifth receptor of this type to be identified we suggest that it be named P2Y5.

Search for the signature of a halo structure in the p(6He,6Li)n reaction

International Nuclear Information System (INIS)

Cortina-Gil, M.D.; Roussel-Chomaz, P.; Mittig, W.; Casandjian, J.M.; Chartier, M.; Alamanos, N.; Auger, F.; Fekou-Youmbi, V.; Blumenfeld, Y.; and others.

1995-01-01

The elastic scattering p( 6 He, 6 He)p and charge exchange reaction p( 6 He, 6 Li)n have been measured in reverse kinematics with a secondary 6 He beam. The angular distributions for these reactions were obtained. In the case of the charge exchange reaction, the ratio of the cross section for the Gamow-Teller transition to the ground state, and for the Fermi transition to the isobaric analog state is a measure of the relative strength of the two components of the exchange interaction. This ratio is found compatible with existing systematics for stable T=1 nuclei, and no clear signature of a halo structure was found in the present data. (author)

Generation and characterization of polyclonal antibodies specific to N-terminal extension of p85 isoform of ribosomal protein S6 kinase 1 (p85 S6K1

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Savinska L. O.

2015-08-01

Full Text Available Aim. Generation of polyclonal antibodies specific to the ribosomal protein S6 kinase isoform – p85S6K1 and directed to the N-terminal (1–23 aa extension of p85S6K1. Methods. Animal immunization with synthetic (1–23 aa peptide, ELISA, Western blot, Immunoprecipitation, immunofluorescent analysis. Results. Polyclonal antibodies have been generated, which specifically recognize only p85 but not p70 isoform of S6K1 in western blot, immunoprecipitation and immunofluorescence analysis. Conclusions. The obtained antibodies can be recommended for studies on the p85S6K1 and other S6K1 isoforms possessing the N-terminal extension – the identification of binding protein partners, analysis of subcellular localization under different physiological conditions, elucidation of the signal transduction pathways involving different S6K1 isoforms.

Diversion of phagosome trafficking by pathogenic Rhodococcus equi depends on mycolic acid chain length.

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Sydor, Tobias; von Bargen, Kristine; Hsu, Fong-Fu; Huth, Gitta; Holst, Otto; Wohlmann, Jens; Becken, Ulrike; Dykstra, Tobias; Söhl, Kristina; Lindner, Buko; Prescott, John F; Schaible, Ulrich E; Utermöhlen, Olaf; Haas, Albert

2013-03-01

Rhodococcus equi is a close relative of Mycobacterium spp. and a facultative intracellular pathogen which arrests phagosome maturation in macrophages before the late endocytic stage. We have screened a transposon mutant library of R. equi for mutants with decreased capability to prevent phagolysosome formation. This screen yielded a mutant in the gene for β-ketoacyl-(acyl carrier protein)-synthase A (KasA), a key enzyme of the long-chain mycolic acid synthesizing FAS-II system. The longest kasA mutant mycolic acid chains were 10 carbon units shorter than those of wild-type bacteria. Coating of non-pathogenic E. coli with purified wild-type trehalose dimycolate reduced phagolysosome formation substantially which was not the case with shorter kasA mutant-derived trehalose dimycolate. The mutant was moderately attenuated in macrophages and in a mouse infection model, but was fully cytotoxic.Whereas loss of KasA is lethal in mycobacteria, R. equi kasA mutant multiplication in broth was normal proving that long-chain mycolic acid compounds are not necessarily required for cellular integrity and viability of the bacteria that typically produce them. This study demonstrates a central role of mycolic acid chain length in diversion of trafficking by R. equi. © 2012 Blackwell Publishing Ltd.

Prediction of pKa values using the PM6 semiempirical method

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Jimmy C. Kromann

2016-08-01

Full Text Available The PM6 semiempirical method and the dispersion and hydrogen bond-corrected PM6-D3H+ method are used together with the SMD and COSMO continuum solvation models to predict pKa values of pyridines, alcohols, phenols, benzoic acids, carboxylic acids, and phenols using isodesmic reactions and compared to published ab initio results. The pKa values of pyridines, alcohols, phenols, and benzoic acids considered in this study can generally be predicted with PM6 and ab initio methods to within the same overall accuracy, with average mean absolute differences (MADs of 0.6–0.7 pH units. For carboxylic acids, the accuracy (0.7–1.0 pH units is also comparable to ab initio results if a single outlier is removed. For primary, secondary, and tertiary amines the accuracy is, respectively, similar (0.5–0.6, slightly worse (0.5–1.0, and worse (1.0–2.5, provided that di- and tri-ethylamine are used as reference molecules for secondary and tertiary amines. When applied to a drug-like molecule where an empirical pKa predictor exhibits a large (4.9 pH unit error, we find that the errors for PM6-based predictions are roughly the same in magnitude but opposite in sign. As a result, most of the PM6-based methods predict the correct protonation state at physiological pH, while the empirical predictor does not. The computational cost is around 2–5 min per conformer per core processor, making PM6-based pKa prediction computationally efficient enough to be used for high-throughput screening using on the order of 100 core processors.

Accumulation of a poly(hydroxyalkanoate) copolymer containing primarily 3-hydroxyvalerate from simple carbohydrate substrates by Rhodococcus sp. NCIMB 40126.

Science.gov (United States)

Haywood, G W; Anderson, A J; Williams, D R; Dawes, E A; Ewing, D F

1991-04-01

A number of taxonomically-related bacteria have been identified which accumulate poly(hydroxyalkanoate) (PHA) copolymers containing primarily 3-hydroxyvalerate (3HV) monomer units from a range of unrelated single carbon sources. One of these, Rhodococcus sp. NCIMB 40126, was further investigated and shown to produce a copolymer containing 75 mol% 3HV and 25 mol% 3-hydroxybutyrate (3HB) from glucose as sole carbon source. Polyesters containing both 3HV and 3HB monomer units, together with 4-hydroxybutyrate (4HB), 5-hydroxyvalerate (5HV) or 3-hydroxyhexanoate (3HHx), were also produced by this organism from certain accumulation substrates. With valeric acid as substrate, almost pure (99 mol% 3HV) poly(3-hydroxyvalerate) was produced. N.m.r. analysis confirmed the composition of these polyesters. The thermal properties and molecular weight of the copolymer produced from glucose were comparable to those of PHB produced by Alcaligenes eutrophus.

REVIEW ON CONSTRUCTION PROJECT MANAGEMENT SOFTWARE PRIMAVERA P6

OpenAIRE

Piyush Pramod Bagade* & Prof. Abhijit Bhirud

2018-01-01

Planning, scheduling and Resource levelling plays an important role in any construction project maybe it is construction of building or construction of road. In absence of proper planning, scheduling and resource levelling construction industry does not get profit over the project. For this purpose, proper software’s and techniques must be utilize. This paper is focussing on the advantages of Oracle Primavera P6 Software. The latest version of Primavera is P6-17. In any construction work huge...

Purification and characterization of lignin peroxidases from Penicillium decumbens P6

Energy Technology Data Exchange (ETDEWEB)

Yang, J.S.; Yuan, H.L.; Wang, H.X.; Chen, W.X. [China Agricultural University, Beijing (China). College of Biological Science

2005-06-01

Peroxidases are essential enzymes in biodegradation of lignin and lignite which have been investigated intensively in the white-rot fungi. This is the first report of purification and characterization of lignin peroxidase from Penicillium sp. P6 as lignite degradation fungus. The results indicated that the lignin peroxidase of Penicillium decumbens P6 had physical and chemical properties and a N-terminal amino acid sequence different from the lignin peroxidases of white-rot fungi. The lignin peroxidase was isolated from a liquid culture of P. decumbens P6. This enzyme had a molecular weight of 46.3 KDa in SDS-PAGE and exhibited greater activity, temperature stability and wider pH range than those previously reported. The isolation procedure involved (NH{sub 4}){sub 2}SO{sub 4} precipitation, ion-exchange chromatography on DEAE-cellulose and CM-cellulose, gel filtration on Sephadex G-100, and non-denaturing, discontinuous polyacrylamide gel electrophoresis. The K{sub m} and V{sub max} values of this enzyme using veratryl alcohol as substrate were 0.565 mmol L{sup -1} and 0.088 mmol (mg protein){sup -1} min{sup -1} respectively. The optimum pH of P6 lignin peroxidase was 4.0, and 70.6% of the relative activity was remained at pH 9.0. The optimum temperature of the enzyme was 45{sup o}C.

Disseminated Rhodococcus equi infection in a patient with Hodgkin’s lymphoma

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Mikić Dragan

2014-01-01

Full Text Available Introduction. Rhodococcus (R equi is an opportunistic, uncommon human pathogen that causes mainly infection in immunocompromised hosts. The disease is usually presented as subacute pneumonia that is mostly cavitary and sometimes bacteremic. Case report. We reported the extremly rare case of a 43-year-old woman with Hodgkin's lymphoma, who developed R. equi pulmonary infection after recieving multiple courses of chemotherapy. Secondary, the patient developed bacteremia, leading to sepsis and dissemination of R. equi infection in many extrapulmonary sites. At addmission the patient was febrile, tachypnoic, tachycardic, hypotensive, with facial edema, splenomegaly, positive meningeal signs, left hemiparesis and paraparesis. Laboratory data included erythrocyte sedimentation rate (ESR > 140 mm/h, C-reactive protein (CRP 143.0 mg/L, red blood cells (RBC 2.14 × 1012/L, whyite blood cells (WBC 2.8 × 109/L, lactate dehydrogenase (LDH 706 U/L, serum albumin 26 g/L, sodium 127 mmol/L and potassium 2.7 mmol/L. Blood culture and culture of sputum and empyema were positive for R. equi. Imaging studies demonstrated a large right cavitary pneumonia and abscess, empyema, pericarditis, mediastinal and intra-abdominal lymphadenopathy, brain and psoas abscesses, osteomyelitis and spondylodiscitis. The patient recovered completely after a 12-month treatment with combinations of parenteral and oral antibiotics (meropenem, vancomycin, teicoplanin, ciprofloxacin, rifampicin, macrolides etc, including drainage of abscesses and empyema. Eight years after completition of the treatment the patient was without recurrence of R. equi infection and lymphoma. Conclusion. Since the eradication od R. equi is very difficult, it is very important to make the diagnosis and initiate appropriate antibiotic therapy as soon as possible.

GRIM-19 disrupts E6/E6AP complex to rescue p53 and induce apoptosis in cervical cancers.

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Ying Zhou

Full Text Available BACKGROUND: Our previous studies showed a down-regulation of GRIM-19 in primary human cervical cancers, and restoration of GRIM-19 induced tumor regression. The induction of tumor suppressor protein p53 ubiquitination and degradation by E6 oncoportein of high risk-HPV through forming a stable complex with E6AP is considered as a critical mechanism for cervical tumor development. The aims of this study were to determine the potential role of GRIM-19 in rescuing p53 protein and inducing cervical cancer cell apoptosis. METHODOLOGY/PRINCIPAL FINDINGS: The protein levels of GRIM-19 and p53 were detected in normal cervical tissues from 45 patients who underwent hysterectomy for reasons other than neoplasias of either the cervix or endometrium, and cervical cancer tissues from 60 patients with non-metastatic squamous epithelial carcinomas. Coimmunoprecipitation and GST pull-down assay were performed to examine the interaction of GRIM-19 with 18E6 and E6AP in vivo and in vitro respectively. The competition of 18E6 with E6AP in binding GRIM-19 by performing competition pull-down assays was designed to examine the disruption of E6/E6AP complex by GRIM-19. The augment of E6AP ubiquitination by GRIM-19 was detected in vivo and in vitro ubiquitination assay. The effects of GRIM-19-dependent p53 accumulation on cell proliferation, cell cycle, apoptosis were explored by MTT, flow cytometry and transmission electron microscopy respectively. The tumor suppression was detected by xenograft mouse model. CONCLUSION/SIGNIFICANCE: The levels of GRIM-19 and p53 were concurrently down regulated in cervical cancers. The restoration of GRIM-19 can induce ubiquitination and degradation of E6AP, and disrupt the E6/E6AP complex through the interaction of N-terminus of GRIM-19 with both E6 and E6AP, which protected p53 from degradation and promoted cell apoptosis. Tumor xenograft studies also revealed the suppression of p53 degradation in presence of GRIM-19. These data

LKM-1 autoantibodies recognize a short linear sequence in P450IID6, a cytochrome P-450 monooxygenase.

OpenAIRE

Manns, M P; Griffin, K J; Sullivan, K F; Johnson, E F

1991-01-01

LKM-1 autoantibodies, which are associated with autoimmune chronic active hepatitis, recognize P450IID6, a cytochrome P-450 monooxygenase. The reactivities of 26 LKM-1 antisera were tested with a panel of deletion mutants of P450IID6 expressed in Escherichia coli. 22 sera recognize a 33-amino acid segment of P450IID6, and 11 of these recognize a shorter segment, DPAQPPRD. PAQPPR is also found in IE175 of herpes simplex virus type 1 (HSV-1). Antibodies for HSV-1 proteins were detected by ELISA...

Alignment creation in atomic ensembles by elastic electron scattering; the case of 138Ba(...6s6p 1P1) atoms

International Nuclear Information System (INIS)

Trajmar, S.; Kanik, I.; LeClair, L.R.; Khakoo, M.A.; Bray, I.; Fursa, D.; Csanak, G.

1998-01-01

We describe some of our results from a joint experimental and theoretical program concerning elastic electron scattering by 138 Ba(...6s6p 1 P 1 ) atoms. From the experimental results, we derived various scattering parameters and magnetic sublevel specific differential elastic scattering cross sections at impact energy (E 0 ) of 20.0 eV and at scattering angles (θ) of 10deg, 15deg, and 20deg. The same parameters and cross sections were calculated by the convergent close coupling (CCC) approximation and compared to the experimental results. An excellent agreement, found for the two sets of data, gave us confidence in the CCC method and allowed us to extend the angular and energy ranges for the purpose of generating integral elastic scattering cross sections needed for the deduction of the alignment creation cross sections. (J.P.N.)

Biological and physicochemical properties of biosurfactants produced by Lactobacillus jensenii P6A and Lactobacillus gasseri P65.

Science.gov (United States)

Morais, I M C; Cordeiro, A L; Teixeira, G S; Domingues, V S; Nardi, R M D; Monteiro, A S; Alves, R J; Siqueira, E P; Santos, V L

2017-09-19

Lactobacillus species produce biosurfactants that can contribute to the bacteria's ability to prevent microbial infections associated with urogenital and gastrointestinal tracts and the skin. Here, we described the biological and physicochemical properties of biosurfactants produced by Lactobacillus jensenii P 6A and Lactobacillus gasseri P 65 . The biosurfactants produced by L. jensenii P 6A and L. gasseri P 65 reduced the water surface tension from 72 to 43.2 mN m -1 and 42.5 mN m -1 as their concentration increased up to the critical micelle concentration (CMC) values of 7.1 and 8.58 mg mL -1 , respectively. Maximum emulsifying activity was obtained at concentrations of 1 and 5 mg mL -1 for the P 6A and P 65 strains, respectively. The Fourier transform infrared spectroscopy data revealed that the biomolecules consist of a mixture of carbohydrates, lipids and proteins. The gas chromatography-mass spectrum analysis of L. jensenii P 6A biosurfactant showed a major peak for 14-methypentadecanoic acid, which was the main fatty acid present in the biomolecule; conversely, eicosanoic acid dominated the biosurfactant produced by L. gasseri P 65 . Although both biosurfactants contain different percentages of the sugars galactose, glucose and ribose; rhamnose was only detected in the biomolecule produced by L. jensenii P 6A . Emulsifying activities were stable after a 60-min incubation at 100 °C, at pH 2-10, and after the addition of potassium chloride and sodium bicarbonate, but not in the presence of sodium chloride. The biomolecules showed antimicrobial activity against clinical isolates of Escherichia coli and Candida albicans, with MIC values of 16 µg mL -1 , and against Staphylococcus saprophyticus, Enterobacter aerogenes and Klebsiella pneumoniae at 128 µg mL -1 . The biosurfactants also disrupted preformed biofilms of microorganisms at varying concentrations, being more efficient against E. aerogenes (64%) (P 6A biosurfactant), and E. coli (46

The steroid catabolic pathway of the intracellular pathogen Rhodococcus equi is important for pathogenesis and a target for vaccine development.

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R van der Geize

2011-08-01

Full Text Available Rhodococcus equi causes fatal pyogranulomatous pneumonia in foals and immunocompromised animals and humans. Despite its importance, there is currently no effective vaccine against the disease. The actinobacteria R. equi and the human pathogen Mycobacterium tuberculosis are related, and both cause pulmonary diseases. Recently, we have shown that essential steps in the cholesterol catabolic pathway are involved in the pathogenicity of M. tuberculosis. Bioinformatic analysis revealed the presence of a similar cholesterol catabolic gene cluster in R. equi. Orthologs of predicted M. tuberculosis virulence genes located within this cluster, i.e. ipdA (rv3551, ipdB (rv3552, fadA6 and fadE30, were identified in R. equi RE1 and inactivated. The ipdA and ipdB genes of R. equi RE1 appear to constitute the α-subunit and β-subunit, respectively, of a heterodimeric coenzyme A transferase. Mutant strains RE1ΔipdAB and RE1ΔfadE30, but not RE1ΔfadA6, were impaired in growth on the steroid catabolic pathway intermediates 4-androstene-3,17-dione (AD and 3aα-H-4α(3'-propionic acid-5α-hydroxy-7aβ-methylhexahydro-1-indanone (5α-hydroxy-methylhexahydro-1-indanone propionate; 5OH-HIP. Interestingly, RE1ΔipdAB and RE1ΔfadE30, but not RE1ΔfadA6, also displayed an attenuated phenotype in a macrophage infection assay. Gene products important for growth on 5OH-HIP, as part of the steroid catabolic pathway, thus appear to act as factors involved in the pathogenicity of R. equi. Challenge experiments showed that RE1ΔipdAB could be safely administered intratracheally to 2 to 5 week-old foals and oral immunization of foals even elicited a substantial protective immunity against a virulent R. equi strain. Our data show that genes involved in steroid catabolism are promising targets for the development of a live-attenuated vaccine against R. equi infections.

Genome and Proteome Analysis of Rhodococcus erythropolis MI2: Elucidation of the 4,4´-Dithiodibutyric Acid Catabolism

Science.gov (United States)

Khairy, Heba; Meinert, Christina; Wübbeler, Jan Hendrik; Poehlein, Anja; Daniel, Rolf; Voigt, Birgit; Riedel, Katharina; Steinbüchel, Alexander

2016-01-01

Rhodococcus erythropolis MI2 has the extraordinary ability to utilize the xenobiotic 4,4´-dithiodibutyric acid (DTDB). Cleavage of DTDB by the disulfide-reductase Nox, which is the only verified enzyme involved in DTDB-degradation, raised 4-mercaptobutyric acid (4MB). 4MB could act as building block of a novel polythioester with unknown properties. To completely unravel the catabolism of DTDB, the genome of R. erythropolis MI2 was sequenced, and subsequently the proteome was analyzed. The draft genome sequence consists of approximately 7.2 Mbp with an overall G+C content of 62.25% and 6,859 predicted protein-encoding genes. The genome of strain MI2 is composed of three replicons: one chromosome and two megaplasmids with sizes of 6.45, 0.4 and 0.35 Mbp, respectively. When cells of strain MI2 were cultivated with DTDB as sole carbon source and compared to cells grown with succinate, several interesting proteins with significantly higher expression levels were identified using 2D-PAGE and MALDI-TOF mass spectrometry. A putative luciferase-like monooxygenase-class F420-dependent oxidoreductase (RERY_05640), which is encoded by one of the 126 monooxygenase-encoding genes of the MI2-genome, showed a 3-fold increased expression level. This monooxygenase could oxidize the intermediate 4MB into 4-oxo-4-sulfanylbutyric acid. Next, a desulfurization step, which forms succinic acid and volatile hydrogen sulfide, is proposed. One gene coding for a putative desulfhydrase (RERY_06500) was identified in the genome of strain MI2. However, the gene product was not recognized in the proteome analyses. But, a significant expression level with a ratio of up to 7.3 was determined for a putative sulfide:quinone oxidoreductase (RERY_02710), which could also be involved in the abstraction of the sulfur group. As response to the toxicity of the intermediates, several stress response proteins were strongly expressed, including a superoxide dismutase (RERY_05600) and an osmotically induced

Genome and Proteome Analysis of Rhodococcus erythropolis MI2: Elucidation of the 4,4´-Dithiodibutyric Acid Catabolism.

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Heba Khairy

Full Text Available Rhodococcus erythropolis MI2 has the extraordinary ability to utilize the xenobiotic 4,4´-dithiodibutyric acid (DTDB. Cleavage of DTDB by the disulfide-reductase Nox, which is the only verified enzyme involved in DTDB-degradation, raised 4-mercaptobutyric acid (4MB. 4MB could act as building block of a novel polythioester with unknown properties. To completely unravel the catabolism of DTDB, the genome of R. erythropolis MI2 was sequenced, and subsequently the proteome was analyzed. The draft genome sequence consists of approximately 7.2 Mbp with an overall G+C content of 62.25% and 6,859 predicted protein-encoding genes. The genome of strain MI2 is composed of three replicons: one chromosome and two megaplasmids with sizes of 6.45, 0.4 and 0.35 Mbp, respectively. When cells of strain MI2 were cultivated with DTDB as sole carbon source and compared to cells grown with succinate, several interesting proteins with significantly higher expression levels were identified using 2D-PAGE and MALDI-TOF mass spectrometry. A putative luciferase-like monooxygenase-class F420-dependent oxidoreductase (RERY_05640, which is encoded by one of the 126 monooxygenase-encoding genes of the MI2-genome, showed a 3-fold increased expression level. This monooxygenase could oxidize the intermediate 4MB into 4-oxo-4-sulfanylbutyric acid. Next, a desulfurization step, which forms succinic acid and volatile hydrogen sulfide, is proposed. One gene coding for a putative desulfhydrase (RERY_06500 was identified in the genome of strain MI2. However, the gene product was not recognized in the proteome analyses. But, a significant expression level with a ratio of up to 7.3 was determined for a putative sulfide:quinone oxidoreductase (RERY_02710, which could also be involved in the abstraction of the sulfur group. As response to the toxicity of the intermediates, several stress response proteins were strongly expressed, including a superoxide dismutase (RERY_05600 and an

Measurement of A and A/sub nn/ in p + p → p + p at 6 GeV/c

International Nuclear Information System (INIS)

Crosbie, E.A.; Ratner, L.G.; Schultz, P.F.

1980-01-01

Previous measurements of the 6 GeV/c p - p elastic scattering rates have been extended with beam and target polarizations normal to the scattering plane. Improvements to the measuring technique include additional momentum resolution added to the recoil arm and increased solid angle resulting from two-element hodoscopes. Beam intensity was increased by a factor of 3 and additional shielding was installed. Along with the usual statistical uncertainty and errors in measurement of the target polarization, a term was added in quadrature due to a systematic difference between target and beam asymmetries (A/sub T/ and A/sub B/)

Identification of genomic loci associated with Rhodococcus equi susceptibility in foals.

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Cole M McQueen

Full Text Available Pneumonia caused by Rhodococcus equi is a common cause of disease and death in foals. Although agent and environmental factors contribute to the incidence of this disease, the genetic factors influencing the clinical outcomes of R. equi pneumonia are ill-defined. Here, we performed independent single nucleotide polymorphism (SNP- and copy number variant (CNV-based genome-wide association studies to identify genomic loci associated with R. equi pneumonia in foals. Foals at a large Quarter Horse breeding farm were categorized into 3 groups: 1 foals with R. equi pneumonia (clinical group [N = 43]; 2 foals with ultrasonographic evidence of pulmonary lesions that never developed clinical signs of pneumonia (subclinical group [N = 156]; and, 3 foals without clinical signs or ultrasonographic evidence of pneumonia (unaffected group [N = 49]. From each group, 24 foals were randomly selected and used for independent SNP- and CNV-based genome-wide association studies (GWAS. The SNP-based GWAS identified a region on chromosome 26 that had moderate evidence of association with R. equi pneumonia when comparing clinical and subclinical foals. A joint analysis including all study foals revealed a 3- to 4-fold increase in odds of disease for a homozygous SNP within the associated region when comparing the clinical group with either of the other 2 groups of foals or their combination. The region contains the transient receptor potential cation channel, subfamily M, member 2 (TRPM2 gene, which is involved in neutrophil function. No associations were identified in the CNV-based GWAS. Collectively, these data identify a region on chromosome 26 associated with R. equi pneumonia in foals, providing evidence that genetic factors may indeed contribute to this important disease of foals.

Improvement of Biodesulfurization Rate of Alginate Immobilized Rhodococcus erythropolis R1.

Science.gov (United States)

Derikvand, Peyman; Etemadifar, Zahra

2014-03-01

Sulfur oxides released from the burning of oil causes severe environmental pollution. The sulfur can be removed via the 4S pathway in biodesulfurization (BDS). Immobilization approaches have been developed to prevent cell contamination of oil during the BDS process. The encapsulation of Rhodococcus erythropolis R1 in calcium alginate beads was studied in order to enhance conversion of dibenzothiophene (DBT) to 2-hydroxy biphenyl (2-HBP) as the final product. Also the effect of different factors on the BDS process was investigated. Calcium alginate capsules were prepared using peristaltic pumps with different needle sizes to control the beads sizes. Scanning electron microscopy and flow cytometry methods were used to study the distribution and viability of encapsulated cells, respectively. Two non-ionic surfactants and also nano Ƴ-Al2O3were used with the ratio of 0.5% (v/v) and 1:5 (v/v) respectively to investigate their BDS efficiency. In addition, the effect of different bead sizes and different concentrations of sodium alginate in BDS activity was studied. The 2% (w/v) sodium alginate beads with 1.5mm size were found to be the optimum for beads stability and efficient 2-HBP production. The viability of encapsulated cells decreased by 12% after 20 h of desulfurization, compared to free cells. Adding the non-ionic surfactants markedly enhanced the rate of BDS, because of increasing mass transfer of DBT to the gel matrix. In addition, Span 80 was more effective than Tween 80. The nanoƳ-Al2O3 particles could increase BDS rate by up to two-folds greater than that of the control beads. The nano Ƴ-Al2O3 can improve the immobilized biocatalyst for excellent efficiency of DBT desulfurization. Also the BDS activity can be enhanced by setting the other explained factors at optimum levels.

Biochemical and functional analysis of two Plasmodium falciparum blood-stage 6-cys proteins: P12 and P41.

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Tana Taechalertpaisarn

Full Text Available The genomes of Plasmodium parasites that cause malaria in humans, other primates, birds, and rodents all encode multiple 6-cys proteins. Distinct 6-cys protein family members reside on the surface at each extracellular life cycle stage and those on the surface of liver infective and sexual stages have been shown to play important roles in hepatocyte growth and fertilization respectively. However, 6-cys proteins associated with the blood-stage forms of the parasite have no known function. Here we investigate the biochemical nature and function of two blood-stage 6-cys proteins in Plasmodium falciparum, the most pathogenic species to afflict humans. We show that native P12 and P41 form a stable heterodimer on the infective merozoite surface and are secreted following invasion, but could find no evidence that this complex mediates erythrocyte-receptor binding. That P12 and P41 do not appear to have a major role as adhesins to erythrocyte receptors was supported by the observation that antisera to these proteins did not substantially inhibit erythrocyte invasion. To investigate other functional roles for these proteins their genes were successfully disrupted in P. falciparum, however P12 and P41 knockout parasites grew at normal rates in vitro and displayed no other obvious phenotypic changes. It now appears likely that these blood-stage 6-cys proteins operate as a pair and play redundant roles either in erythrocyte invasion or in host-immune interactions.

Polymorphisms of cytochrome P450 2B6 (CYP2B6) in cynomolgus and rhesus macaques.

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Uno, Yasuhiro; Uehara, Shotaro; Yamazaki, Hiroshi

2018-02-22

Cytochrome P450 2B6 (CYP2B6) is an important drug-metabolizing enzyme and is expressed in liver. Although human CYP2B6 variants account for variable enzyme properties among individuals and populations, CYP2B6 genetic variants have not been investigated in cynomolgus macaques, widely used in drug metabolism studies. CYP2B6 was resequenced in 120 cynomolgus macaques and 23 rhesus macaques by direct sequencing. Twenty-three non-synonymous variants were found, of which 12 and 3 were unique to cynomolgus macaques and rhesus macaques, respectively. By functional characterization using the 14 variant proteins, 8 variants (V114I, R253C, M435I, V459M, L465P, C475S, R487C, and R487H) showed different rate (>1.5-fold) of testosterone 16β-hydroxylation to wild type. However, the four variants (M435I, L465P, C475S, and R487H) were analyzed in liver microsomes, and the catalytic rates were not substantially different from wild type. Macaque CYP2B6 was polymorphic, and the genotype could partly account for variable enzyme activities of macaque CYP2B6. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Customer Decision Making in Web Services with an Integrated P6 Model

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Sun, Zhaohao; Sun, Junqing; Meredith, Grant

Customer decision making (CDM) is an indispensable factor for web services. This article examines CDM in web services with a novel P6 model, which consists of the 6 Ps: privacy, perception, propensity, preference, personalization and promised experience. This model integrates the existing 6 P elements of marketing mix as the system environment of CDM in web services. The new integrated P6 model deals with the inner world of the customer and incorporates what the customer think during the DM process. The proposed approach will facilitate the research and development of web services and decision support systems.

P2Y6 Receptor Activation Promotes Inflammation and Tissue Remodeling in Pulmonary Fibrosis

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Müller, Tobias; Fay, Susanne; Vieira, Rodolfo Paula; Karmouty-Quintana, Harry; Cicko, Sanja; Ayata, Cemil Korcan; Zissel, Gernot; Goldmann, Torsten; Lungarella, Giuseppe; Ferrari, Davide; Di Virgilio, Francesco; Robaye, Bernard; Boeynaems, Jean-Marie; Lazarowski, Eduardo R.; Blackburn, Michael R.; Idzko, Marco

2017-01-01

Idiopathic pulmonary fibrosis (IPF) is a disease with a poor prognosis and very few available treatment options. The involvement of the purinergic receptor subtypes P2Y2 and P2X7 in fibrotic lung disease has been demonstrated recently. In this study, we investigated the role of P2Y6 receptors in the pathogenesis of IPF in humans and in the animal model of bleomycin-induced lung injury. P2Y6R expression was upregulated in lung structural cells but not in bronchoalveolar lavage (BAL) cells derived from IPF patients as well as in animals following bleomycin administration. Furthermore, BAL fluid levels of the P2Y6R agonist uridine-5′-diphosphate were elevated in animals with bleomycin-induced pulmonary fibrosis. Inflammation and fibrosis following bleomycin administration were reduced in P2Y6R-deficient compared to wild-type animals confirming the pathophysiological relevance of P2Y6R subtypes for fibrotic lung diseases. Experiments with bone marrow chimeras revealed the importance of P2Y6R expression on lung structural cells for pulmonary inflammation and fibrosis. Similar effects were obtained when animals were treated with the P2Y6R antagonist MRS2578. In vitro studies demonstrated that proliferation and secretion of the pro-inflammatory/pro-fibrotic cytokine IL-6 by lung fibroblasts are P2Y6R-mediated processes. In summary, our results clearly demonstrate the involvement of P2Y6R subtypes in the pathogenesis of pulmonary fibrosis. Thus, blocking pulmonary P2Y6 receptors might be a new target for the treatment of IPF. PMID:28878780

P2Y6 Receptor Activation Promotes Inflammation and Tissue Remodeling in Pulmonary Fibrosis

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Tobias Müller

2017-08-01

Full Text Available Idiopathic pulmonary fibrosis (IPF is a disease with a poor prognosis and very few available treatment options. The involvement of the purinergic receptor subtypes P2Y2 and P2X7 in fibrotic lung disease has been demonstrated recently. In this study, we investigated the role of P2Y6 receptors in the pathogenesis of IPF in humans and in the animal model of bleomycin-induced lung injury. P2Y6R expression was upregulated in lung structural cells but not in bronchoalveolar lavage (BAL cells derived from IPF patients as well as in animals following bleomycin administration. Furthermore, BAL fluid levels of the P2Y6R agonist uridine-5′-diphosphate were elevated in animals with bleomycin-induced pulmonary fibrosis. Inflammation and fibrosis following bleomycin administration were reduced in P2Y6R-deficient compared to wild-type animals confirming the pathophysiological relevance of P2Y6R subtypes for fibrotic lung diseases. Experiments with bone marrow chimeras revealed the importance of P2Y6R expression on lung structural cells for pulmonary inflammation and fibrosis. Similar effects were obtained when animals were treated with the P2Y6R antagonist MRS2578. In vitro studies demonstrated that proliferation and secretion of the pro-inflammatory/pro-fibrotic cytokine IL-6 by lung fibroblasts are P2Y6R-mediated processes. In summary, our results clearly demonstrate the involvement of P2Y6R subtypes in the pathogenesis of pulmonary fibrosis. Thus, blocking pulmonary P2Y6 receptors might be a new target for the treatment of IPF.

Refined study of the interaction between HIV-1 p6 late domain and ALIX

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Gerlier Denis

2008-05-01

Full Text Available Abstract The interaction between the HIV-1 p6 late budding domain and ALIX, a class E vacuolar protein sorting factor, was explored by using the yeast two-hybrid approach. We refined the ALIX binding site of p6 as being the leucine triplet repeat sequence (Lxx4 (LYPLTSLRSLFG. Intriguingly, the deletion of the C-terminal proline-rich region of ALIX prevented detectable binding to p6. In contrast, a four-amino acid deletion in the central hinge region of p6 increased its association with ALIX as shown by its ability to bind to ALIX lacking the proline rich domain. Finally, by using a random screening approach, the minimal ALIX391–510 fragment was found to specifically interact with this p6 deletion mutant. A parallel analysis of ALIX binding to the late domain p9 from EIAV revealed that p6 and p9, which exhibit distinct ALIX binding motives, likely bind differently to ALIX. Altogether, our data support a model where the C-terminal proline-rich domain of ALIX allows the access of its binding site to p6 by alleviating a conformational constraint resulting from the presence of the central p6 hinge.

Molybdenum (6) determination by the method of high-frequency and pH titration

International Nuclear Information System (INIS)

Lukianets, I.G.; Kulish, N.G.

1981-01-01

A possibility to determine Mo(6) using the method of high frequency and pH-metric titration is investigated. By means of the high frequency contact R-cell Mo(6) titration is performed using different precipitators: 8-oxiquinoline, lead nitrate and acetate and silver nitrate. It is established that the best conditions are achieved during titration of 67.2-1343 mg/10 ml Mo(6) with lead acetate (pH 4.5-6.5). Relative standard deviation Ssub(r) constitutes 0.005 pH-metric titration of Mo(6) with complexone 3 is studied. The range of Mo(6) concentrations determined constitutes 13.43-134.3 mg at pH initial value of 3.5-5.5. Relative standard deviation Ssub(r) constitutes 0.003. Techniques of molybdenum determination in ferromolybdenum using the method of high frequency and pH-metric titration are developed [ru

Rigid-rod polyamides and polyimides derived from 4,3 ''-diamino-2 ',6 '-diphenyl- or di(4-biphenylyl)-p-terphenyl and 4-amino-4 ''-carboxy-2 ',6 '-diphenyl-p-terphenyl

NARCIS (Netherlands)

Spiliopoulos, IK; Mikroyannidis, JA; Tsivgoulis, GM

1998-01-01

4,3 "-Diamino-2',6'-diphenyl- or di(4-biphenylyl)p-terphenyl (3a or 3b) and 4-amino-4 "-carboxy-2',6'-diphenyl-p-terphenyl (6) were synthesized through pyrylium salts and used for the preparation of rigid-rod polyamides and polyimides. The polymers were characterized by inherent viscosity, elemental

High-resolution two-photon spectroscopy of a 5 p56 p ←5 p6 transition of xenon

Science.gov (United States)

Altiere, Emily; Miller, Eric R.; Hayamizu, Tomohiro; Jones, David J.; Madison, Kirk W.; Momose, Takamasa

2018-01-01

We report high-resolution Doppler-free two-photon excitation spectroscopy of Xe from the ground state to the 5 p5(P 3 /2 2 ) 6 p [3 /2 ] 2 2 electronic excited state. This is a first step to developing a comagnetometer using polarized 129Xe atoms for planned neutron electric dipole moment measurements at TRIUMF. Narrow linewidth radiation at 252.5 nm produced by a continuous wave laser was built up in an optical cavity to excite the two-photon transition, and the near-infrared emission from the 5 p56 p excited state to the 5 p56 s intermediate electronic state was used to detect the two-photon transition. Hyperfine constants and isotope shift parameters were evaluated and compared with previously reported values. In addition, the detected photon count rate was estimated from the observed intensities.

Improvement of phytoremediation of an aged petroleum hydrocarbon-contaminated soil by Rhodococcus erythropolis CD 106 strain.

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Płociniczak, Tomasz; Fic, Ewa; Pacwa-Płociniczak, Magdalena; Pawlik, Małgorzata; Piotrowska-Seget, Zofia

2017-07-03

The aim of this study was to assess the impact of soil inoculation with the Rhodococcus erythropolis CD 106 strain on the effectiveness of the phytoremediation of an aged hydrocarbon-contaminated [approx. 1% total petroleum hydrocarbon (TPH)] soil using ryegrass (Lolium perenne). The introduction of CD 106 into the soil significantly increased the biomass of ryegrass and the removal of hydrocarbons in planted soil. The fresh weight of the shoots and roots of plants inoculated with CD 106 increased by 49% and 30%, respectively. After 210 days of the experiment, the concentration of TPH was reduced by 31.2%, whereas in the planted, non-inoculated soil, it was reduced by 16.8%. By contrast, the concentration of petroleum hydrocarbon decreased by 18.7% in non-planted soil bioaugmented with the CD 106 strain. The rifampicin-resistant CD 106 strain survived after inoculation into soil and was detected in the soil during the entire experimental period, but the number of CD 106 cells decreased constantly during the enhanced phytoremediation and bioaugmentation experiments. The plant growth-promoting and hydrocarbon-degrading properties of CD 106, which are connected with its long-term survival and limited impact on autochthonous microflora, make this strain a good candidate for improving the phytoremediation efficiency of soil contaminated with hydrocarbons.

Recovery and subsequent characterization of polyhydroxybutyrate from Rhodococcus equi cells grown on crude palm kernel oil

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Nadia Altaee

2016-07-01

Full Text Available The gram-positive bacterium Rhodococcus equi was isolated from fertile soil, and mineral salt media (MM and trace elements were used to provide the necessary elements for its growth and PHB production in addition to using crude palm kernel oil (CPKO 1% as the carbon source. Gas chromatography (GC demonstrated that the composition of the recovered biopolymer was homopolymer polyhydroxybutyrate (PHB. The strain of the present study has a dry biomass of 1.43 (g/l with 38% PHB, as determined by GC. The recovered PHB was characterized by NMR to study the chemical structure. In addition, DSC and TGA were used to study the thermal properties of the recovered polymer, where the melting temperature (Tm was 173 °C, the glass transition temperature (Tg was 2.79 °C, and the decomposition temperature (Td was 276 °C. Gel permeation chromatography (GPC was used to study the molecular mass of the recovered PHB in addition to comparing the results with other studies using different bacteria and substrates, where the molecular weight was 642 kDa, to enable its usage in many applications. The present study demonstrated the use of an inexpensive substrate for PHB production, i.e., using gram-positive bacteria to produce PHB polymer with characterization.

Structural evolution of the P22-like phages: Comparison of Sf6 and P22 procapsid and virion architectures

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Parent, Kristin N. [University of California, San Diego, Department of Chemistry and Biochemistry, La Jolla, CA 92093 (United States); Gilcrease, Eddie B. [University of Utah School of Medicine, Division of Microbiology and Immunology, Department of Pathology, Salt Lake City, UT 84112 (United States); Casjens, Sherwood R., E-mail: sherwood.casjens@path.utah.edu [University of Utah School of Medicine, Division of Microbiology and Immunology, Department of Pathology, Salt Lake City, UT 84112 (United States); Baker, Timothy S., E-mail: tsb@ucsd.edu [University of California, San Diego, Department of Chemistry and Biochemistry, La Jolla, CA 92093 (United States); University of California, San Diego, Division of Biological Sciences, La Jolla, CA 92093 (United States)

2012-06-05

Coat proteins of tailed, dsDNA phages and in herpesviruses include a conserved core similar to the bacteriophage HK97 subunit. This core is often embellished with other domains such as the telokin Ig-like domain of phage P22. Eighty-six P22-like phages and prophages with sequenced genomes share a similar set of virion assembly genes and, based on comparisons of twelve viral assembly proteins (structural and assembly/packaging chaperones), these phages are classified into three groups (P22-like, Sf6-like, and CUS-3-like). We used cryo-electron microscopy and 3D image reconstruction to determine the structures of Sf6 procapsids and virions ({approx} 7 A resolution), and the structure of the entire, asymmetric Sf6 virion (16-A resolution). The Sf6 coat protein is similar to that of P22 yet it has differences in the telokin domain and in its overall quaternary organization. Thermal stability and agarose gel experiments show that Sf6 virions are slightly less stable than those of P22. Finally, bacterial host outer membrane proteins A and C were identified in lipid vesicles that co-purify with Sf6 particles, but are not components of the capsid.

Fasciola spp: Mapping of the MF6 epitope and antigenic analysis of the MF6p/HDM family of heme-binding proteins.

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Victoria Martínez-Sernández

Full Text Available MF6p/FhHDM-1 is a small cationic heme-binding protein which is recognized by the monoclonal antibody (mAb MF6, and abundantly present in parenchymal cells and secreted antigens of Fasciola hepatica. Orthologs of this protein (MF6p/HDMs also exist in other causal agents of important foodborne trematodiasis, such as Clonorchis sinensis, Opisthorchis viverrini and Paragonimus westermani. Considering that MF6p/FhHDM-1 is relevant for heme homeostasis in Fasciola and was reported to have immunomodulatory properties, this protein is expected to be a useful target for vaccination. Thus, in this study we mapped the epitope recognized by mAb MF6 and evaluated its antigenicity in sheep. The sequence of the MF6p/FhHDM-1 ortholog from F. gigantica (MF6p/FgHDM-1 was also reported. By means of ELISA inhibitions with overlapping synthetic peptides, we determined that the epitope recognized by mAb MF6 is located within the C-terminal moiety of MF6p/FhHDM-1, which is the most conserved region of MF6p/HDMs. By immunoblotting analysis of parasite extracts and ELISA inhibitions with synthetic peptides we also determined that mAb MF6 reacted with the same intensity with F. hepatica and F. gigantica, and in decreasing order of intensity with C. sinensis, O.viverrini and P. westermani orthologs. On the contrary, mAb MF6 showed no reactivity against Dicrocoelium dendriticum and Schistosoma mansoni. The study of the recognition of peptides covering different regions of MF6p/FhHDM-1 by sera from immunized sheep revealed that the C-terminal moiety is the most antigenic, thus being of potential interest for vaccination. We also demonstrated that the production of antibodies to MF6p/FhHDM-1 in sheep infected by F. hepatica occurs relatively early and follows the same pattern as those produced against L-cathepsins.

WARM SPITZER OBSERVATIONS OF THREE HOT EXOPLANETS: XO-4b, HAT-P-6b, AND HAT-P-8b

Energy Technology Data Exchange (ETDEWEB)

Todorov, Kamen O. [Department of Astronomy and Astrophysics, The Pennsylvania State University, University Park, PA 16802 (United States); Deming, Drake [Planetary Systems Laboratory, NASA' s Goddard Space Flight Center, Greenbelt, MD 20771 (United States); Knutson, Heather A. [Division of Geological and Planetary Sciences, California Institute of Technology, Pasadena, CA 91125 (United States); Burrows, Adam [Department of Astrophysical Sciences, Princeton University, Princeton, NJ 05844 (United States); Sada, Pedro V. [Department of Physics and Mathematics, University of Monterrey, Monterrey (Mexico); Cowan, Nicolas B. [Department of Physics and Astronomy, Northwestern University, Evanston, IL 60208 (United States); Agol, Eric [Department of Astronomy, University of Washington, Box 351580, Seattle, WA 98195 (United States); Desert, Jean-Michel; Charbonneau, David [Harvard-Smithsonian Center for Astrophysics, Cambridge, MA 02138 (United States); Fortney, Jonathan J.; Laughlin, Gregory [Department of Astronomy and Astrophysics, University of California at Santa Cruz, Santa Cruz, CA 95064 (United States); Langton, Jonathan [Department of Physics, Principia College, Elsah, IL 62028 (United States); Showman, Adam P.; Lewis, Nikole K. [Lunar and Planetary Laboratory, University of Arizona, Tucson, AZ 85721 (United States)

2012-02-10

We analyze Warm Spitzer/Infrared Array Camera observations of the secondary eclipses of three planets, XO-4b, HAT-P-6b, and HAT-P-8b. We measure secondary eclipse amplitudes at 3.6 {mu}m and 4.5 {mu}m for each target. XO-4b exhibits a stronger eclipse depth at 4.5 {mu}m than at 3.6 {mu}m, which is consistent with the presence of a temperature inversion. HAT-P-8b shows a stronger eclipse amplitude at 3.6 {mu}m and is best described by models without a temperature inversion. The eclipse depths of HAT-P-6b can be fitted with models with a small or no temperature inversion. We consider our results in the context of a postulated relationship between stellar activity and temperature inversion and a relationship between irradiation level and planet dayside temperature, as discussed by Knutson et al. and Cowan and Agol, respectively. Our results are consistent with these hypotheses, but do not significantly strengthen them. To measure accurate secondary eclipse central phases, we require accurate ephemerides. We obtain primary transit observations and supplement them with publicly available observations to update the orbital ephemerides of the three planets. Based on the secondary eclipse timing, we set upper boundaries for ecos ({omega}) for HAT-P-6b, HAT-P-8b, and XO-4b and find that the values are consistent with circular orbits.

6-REXOS: Upper Limb Exoskeleton Robot with Improved pHRI

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Malin Gunasekara

2015-04-01

Full Text Available Close interaction can be observed between an exoskeleton robot and its wearer. Therefore, appropriate physical human-robot interaction (pHRI should be considered when designing an exoskeleton robot to provide safe and comfortable motion assistance. Different features have been used in recent studies to enhance the pHRI in upper-limb exoskeleton robots. However, less attention has been given to integrating kinematic redundancy into upper-limb exoskeleton robots to improve the pHRI. In this context, this paper proposes a six-degrees-of-freedom (DoF upper-limb exoskeleton robot (6-REXOS for the motion assistance of physically weak individuals. The 6-REXOS uses a kinematically different structure to that of the human lower arm, where the exoskeleton robot is worn. The 6-REXOS has four active DoFs to generate the motion of the human lower arm. Furthermore, two flexible bellow couplings are attached to the wrist and elbow joints to generate two passive DoFs. These couplings not only allow translational motion in wrist and elbow joints but also a redundancy in the robot. Furthermore, the compliance of the flexible coupling contributes to avoiding misalignments between human and robot joint axes. The redundancy in the 6-REXOS is verified based on manipulability index, minimum singular value, condition number and manipulability ellipsoids. The 6-REXOS and a four-DoF exoskeleton robot are compared to verify the manipulation advantage due to the redundancy. The four-DoF exoskeleton robot is designed by excluding the two passive DoFs of the 6-REXOS. In addition, a kinematic model is proposed for the human lower arm to validate the performance of the 6-REXOS. Kinematic analysis and simulations are carried out to validate the 6-REXOS and human-lower-arm model.

Microglia P2Y6 receptor is related to Parkinson’s disease through neuroinflammatory process

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Xiaodong Yang

2017-02-01

Full Text Available Abstract Background Microglia in the central nervous system (CNS were reported to play crucial role in neurodegeneration. Previous studies showed that P2Y6 receptor (P2Y6R mainly contributed to microglia activation and phagocytosis in CNS. However, the level of P2Y6R in Parkinson’s disease (PD patients is unclear. Therefore, we measured the level of P2Y6R in PD patients and speculated whether it could be a potential biomarker for PD. Given on the basis that P2Y6R was higher in PD patients, we further explored the mechanisms underlying P2Y6R in the pathogenesis of PD. Methods We tested the expression level of P2Y6R in the peripheral blood mononuclear cells (PBMCs among 145 PD patients, 170 healthy controls, and 30 multiple system atrophy (MSA patients. We also used a lipopolysaccharide (LPS-stimulated microglial cell culture model to investigate (i the effects of LPS on P2Y6R expression with western blot and RT-PCR, (ii the effects of LPS on UDP expression using HPLC, (iii the effects of UDP/P2Y6R signaling on cytokine expression using western blot, RT-PCR, and ELISA, and (iv the signaling pathways activated by the P2Y6R involved in the neuroinflammation. Results Expression levels of P2Y6R in PD patients were higher than healthy controls and MSA patients. P2Y6R could be a good biomarker of PD. P2Y6R was also upregulated in LPS-treated BV-2 cells and involved in proinflammatory cytokine release through an autocrine loop based on LPS-triggered UDP secretion and accelerated neuroinflammatory responses through the ERK1/2 pathway. Importantly, blocking UDP/P2Y6R signaling could reverse these pathological processes. Conclusions P2Y6R may be a potential clinical biomarker of PD. Blocking P2Y6R may be a potential therapeutic approach to the treatment of PD patients through inhibition of microglia-activated neuroinflammation.

Spectrofluorimetric determination of benzoimidazolic pesticides: Effect of p-sulfonatocalix[6]arene and cyclodextrins

International Nuclear Information System (INIS)

Pacioni, Natalia L.; Sueldo Occello, Valeria N.; Lazzarotto, Marcio; Veglia, Alicia V.

2008-01-01

The effect of the addition of a macrocyclic host (H) such as p-sulfonatocalix[6]arene (C6S), native and modified cyclodextrins (CDs), on the fluorescence of benzoimidazolic fungicides (P), like Benomyl (BY) and Carbendazim (CZ), has been studied. The fluorescence of BY in water at pH 1.000 and 25.0 deg. C was increased in the presence of C6S, αCD and hydroxypropyl-β-CD (HPCD). The association constants determined by fluorescence enhancement showed weak interactions (K A ∼ 10 1 to 10 2 M -1 ) between the fungicide with both CDs, whereas they were stronger with C6S (K A ∼ 10 5 M -1 ). Molecular recognition of BY for C6S was mainly attributed to electrostatic interactions, and for CDs to the hydrophobic effect and hydrogen bond formation. On the other hand, the fluorescent behaviour of CZ in the presence of C6S at pH 6.994 was interpreted as the formation of two complexes with 1:1 (P:H) and 1:2 (P:H 2 ) stoichiometry, the latter being less fluorescent than the free analyte. Relative fluorescence quantum yield ratios between the complexed and free BY (φ P:H /φ P ) were 2.00 ± 0.05, 1.40 ± 0.03 and 2.8 ± 0.4 for C6S, αCD and HPCD, respectively. The analytical parameters improved in the presence of C6S and CDs. The best limit of detection (L D , ng mL -1 ) was 17.4 ± 0.8 with HPCD. The proposed method with C6S and HPCD was successfully applied to fortified samples of tap water and orange flesh extract with good recoveries (91-106%) and R.S.D. (≤2%) by triplicate analysis. The method is rapid, direct and simple and needs no previous degradation or derivatization reaction

Spectrofluorimetric determination of benzoimidazolic pesticides: Effect of p-sulfonatocalix[6]arene and cyclodextrins

Energy Technology Data Exchange (ETDEWEB)

Pacioni, Natalia L.; Sueldo Occello, Valeria N. [Instituto de Investigaciones en Fisico Quimica de Cordoba (INFIQC), Departamento de Quimica Organica, Facultad de Ciencias Quimicas, Universidad Nacional de Cordoba, Ciudad Universitaria, 5000 Cordoba (Argentina); Lazzarotto, Marcio [Departamento de Quimica Organica, Instituto de Quimica, Universidade Federal do Rio Grande do Sul-UFRGS, 15003 Porto Alegre, R.S. (Brazil); Veglia, Alicia V. [Instituto de Investigaciones en Fisico Quimica de Cordoba (INFIQC), Departamento de Quimica Organica, Facultad de Ciencias Quimicas, Universidad Nacional de Cordoba, Ciudad Universitaria, 5000 Cordoba (Argentina)], E-mail: aveglia@fcq.unc.edu.ar

2008-08-22

The effect of the addition of a macrocyclic host (H) such as p-sulfonatocalix[6]arene (C6S), native and modified cyclodextrins (CDs), on the fluorescence of benzoimidazolic fungicides (P), like Benomyl (BY) and Carbendazim (CZ), has been studied. The fluorescence of BY in water at pH 1.000 and 25.0 deg. C was increased in the presence of C6S, {alpha}CD and hydroxypropyl-{beta}-CD (HPCD). The association constants determined by fluorescence enhancement showed weak interactions (K{sub A} {approx} 10{sup 1} to 10{sup 2} M{sup -1}) between the fungicide with both CDs, whereas they were stronger with C6S (K{sub A} {approx} 10{sup 5} M{sup -1}). Molecular recognition of BY for C6S was mainly attributed to electrostatic interactions, and for CDs to the hydrophobic effect and hydrogen bond formation. On the other hand, the fluorescent behaviour of CZ in the presence of C6S at pH 6.994 was interpreted as the formation of two complexes with 1:1 (P:H) and 1:2 (P:H{sub 2}) stoichiometry, the latter being less fluorescent than the free analyte. Relative fluorescence quantum yield ratios between the complexed and free BY ({phi}{sup P:H}/{phi}{sup P}) were 2.00 {+-} 0.05, 1.40 {+-} 0.03 and 2.8 {+-} 0.4 for C6S, {alpha}CD and HPCD, respectively. The analytical parameters improved in the presence of C6S and CDs. The best limit of detection (L{sub D}, ng mL{sup -1}) was 17.4 {+-} 0.8 with HPCD. The proposed method with C6S and HPCD was successfully applied to fortified samples of tap water and orange flesh extract with good recoveries (91-106%) and R.S.D. ({<=}2%) by triplicate analysis. The method is rapid, direct and simple and needs no previous degradation or derivatization reaction.

Corrosion of carbon steel in the [P_1_4_6_6_6][Br] ionic liquid: The effects of γ-radiation and cover gas

International Nuclear Information System (INIS)

Morco, Ryan P.; Musa, Ahmed Y.; Momeni, Mojtaba; Wren, J.C.

2016-01-01

Highlights: • Carbon steel corrosion in non-aqueous ionic liquid ([P_1_4_6_6_6] [Br]) electrolyte. • Gamma-irradiation results to less corrosion, forming protective oxides. • Substantial corrosion is seen in the absence of gamma-radiation. • A corrosion mechanism is proposed for the observed results. - Abstract: The corrosion of carbon steel in the ionic liquid (IL) [P_1_4_6_6_6] [Br] was studied with the IL in contact with an inert (Ar) or oxidizing (air) cover gas in the presence and absence of γ-radiation. Significant corrosion was observed for the tests performed in the absence of γ-radiation while a protective oxide layer is formed in the presence of γ-radiation. The corrosion is attributed to the presence of impurity H_2O and O_2 dissolved in the IL, and a corrosion mechanism is proposed.

pH dependent green synthesis of gold nanoparticles by completely C6-carboxylated curdlan under high temperature and various pH conditions.

Science.gov (United States)

Qiu, Wen-Yi; Wang, Kai; Wang, Yao-Yao; Ding, Zhi-Chao; Wu, Li-Xia; Cai, Wu-Dan; Yan, Jing-Kun

2018-01-01

A C6-carboxylated curdlan (C6-Cc) obtained from 4-acetamido-TEMPO-mediated oxidation of curdlan was used both as a reducing and stabilizing agent for green synthesis of pH-responsive AuNPs, which was carried out by controlling the pH of the C6-Cc solution at a high temperature (100°C). C6-Cc presented a semi-flexible random coil chain in the aqueous medium at pH 5.5 and became more expanded and rigid in alkaline conditions (pH 7.1-12.0), though the primary chemical structure of C6-Cc was virtually unchanged with the pH variation. The AuNPs prepared with C6-Cc at various pHs were characterized by various instrumental measurements. The shapes and sizes of AuNPs were found to be strongly dependent on the pH of the C6-Cc solution. The C6-Cc-decorated AuNPs exhibited a more well-dispersed spherical morphology with smaller particle sizes under alkaline conditions (pH 7.1-12.0). Through this study, a facile, simple, and green method has been demonstrated for preparation of stimuli-sensitive AuNPs using biocompatible polyanionic polysaccharides. Copyright © 2017 Elsevier B.V. All rights reserved.

The Impact of Polymorphic Variations in the 5p15, 6p12, 6p21 and 15q25 Loci on the Risk and Prognosis of Portuguese Patients with Non-Small Cell Lung Cancer

Science.gov (United States)

de Mello, Ramon Andrade; Ferreira, Mónica; Soares-Pires, Filipa; Costa, Sandra; Cunha, João; Oliveira, Pedro; Hespanhol, Venceslau; Reis, Rui Manuel

2013-01-01

Introduction Polymorphic variants in the 5p15, 6p12, 6p21, and 15q25 loci were demonstrated to potentially contribute to lung cancer carcinogenesis. Therefore, this study was performed to assess the role of those variants in non-small cell lung cancer (NSCLC) risk and prognosis in a Portuguese population. Materials and Methods Blood from patients with NSCLC was prospectively collected. To perform an association study, DNA from these patients and healthy controls were genotyped for a panel of 19 SNPs using a Sequenom® MassARRAY platform. Kaplan-Meier curves were used to assess the overall survival (OS) and progression-free survival (PFS). Results One hundred and forty-four patients with NSCLC were successfully consecutively genotyped for the 19 SNPs. One SNP was associated with NSCLC risk: rs9295740 G/A. Two SNPs were associated with non-squamous histology: rs3024994 (VEGF intron 2) T/C and rs401681 C/T. Three SNPs were associated with response rate: rs3025035 (VEGF intron 7) C/T, rs833061 (VEGF –460) C/T and rs9295740 G/A. One SNP demonstrated an influence on PFS: rs401681 C/T at 5p15, p = 0.021. Four SNPs demonstrated an influence on OS: rs2010963 (VEGF +405 G/C), p = 0.042; rs3025010 (VEGF intron 5 C/T), p = 0.047; rs401681 C/T at 5p15, p = 0.046; and rs31489 C/A at 5p15, p = 0.029. Conclusions Our study suggests that SNPs in the 6p12, 6p21, and 5p15 loci may serve as risk, predictive and prognostic NSCLC biomarkers. In the future, SNPs identified in the genomes of patients may improve NSCLC screening strategies and therapeutic management as well. PMID:24039754

Síntesis del 3-p-metoxibenzoil-4-hidroxi-6-p-metoxifenil-2H-piran-2-ona

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María Soledad Ortiz

2013-05-01

Full Text Available During the study of conditions to prepare p-methoxybenzoylacetic acid methyl ester fromp-methoxyacetophenone andmethyl carbonate using sodium hydride in refluxing ethyl ether, the desired product was obtained with 82% yield. Moreover, a slight variation in reaction conditions, such as solvent and temperature, the 3-p-methoxy benzoyl-4-hydroxi-6-p-methoxyphenyl-2H-pyran-2-one was obtained as an unexpected major product.

One-dimensional zinc selenophosphates: A_2ZnP_2Se_6 (A = K, Rb, Cs)

International Nuclear Information System (INIS)

Haynes, Alyssa S.; Lee, Katherine; Kanatzidis, Mercouri G.

2016-01-01

The new compounds A_2ZnP_2Se_6 (A = K, Rb, Cs) were synthesized via molten salt flux syntheses. The crystals feature one-dimensional "1/_∞[ZnP_2Se_6]"2"- chains charge balanced by alkali metal ions between the chains. K_2ZnP_2Se_6 crystallizes in the monoclinic space group P2_1/c; cell parameters a = 12.537(3) Aa, b = 7.2742(14) Aa, c = 14.164(3) Aa, β = 109.63(3) , Z = 4, and V = 1216.7(4) Aa"3. Rb_2ZnP_2Se_6 and Cs_2ZnP_2Se_6 are isotypic, crystallizing in the triclinic space group P anti 1. Rb_2ZnP_2Se_6 has cell parameters of a = 7.4944(15) Aa, b = 7.6013(15) Aa, c = 12.729(3) Aa, α = 96.57(3) , β = 105.52(3) , γ = 110.54(3) , Z = 2, and V = 636.6(2) Aa"3. Cs_2ZnP_2Se_6 has cell parameters of a = 7.6543(6) Aa, b = 7.7006(6) Aa, c = 12.7373(11) Aa, α = 97.007(7) , β = 104.335(7) , γ = 109.241(6) , Z = 2, and V = 669.54(10) Aa"3. (Copyright copyright 2016 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

Synthesis and reactions of 4-(p-methoxybenzyl-6-[5,6,7,8-tetrahydro-2-naphthyl]-pyridazin-3(2H-one

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NAGWA M.S. EL-DIN HARB

1999-11-01

Full Text Available The condensation of 4-(p-methoxybenzyl-6-[5,6,7,8-tetrahydro-2-naphthyl]-pyridazin-3(2H-one (3, prepared by the reaction of 6-[5,6,7,8-tetrahydro-2-naphthyl]-4,5-dihydropyridazin-3(2H-one (1 and anisaldehyde, with dimethyl sulphate, formaldehyde and acrylonitrile, and also the formation of the Mannich base, proceeded smoothly at the 2-position to give compounds 4,5,6,7, respectively. 4-p-Methoxybenzyl-3-chloro-6-[5,6,7,8-tetrahydro-2-naphthyl]-pyridazine (9 was prepared in law yield by the action of phosphorus oxychloride on 3. The reaction of 9 with benzylamine, aniline and piperidine gave 10a,b,c, respectively. 4-p-Methoxybenzyl-6-[5,6,7,8-tetrahydro-2-napthyl]pyridazine-3(2H-thione (12 was prepared either by the action of thiourea on 9, or by the reaction of 3 with phosphorus pentasulphide. The reaction of these thiones with acrylonitrile, morpholine and piperidine to give 13 and 14 a,b, respectively, were also investigated.

Parity nonconserving asymmetry in p-p scattering up to 6 GeV/c

International Nuclear Information System (INIS)

Oka, Takamitsu.

1981-03-01

A parity nonconserving asymmetry in proton-proton scattering is studied. We calculate the longitudinal analyzing power for the momentum of incident proton up to P sub(L) -- 6 GeV/c. For the weak matrix element, we adopt one-boson-exchange interactions coming from vector mesons (rho 0 and ω) as well as Z-boson. (author)

EVALUACIÓN DE LA BIOTRANSFORMACIÓN DE GERANIOL Y (R-(+-α-PINENO EMPLEANDO CÉLULAS DE Rhodococcus opacus DSM 44313

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JENNIFER PILAR ROJAS

Full Text Available La alta biodisponibilidad de los monoterpenos los hace precursores promisorios en los procesos de biotransformación, mediante los que se producen compuestos de valor agregado que pueden considerarse naturales, debido a que se obtienen por métodos enzimáticos a partir de precursores aislados de la naturaleza. En el presente estudio se evaluó la biotransformación de geraniol y (R-(+-α-pineno empleando la cepa bacteriana Rhodococcus opacus DSM 44313, para ello se determinó la influencia del tiempo de crecimiento de la bacteria, tomando suspensiones celulares en la mitad y finalizando la fase exponencial. También se evaluaron tres tiempos de reacción (12, 24 y 48 h y el efecto de un cosolvente, agregando los sustratos puros y disueltos en etanol al 10%. A partir del geraniol se produjeron geranial, ácido geránico y 6-metil-5-hepten-2-ona, sólo cuando el sustrato se adicionó puro, y se formaron en mayor concentración cuando la bacteria estaba finalizando su fase de crecimiento exponencial. Con el (R-(+-α-pineno se produjo como compuesto principal el (R-(+-cis-verbenol, siendo mayor su concentración agregando el pineno disuelto en etanol, y cuando la bacteria se encontraba en la mitad de su fase de crecimiento exponencial. Los resultados indican que el comportamiento de la bacteria cambia según el sustrato adicionado, debido a las propiedades de cada monoterpeno, pero los compuestos obtenidos con ambos sustratos tienen importantes aplicaciones en las industrias farmacéutica, alimenticia y de perfumería

Photodynamic inactivation of bacteria using polyethylenimine-chlorin(e6) conjugates: Effect of polymer molecular weight, substitution ratio of chlorin(e6) and pH.

Science.gov (United States)

Huang, Liyi; Zhiyentayev, Timur; Xuan, Yi; Azhibek, Dulat; Kharkwal, Gitika B; Hamblin, Michael R

2011-04-01

Antimicrobial photodynamic therapy (APDT) is a novel technique to treat local infections. Previously we reported that the attachment of chlorin(e6) to polyethylenimine (PEI) polymers to form PEI-ce6 conjugates is an effective way to improve ce6 PDT activity against bacteria. The aim of this work was to explore how the polymer molecular weight, substitution ratio (SR) of ce6 and pH value affect the PDT efficacy. We have synthesized PEI-ce6(10) (MW = 60,000, SR = 1) and PEI-ce6(11) (MW = 60,000, SR = 5) and compared these with the previous PEI-ce6(9) (MW = 10,000, SR = 1). We tested the PDT efficacy of these three conjugates against Gram-negative E. coli and Gram-positive bacteria (S. aureus and E. fecalis) at three different pH values (5.0, 7.4, 10.0) that may affect the charge on both the bacterial cells and on the conjugate (that has both basic and acidic groups). PEI-ce6(9) and PEI-ce6(10) were the most effective against these tested bacteria. The PDT effect of all three conjugates depended on pH values. The effective order was pH = 10.0 > pH = 7.4 > pH = 5.0 on E. coli. For S. aureus and E. fecalis the order was pH = 5.0 > pH = 10.0 > pH = 7.4. PEI-ce6(11) PDT activity was worse than PEI-ce6(10) activity which is probably connected to the fact that ce6 molecules are self-quenched within the PEI-ce6(11) molecule. Ce6 quenching within the PEI-ce6 molecules was proved by analyzing fluorescence spectra of PEI-ce6 conjugates at different pH values. There were no differences in bacterial uptake between different pH values in three PEI-ce6 conjugates. We assume high pH (rather than low pH as was hypothesized) disaggregates the conjugates, so the higher pH was more effective than the lower pH against E. coli. But for Gram-positive bacteria, low pH was more effective possibly due to more overall positive charge on the conjugate. Copyright © 2011 Wiley-Liss, Inc.

Improvement of Biodesulfurization Rate of Alginate Immobilized Rhodococcus erythropolis R1

Science.gov (United States)

Derikvand, Peyman; Etemadifar, Zahra

2014-01-01

Background: Sulfur oxides released from the burning of oil causes severe environmental pollution. The sulfur can be removed via the 4S pathway in biodesulfurization (BDS). Immobilization approaches have been developed to prevent cell contamination of oil during the BDS process. Objectives: The encapsulation of Rhodococcus erythropolis R1 in calcium alginate beads was studied in order to enhance conversion of dibenzothiophene (DBT) to 2-hydroxy biphenyl (2-HBP) as the final product. Also the effect of different factors on the BDS process was investigated. Materials and Methods: Calcium alginate capsules were prepared using peristaltic pumps with different needle sizes to control the beads sizes. Scanning electron microscopy and flow cytometry methods were used to study the distribution and viability of encapsulated cells, respectively. Two non-ionic surfactants and also nano Ƴ-Al2O3were used with the ratio of 0.5% (v/v) and 1:5 (v/v) respectively to investigate their BDS efficiency. In addition, the effect of different bead sizes and different concentrations of sodium alginate in BDS activity was studied. Results: The 2% (w/v) sodium alginate beads with 1.5mm size were found to be the optimum for beads stability and efficient 2-HBP production. The viability of encapsulated cells decreased by 12% after 20 h of desulfurization, compared to free cells. Adding the non-ionic surfactants markedly enhanced the rate of BDS, because of increasing mass transfer of DBT to the gel matrix. In addition, Span 80 was more effective than Tween 80. The nanoƳ-Al2O3 particles could increase BDS rate by up to two-folds greater than that of the control beads. Conclusions: The nano Ƴ-Al2O3 can improve the immobilized biocatalyst for excellent efficiency of DBT desulfurization. Also the BDS activity can be enhanced by setting the other explained factors at optimum levels. PMID:25147685

Evidence for presumable feline origin of sporadic G6P[9] rotaviruses in humans.

Science.gov (United States)

Pietsch, Corinna; Liebert, Uwe G

2018-05-31

Species A rotaviruses are highly diverse and impose a substantial burden to human and animal health. Interspecies transmission between livestock, domestic animals and humans is commonly observed, but spread of animal-like rotaviruses within the human population is limited. During the continued monitoring of rotavirus strains in Germany, an unusual G6P[9] rotavirus strain was detected in feces of a child. The complete rotavirus coding sequences revealed a unique G6-P[9]-I2-R2-C2-M2-A3-N2-T3-E2-H3 genotype constellation. The virus was phylogenetically related to feline G3P[9] strains and other human G6P[9] rotaviruses of presumable zoonotic origin. Analysis of primer binding sites of G6 specific genotyping revealed further evidence of a G6P[9] feline reservoir. Moreover, substantial deficits of conventional semi-nested PCR genotyping approaches in detecting contemporary G6P[9] were revealed. Rotavirus strain GER29-14 most likely resulted from a direct or recent interspecies transmission from a cat to human. Further studies could assess nucleic acid sequences and genotype constellations of feline rotavirus to confirm the likely feline origin of sporadic human G6P[9] strains. Copyright © 2018 Elsevier B.V. All rights reserved.

P450 reductase and cytochrome b5 interactions with cytochrome P450: Effects on house fly CYP6A1 catalysis

OpenAIRE

Murataliev, Marat B.; Guzov, Victor M.; Walker, F. Ann; Feyereisen, René

2008-01-01

The interactions of protein components of the xenobiotic-metabolizing cytochrome P450 system, CYP6A1, P450 reductase, and cytochrome b5 from the house fly (Musca domestica) have been characterized. CYP6A1 activity is determined by the concentration of the CYP6A1-P450 reductase complex, regardless of which protein is present in excess. Both holo- and apo-b5 stimulated CYP6A1 heptachlor epoxidase and steroid hydroxylase activities and influenced the regioselectivity of testosterone hydroxylatio...

Distribution of a Nocardia brasiliensis Catalase Gene Fragment in Members of the Genera Nocardia, Gordona, and Rhodococcus

Science.gov (United States)

Vera-Cabrera, Lucio; Johnson, Wendy M.; Welsh, Oliverio; Resendiz-Uresti, Francisco L.; Salinas-Carmona, Mario C.

1999-01-01

An immunodominant protein from Nocardia brasiliensis, P61, was subjected to amino-terminal and internal sequence analysis. Three sequences of 22, 17, and 38 residues, respectively, were obtained and compared with the protein database from GenBank by using the BLAST system. The sequences showed homology to some eukaryotic catalases and to a bromoperoxidase-catalase from Streptomyces violaceus. Its identity as a catalase was confirmed by analysis of its enzymatic activity on H2O2 and by a double-staining method on a nondenaturing polyacrylamide gel with 3,3′-diaminobenzidine and ferricyanide; the result showed only catalase activity, but no peroxidase. By using one of the internal amino acid sequences and a consensus catalase motif (VGNNTP), we were able to design a PCR assay that generated a 500-bp PCR product. The amplicon was analyzed, and the nucleotide sequence was compared to the GenBank database with the observation of high homology to other bacterial and eukaryotic catalases. A PCR assay based on this target sequence was performed with primers NB10 and NB11 to confirm the presence of the NB10-NB11 gene fragment in several N. brasiliensis strains isolated from mycetoma. The same assay was used to determine whether there were homologous sequences in several type strains from the genera Nocardia, Rhodococcus, Gordona, and Streptomyces. All of the N. brasiliensis strains presented a positive result but only some of the actinomycetes species tested were positive in the PCR assay. In order to confirm these findings, genomic DNA was subjected to Southern blot analysis. A 1.7-kbp band was observed in the N. brasiliensis strains, and bands of different molecular weight were observed in cross-reacting actinomycetes. Sequence analysis of the amplicons of selected actinomycetes showed high homology in this catalase fragment, thus demonstrating that this protein is highly conserved in this group of bacteria. PMID:10325357

IL-6 Impairs Myogenic Differentiation by Downmodulation of p90RSK/eEF2 and mTOR/p70S6K Axes, without Affecting AKT Activity

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Michele Pelosi

2014-01-01

Full Text Available IL-6 is a multifaceted pleiotropic cytokine, which is produced by a variety of cell types and targets different cells and tissues. In physiological conditions, IL-6 can be locally and transiently produced by skeletal muscle and plays an important role in muscle homeostasis. Circulating IL-6 levels are normally very low or undetectable but are dramatically increased in several pathologic conditions. In this study, we aimed to define the potential molecular mechanisms underlying the effects of IL-6 on myogenic program. We explored the molecular mechanisms through which exogenous IL-6, or the conditioned medium from the murine C-26 adenocarcinoma cells (a cellular model that secretes high levels of IL-6 and induces cancer cachexia in mice, interferes with the myogenic program. Our study revealed that IL-6 induces the activation of the Stat3 signaling and promotes the downmodulation of the p90RSK/eEF2 and mTOR/p70S6K axes, while it does not affect the activation of AKT. We thus identified potential molecular mediators of the inhibitory effects of IL-6 on myogenic program.

Chloromuconolactone dehalogenase ClcF of actinobacteria.

Science.gov (United States)

Solyanikova, Inna P; Plotnikova, Elena G; Shumkova, Ekaterina S; Robota, Irina V; Prisyazhnaya, Natalya V; Golovleva, Ludmila A

2014-01-01

This work investigated the distribution of the clcF gene in actinobacteria isolated from different ecotopes. The gene encodes chloromuconolactone dehalogenase (CMLD) ClcF, the enzyme found to date in only one representative of Gram-positive bacteria, Rhodococcus opacus 1CP, adapted to 2-chlorophenol (2CP). Using primers specific to the clcF gene, from the DNA matrix of rhodococcal strains closely related to species Rhodococcus wratislaviensis (P1, P12, P13, P20, G10, KT112, KT723, BO1) we obtained PCR products whose nucleotide sequences were 100% identical to that of the clcF gene from strain R. opacus 1CP. CMLDs isolated from the biomass of strains Rhodococcus spp. G10 and P1 grown on 2CP did not differ by their subunit molecular mass deduced from the known amino acid sequence of the clcF gene from the ClcF of strain R. opacus 1CP. Matrix-assisted laser dissociation/ionization time-of-flight mass spectrometry showed the presence of a peak with m/z 11,194-11,196 Da both in whole cells and in protein solutions with a ClcF activity. Thus, we have first time shown the distribution of ClcF among actinobacteria isolated from geographically distant habitats.

Measurements of spin parameters in p-p elastic scattering at 6 GeV/c

International Nuclear Information System (INIS)

Linn, S.L.; Perlmutter, A.; Crosbie, E.A.; Ratner, L.G.; Schultz, P.F.; O'Fallon, J.R.; Cameron, P.R.; Crabb, D.G.; Fernow, R.C.; Hansen, P.H.; Krisch, A.D.; Salthouse, A.J.; Sandler, B.; Shima, T.; Terwilliger, K.M.

1982-01-01

We measured the differential cross section for proton-proton elastic scattering in 6 GeV/c, with both initial spins oriented normal to the scattering plane. The analyzing power A shows significant structure with a large broad peak reaching about 24% near P/sub perpendicular/ 2 = 1.6 (GeV/c) 2 . The spin-spin correlation parameter A/sub n/n exhibits more dramatic structure, with a small but very sharp peak rising rapidly to about 13% at 90 0 /sub tsc.m./. This sharp peak may be caused by particle-identity effects

Rational evolution of the unusual Y-type oxyanion hole of Rhodococcus sp. CR53 lipase LipR.

Science.gov (United States)

Infanzón, Belén; Sotelo, Pablo H; Martínez, Josefina; Diaz, Pilar

2018-01-01

Rhodococcus sp CR-53 lipase LipR was the first characterized member of bacterial lipase family X. Interestingly, LipR displays some similarity with α/β-hydrolases of the C. antartica lipase A (CAL-A)-like superfamily (abH38), bearing a Y-type oxyanion hole, never found before among bacterial lipases. In order to explore this unusual Y-type oxyanion hole, and to improve LipR performance, two modification strategies based on site directed or saturation mutagenesis were addressed. Initially, a small library of mutants was designed to convert LipR Y-type oxyanion hole (YDS) into one closer to those most frequently found in bacteria (GGG(X)). However, activity was completely lost in all mutants obtained, indicating that the Y-type oxyanion hole of LipR is required for activity. A second approach was addressed to modify the two main oxyanion hole residues Tyr 110 and Asp 111 , previously described for CAL-A as the most relevant amino acids involved in stabilization of the enzyme-substrate complex. A saturation mutagenesis library was prepared for each residue (Tyr 110 and Asp 111 ), and activity of the resulting variants was assayed on different chain length substrates. No functional LipR variants could be obtained when Tyr 110 was replaced by any other amino acids, indicating that this is a crucial residue for catalysis. However, among the Asp 111 variants obtained, LipR D111G produced a functional enzyme. Interestingly, this LipR-YGS variant showed less activity than wild type LipR on short- or mid- chain substrates but displayed a 5.6-fold increased activity on long chain length substrates. Analysis of the 3D model and in silico docking studies of this enzyme variant suggest that substitution of Asp by Gly produces a wider entrance tunnel that would allow for a better and tight accommodation of larger substrates, thus justifying the experimental results obtained. Copyright © 2017 Elsevier Inc. All rights reserved.

Optical frequency measurements of 6s 2S1/2-6p 2P3/2 transition in a 133Cs atomic beam using a femtosecond laser frequency comb

International Nuclear Information System (INIS)

Gerginov, V.; Tanner, C.E.; Diddams, S.; Bartels, A.; Hollberg, L.

2004-01-01

Optical frequencies of the hyperfine components of the D 2 line in 133 Cs are determined using high-resolution spectroscopy and a femtosecond laser frequency comb. A narrow-linewidth probe laser excites the 6s 2 S 1/2 (F=3,4)→6p 2 P 3/2 (F=2,3,4,5) transition in a highly collimated atomic beam. Fluorescence spectra are taken by scanning the laser frequency over the excited-state hyperfine structure. The laser optical frequency is referenced to a Cs fountain clock via a reference laser and a femtosecond laser frequency comb. A retroreflected laser beam is used to estimate and minimize the Doppler shift due to misalignment between the probe laser and the atomic beam. We achieve an angular resolution on the order of 5x10 -6 rad. The final uncertainties (∼±5 kHz) in the frequencies of the optical transitions are a factor of 20 better than previous results [T. Udem et al., Phys. Rev. A 62, 031801 (2000).]. We find the centroid of the 6s 2 S 1/2 →6p 2 P 3/2 transition to be f D2 =351 725 718.4744(51) MHz

Associations between physical examination, laboratory, and radiographic findings and outcome and subsequent racing performance of foals with Rhodococcus equi infection: 115 cases (1984-1992)

International Nuclear Information System (INIS)

Ainsworth, D.M.; Eicker, S.W.; Yeager, A.E.; Sweeney, C.R.; Viel, L.; Tesarowski, D.; Lavoie, J.P.; Hoffman, A.; Paradis, M.R.; Reed, S.M.

1998-01-01

Objective-To determine whether physical examination, laboratory, or radiographic abnormalities in foals with Rhodococcus equi infection were associated with survival, ability to race at least once after recovery, or, for foals that survived and went on to race, subsequent Facing performance. Design-Retrospective study. Animals-49 Thoroughbreds and 66 Standardbreds admitted to 1 of 6 veterinary teaching hospitals between 1984 and 1992 in which R equi infection was positively diagnosed. Procedure-Results of physical examination, laboratory testing, and thoracic radiography were reviewed. Indices of Facing performance were obtained for feats that recovered and eventually raced and compared with values for the US racing population. Results-83 (72%) feats survived. Foals that did not survive were more likely to have extreme tachycardia (heart rate > 100 beats/min), be in respiratory distress, and have severe radiographic abnormalities on thoracic radiographs at the time of initial examination than were foals that survived. Clinicopathologic abnormalities were not associated with whether feats did or did not survive. Forty-five of the 83 surviving foals (54%) eventually raced at least once, but none of the factors examined was associated with whether foals went on to race. Racing performance of foals that raced as adults was not significantly different from that of the US racing population. Clinical Implications-R equi infection in foals is associated with a decreased chance of racing as an adult, however, foals that eventually go on to race perform comparably to the US racing population

MSH6 Mutations are Frequent in Hereditary Nonpolyposis Colorectal Cancer Families With Normal pMSH6 Expression as Detected by Immunohistochemistry

DEFF Research Database (Denmark)

Okkels, Henrik; Larsen, K.L.; Thorlacius-Ussing, O.

2012-01-01

INTRODUCTION:: Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal dominant condition accounting for 2% to 4% of all colorectal cancer cases worldwide. Families with germ line mutations in 1 of 6 mismatch repair genes are known as Lynch syndrome families. The largest number...... this approach in Lynch families carrying mutations in MSH6. MATERIALS AND METHODS:: Results of the screening of the MSH6 gene in HNPCC families were compared with those obtained on immunohistochemical protein analysis. RESULTS:: In 56 (7%) of 815 families, at least 1 MSH6 mutation, 23 definitively pathogenic...... be detected, whereas in 34.5% pMSH6 was present and pMLH1/pPMS2 was absent. CONCLUSIONS:: If genetic screening of HNPCC families depended on immunohistochemical results, a substantial number of families harboring a pathogenic mutation in MSH6 and the vast majority of families harboring an MSH6 unclassified...

Characterization of P1 promoter activity of the β-galactoside α2,6 ...

Indian Academy of Sciences (India)

2012-04-05

Apr 5, 2012 ... The level of β-galactoside α2,6-sialyltransferase I (ST6Gal I) mRNA, encoded by the gene siat1, is increased in malignant tissues. Expression is regulated by different promoters – P1, P2 and P3 – generating three mRNA isoforms. H, X and YZ. In cervical cancer tissue the mRNA isoform H, which results ...

Characterization of the Rhodococcus sp. MK1 strain and its pilot application for bioremediation of diesel oil-contaminated soil.

Science.gov (United States)

Kis, Ágnes Erdeiné; Laczi, Krisztián; Zsíros, Szilvia; Kós, Péter; Tengölics, Roland; Bounedjoum, Naila; Kovács, Tamás; Rákhely, Gábor; Perei, Katalin

2017-12-01

Petroleum hydrocarbons and derivatives are widespread contaminants in both aquifers and soil, their elimination is in the primary focus of environmental studies. Microorganisms are key components in biological removal of pollutants. Strains capable to utilize hydrocarbons usually appear at the contaminated sites, but their metabolic activities are often restricted by the lack of nutrients and/or they can only utilize one or two components of a mixture. We isolated a novel Rhodococcus sp. MK1 strain capable to degrade the components of diesel oil simultaneously. The draft genome of the strain was determined and besides the chromosome, the presence of one plasmid could be revealed. Numerous routes for oxidation of aliphatic and aromatic compounds were identified. The strain was tested in ex situ applications aiming to compare alternative solutions for microbial degradation of hydrocarbons. The results of bioaugmentation and biostimulation experiments clearly demonstrated that - in certain cases - the indigenous microbial community could be exploited for bioremediation of oil-contaminated soils. Biostimulation seems to be efficient for removal of aged contaminations at lower concentration range, whereas bioaugmentation is necessary for the treatment of freshly and highly polluted sites.

Synthesis and crystal structure of Bi6.4Pb0.6P2O15.2

International Nuclear Information System (INIS)

Arumugam, N.; Lynch, V.; Steinfink, H.

2007-01-01

Bi 6.4 Pb 0.6 P 2 O 15.2 is a polymorph of structures with the general stoichiometry Bi 6+x M 1-x P 2 O 15+y . However, unlike previously published structures that consist of layers formed by edge sharing OBi 4 tetrahedra bridged by PO 4 and TO 6 (T=transition metal) tetrahedra and octahedra the title compound's structure is more complex. It is monoclinic, C2, a=19.4698(4) A, b=11.3692(3) A, c=16.3809(5) A, β=101.167(1) o , Z=10. Single-crystal X-ray diffraction data were refined by least squares on F 2 converging to R 1 =0.0387, wR 2 =0.0836 for 7023 intensities. The crystal twins by mirror reflection across (001) as the twin plane and twin component 1 equals 0.74(1). Oxygen ions are in tetrahedral coordination to four metal ions and the O(BiPb) 4 units share corners to form layers that are part of the three-dimensional framework. Eight oxygen ions form a cube around the two crystallographically independent Pb ions. Pb-O bond lengths vary from 2.265(14) to 2.869(14) A. Pairs of such cubes share an edge to form a Pb 3 O 20 unit. The two oxygen ions from the unshared edges are part of irregular Bi polyhedra. Other oxygen ions of Bi polyhedra are part only of O(BiPb) 4 units, and some oxygen ions of the polyhedra are also part of PO 4 tetrahedra. One, two, three and or four PO 4 moieties are connected to the Bi polyhedra. Bi-O bond lengths ≤3.1 A vary from 2.090(12) to 3.07(3) A. The articulations of Pb cubes, Bi polyhedra and PO 4 tetrahedra link into the three-dimensional structure. - Graphical abstract: View of the structure of Bi 6.4 Pb 0.6 P 2 O 15.2 parallel to the b-axis

Hsp60 and p70S6K form a complex in human cardiomyocytes

Directory of Open Access Journals (Sweden)

Kroupskaya I. V.

2011-02-01

Full Text Available Molecular chaperon Hsp60 and protein kinase p70S6K play an important functional role in the regulation of cardiomyocytes vital function or apoptosis. Aim. To study a possibility of in vivo complex formation between Hsp60 and p70S6K in cardiomyocytes. Methods. Co-immunoprecipitation, Western-blot analysis. Results. We have identified in vivo interaction between molecular chaperone Hsp60 and two isoforms of proteinkinase p70S6K in human myocardium, normal and affected by cardiomyopathy. Conclusions. The results obtained suggest a possible participation of molecular chaperon Hsp60 in regulation of p70S6K activity in stressinduced apoptotic signaling pathway in cardiomyocytes.

Rice black-streaked dwarf virus P6 self-interacts to form punctate, viroplasm-like structures in the cytoplasm and recruits viroplasm-associated protein P9-1

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Yu Jialin

2011-01-01

Full Text Available Abstract Background Rice black-streaked dwarf virus (RBSDV, a member of the genus Fijivirus within the family Reoviridae, can infect several graminaceous plant species including rice, maize and wheat, and is transmitted by planthoppers. Although several RBSDV proteins have been studied in detail, functions of the nonstructural protein P6 are still largely unknown. Results In the current study, we employed yeast two-hybrid assays, bimolecular fluorescence complementation and subcellular localization experiments to show that P6 can self-interact to form punctate, cytoplasmic viroplasm-like structures (VLS when expressed alone in plant cells. The region from residues 395 to 659 is necessary for P6 self-interaction, whereas two polypeptides (residues 580-620 and 615-655 are involved in the subcellular localization of P6. Furthermore, P6 strongly interacts with the viroplasm-associated protein P9-1 and recruits P9-1 to localize in VLS. The P6 395-659 region is also important for the P6-P9-1 interaction, and deleting any region of P9-1 abolishes this heterologous interaction. Conclusions RBSDV P6 protein has an intrinsic ability to self-interact and forms VLS without other RBSDV proteins or RNAs. P6 recruits P9-1 to VLS by direct protein-protein interaction. This is the first report on the functionality of RBSDV P6 protein. P6 may be involved in the process of viroplasm nucleation and virus morphogenesis.

Human herpesvirus 6B inhibits cell proliferation by a p53-independent pathway

DEFF Research Database (Denmark)

Øster, Bodil; Kaspersen, M.D.; Kofod-Olsen, Emil

2006-01-01

BACKGROUND: Various forms of cellular stress can activate the tumour suppressor protein p53, an important regulator of cell cycle arrest, apoptosis, and cellular senescence. Cells infected by human herpesvirus 6B (HHV-6B) accumulate aberrant amounts of p53. OBJECTIVES: The aim of this study...

Specific binding of a Pop6/Pop7 heterodimer to the P3 stem of the yeast RNase MRP and RNase P RNAs.

Science.gov (United States)

Perederina, Anna; Esakova, Olga; Koc, Hasan; Schmitt, Mark E; Krasilnikov, Andrey S

2007-10-01

Pop6 and Pop7 are protein subunits of Saccharomyces cerevisiae RNase MRP and RNase P. Here we show that bacterially expressed Pop6 and Pop7 form a soluble heterodimer that binds the RNA components of both RNase MRP and RNase P. Footprint analysis of the interaction between the Pop6/7 heterodimer and the RNase MRP RNA, combined with gel mobility assays, demonstrates that the Pop6/7 complex binds to a conserved region of the P3 domain. Binding of these proteins to the MRP RNA leads to local rearrangement in the structure of the P3 loop and suggests that direct interaction of the Pop6/7 complex with the P3 domain of the RNA components of RNases MRP and P may mediate binding of other protein components. These results suggest a role for a key element in the RNase MRP and RNase P RNAs in protein binding, and demonstrate the feasibility of directly studying RNA-protein interactions in the eukaryotic RNases MRP and P complexes.

ARF6-dependent regulation of P2Y receptor traffic and function in human platelets.

Science.gov (United States)

Kanamarlapudi, Venkateswarlu; Owens, Sian E; Saha, Keya; Pope, Robert J; Mundell, Stuart J

2012-01-01

Adenosine diphosphate (ADP) is a critical regulator of platelet activation, mediating its actions through two G protein-coupled receptors, the P2Y(1) and P2Y(12) purinoceptors. Recently, we demonstrated that P2Y(1) and P2Y(12) purinoceptor activities are rapidly and reversibly modulated in human platelets, revealing that the underlying mechanism requires receptor internalization and subsequent trafficking as an essential part of this process. In this study we investigated the role of the small GTP-binding protein ADP ribosylation factor 6 (ARF6) in the internalization and function of P2Y(1) and P2Y(12) purinoceptors in human platelets. ARF6 has been implicated in the internalization of a number of GPCRs, although its precise molecular mechanism in this process remains unclear. In this study we show that activation of either P2Y(1) or P2Y(12) purinoceptors can stimulate ARF6 activity. Further blockade of ARF6 function either in cell lines or human platelets blocks P2Y purinoceptor internalization. This blockade of receptor internalization attenuates receptor resensitization. Furthermore, we demonstrate that Nm23-H1, a nucleoside diphosphate (NDP) kinase regulated by ARF6 which facilitates dynamin-dependent fission of coated vesicles during endocytosis, is also required for P2Y purinoceptor internalization. These data describe a novel function of ARF6 in the internalization of P2Y purinoceptors and demonstrate the integral importance of this small GTPase upon platelet ADP receptor function.

ARF6-dependent regulation of P2Y receptor traffic and function in human platelets.

Directory of Open Access Journals (Sweden)

Venkateswarlu Kanamarlapudi

Full Text Available Adenosine diphosphate (ADP is a critical regulator of platelet activation, mediating its actions through two G protein-coupled receptors, the P2Y(1 and P2Y(12 purinoceptors. Recently, we demonstrated that P2Y(1 and P2Y(12 purinoceptor activities are rapidly and reversibly modulated in human platelets, revealing that the underlying mechanism requires receptor internalization and subsequent trafficking as an essential part of this process. In this study we investigated the role of the small GTP-binding protein ADP ribosylation factor 6 (ARF6 in the internalization and function of P2Y(1 and P2Y(12 purinoceptors in human platelets. ARF6 has been implicated in the internalization of a number of GPCRs, although its precise molecular mechanism in this process remains unclear. In this study we show that activation of either P2Y(1 or P2Y(12 purinoceptors can stimulate ARF6 activity. Further blockade of ARF6 function either in cell lines or human platelets blocks P2Y purinoceptor internalization. This blockade of receptor internalization attenuates receptor resensitization. Furthermore, we demonstrate that Nm23-H1, a nucleoside diphosphate (NDP kinase regulated by ARF6 which facilitates dynamin-dependent fission of coated vesicles during endocytosis, is also required for P2Y purinoceptor internalization. These data describe a novel function of ARF6 in the internalization of P2Y purinoceptors and demonstrate the integral importance of this small GTPase upon platelet ADP receptor function.

Rhodococcus erythropolis and Its γ-Lactone Catabolic Pathway: An Unusual Biocontrol System That Disrupts Pathogen Quorum Sensing Communication

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Xavier Latour

2013-12-01

Full Text Available Rhodococcus erythropolis is an environmental Gram-positive Actinobacterium with a versatile metabolism involved in various bioconversions and degradations. Rhodococci are best known for their great potential in numerous decontamination and industrial processes. However, they can also prevent plant disease by disrupting quorum sensing-based communication of Gram-negative soft-rot bacteria, by degrading N-acyl-homoserine lactone signaling molecules. Such biocontrol activity results partly from the action of the γ-lactone catabolic pathway. This pathway is responsible for cleaving the lactone bond of a wide range of compounds comprising a γ-butyrolactone ring coupled to an alkyl or acyl chain. The aliphatic products of this hydrolysis are then activated and enter fatty acid metabolism. This short pathway is controlled by the presence of the γ-lactone, presumably sensed by a TetR-like transcriptional regulator, rather than the presence of the pathogen or the plant-host in the environment of the Rhodococci. Both the density and biocontrol activity of R. erythropolis may be boosted in crop systems. Treatment with a cheap γ-lactone stimulator, for example, the food flavoring γ-caprolactone, induces the activity in the biocontrol agent, R. erythropolis, of the pathway degrading signaling molecules; such treatments thus promote plant protection.

TAF6delta controls apoptosis and gene expression in the absence of p53.

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Emmanuelle Wilhelm

Full Text Available BACKGROUND: Life and death decisions of metazoan cells hinge on the balance between the expression of pro- versus anti-apoptotic gene products. The general RNA polymerase II transcription factor, TFIID, plays a central role in the regulation of gene expression through its core promoter recognition and co-activator functions. The core TFIID subunit TAF6 acts in vitro as an essential co-activator of transcription for the p53 tumor suppressor protein. We previously identified a splice variant of TAF6, termed TAF6delta that can be induced during apoptosis. METHODOLOGY/PRINCIPAL FINDINGS: To elucidate the impact of TAF6delta on cell death and gene expression, we have employed modified antisense oligonucleotides to enforce expression of endogenous TAF6delta. The induction of endogenous TAF6delta triggered apoptosis in tumor cell lines, including cells devoid of p53. Microarray experiments revealed that TAF6delta activates gene expression independently of cellular p53 status. CONCLUSIONS: Our data define TAF6delta as a pivotal node in a signaling pathway that controls gene expression programs and apoptosis in the absence of p53.

One step beyond the electric dipole approximation: An experiment to observe the 5p → 6p forbidden transition in atomic rubidium

Science.gov (United States)

Ponciano-Ojeda, F.; Hernández-Gómez, S.; Mojica-Casique, C.; Ruiz-Martínez, E.; López-Hernández, O.; Colín-Rodríguez, R.; Ramírez-Martínez, F.; Flores-Mijangos, J.; Sahagún, D.; Jáuregui, R.; Jiménez-Mier, J.

2018-01-01

An advanced undergraduate experiment to study the 5 P 3 / 2 → 6 P 3 / 2 electric quadrupole transition in rubidium atoms is presented. The experiment uses two external cavity diode lasers, one operating at the D2 rubidium resonance line and the other built with commercial parts to emit at 911 nm. The lasers produce the 5 s → 5 p → 6 p excitation sequence in which the second step is the forbidden transition. Production of atoms in the 6 P 3 / 2 state is observed by detection of the 420 nm fluorescence that results from electric dipole decay into the ground state. Lines whose widths are significantly narrower than the Doppler width are used to study the hyperfine structure of the 6 P 3 / 2 state in rubidium. The spectra illustrate characteristics unique to electric dipole forbidden transitions, like the electric quadrupole selection rules; they are also used to show general aspects of two-color laser spectroscopy such as velocity selection and hyperfine pumping.

Study on the inclusion behavior of p-sulfonatocalix[6]arene with propranolol by spectrofluorometry

Science.gov (United States)

Li, Hui; Song, Jin-Ping; Chao, Jian-Bin; Shuang, Shao-Min; Dong, Chuan

2012-11-01

The inclusion interaction between propranolol (PPL) and p-sulfonatocalix[6]arene (SCX6) was investigated by fluorescence and 1H NMR spectroscopy. Influences of pH, temperature, ionic strength and the concentration of SCX6 were examined in detail. In phosphate buffer solution with pH 7.5, the fluorescence of PPL dramatically quenched upon addition of SCX6 revealing the formation of inclusion complexes between PPL and SCX6. The stoichiometric ratio was verified to be 1:1 by the continuous variation method. The inclusion constant of PPL-SCX6 complexes was calculated as 2.2 × 104 L/mol by the nonlinear curve fitting method. 1H NMR titration spectra testified that the aliphatic chain of PPL may be partially penetrated into the hydrophobic cavity of SCX6. This was confirmed by molecular dynamics calculations.

A Unique Failure Mechanism in the Nexus 6P Lithium-Ion Battery

OpenAIRE

Saurabh Saxena; Yinjiao Xing; Michael Pecht

2018-01-01

Nexus 6P smartphones have been beset by battery drain issues, which have been causing premature shutdown of the phone even when the charge indicator displays a significant remaining runtime. To investigate the premature battery drain issue, two Nexus 6P smartphones (one new and one used) were disassembled and their batteries were evaluated using computerized tomography (CT) scan analysis, electrical performance (capacity, resistance, and impedance) tests, and cycle life capacity fade tests. T...

Human herpesvirus 6B induces phosphorylation of p53 in its regulatory domain by a CK2- and p38-independent pathway

DEFF Research Database (Denmark)

Øster, Bodil; Bundgaard, B; Hupp, T R

2008-01-01

Here, we demonstrate that human herpesvirus 6B (HHV-6B) infection upregulates the tumour suppressor p53 and induces phosphorylation of p53 at Ser392. Interestingly, phosphorylation at the equivalent site has previously been shown to correlate with p53 tumour suppression in murine models. Although...

Heterologous expression of Helicoverpa armigera cytochrome P450 CYP6B7 in Pichia pastoris and interactions of CYP6B7 with insecticides.

Science.gov (United States)

Zhao, Chunqing; Song, Genmiao; Duan, Hongxia; Tang, Tao; Wang, Chen; Qiu, Lihong

2017-09-01

Previous studies indicated that constitutive over-expression of cytochrome P450 CYP6B7 was involved in fenvalerate resistance in Helicoverpa armigera. In this study, the CYP6B7 gene from H. armigera (namely HaCYP6B7), was heterologously expressed in Pichia pastoris GS115. A vector pPICZA-HaCYP6B7 was constructed and transformed into P. pastoris GS115, the transformant of pPICZA-HaCYP6B7-GS115 was then cultured and induced by 1% (v/v) methanol and the heterologous expression of HaCYP6B7 protein in P. pastoris was confirmed by SDS-PAGE and western blot. Microsomes containing the expressed HaCYP6B7 showed activities against model substrate p-nitroanisole and 7-ethoxycoumarin, with p-nitroanisole O-demethylation (PNOD) and 7-ethoxycoumarin O-deethylation (ECOD) activities of 15.66- and 4.75-fold of the control, respectively. Moreover, it showed degradation activities against the insecticides bifenthrin, fenvalerate and chlorpyrifos, with clearance activities of 6.88-, 1.49- and 2.27-fold of the control, respectively. The interactions of HaCYP6B7 with insecticides were further confirmed by molecular docking in silico with binding scores of 5.450, 5.295 and 2.197 between putative HaCYP6B7 protein and bifenthrin, fenvalerate and chlorpyrifos, respectively. The results of present study provided more direct and important evidence on the role of HaCYP6B7 conferring pyrethroid resistance in H. armigera. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Chapter A6. Section 6.4. pH

Science.gov (United States)

Wilde, Franceska D.; Busenberg, Eurybiades; Radtke, Dean B.

2006-01-01

Measurement of pH is critical to the understanding of the viability and vulnerability of environmental waters and is considered a master variable in determining the aqueous geochemistry of an aqueous system. pH is a measure that represents the hydrogen-ion concentration (activity) of a solution. This section of the National Field Manual (NFM) describes U.S. Geological Survey (USGS) guidance and protocols for measurement of pH in ground and surface waters.

The E7 protein of the cottontail rabbit papillomavirus immortalizes normal rabbit keratinocytes and reduces pRb levels, while E6 cooperates in immortalization but neither degrades p53 nor binds E6AP

International Nuclear Information System (INIS)

Ganzenmueller, Tina; Matthaei, Markus; Muench, Peter; Scheible, Michael; Iftner, Angelika; Hiller, Thomas; Leiprecht, Natalie; Probst, Sonja; Stubenrauch, Frank; Iftner, Thomas

2008-01-01

Human papillomaviruses (HPVs) cause cervical cancer and are associated with the development of non-melanoma skin cancer. A suitable animal model for papillomavirus-associated skin carcinogenesis is the infection of domestic rabbits with the cottontail rabbit papillomavirus (CRPV). As the immortalizing activity of CRPV genes in the natural target cells remains unknown, we investigated the properties of CRPV E6 and E7 in rabbit keratinocytes (RK) and their influence on the cell cycle. Interestingly, CRPV E7 immortalized RK after a cellular crisis but showed no such activity in human keratinocytes. Co-expressed CRPV E6 prevented cellular crisis. The HPV16 or CRPV E7 protein reduced rabbit pRb levels thereby causing rabbit p19 ARF induction and accumulation of p53 without affecting cellular proliferation. Both CRPV E6 proteins failed to degrade rabbit p53 in vitro or to bind E6AP; however, p53 was still inducible by mitomycin C. In summary, CRPV E7 immortalizes rabbit keratinocytes in a species-specific manner and E6 contributes to immortalization without directly affecting p53

Synthesis and luminescent properties of Sr{sub 2}Gd{sub 6.8}Eu{sub 1.2}Si{sub 6(1−x)}P{sub 6x}O{sub 26} oxyapatites

Energy Technology Data Exchange (ETDEWEB)

Ishchenko, A.V., E-mail: a-v-i@mail.ru [Ural Federal University, 620002 Ekaterinburg (Russian Federation); Zuev, M.G. [Ural Federal University, 620002 Ekaterinburg (Russian Federation); Institute of Solid State Chemistry, Ural Branch of the Russian Academy of Sciences, 620990 Ekaterinburg (Russian Federation); Vasin, A.A. [Institute of Solid State Chemistry, Ural Branch of the Russian Academy of Sciences, 620990 Ekaterinburg (Russian Federation); Yagodin, V.V.; Viktorov, L.V.; Shulgin, B.V. [Ural Federal University, 620002 Ekaterinburg (Russian Federation)

2016-01-15

The solid solutions Sr{sub 2}Gd{sub 6.8}Eu{sub 1.2}Si{sub 6(1−x)}P{sub 6x}O{sub 26−δ} (where x=0–0.15 and δ is oxygen nonstoichiometry) were synthesized. The structural properties of the crystal lattice of the solid solutions and the peculiarities of Eu{sup 3+} and P{sup 5+} dopants substitution for matrix ions have been considered. The photo-, X-ray and pulsed cathode luminescence properties have been studied. It has been found that substitution of (SiO{sub 4}){sup 4−} by (PO{sub 4}){sup 3−} tetrahedron in Eu{sup 3+}-doped oxyapatites does not bring significant changes to bands structure Eu{sup 3+} in luminescence spectra under different excitation (UV, X-ray, pulse cathode beam). However, the increase of P{sup 5+} concentration in Sr{sub 2}Gd{sub 6.8}Eu{sub 1.2}Si{sub 6(1−x)}P{sub 6x}O{sub 26–δ} compounds leads to a decrease of integral intensity of Eu{sup 3+} luminescence bands due to local environment symmetry modifications and covalency degree changes. Two nonequivalent optical Eu{sup 3+} centers have been found. These compounds are of interest for efficient X-ray phosphors, display devices and LED engineering material creation. - Highlights: • The luminescence properties were studied upon UV, X-ray and pulse cathode beam. • P{sup 5+} doping of Sr{sub 2}Gd{sub 6.8}Eu{sub 1.2}Si{sub 6}O{sub 26} leads to luminescence intensity reduction. • At least two types of optical centers formed by Eu{sup 3+} ions were found. • The structural features of Sr{sub 2}Gd{sub 6.8}Eu{sub 1.2}Si{sub 6(1−x)}P{sub 6x}O{sub 26} were reported. • Partial replacement of Si by P does not change the Sr{sub 2}Gd{sub 6.8}Eu{sub 1.2}Si{sub 6}O{sub 26} structure.

Targeting Aberrant p70S6K Activation for Estrogen Receptor-Negative Breast Cancer Prevention.

Science.gov (United States)

Wang, Xiao; Yao, Jun; Wang, Jinyang; Zhang, Qingling; Brady, Samuel W; Arun, Banu; Seewaldt, Victoria L; Yu, Dihua

2017-11-01

The prevention of estrogen receptor-negative (ER-) breast cancer remains a major challenge in the cancer prevention field, although antiestrogen and aromatase inhibitors have shown adequate efficacy in preventing estrogen receptor-positive (ER + ) breast cancer. Lack of commonly expressed, druggable targets is a major obstacle for meeting this challenge. Previously, we detected the activation of Akt signaling pathway in atypical hyperplasic early-stage lesions of patients. In the current study, we found that Akt and the downstream 70 kDa ribosomal protein S6 kinase (p70S6K) signaling pathway was highly activated in ER - premalignant breast lesions and ER - breast cancer. In addition, p70S6K activation induced transformation of ER - human mammary epithelial cells (hMEC). Therefore, we explored the potential of targeting Akt/p70S6K in the p70S6K activated, ER - hMEC models and mouse mammary tumor models for the prevention of ER - breast cancer. We found that a clinically applicable Akt/p70S6K dual inhibitor, LY2780301, drastically decreased proliferation of hMECs with ErbB2-induced p70S6K activation via Cyclin B1 inhibition and cell-cycle blockade at G 0 -G 1 phase, while it did not significantly reverse the abnormal acinar morphology of these hMECs. In addition, a brief treatment of LY2780301 in MMTV- neu mice that developed atypical hyperplasia (ADH) and mammary intraepithelial neoplasia (MIN) lesions with activated p70S6K was sufficient to suppress S6 phosphorylation and decrease cell proliferation in hyperplasic MECs. In summary, targeting the aberrant Akt/p70S6K activation in ER - hMEC models in vitro and in the MMTV- neu transgenic mouse model in vivo effectively inhibited Akt/S6K signaling and reduced proliferation of hMECs in vitro and ADH/MIN lesions in vivo , indicating its potential in prevention of p70S6K activated ER - breast cancer. Cancer Prev Res; 10(11); 641-50. ©2017 AACR . ©2017 American Association for Cancer Research.

Influence of the solution pH in the 6-mercaptopurine self-assembled monolayer (6MP-SAM) on a Au(111) single-crystal electrode.

Science.gov (United States)

Madueño, Rafael; García-Raya, Daniel; Viudez, Alfonso J; Sevilla, José M; Pineda, Teresa; Blázquez, Manuel

2007-10-23

Self-assembled monolayers (SAMs) of 6-mercaptopurine (6MP) have been prepared on a Au(111) single-crystal electrode by immersion of the metal surface in a 100 microM 6MP and 0.01 M HClO4 solution. The 6MP-SAM Au(111) single-crystal electrodes were transferred to the cell and allowed to equilibrate with the different aqueous working solutions before the electrochemical experiments. The influence of the solution pH was studied by cyclic voltammetry, double layer capacitance curves, and electrochemical impedance spectroscopy. The electrochemical behavior of the 6MP-SAM in acetic acid at pH 4 presents important differences in comparison to that obtained in 0.1 M KOH solutions. Cyclic voltammograms for the reductive desorption process in acid medium are broad and show some features that can be explained by a phase transition between a chemisorbed and a physisorbed state of the 6MP molecules. The low solubility of these molecules in acid medium could explain this phenomenon and the readsorption of the complete monolayer when the potential is scanned in the positive direction. The variation of the double-layer capacitance values in the potential range of monolayer stability with the pH suggests that the acid-base chemistry of the 6MP molecules is playing a role. This fact has been studied by following the variations of the electron-transfer rate constant of the highly charged redox probes as are Fe(CN)(6)-3/-4 and Ru(NH3)(6)+3/+2 as a function of solution pH. The apparent surface pKa value for the 6MP-SAM (pKa approximately 8) is explained by the total conversion of the different 6MP tautomers that exist in solution to the thiol species in the adsorbed state.

The Cytochrome P450 gene CYP6P12 confers pyrethroid resistance in kdr-free Malaysian populations of the dengue vector Aedes albopictus.

Science.gov (United States)

Ishak, Intan H; Riveron, Jacob M; Ibrahim, Sulaiman S; Stott, Rob; Longbottom, Joshua; Irving, Helen; Wondji, Charles S

2016-04-20

Control of Aedes albopictus, major dengue and chikungunya vector, is threatened by growing cases of insecticide resistance. The mechanisms driving this resistance remain poorly characterised. This study investigated the molecular basis of insecticide resistance in Malaysian populations of Ae. albopictus. Microarray-based transcription profiling revealed that metabolic resistance (cytochrome P450 up-regulation) and possibly a reduced penetration mechanism (consistent over-expression of cuticular protein genes) were associated with pyrethroid resistance. CYP6P12 over-expression was strongly associated with pyrethroid resistance whereas CYP6N3 was rather consistently over-expressed across carbamate and DDT resistant populations. Other detoxification genes also up-regulated in permethrin resistant mosquitoes included a glucuronosyltransferase (AAEL014279-RA) and the glutathione-S transferases GSTS1 and GSTT3. Functional analyses further supported that CYP6P12 contributes to pyrethroid resistance in Ae. albopictus as transgenic expression of CYP6P12 in Drosophila was sufficient to confer pyrethroid resistance in these flies. Furthermore, molecular docking simulations predicted CYP6P12 possessing enzymatic activity towards pyrethroids. Patterns of polymorphism suggested early sign of selection acting on CYP6P12 but not on CYP6N3. The major role played by P450 in the absence of kdr mutations suggests that addition of the synergist PBO to pyrethroids could improve the efficacy of this insecticide class and overcome resistance in field populations of Ae. albopictus.

Purine receptor P2Y_6 mediates cellular response to γ-ray-induced DNA damage

International Nuclear Information System (INIS)

Ide, Shunta; Nishimaki, Naoko; Tsukimoto, Mitsutoshi; Kojima, Shuji

2014-01-01

We previously showed that nucleotide P2 receptor agonists such as ATP and UTP amplify γ-ray-induced focus formation of phosphorylated histone H2A variant H2AX (γH2AX), which is considered to be an indicator of DNA damage so far, by activating purine P2Y_6 and P2Y_1_2 receptors. Therefore, we hypothesized that these P2 receptors play a role in inducing the repair response to γ-ray-induced DNA damage. In the present study, we tested this idea by using human lung cancer A549 cells. First, reverse-transcription polymerase chain reaction (RT-PCR) showed that P2Y_6 receptor is highly expressed in A549 cells, but P2Y_1_2 receptor is only weakly expressed. Next, colony formation assay revealed that P2Y_6 receptor antagonist MRS2578 markedly reduced the survival rate of γ-ray-exposed A549 cells. The survival rate was also significantly reduced in P2Y_6-knock-down cells, compared with scramble siRNA-transfected cells. Since it has reported that phosphorylation of ERK1/2 after activation of EGFR via P2Y_6 and P2Y_1_2 receptors is involved in the repair response to γ-ray-induced DNA damage, we next examined whether γ-ray-induced phosphorylation of ERK1/2 was also inhibited by MRS2578 in A549 cells. We found that it was. Taken together, these findings indicate that purinergic signaling through P2Y_6 receptor, followed by ERK1/2 activation, promotes the cellular repair response to γ-ray-induced DNA damage. (author)

P-wave Feshbach resonances of ultracold 6Li

International Nuclear Information System (INIS)

Zhang, J.; Kempen, E.G.M. van; Bourdel, T.; Cubizolles, J.; Chevy, F.; Teichmann, M.; Tarruell, L.; Salomon, C.; Khaykovich, L.; Kokkelmans, S.J.J.M.F.

2004-01-01

We report the observation of three p-wave Feshbach resonances of 6 Li atoms in the lowest hyperfine state f=1/2. The positions of the resonances are in good agreement with theory. We study the lifetime of the cloud in the vicinity of the Feshbach resonances and show that, depending on the spin states, two- or three-body mechanisms are at play. In the case of dipolar losses, we observe a nontrivial temperature dependence that is well explained by a simple model

Hinder för utomhusundervisning med fokus på årskurs 4-6

OpenAIRE

Petersson, Essi; Carstensen, Linnea

2018-01-01

I den här studien har vi tagit fram de hinder som begränsar möjligheterna för utomhusundervisning för elever i grundskolan, med fokus på årskurs 4-6 och middle school. Syftet med studien var att synliggöra vilka hinder som kan stå i vägen för utomhusundervisning samt visa på behov av vidare forskning i området. Forskningsfrågan som vi har försökt svara på är: Vad utgör hinder för utomhusundervisning med elever i grundskolan, med fokus på årskurs 4-6 och middle school? Studien bygger på den pr...

Cauliflower mosaic virus protein P6 inhibits signaling responses to salicylic acid and regulates innate immunity.

Directory of Open Access Journals (Sweden)

Andrew J Love

Full Text Available Cauliflower mosaic virus (CaMV encodes a multifunctional protein P6 that is required for translation of the 35S RNA and also acts as a suppressor of RNA silencing. Here we demonstrate that P6 additionally acts as a pathogenicity effector of an unique and novel type, modifying NPR1 (a key regulator of salicylic acid (SA- and jasmonic acid (JA-dependent signaling and inhibiting SA-dependent defence responses We find that that transgene-mediated expression of P6 in Arabidopsis and transient expression in Nicotiana benthamiana has profound effects on defence signaling, suppressing expression of representative SA-responsive genes and increasing expression of representative JA-responsive genes. Relative to wild-type Arabidopsis P6-expressing transgenics had greatly reduced expression of PR-1 following SA-treatment, infection by CaMV or inoculation with an avirulent bacterial pathogen Pseudomonas syringae pv tomato (Pst. Similarly transient expression in Nicotiana benthamiana of P6 (including a mutant form defective in translational transactivation activity suppressed PR-1a transcript accumulation in response to Agrobacterium infiltration and following SA-treatment. As well as suppressing the expression of representative SA-regulated genes, P6-transgenic Arabidopsis showed greatly enhanced susceptibility to both virulent and avirulent Pst (titres elevated 10 to 30-fold compared to non-transgenic controls but reduced susceptibility to the necrotrophic fungus Botrytis cinerea. Necrosis following SA-treatment or inoculation with avirulent Pst was reduced and delayed in P6-transgenics. NPR1 an important regulator of SA/JA crosstalk, was more highly expressed in the presence of P6 and introduction of the P6 transgene into a transgenic line expressing an NPR1:GFP fusion resulted in greatly increased fluorescence in nuclei even in the absence of SA. Thus in the presence of P6 an inactive form of NPR1 is mislocalized in the nucleus even in uninduced plants

Observation of the Forbidden Magnetic Dipole Transition 6{sup 2}P{sub ½} --> 7{sup 2}P{sub ½} in Atomic Thallium

Science.gov (United States)

Chu, S.

1976-10-01

A measurement of the 6{sup 2}P{sub ½} --> 7{sup 2}P{sub ½} forbidden magnetic dipole matrix element in atomic thallium is described. A pulsed, linearly polarized dye laser tuned to the transition frequency is used to excite the thallium vapor from the 6{sup 2}P{sub ½} ground state to the 7{sup 2}P{sub ½} excited state. Interference between the magnetic dipole M1 amplitude and a static electric field induced E1 amplitude results in an atomic polarization of the 7{sup 2}P{sub ½} state, and the subsequent circular polarization of 535 nm fluorescence. The circular polarization is seen to be proportional to / as expected, and measured for several transitions between hyperfine levels of the 6{sup 2}P{sub ½} and 7{sup 2}P{sub ½} states. The result is = -(2.11 +- 0.30) x 10{sup -5} parallel bar e parallel bar dirac constant/2mc, in agreement with theory.

Small supernumerary marker chromosome causing partial trisomy 6p in a child with craniosynostosis.

Science.gov (United States)

Villa, Olaya; Del Campo, Miguel; Salido, Marta; Gener, Blanca; Astier, Laura; Del Valle, Jesús; Gallastegui, Fátima; Pérez-Jurado, Luis A; Solé, Francesc

2007-05-15

We report on a child with a small supernumerary marker chromosome (sSMC) causing partial trisomy 6p. The child showed a phenotype consisting of neonatal craniosynostosis, microcephaly, and borderline developmental delay. By molecular techniques the sSMC has been shown to contain approximately 16 Mb of genomic DNA from 6p21.1 to 6cen, being de novo and of maternal origin.

Clustering by neurocognition for fine-mapping of the schizophrenia susceptibility loci on chromosome 6p

Science.gov (United States)

Lin, Sheng-Hsiang; Liu, Chih-Min; Liu, Yu-Li; Fann, Cathy Shen-Jang; Hsiao, Po-Chang; Wu, Jer-Yuarn; Hung, Shuen-Iu; Chen, Chun-Houh; Wu, Han-Ming; Jou, Yuh-Shan; Liu, Shi K.; Hwang, Tzung J.; Hsieh, Ming H.; Chang, Chien-Ching; Yang, Wei-Chih; Lin, Jin-Jia; Chou, Frank Huang-Chih; Faraone, Stephen V.; Tsuang, Ming T.; Hwu, Hai-Gwo; Chen, Wei J.

2009-01-01

Chromosome 6p is one of the most commonly implicated regions in the genome-wide linkage scans of schizophrenia, whereas further association studies for markers in this region were inconsistent likely due to heterogeneity. This study aimed to identify more homogeneous subgroups of families for fine mapping on regions around markers D6S296 and D6S309 (both in 6p24.3) as well as D6S274 (in 6p22.3) by means of similarity in neurocognitive functioning. A total of 160 families of patients with schizophrenia comprising at least two affected siblings who had data for 8 neurocognitive test variables of the Continuous Performance Test (CPT) and the Wisconsin Card Sorting Test (WCST) were subjected to cluster analysis with data visualization using the test scores of both affected siblings. Family clusters derived were then used separately in family-based association tests for 64 single nucleotide polymorphisms covering the region of 6p24.3 and 6p22.3. Three clusters were derived from the family-based clustering, with deficit cluster 1 representing deficit on the CPT, deficit cluster 2 representing deficit on both the CPT and the WCST, and a third cluster of non-deficit. After adjustment using false discovery rate for multiple testing, SNP rs13873 and haplotype rs1225934-rs13873 on BMP6-TXNDC5 genes were significantly associated with schizophrenia for the deficit cluster 1 but not for the deficit cluster 2 or non-deficit cluster. Our results provide further evidence that the BMP6-TXNDC5 locus on 6p24.3 may play a role in the selective impairments on sustained attention of schizophrenia. PMID:19694819

Nonsyndromic cleft lip and palate: Evidence of linkage to a microsatellite marker on 6p23

Energy Technology Data Exchange (ETDEWEB)

Carinci, F.; Pezzetti, F.; Scapoli, L.; Padula, E.; Baciliero, U.; Curioni, C.; Tognon, M.

1995-01-01

Nonsydromic cleft lip with or without secondary clefting of the palate (CL+/{minus}P) is one of the most common birth defects. A previous linkage study concerning CL+/{minus}P and cleft palate (CP) families indicated chromosome 6p, near F13A locus, as a possible region for the presence of a clefting gene. More recently, another linkage study performed on a sample of 12 families with nonsyndromic CL+/{minus}P seemed to exclude this association. To test the hypothesis on the possible presence of a major gene on chromosome 6p, we carried out a study on a large sample (21) of CL+/{minus}P families from northeastern Italy. In conclusion, our investigation can be summarized as follows: (i) CL+/{minus}P disease appears to be heterogeneous; (ii) {approximately}66% of the pedigrees showed an autosomal dominant inheritance with incomplete penetrance; and (iii) CL+/{minus}P locus maps on 6p23 very close to or at the microsatellite marker D6S89. To verify whether the D6S89 is the closest marker to the CL+/{minus}P locus, additional examinations with new markers are underway. 19 refs., 1 fig., 1 tab.

Electrical resistivity tomography to monitor enhanced biodegradation of hydrocarbons with Rhodococcus erythropolis T902.1 at a pilot scale.

Science.gov (United States)

Masy, Thibaut; Caterina, David; Tromme, Olivier; Lavigne, Benoît; Thonart, Philippe; Hiligsmann, Serge; Nguyen, Frédéric

2016-01-01

Petroleum hydrocarbons (HC) represent the most widespread contaminants and in-situ bioremediation remains a competitive treatment in terms of cost and environmental concerns. However, the efficiency of such a technique (by biostimulation or bioaugmentation) strongly depends on the environment affected and is still difficult to predict a priori. In order to overcome these uncertainties, Electrical Resistivity Tomography (ERT) appears as a valuable non-invasive tool to detect soil heterogeneities and to monitor biodegradation. The main objective of this study was to isolate an electrical signal linked to an enhanced bacterial activity with ERT, in an aged HC-contaminated clay loam soil. To achieve this, a pilot tank was built to mimic field conditions. Compared to a first insufficient biostimulation phase, bioaugmentation with Rhodococcus erythropolis T902.1 led to a HC depletion of almost 80% (6900 to 1600ppm) in 3months in the center of the contaminated zone, where pollutants were less bioavailable. In the meantime, lithological heterogeneities and microbial activities (growth and biosurfactant production) were successively discriminated by ERT images. In the future, this cost-effective technique should be more and more transferred to the field in order to monitor biodegradation processes and assist in selecting the most appropriate remediation technique. Copyright © 2015 Elsevier B.V. All rights reserved.

Electrical resistivity tomography to monitor enhanced biodegradation of hydrocarbons with Rhodococcus erythropolis T902.1 at a pilot scale

Science.gov (United States)

Masy, Thibaut; Caterina, David; Tromme, Olivier; Lavigne, Benoît; Thonart, Philippe; Hiligsmann, Serge; Nguyen, Frédéric

2016-01-01

Petroleum hydrocarbons (HC) represent the most widespread contaminants and in-situ bioremediation remains a competitive treatment in terms of cost and environmental concerns. However, the efficiency of such a technique (by biostimulation or bioaugmentation) strongly depends on the environment affected and is still difficult to predict a priori. In order to overcome these uncertainties, Electrical Resistivity Tomography (ERT) appears as a valuable non-invasive tool to detect soil heterogeneities and to monitor biodegradation. The main objective of this study was to isolate an electrical signal linked to an enhanced bacterial activity with ERT, in an aged HC-contaminated clay loam soil. To achieve this, a pilot tank was built to mimic field conditions. Compared to a first insufficient biostimulation phase, bioaugmentation with Rhodococcus erythropolis T902.1 led to a HC depletion of almost 80% (6900 to 1600 ppm) in 3 months in the center of the contaminated zone, where pollutants were less bioavailable. In the meantime, lithological heterogeneities and microbial activities (growth and biosurfactant production) were successively discriminated by ERT images. In the future, this cost-effective technique should be more and more transferred to the field in order to monitor biodegradation processes and assist in selecting the most appropriate remediation technique.

A common soil flagellate (Cercomonas sp.) grows slowly when feeding on the bacterium Rhodococcus fascians in isolation, but does not discriminate against it in a mixed culture with Sphingopyxis witflariensis

DEFF Research Database (Denmark)

Lekfeldt, Jonas D S; Rønn, Regin

2008-01-01

Flagellates are very important predators on bacteria in soil. Because of their high growth rates, flagellate populations respond rapidly to changes in bacterial numbers. Previous results indicate that actinobacteria are generally less suitable than proteobacteria as food for flagellates. In this ......Flagellates are very important predators on bacteria in soil. Because of their high growth rates, flagellate populations respond rapidly to changes in bacterial numbers. Previous results indicate that actinobacteria are generally less suitable than proteobacteria as food for flagellates....... In this study, we investigated the growth of the flagellate Cercomonas sp. (ATCC 50334) on each of the two bacteria Sphingopyxis witflariensis (Alphaproteobacteria) and Rhodococcus fascians (actinobacteria) separately and in combination. The growth rate of the flagellate was lower and the lag phase was longer...

Crystallization and preliminary X-ray analysis of coagulation factor IX-binding protein from habu snake venom at pH 6.5 and 4.6

International Nuclear Information System (INIS)

Suzuki, Nobuhiro; Shikamoto, Yasuo; Fujimoto, Zui; Morita, Takashi; Mizuno, Hiroshi

2004-01-01

Crystals of habu coagulation factor IX-binding protein have been obtained at pH 6.5 and 4.6 and characterized by X-ray diffraction. Coagulation factor IX-binding protein isolated from Trimeresurus flavoviridis (IX-bp) is a C-type lectin-like protein. It is an anticoagulant protein consisting of homologous subunits A and B. The subunits both contain a Ca 2+ -binding site with differing affinity (K d values of 14 and 130 µM at pH 7.5). These binding characteristics are pH-dependent; under acidic conditions, the affinity of the low-affinity site was reduced considerably. In order to identify which site has high affinity and also to investigate the Ca 2+ -releasing mechanism, IX-bp was crystallized at pH 6.5 and 4.6. The crystals at pH 6.5 and 4.6 diffracted to 1.72 and 2.29 Å resolution, respectively; the former crystals belong to the monoclinic space group P2 1 , with unit-cell parameters a = 60.7, b = 63.5, c = 66.9 Å, β = 117.0°, while the latter belong to the monoclinic space group C2, with a = 134.1, b = 37.8, c = 55.8 Å, β = 110.4°

Energy-Transfer Kinetics for Xe (6p[1/2]0) Atoms in Kr, Ar, Ne, and He.

Science.gov (United States)

He, Shan; Liu, Dong; Li, Xueyang; Chu, Junzhi; Guo, Jingwei; Liu, Jinbo; Hu, Shu; Sang, Fengting; Jin, Yuqi

2018-06-11

The kinetic processes for the Xe (6p[1/2] 0 ) atoms in Kr, Ar, Ne, and He buffer gases were studied. We found that Kr, Ar, and Ne atoms can be used to switch the amplified spontaneous emission (ASE) channel from 3408 nm (6p[1/2] 0 -6s'[1/2] 1 ) to 3680 nm (5d[1/2] 1 -6p[1/2] 1 ), while Xe and He atoms do not show such a phenomenon. This ASE channel switch is mainly ascribed to the fast transfer of 6p[1/2] 0 → 5d[1/2] 1 . On the basis of the rate equations for two-state coupling (energy-transfer processes between the two states are very rapid), the reason why the ASE channel switch effect normally coincides with a double exponential decay of the spontaneous emission at 828 nm (6p[1/2] 0 -6s[3/2] 1 ) is explained. The actual situations in Xe, Ar, Ne, and He follow this rule. However, the strictly single exponential decay of the spontaneous emission at 828 nm and strong ASE channel switch effect simultaneously emerge in Kr. This indicates that the transfer of 6p[1/2] 0 → 5d[1/2] 1 in Kr does not occur via two-state coupling, but via two steps of near-resonance collision through the 5s[3/2] 2 (Kr) state as the intermediate state (6p[1/2] 0 → 5s[3/2] 2 (Kr) → 5d[1/2] 1 ). In addition, we found Xe (6p[1/2] 0 ) atoms strongly tend to reach the 6p[3/2] 2 , 6p[3/2] 1 , and 6p[5/2] 2 states through the 5s[3/2] 2 (Kr) state as the intermediate state in Kr. The 5s[3/2] 2 (Kr) state plays a very important role in the energy-transfer kinetics for the Xe (6p[1/2] 0 ) atoms. Kr is probably an excellent buffer gas for laser systems based on Xe.

Structural analysis of the yeast exosome Rrp6p–Rrp47p complex by small-angle X-ray scattering

Energy Technology Data Exchange (ETDEWEB)

Dedic, Emil; Seweryn, Paulina; Jonstrup, Anette Thyssen; Flygaard, Rasmus Koch [Centre for mRNP Biogenesis and Metabolism, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C (Denmark); Department of Molecular Biology and Genetics, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C (Denmark); Fedosova, Natalya U. [Department of Biomedicine, Ole Worms Allé 6, Aarhus University, DK-8000 Aarhus C (Denmark); Hoffmann, Søren Vrønning [Institute for Storage Ring Facilities (ISA), Department of Physics and Astronomy, Ny Munkegade 120, Aarhus University, DK-8000 Aarhus C (Denmark); Boesen, Thomas [Centre for Membrane Pumps in Cells and Disease – PUMPKIN, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C (Denmark); Department of Molecular Biology and Genetics, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C (Denmark); Brodersen, Ditlev Egeskov, E-mail: deb@mb.au.dk [Centre for mRNP Biogenesis and Metabolism, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C (Denmark); Department of Molecular Biology and Genetics, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C (Denmark)

2014-07-18

Highlights: • We show that S. cerevisiae Rrp6p and Rrp47p stabilise each other in vitro. • We determine molecular envelopes of the Rrp6p–Rrp47p complex by SAXS. • Rrp47p binds at the top of the Rrp6p exonuclease domain. • Rrp47p modulates the activity of Rrp6p on a variety of RNA substrates. • Rrp47p does not affect RNA affinity by Rrp6p. - Abstract: The RNase D-type 3′–5′ exonuclease Rrp6p from Saccharomyces cerevisiae is a nuclear-specific cofactor of the RNA exosome and associates in vivo with Rrp47p (Lrp1p). Here, we show using biochemistry and small-angle X-ray scattering (SAXS) that Rrp6p and Rrp47p associate into a stable, heterodimeric complex with an elongated shape consistent with binding of Rrp47p to the nuclease domain and opposite of the HRDC domain of Rrp6p. Rrp47p reduces the exonucleolytic activity of Rrp6p on both single-stranded and structured RNA substrates without significantly altering the affinity towards RNA or the ability of Rrp6p to degrade RNA secondary structure.

Structural analysis of the yeast exosome Rrp6p–Rrp47p complex by small-angle X-ray scattering

International Nuclear Information System (INIS)

Dedic, Emil; Seweryn, Paulina; Jonstrup, Anette Thyssen; Flygaard, Rasmus Koch; Fedosova, Natalya U.; Hoffmann, Søren Vrønning; Boesen, Thomas; Brodersen, Ditlev Egeskov

2014-01-01

Highlights: • We show that S. cerevisiae Rrp6p and Rrp47p stabilise each other in vitro. • We determine molecular envelopes of the Rrp6p–Rrp47p complex by SAXS. • Rrp47p binds at the top of the Rrp6p exonuclease domain. • Rrp47p modulates the activity of Rrp6p on a variety of RNA substrates. • Rrp47p does not affect RNA affinity by Rrp6p. - Abstract: The RNase D-type 3′–5′ exonuclease Rrp6p from Saccharomyces cerevisiae is a nuclear-specific cofactor of the RNA exosome and associates in vivo with Rrp47p (Lrp1p). Here, we show using biochemistry and small-angle X-ray scattering (SAXS) that Rrp6p and Rrp47p associate into a stable, heterodimeric complex with an elongated shape consistent with binding of Rrp47p to the nuclease domain and opposite of the HRDC domain of Rrp6p. Rrp47p reduces the exonucleolytic activity of Rrp6p on both single-stranded and structured RNA substrates without significantly altering the affinity towards RNA or the ability of Rrp6p to degrade RNA secondary structure

Expression control of nitrile hydratase and amidase genes in Rhodococcus erythropolis and substrate specificities of the enzymes.

Science.gov (United States)

Rucká, Lenka; Volkova, Olga; Pavlík, Adam; Kaplan, Ondřej; Kracík, Martin; Nešvera, Jan; Martínková, Ludmila; Pátek, Miroslav

2014-06-01

Bacterial amidases and nitrile hydratases can be used for the synthesis of various intermediates and products in the chemical and pharmaceutical industries and for the bioremediation of toxic pollutants. The aim of this study was to analyze the expression of the amidase and nitrile hydratase genes of Rhodococcus erythropolis and test the stereospecific nitrile hydratase and amidase activities on chiral cyanohydrins. The nucleotide sequences of the gene clusters containing the oxd (aldoxime dehydratase), ami (amidase), nha1, nha2 (subunits of the nitrile hydratase), nhr1, nhr2, nhr3 and nhr4 (putative regulatory proteins) genes of two R. erythropolis strains, A4 and CCM2595, were determined. All genes of both of the clusters are transcribed in the same direction. RT-PCR analysis, primer extension and promoter fusions with the gfp reporter gene showed that the ami, nha1 and nha2 genes of R. erythropolis A4 form an operon transcribed from the Pami promoter and an internal Pnha promoter. The activity of Pami was found to be weakly induced when the cells grew in the presence of acetonitrile, whereas the Pnha promoter was moderately induced by both the acetonitrile or acetamide used instead of the inorganic nitrogen source. However, R. erythropolis A4 cells showed no increase in amidase and nitrile hydratase activities in the presence of acetamide or acetonitrile in the medium. R. erythropolis A4 nitrile hydratase and amidase were found to be effective at hydrolysing cyanohydrins and 2-hydroxyamides, respectively.

In Planta Biocontrol of Pectobacterium atrosepticum by Rhodococcus erythropolis Involves Silencing of Pathogen Communication by the Rhodococcal Gamma-Lactone Catabolic Pathway.

Directory of Open Access Journals (Sweden)

Corinne Barbey

Full Text Available The virulence of numerous Gram-negative bacteria is under the control of a quorum sensing process based on synthesis and perception of N-acyl homoserine lactones. Rhodococcus erythropolis, a Gram-positive bacterium, has recently been proposed as a biocontrol agent for plant protection against soft-rot bacteria, including Pectobacterium. Here, we show that the γ-lactone catabolic pathway of R. erythropolis disrupts Pectobacterium communication and prevents plant soft-rot. We report the first characterization and demonstration of N-acyl homoserine lactone quenching in planta. In particular, we describe the transcription of the R. erythropolis lactonase gene, encoding the key enzyme of this pathway, and the subsequent lactone breakdown. The role of this catabolic pathway in biocontrol activity was confirmed by deletion of the lactonase gene from R. erythropolis and also its heterologous expression in Escherichia coli. The γ-lactone catabolic pathway is induced by pathogen communication rather than by pathogen invasion. This is thus a novel and unusual biocontrol pathway, differing from those previously described as protecting plants from phytopathogens. These findings also suggest the existence of an additional pathway contributing to plant protection.

p62 regulates CD40-mediated NFκB activation in macrophages through interaction with TRAF6

Energy Technology Data Exchange (ETDEWEB)

Seibold, Kristina; Ehrenschwender, Martin, E-mail: martin.ehrenschwender@ukr.de

2015-08-14

CD40 is a member of the tumor necrosis factor (TNF) receptor family. Activation-induced recruitment of adapter proteins, so-called TNF-receptor-associated factors (TRAFs) to the cytoplasmic tail of CD40 triggers signaling cascades important in the immune system, but has also been associated with excessive inflammation in diseases such as atherosclerosis and rheumatoid arthritis. Especially, pro-inflammatory nuclear factor κB (NFκB) signaling emanating from CD40-associated TRAF6 appears to be a key pathogenic driving force. Consequently, targeting the CD40-TRAF6 interaction is emerging as a promising therapeutic strategy, but the underlying molecular machinery of this signaling axis is to date poorly understood. Here, we identified the multifunctional adaptor protein p62 as a critical regulator in CD40-mediated NFκB signaling via TRAF6. CD40 activation triggered formation of a TRAF6-p62 complex. Disturbing this interaction tremendously reduced CD40-mediated NFκB signaling in macrophages, while TRAF6-independent signaling pathways remained unaffected. This highlights p62 as a potential target in hyper-inflammatory, CD40-associated pathologies. - Highlights: • CD40 activation triggers interaction of the adapter protein TRAF6 with p62. • TRAF6-p62 interaction regulates CD40-mediated NFκB signaling in macrophages. • Defective TRAF6-p62 interaction reduces CD40-mediated NFκB activation in macrophages.

The anomalous chiral Lagrangian of order p6

International Nuclear Information System (INIS)

Bijnens, J.; Talavera, P.

2002-01-01

We construct the effective chiral Lagrangian for chiral perturbation theory in the mesonic odd-intrinsic-parity sector at order p 6 . The Lagrangian contains 24 in principle measurable terms and no contact terms for the general case of N f light flavors, 23 terms for three and 5 for two flavors. In the two flavor case we need a total of 13 terms if an external singlet vector field is included. We discuss and implement the methods used to reduce to a minimal set. The infinite parts needed for renormalization are calculated and presented as well. (orig.)

Inositol-P6 induced structural changes in duck major haemoglobin

African Journals Online (AJOL)

Bamiji Babalola

2011-11-02

Nov 2, 2011 ... of duck (Anas platyrhinchos) have been carried out in the presence of ... dependence profile of the apparent second-order rate constant of both oxy- and ... Key words: 5,5‟-Dithiobis (2-nitrobenzoate), duck, haemoglobin, inositol-P6, sulphydryl group. ... Saigo, 1999), thereby making it possible to detect the.

The change of p16 gene expression in glioma cell line C6 after radiation with gamma knife

International Nuclear Information System (INIS)

Zhao Xingli; Zhao Conghai; Tian Yu

2002-01-01

Objective: T observe the change of expression of p16 gene product, P16 protein, after treated by gamma knife on glioma cell line C6. Methods: Glioma C6 cells proliferated in vitro, treated by γ-knife in dose of 5.00 and 6.22 Gy, respectively. P16 protein was detected by immunohistochemical technique and image analysis. Results: The P16 protein in glioma C6 cells was notably increased after treatment with γ knife (P < 0.01). The grey number in C6 group (control group) was 167.1 +- 6.2 and was 155.4 +- 2.0 and 124.9 +- 7.1, respectively, in 5.00 Gy and 6.22 Gy gamma knife treated group. Conclusion: It is suggests that one of the mechanisms of glioma cell C6 apoptosis induced by γ-knife radiation may be associated with activation of p16 gene and increase of P16 protein expression

The energy structure and decay channels of the 4p6-shell excited states in Sr

Science.gov (United States)

Kupliauskienė, A.; Kerevičius, G.; Borovik, V.; Shafranyosh, I.; Borovik, A.

2017-11-01

The ejected-electron spectra arising from the decay of the 4p{}5{{nln}}{\\prime }{l}{\\prime }{n}{\\prime\\prime }{l}{\\prime\\prime } autoionizing states in Sr atoms have been studied precisely at the incident-electron energies close to excitation and ionization thresholds of the 4{{{p}}}6 subshell. The excitation behaviors for 58 lines observed between 12 and 21 eV ejected-electron kinetic energy have been investigated. Also, the ab initio calculations of excitation energies, autoionization probabilities and electron-impact excitation cross sections of the states 4p{}5{{nln}}{\\prime }{l}{\\prime }{n}{\\prime\\prime }{l}{\\prime\\prime } (nl = 4d, 5s, 5p; {n}{\\prime }{l}{\\prime } = 4d, 5s, 5p; {n}{\\prime\\prime }{l}{\\prime\\prime } = 5s, 6s, 7s, 8s, 9s, 5p, 6p, 5d, 6d, 7d, 8d, 4f, 5g) have been performed by employing the large-scale configuration-interaction method in the basis of the solutions of Dirac-Fock-Slater equations. The obtained experimental and theoretical data have been used for the accurate identification of the 60 lines in ejected-electron spectra and the 68 lines observed earlier in photoabsorption spectra. The excitation and decay processes for 105 classified states in the 4p55s{}2{nl}, 4p54d{}2{nl} and 4p55s{{nln}}{\\prime }{l}{\\prime } configurations have been considered in detail. In particular, most of the states lying below the ionization threshold of the 4p6 subshell at 26.92 eV possess up to four decay channels with formation of Sr+ in 5s{}1/2, 4d{}3/{2,5/2} and 5p{}1/{2,3/2} states. Two-step autoionization and two-electron Auger transitions with formation of Sr2+ in the 4p6 {}1{{{S}}}0 ground state are the main decay paths for high-lying autoionizing states. The excitation threshold of the 4{{{p}}}6 subshell in Sr has been established at 20.98 ± 0.05 eV.

Ab initio investigations of the electronic structures and chemical bonding in LiCo{sub 6}P{sub 4} and Li{sub 2}Co{sub 12}P{sub 7}

Energy Technology Data Exchange (ETDEWEB)

Matar, Samir F. [CNRS, ICMCB, UPR 9048, F‐33600 Pessac (France); Université de Bordeaux, ICMCB, UPR 9048, F‐33600 Pessac (France); Al-Alam, Adel; Ouaini, Naïm [Université Saint-Esprit de Kaslik (USEK), Groupe OCM (Optimization et Caractérisation des Matériaux), CSR-USEK, CNRS-L, Jounieh (Lebanon); Pöttgen, Rainer, E-mail: pottgen@uni-muenster.de [Institut für Anorganische und Analytische Chemie, Universität Münster, Corrensstraße 30, D-48149 Münster (Germany)

2013-06-15

The electronic structures of the metal-rich phosphides LiCo{sub 6}P{sub 4} and Li{sub 2}Co{sub 12}P{sub 7} were studied by DFT calculations. Both phosphides consist of three-dimensional [Co{sub 6}P{sub 4}] and [Co{sub 12}P{sub 7}] polyanionic networks which leave hexagonal channels for the lithium atoms. COOP data show strong Co–P and Co–Co bonding within the polyanions. The lithium atoms have trigonal prismatic phosphorus coordination. Total energy calculations indicate stability upon de-lithiation towards the Co{sub 6}P{sub 4} and Co{sub 12}P{sub 7} substructures - Graphical abstract: The cobalt–phosphorus networks in LiCo{sub 6}P{sub 4} and Li{sub 2}Co{sub 12}P{sub 7}. - Highlights: • Chemical bonding resolved in the metal-rich phosphides LiCo{sub 6}P{sub 4} and Li{sub 2}Co{sub 12}P{sub 7}. • Strong covalent Co–P bonding character in the [Co{sub 6}P{sub 4}] and [Co{sub 12}P{sub 7}] substructures. • Total energy calculations indicate stability of the de-lithiated substructures.

Genetic rearrangements of six wheat-agropyron cristatum 6P addition lines revealed by molecular markers.

Directory of Open Access Journals (Sweden)

Haiming Han

Full Text Available Agropyron cristatum (L. Gaertn. (2n = 4x = 28, PPPP not only is cultivated as pasture fodder but also could provide many desirable genes for wheat improvement. It is critical to obtain common wheat-A. cristatum alien disomic addition lines to locate the desired genes on the P genome chromosomes. Comparative analysis of the homoeologous relationships between the P genome chromosome and wheat genome chromosomes is a key step in transferring different desirable genes into common wheat and producing the desired alien translocation line while compensating for the loss of wheat chromatin. In this study, six common wheat-A. cristatum disomic addition lines were produced and analyzed by phenotypic examination, genomic in situ hybridization (GISH, SSR markers from the ABD genomes and STS markers from the P genome. Comparative maps, six in total, were generated and demonstrated that all six addition lines belonged to homoeologous group 6. However, chromosome 6P had undergone obvious rearrangements in different addition lines compared with the wheat chromosome, indicating that to obtain a genetic compensating alien translocation line, one should recombine alien chromosomal regions with homoeologous wheat chromosomes. Indeed, these addition lines were classified into four types based on the comparative mapping: 6PI, 6PII, 6PIII, and 6PIV. The different types of chromosome 6P possessed different desirable genes. For example, the 6PI type, containing three addition lines, carried genes conferring high numbers of kernels per spike and resistance to powdery mildew, important traits for wheat improvement. These results may prove valuable for promoting the development of conventional chromosome engineering techniques toward molecular chromosome engineering.

D-Glucosamine inhibits proliferation of human cancer cells through inhibition of p70S6K

International Nuclear Information System (INIS)

Oh, Hyun-Ji; Lee, Jason S.; Song, Dae-Kyu; Shin, Dong-Hoon; Jang, Byeong-Churl; Suh, Seong-Il; Park, Jong-Wook; Suh, Min-Ho; Baek, Won-Ki

2007-01-01

Although D-glucosamine has been reported as an inhibitor of tumor growth both in vivo and in vitro, the mechanism for the anticancer effect of D-glucosamine is still unclear. Since there are several reports suggesting D-glucosamine inhibits protein synthesis, we examined whether D-glucosamine affects p70S6 K activity, an important signaling molecule involved in protein translation. In the present study, we found D-glucosamine inhibited the activity of p70S6K and the proliferation of DU145 prostate cancer cells and MDA-MB-231 breast cancer cells. D-Glucosamine decreased phosphorylation of p70S6K, and its downstream substrates RPS6, and eIF-4B, but not mTOR and 4EBP1 in DU145 cells, suggesting that D-glucosamine induced inhibition of p70S6K is not through the inhibition of mTOR. In addition, D-glucosamine enhanced the growth inhibitory effects of rapamycin, a specific inhibitor of mTOR. These findings suggest that D-glucosamine can inhibit growth of cancer cells through dephosphorylation of p70S6K

One-dimensional zinc selenophosphates: A{sub 2}ZnP{sub 2}Se{sub 6} (A = K, Rb, Cs)

Energy Technology Data Exchange (ETDEWEB)

Haynes, Alyssa S.; Lee, Katherine; Kanatzidis, Mercouri G. [Department of Chemistry, Northwestern University, Evanston, IL (United States)

2016-09-15

The new compounds A{sub 2}ZnP{sub 2}Se{sub 6} (A = K, Rb, Cs) were synthesized via molten salt flux syntheses. The crystals feature one-dimensional {sup 1}/{sub ∞}[ZnP{sub 2}Se{sub 6}]{sup 2-} chains charge balanced by alkali metal ions between the chains. K{sub 2}ZnP{sub 2}Se{sub 6} crystallizes in the monoclinic space group P2{sub 1}/c; cell parameters a = 12.537(3) Aa, b = 7.2742(14) Aa, c = 14.164(3) Aa, β = 109.63(3) , Z = 4, and V = 1216.7(4) Aa{sup 3}. Rb{sub 2}ZnP{sub 2}Se{sub 6} and Cs{sub 2}ZnP{sub 2}Se{sub 6} are isotypic, crystallizing in the triclinic space group P anti 1. Rb{sub 2}ZnP{sub 2}Se{sub 6} has cell parameters of a = 7.4944(15) Aa, b = 7.6013(15) Aa, c = 12.729(3) Aa, α = 96.57(3) , β = 105.52(3) , γ = 110.54(3) , Z = 2, and V = 636.6(2) Aa{sup 3}. Cs{sub 2}ZnP{sub 2}Se{sub 6} has cell parameters of a = 7.6543(6) Aa, b = 7.7006(6) Aa, c = 12.7373(11) Aa, α = 97.007(7) , β = 104.335(7) , γ = 109.241(6) , Z = 2, and V = 669.54(10) Aa{sup 3}. (Copyright copyright 2016 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

P2Y6 receptor potentiates pro-inflammatory responses in macrophages and exhibits differential roles in atherosclerotic lesion development.

Directory of Open Access Journals (Sweden)

Ricardo A Garcia

Full Text Available BACKGROUND: P2Y(6, a purinergic receptor for UDP, is enriched in atherosclerotic lesions and is implicated in pro-inflammatory responses of key vascular cell types and macrophages. Evidence for its involvement in atherogenesis, however, has been lacking. Here we use cell-based studies and three murine models of atherogenesis to evaluate the impact of P2Y(6 deficiency on atherosclerosis. METHODOLOGY/PRINCIPAL FINDINGS: Cell-based studies in 1321N1 astrocytoma cells, which lack functional P2Y(6 receptors, showed that exogenous expression of P2Y(6 induces a robust, receptor- and agonist-dependent secretion of inflammatory mediators IL-8, IL-6, MCP-1 and GRO1. P2Y(6-mediated inflammatory responses were also observed, albeit to a lesser extent, in macrophages endogenously expressing P2Y(6 and in acute peritonitis models of inflammation. To evaluate the role of P2Y(6 in atherosclerotic lesion development, we used P2Y(6-deficient mice in three mouse models of atherosclerosis. A 43% reduction in aortic arch plaque was observed in high fat-fed LDLR knockout mice lacking P2Y(6 receptors in bone marrow-derived cells. In contrast, no effect on lesion development was observed in fat-fed whole body P2Y(6xLDLR double knockout mice. Interestingly, in a model of enhanced vascular inflammation using angiotensin II, P2Y(6 deficiency enhanced formation of aneurysms and exhibited a trend towards increased atherosclerosis in the aorta of LDLR knockout mice. CONCLUSIONS: P2Y(6 receptor augments pro-inflammatory responses in macrophages and exhibits a pro-atherogenic role in hematopoietic cells. However, the overall impact of whole body P2Y(6 deficiency on atherosclerosis appears to be modest and could reflect additional roles of P2Y(6 in vascular disease pathophysiologies, such as aneurysm formation.

The supplementation of low-P diets with microbial 6-phytase expressed in Aspergillus oryzae increases P and Ca digestibility in growing pigs.

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Torrallardona, D; Salvadó, R; Broz, J

2012-12-01

A trial was conducted to evaluate the dose response of a novel microbial 6-phytase expressed in Aspergillus oryzae (Ronozyme HiPhos; DSM Nutritional Products, Basel, Switzerland) in pigs. Forty-eight individually housed pigs (Landrace × Pietrain; 52 kg BW; 24 males and 24 females) were distributed among 6 experimental treatments consisting of a low-P diet (3.5 g P/kg; 1.1 g digestible P/kg), which was supplemented with 500, 1000, 2000, or 4000 units of phytase activity/kg, and a standard-P diet (4.5 g P/kg; 1.8 g digestible P/kg) that was supplemented with CaHPO(4). After 17 d, fresh feces were sampled from all pigs and the apparent total tract digestibility of DM, OM, ash, P, and Ca was measured using TiO(2) as indigestible marker. Blood samples were also obtained from each pig and serum was analyzed for P and Ca concentrations. The nonsupplemented low-P diet increased Ca and reduced P blood serum concentrations (P Phytase supplementation of the low-P diet reduced Ca (from 10.8 to 9.9 mg/dL; linear, P Phytase improved the total tract digestibility of P (from 29.0 to 62.3%; linear and quadratic, P phytase tested improves the apparent total tract digestibility of P in growing pigs and reduces P excretion in feces in a dose-dependent manner.

The cytochrome P450 CYP6P4 is responsible for the high pyrethroid resistance in knockdown resistance-free Anopheles arabiensis.

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Ibrahim, Sulaiman S; Riveron, Jacob M; Stott, Robert; Irving, Helen; Wondji, Charles S

2016-01-01

Pyrethroid insecticides are the front line vector control tools used in bed nets to reduce malaria transmission and its burden. However, resistance in major vectors such as Anopheles arabiensis is posing a serious challenge to the success of malaria control. Herein, we elucidated the molecular and biochemical basis of pyrethroid resistance in a knockdown resistance-free Anopheles arabiensis population from Chad, Central Africa. Using heterologous expression of P450s in Escherichia coli coupled with metabolism assays we established that the over-expressed P450 CYP6P4, located in the major pyrethroid resistance (rp1) quantitative trait locus (QTL), is responsible for resistance to Type I and Type II pyrethroid insecticides, with the exception of deltamethrin, in correlation with field resistance profile. However, CYP6P4 exhibited no metabolic activity towards non-pyrethroid insecticides, including DDT, bendiocarb, propoxur and malathion. Combining fluorescent probes inhibition assays with molecular docking simulation, we established that CYP6P4 can bind deltamethrin but cannot metabolise it. This is possibly due to steric hindrance because of the large vdW radius of bromine atoms of the dihalovinyl group of deltamethrin which docks into the heme catalytic centre. The establishment of CYP6P4 as a partial pyrethroid resistance gene explained the observed field resistance to permethrin, and its inability to metabolise deltamethrin probably explained the high mortality from deltamethrin exposure in the field populations of this Sudano-Sahelian An. arabiensis. These findings describe the heterogeneity in resistance towards insecticides, even from the same class, highlighting the need to thoroughly understand the molecular basis of resistance before implementing resistance management/control tools. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

IL-3 Maintains Activation of the p90S6K/RPS6 Pathway and Increases Translation in Human Eosinophils.

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Esnault, Stephane; Kelly, Elizabeth A B; Shen, Zhong-Jian; Johansson, Mats W; Malter, James S; Jarjour, Nizar N

2015-09-15

IL-5 is a major therapeutic target to reduce eosinophilia. However, all of the eosinophil-activating cytokines, such as IL-5, IL-3, and GM-CSF, are typically present in atopic diseases, including allergic asthma. As a result of the functional redundancy of these three cytokines on eosinophils and the loss of IL-5R on airway eosinophils, it is important to take IL-3 and GM-CSF into account to efficiently reduce tissue eosinophil functions. Moreover, these three cytokines signal through a common β-chain receptor but yet differentially affect protein production in eosinophils. Notably, the increased ability of IL-3 to induce the production of proteins, such as semaphorin-7A, without affecting mRNA levels suggests a unique influence of IL-3 on translation. The purpose of this study was to identify the mechanisms by which IL-3 distinctively affects eosinophil function compared with IL-5 and GM-CSF, with a focus on protein translation. Peripheral blood eosinophils were used to study intracellular signaling and protein translation in cells activated with IL-3, GM-CSF, or IL-5. We establish that, unlike GM-CSF or IL-5, IL-3 triggers prolonged signaling through activation of ribosomal protein S6 (RPS6) and the upstream kinase 90-kDa ribosomal S6 kinase (p90S6K). Blockade of p90S6K activation inhibited phosphorylation of RPS6 and IL-3-enhanced semaphorin-7A translation. Furthermore, in an allergen-challenged environment, in vivo phosphorylation of RPS6 and p90S6K was enhanced in human airway compared with circulating eosinophils. Our findings provide new insights into the mechanisms underlying differential activation of eosinophils by IL-3, GM-CSF, and IL-5. These observations identify IL-3 and its downstream intracellular signals as novel targets that should be considered to modulate eosinophil functions. Copyright © 2015 by The American Association of Immunologists, Inc.

Interaction of actinide ions with [NaP5W30O110]14- and [P2W18O62]6-

International Nuclear Information System (INIS)

Choppin, G.R.; Wall, D.E.

2003-01-01

Stability constants (log β 101 ) of Th 4+ , UO 2 2+ , NpO 2 + and Am 3+ with [NaP 5 W 30 O 110 ] 14- were determined by solvent extraction (μ = 0.1M NaCl) and found to be 6.18 ± 0.07, 3.80 ± 0.06, 2.98 ± 0.04, and 5.85 ± 0.05, respectively. The order of stability constants: Th 4+ > Am 3+ > UO 2 2+ > NpO 2 + is due to electrostatic repulsion between the actinyl oxygens and oxygens on the polyoxometalate surface. The order of stability constants for metal complexes with [P 2 W 18 O 62 ] 6- is Th 4+ > UO 2 2+ > Eu 3+ >NpO 2 + because the steric repulsion between actinyl oxygens and oxygens on polyoxometalate are less important. Enthalpies of complexation were measured by calorimetric titration of Th 4+ , UO 2 2+ , Nd 3+ with [NaP 5 W 30 O 110 ] 14- and [P 2 W 18 O 62 ] 6- . The results indicate that the conformation and charge distribution of the microscopic surface structures are important factors in the formation of pseudocolloids. (author)

Initiation of proteolysis of yeast fructose-1,6-bisphosphatase by pH-control of adenylate cyclase

International Nuclear Information System (INIS)

Holzer, H.; Purwin, C.; Pohlig, G.; Scheffers, W.A.; Nicolay, K.

1986-01-01

Addition of fermentable sugars or uncouplers such as CCCP to resting yeast cells grown on glucose initiates phosphorylation of fructose-1,6-bisphosphatase (FBPase). There is good evidence that phosphorylation marks FBPase for proteolytic degradation. 31 P-NMR measurements of the cytosolic pH of yeast cells demonstrated a decrease of the cytosolic pH from 7.0 to 6.5 after addition of glucose or CCCP to starved yeast. Activity of adenylate cyclase in permeabilized yeast cells increases 2-3-fold when the pH is lowered from 7.0 to 6.5. It is concluded that pH controlled activation of adenylate cyclase causes the previously described increase in cyclic AMP which leads to phosphorylation of FBPase and finally to proteolysis of FBPase

Expression and subcellular localization of p70S6 kinase under heart failure

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Usenko V. S.

2010-11-01

Full Text Available The PI3K/PDK/Akt/mTOR/p70S6K signaling pathway is primary associated with the activation of insulin receptors and is important for cardiomyocytes survival. p70S6K is a key regulator of the speed and efficiency of protein biosynthesis within the cell. Recently the pro-apoptotic protein BAD has been identified as a new target for р70S6K1. BAD is inactivated in normal cardiomyocytes by р70S6K1 phosphorylation which prevents the cardiomyocytes apoptosis. Aim. To study possible changes in р70S6K1 expression and/or cellular localization at heart failure progression – in DCM- affected human myocardia and murine hearts with experimental DCM-like pathology. Methods. Western-blot analysis and immunohystochemistry. Results. The substantial decrease in р70S6K1 level was observed at the final stage of pathology progression and in the dynamics of DCM pathogenesis as well. For the first time relocalization of the protein to the connective tissue was shown according to the Western-blot results. Conclusions. The data obtained allow us to understand a possible role of р70S6K1 in the regulation of stress-induced apoptotic signaling in cardiomyocytes.

Pathogenicity of Rhodococcus equi in mice, isolated from environment, human and horse clinical samples Patogenicidade atogenicidade em camundongos de isolados clí- clínicos, nicos, ambientais e humanos de Rhodococcus equi

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Mateus M. Costa

2006-09-01

Full Text Available Rhodococcus equi is a facultative intracellular pathogen associated with bronchopneumonia, mesenteric lymphadenitis and enterocolitis in foals. Although R. equi is likely to be found in every horse-breeding farm, the clinical disease is unrecognized in most of them. Capsule components, equi factor, micolic acid and some products encoded by the large 85-90Kb plasmid were described as virulence factors. However, the pathogenesis of R. equi infections and the sensibility of foals are not completely understood. The aim of this study was evaluate the virulence of R. equi isolated from human, horses and environment for mices. Nine strains carrying the 85-90Kb plasmid isolated from foal clinical specimens, one from immunodeficient human patient and six plasmidless strains (four isolated from feces, one from pasture and one from immunodeficient human patient were inoculated in cyclophosphamide immunossuppressed mice. The pathological changes and viability of R. equi cells in the liver of mice was verified after the 3rd, 6th an 10th day after inoculation for horse and environmental isolates and for R. equi isolates from human patients on the 1st, 3rd and 6th day. During the necropsy procedures, infiltrate of macrophages and pyogranulomatous lesions were detected after the sixth pos-inoculation day in the liver and spleen. In horse isolates, only plasmid positive strains were virulent, but in human isolates both strains (plasmid positive e plasmid negative were virulent. Both groups of the immunossupressed mice inoculated with R. equi isolated from environment showed pathological changes. All R. equi strains were unable to kill non imunossuppressed mice.Rhodococ-cus equi é um patógeno intracelular facultativo associado com broncopneumonia, linfadenite mesentérica e enterocolite em potros. Apesar do patógeno ser amplamente distribuído no ambiente equino, a doença não é encontrada em todos os criatórios. Componentes capsulares, "fator equi", �

Immunization by intrabronchial administration to 1-week-old foals of an unmarked double gene disruption strain of Rhodococcus equi strain 103+.

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Pei, Yanlong; Nicholson, Vivian; Woods, Katharine; Prescott, John F

2007-11-15

Rhodococcus equi causes fatal granulomatous pneumonia in foals and immunocompromised animals and humans. However, there is no effective vaccine against this infection. In this study, the chromosomal genes isocitrate lyase (icl) and cholesterol oxidase (choE) were chosen as targets for mutation and assessment of the double mutant as an intrabronchial vaccine in 1-week-old foals. Using a modification of a suicide plasmid previously developed in this laboratory, we developed a choE-icl unmarked deletion mutant of R. equi strain 103+. Five 1-week-old foals were infected intrabronchially with the mutant and challenged intrabronchially with the parent, virulent, strain 2 weeks later. Three of the foals were protected against pneumonia caused by the virulent strain, but the other two foals developed pneumonia caused by the mutant strain during the post-challenge period. Since infection of 3-week-old foals by an icl mutant in an earlier study had shown complete attenuation of the strain, we conclude that a proportion of foals in the 1st week or so of life are predisposed to developing R. equi pneumonia because of an inability to mount an effective immune response. This has been suspected previously but this is the first time that this has been demonstrated experimentally.

Cdt1p, through its interaction with Mcm6p, is required for the formation, nuclear accumulation and chromatin loading of the MCM complex.

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Wu, Rentian; Wang, Jiafeng; Liang, Chun

2012-01-01

Regulation of DNA replication initiation is essential for the faithful inheritance of genetic information. Replication initiation is a multi-step process involving many factors including ORC, Cdt1p, Mcm2-7p and other proteins that bind to replication origins to form a pre-replicative complex (pre-RC). As a prerequisite for pre-RC assembly, Cdt1p and the Mcm2-7p heterohexameric complex accumulate in the nucleus in G1 phase in an interdependent manner in budding yeast. However, the nature of this interdependence is not clear, nor is it known whether Cdt1p is required for the assembly of the MCM complex. In this study, we provide the first evidence that Cdt1p, through its interaction with Mcm6p with the C-terminal regions of the two proteins, is crucial for the formation of the MCM complex in both the cytoplasm and nucleoplasm. We demonstrate that disruption of the interaction between Cdt1p and Mcm6p prevents the formation of the MCM complex, excludes Mcm2-7p from the nucleus, and inhibits pre-RC assembly and DNA replication. Our findings suggest a function for Cdt1p in promoting the assembly of the MCM complex and maintaining its integrity by interacting with Mcm6p.

The supplementation of low-P diets with microbial 6-phytase expressed in Aspergillus oryzae improves P digestibility in sows.

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Torrallardona, D; Llauradó, L; Broz, J

2012-12-01

Two trials were conducted to evaluate a novel microbial 6-phytase expressed in Aspergillus oryzae (Ronozyme HiPhos; DSM Nutritional Products, Basel, Switzerland) in gestating and lactating sows. In the first trial, 24 sows (Duroc × Landrace; 223 kg BW) were offered, at 16 d of gestation, a low-P control diet (formulated to provide 4.0 g total P/kg; 1.5 g digestible P/kg) supplemented with 0, 500, or 1000 phytase activity (FYT)/kg of phytase. Two weeks later, fresh feces were sampled from all sows and the apparent total tract digestibility of P was measured using TiO(2) as indigestible marker. Phytase supplementation did not (P > 0.10) affect the total tract digestibility of P but reduced (P phytase. After 2 wk, fresh feces were sampled from all sows and the apparent total tract digestibility of P was measured using TiO(2) as indigestible marker. Phytase supplementation improved (P phytase tested increased the apparent total tract digestibility of P in sows and reduced P excretion in feces.

The effect of mutation on Rhodococcus equi virulence plasmid gene expression and mouse virulence.

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Ren, Jun; Prescott, John F

2004-11-15

An 81 kb virulence plasmid containing a pathogenicity island (PI) plays a crucial role in the pathogenesis of Rhodococcus equi pneumonia in foals but its specific function in virulence and regulation of plasmid-encoded virulence genes is unclear. Using a LacZ selection marker developed for R. equi in this study, in combination with an apramycin resistance gene, an efficient two-stage homologous recombination targeted gene mutation procedure was used to mutate three virulence plasmid genes, a LysR regulatory gene homologue (ORF4), a ResD-like two-component response regulator homologue (ORF8), and a gene (ORF10) of unknown function that is highly expressed by R. equi inside macrophages, as well as the chromosomal gene operon, phoPR. Virulence testing by liver clearance after intravenous injection in mice showed that the ORF4 and ORF8 mutants were fully attenuated, that the phoPR mutant was hypervirulent, and that virulence of the ORF10 mutant remained unchanged. A virulence plasmid DNA microarray was used to compare the plasmid gene expression profile of each of the four gene-targeted mutants against the parental R. equi strain. Changes were limited to PI genes and gene induction was observed for all mutants, suggesting that expression of virulence plasmid genes is dominated by a negative regulatory network. The finding of attenuation of ORF4 and ORF8 mutants despite enhanced transcription of vapA suggests that factors other than VapA are important for full expression of virulence. ORF1, a putative Lsr antigen gene, was strongly and similarly induced in all mutants, implying a common regulatory pathway affecting this gene for all four mutated genes. ORF8 is apparently the centre of this common pathway. Two distinct highly correlated gene induction patterns were observed, that of the ORF4 and ORF8 mutants, and that of the ORF10 and phoPR mutants. The gene induction pattern distinguishing these two groups paralleled their virulence in mice.

Functional characterization of 3-ketosteroid 9α-hydroxylases in Rhodococcus ruber strain chol-4.

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Guevara, Govinda; Heras, Laura Fernández de Las; Perera, Julián; Llorens, Juana María Navarro

2017-09-01

The 3-Ketosteroid-9α-Hydroxylase, also known as KshAB [androsta-1,4-diene-3,17-dione, NADH:oxygen oxidoreductase (9α-hydroxylating); EC 1.14.13.142)], is a key enzyme in the general scheme of the bacterial steroid catabolism in combination with a 3-ketosteroid-Δ 1 -dehydrogenase activity (KstD), being both responsible of the steroid nucleus (rings A/B) breakage. KshAB initiates the opening of the steroid ring by the 9α-hydroxylation of the C9 carbon of 4-ene-3-oxosteroids (e.g. AD) or 1,4-diene-3-oxosteroids (e.g. ADD), transforming them into 9α-hydroxy-4-androsten-3,17-dione (9OHAD) or 9α-hydroxy-1,4-androstadiene-3,17-dione (9OHADD), respectively. The redundancy of these enzymes in the actinobacterial genomes results in a serious difficulty for metabolic engineering this catabolic pathway to obtain intermediates of industrial interest. In this work, we have identified three homologous kshA genes and one kshB gen in different genomic regions of R. ruber strain Chol-4. We present a set of data that helps to understand their specific roles in this strain, including: i) description of the KshAB enzymes ii) construction and characterization of ΔkshB and single, double and triple ΔkshA mutants in R. ruber iii) growth studies of the above strains on different substrates and iv) genetic complementation and biotransformation assays with those strains. Our results show that KshA2 isoform is needed for the degradation of steroid substrates with short side chain, while KshA3 works on those molecules with longer side chains. KshA1 is a more versatile enzyme related to the cholic acid catabolism, although it also collaborates with KshA2 or KshA3 activities in the catabolism of steroids. Accordingly to what it is described for other Rhodococcus strains, our results also suggest that the side chain degradation is KshAB-independent. Copyright © 2017 Elsevier Ltd. All rights reserved.

UDP/P2Y6 receptor signaling regulates IgE-dependent degranulation in human basophils

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Manabu Nakano

2017-10-01

Conclusions: This study showed that UDP/P2Y6 receptor signaling is involved in the regulation of IgE-dependent degranulation in basophils, which might stimulate the P2Y6 receptor via the autocrine secretion of UTP. Thus, this receptor represents a potential target to regulate IgE-dependent degranulation in basophils during allergic diseases.

Modification of nylon-6 with semi-rigid poly (p-diphenyl oxide terephthalamide)

International Nuclear Information System (INIS)

Lin, M.F.; Wang, H.H.; Shu, Y.C.

1994-01-01

In this study, the flexible nylon-6 was reinforced by the semi-rigid aromatic polyamide, poly (p-diphenyl oxide terephthalamide) (POA), through physical polybending and chemical copolymerization using p-aminophenylacetic acid (P-APA) as a coupling agent. From the DSC measurements, it was shown that Tg of the polyblends was increased with the increase of POA content. The Tg and Tm of multiblock copolyamides were found to be higher than those of polyblends and triblock copolyamides. From the Rheovibron measurements, it was shown that the POA with nylon-6 poy-blends exhibited higher E' and Tg value than that of nylon-6, which increased with the increase of POA content. The Tg and E' values of multiblock copolyamides were higher than those of polyblends triblock copolyamides. Scanning electron microscopy revealed that the polyblends were a dispersed phase structure, although the multiblock copolyamides exhibited a homogeneous texture rather than an aggregated one. From the wide-angle x-ray diffraction pattern, the triblock copolyamides and polyblends had two diffraction peaks, i.e., 2 θ = 20.5 degrees and 24 degrees. However, the multiblock had only one at 2 θ = 20 degrees, which indicates that a different crystal structure of multiblock copolyamides. For the mechanical properties, it was found that the multiblock copolyamides had a more significant reinforcing effect than those of polyblends and triblock copolyamides. 21 refs., 11 figs., 5 tabs

THE EXON 5, 6, 7, 8 OF P53 MUTATIONS IN ORAL SQUAMOUS CELLS CARCINOMA

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Retno P Rahayu

2012-04-01

Full Text Available Genetic instability may underlie the etiology of multistep carcinogenesis. The altered p53 gene observed in tumors may represent the expression of such instability and may allow the accumulation of other gene alterations caused by multiple mechanism. p53 gene is the guardian of the genome, that is why we pay more attention to this gene. In this study, we evaluated the significance of p53 mutation in 55 patient with oral squamous carcinoma. Thirty among them underwent well-differentiated carcinoma, while the remaining 25 patients underwent poorly differentiated carcinoma. The mutations were detected by PCR-SSCP (Single strand Conformational Polymorphism analysis in the region between exon 5 and exon 8. The results indicated that the p53 mutation in exon 5 (40%, exon 6 (28%, exon 7 (24% and exon 8 (8% were associated with poorly differentiated carcinoma, whereas mutation in exon 5 (10%, exon 6 (30%, exon 7 (40% and exon 8 (20% were associated with well-differentiated carcinoma. These observations suggest that p53 mutation in exon 5, 6, and 7 have strong correlation with poorly differentiated in oral squamous carcinoma while well-differentiated level was related with mutation in exon 6,7 and 8.

Dek-can rearrangement in translocation (6;9)(p23;q34)

NARCIS (Netherlands)

Soekarman, D.; von Lindern, M.; van der Plas, D. C.; Selleri, L.; Bartram, C. R.; Martiat, P.; Culligan, D.; Padua, R. A.; Hasper-Voogt, K. P.; Hagemeijer, A.

1992-01-01

The translocation (6;9)(p23;q34) is mainly found in specific subtypes of acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). The diagnosis of this translocation is not easy since the cytogenetic change is quite subtle. The two genes involved in this translocation were recently isolated

PF-4708671, a specific inhibitor of p70 ribosomal S6 kinase 1, activates Nrf2 by promoting p62-dependent autophagic degradation of Keap1

Energy Technology Data Exchange (ETDEWEB)

Park, Jeong Su [Severance Biomedical Science Institute (Korea, Republic of); Yonsei Biomedical Research Institute, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752 (Korea, Republic of); Kang, Dong Hoon [Department of Life Science and Ewha Research Center for Systems Biology (Korea, Republic of); The Research Center for Cell Homeostasis, Ewha Womans University, Seoul 127-750 (Korea, Republic of); Lee, Da Hyun [Severance Biomedical Science Institute (Korea, Republic of); Yonsei Biomedical Research Institute, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752 (Korea, Republic of); Bae, Soo Han, E-mail: soohanbae@yuhs.ac [Severance Biomedical Science Institute (Korea, Republic of); Yonsei Biomedical Research Institute, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752 (Korea, Republic of)

2015-10-23

p70 ribosomal S6 kinase 1 (S6K1) is an important serine/threonine kinase and downstream target of the mechanistic target of rapamycin complex 1 (mTORC1) signaling pathway. PF-4708671 is a specific inhibitor of S6K1, and prevents S6K1-mediated phosphorylation of the S6 protein. PF-4708671 treatment often leads to apoptotic cell death. However, the protective mechanism against PF-4708671-induced cell death has not been elucidated. The nuclear factor erythroid 2-related factor 2 (Nrf2)-Kelch-like ECH-associated protein 1 (Keap1) pathway is essential for protecting cells against oxidative stress. p62, an adaptor protein in the autophagic process, enhances Nrf2 activation through the impairment of Keap1 activity. In this study, we showed that PF-4708671 induces autophagic Keap1 degradation-mediated Nrf2 activation in p62-dependent manner. Furthermore, p62-dependent Nrf2 activation plays a crucial role in protecting cells from PF-4708671-mediated apoptosis. - Highlights: • PF-4708671, a S6K1-specific inhibitor, prevents S6K1-mediated S6 phosphorylation. • However, PF-4708671 treatment often leads to apoptotic cell death. • Protective mechanism against PF-4708671-induced cell death remains to be elucidated. • PF-4708671 induced p62-dependent, autophagic Keap1 degradation-mediated Nrf2 activation. • p62-dependent Nrf2 activation protects cells from PF-4708671-mediated apoptosis.

IL-6 modulates hepatocyte proliferation via induction of HGF/p21cip1: Regulation by SOCS3

International Nuclear Information System (INIS)

Sun Rui; Jaruga, Barbara; Kulkarni, Shailin; Sun Haoyu; Gao Bin

2005-01-01

The precise role of IL-6 in liver regeneration and hepatocyte proliferation is controversial and the role of SOCS3 in liver regeneration remains unknown. Here we show that in vitro treatment with IL-6 inhibited primary mouse hepatocyte proliferation. IL-6 induced p21 cip1 protein expression in primary mouse hepatocytes. Disruption of the p21 cip1 gene abolished the inhibitory effect of IL-6 on cell proliferation. Co-culture with nonparenchymal liver cells diminished IL-6 inhibition of hepatocyte proliferation, which was likely due to IL-6 stimulation of nonparenchymal cells to produce HGF. Finally, IL-6 induced higher levels of p21 cip1 protein expression and a slightly stronger inhibition of cell proliferation in SOCS3 +/- mouse hepatocytes compared to wild-type hepatocytes, while liver regeneration was enhanced and prolonged in SOCS3 +/- mice. Our findings suggest that IL-6 directly inhibits hepatocyte proliferation via a p21 cip1 -dependent mechanism and indirectly enhances hepatocyte proliferation via stimulating nonparenchymal cells to produce HGF. SOCS3 negatively regulates liver regeneration

Electron-impact excitation cross sections of the higher argon 3p5np (n=5,6,7) levels

International Nuclear Information System (INIS)

Weber, Tobin; Boffard, John B.; Lin, Chun C.

2003-01-01

We have measured the electron-impact excitation cross sections for argon into the ten levels of 3p 5 5p configuration, as well as numerous levels of the 3p 5 6p and 3p 5 7p configurations. Fluorescence from the decay of the excited atoms to the levels of 3p 5 4s configuration was used to determine the optical-emission cross sections. These results were combined with transition probabilities to find apparent cross sections into the 3p 5 5p and 3p 5 6p levels. These new cross sections of the Ar(3p 5 np) levels along with the available Ne(2p 5 3p), Kr(4p 5 5p), and Xe(5p 5 6p) data help provide a global view of the excitation behaviors of the np 5 n ' p levels of the rare-gas series

Degradation of p53 by human Alphapapillomavirus E6 proteins shows a stronger correlation with phylogeny than oncogenicity.

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Leiping Fu

2010-09-01

Full Text Available Human Papillomavirus (HPV E6 induced p53 degradation is thought to be an essential activity by which high-risk human Alphapapillomaviruses (alpha-HPVs contribute to cervical cancer development. However, most of our understanding is derived from the comparison of HPV16 and HPV11. These two viruses are relatively distinct viruses, making the extrapolation of these results difficult. In the present study, we expand the tested strains (types to include members of all known HPV species groups within the Alphapapillomavirus genus.We report the biochemical activity of E6 proteins from 27 HPV types representing all alpha-HPV species groups to degrade p53 in human cells. Expression of E6 from all HPV types epidemiologically classified as group 1 carcinogens significantly reduced p53 levels. However, several types not associated with cancer (e.g., HPV53, HPV70 and HPV71 were equally active in degrading p53. HPV types within species groups alpha 5, 6, 7, 9 and 11 share a most recent common ancestor (MRCA and all contain E6 ORFs that degrade p53. A unique exception, HPV71 E6 ORF that degraded p53 was outside this clade and is one of the most prevalent HPV types infecting the cervix in a population-based study of 10,000 women. Alignment of E6 ORFs identified an amino acid site that was highly correlated with the biochemical ability to degrade p53. Alteration of this amino acid in HPV71 E6 abrogated its ability to degrade p53, while alteration of this site in HPV71-related HPV90 and HPV106 E6s enhanced their capacity to degrade p53.These data suggest that the alpha-HPV E6 proteins' ability to degrade p53 is an evolved phenotype inherited from a most recent common ancestor of the high-risk species that does not always segregate with carcinogenicity. In addition, we identified an amino-acid residue strongly correlated with viral p53 degrading potential.

Rescue of p53 function by small-molecule RITA in cervical carcinoma by blocking E6-mediated degradation.

Science.gov (United States)

Zhao, Carolyn Ying; Szekely, Laszlo; Bao, Wenjie; Selivanova, Galina

2010-04-15

Proteasomal degradation of p53 by human papilloma virus (HPV) E6 oncoprotein plays a pivotal role in the survival of cervical carcinoma cells. Abrogation of HPV-E6-dependent p53 destruction can therefore be a good strategy to combat cervical carcinomas. Here, we show that a small-molecule reactivation of p53 and induction of tumor cell apoptosis (RITA) is able to induce the accumulation of p53 and rescue its tumor suppressor function in cells containing high-risk HPV16 and HPV18 by inhibiting HPV-E6-mediated proteasomal degradation. RITA blocks p53 ubiquitination by preventing p53 interaction with E6-associated protein, required for HPV-E6-mediated degradation. RITA activates the transcription of proapoptotic p53 targets Noxa, PUMA, and BAX, and repressed the expression of pro-proliferative factors CyclinB1, CDC2, and CDC25C, resulting in p53-dependent apoptosis and cell cycle arrest. Importantly, RITA showed substantial suppression of cervical carcinoma xenografts in vivo. These results provide a proof of principle for the treatment of cervical cancer in a p53-dependent manner by using small molecules that target p53. (c)2010 AACR.

Cross sections for electron-impact excitation of krypton from the levels of 4p6, 4p55s, and 4p55p configurations

International Nuclear Information System (INIS)

Zeng Jiaolong; Yuan Jianmin; Wu Jianhua; Jin Fengtao; Zhao Gang

2005-01-01

The electron-impact excitation cross sections at low electron energies have been calculated using a fully relativistic R-matrix method for transitions between levels of 4p 6 , 4p 5 5s, and 4p 5 5p configurations. To ensure the convergence of results, we have paid special attention to the factors that may affect the convergence of cross sections. For examples, we have included extensive configuration interactions in the wave-function expansion of the target states. A large enough R-matrix boundary has been taken to ensure the convergence of atomic wave functions. Contributions to cross sections from a large number of partial waves (up to J=39.5) have been explicitly calculated. The final results are in good agreement with recent experimental data by Jung et al. [Phys. Rev. Lett. 94, 163202 (2005)] after shifting the position of electron energy. The relative difference is about 10% for four transitions out of the metastable levels. The results eliminated the significant discrepancies between theory and experimental work on excitation cross sections out of the metastable levels reported in the literature

The 4p6 autoionization cross section of Rb atoms excited by low-energy electron impact

International Nuclear Information System (INIS)

Borovik, A; Roman, V; Kupliauskienė, A

2012-01-01

The autoionization cross section of rubidium atoms was obtained by measuring the total normalized intensities of ejected-electron spectra arising from the decay of the 4p 5 n 1 l 1 n 2 l 2 autoionizing levels. The electron impact energy range from the 4p 6 excitation threshold at 15.31 up to 50 eV was investigated. The cross section reaches the maximum value of (2.9 ± 0.6) × 10 −16  cm 2 at 21.8 eV impact energy. The general behaviours of the cross section and the role of particular autoionizing configurations in its formation were considered on the basis of large-scale configuration interaction calculations of energies, cross sections, autoionization probabilities in 5snl(n ⩽ 7; l ⩽ 4) and 4d nl(n ⩽ 5; l ⩽ 2) configurations as well as the measured excitation functions for the lowest levels in 5s 2 and 4d5s configurations. The resonance behaviour of the cross section between 15.3 and 18.5 eV impact energy is caused exclusively by the negative-ion resonances present close to the excitation thresholds of the (5s 2 ) 2 P and (4d5s) 4 P autoionizing levels. At higher impact energies, the autoionization cross section is composed of contributions from the high-lying quartet and doublet levels in 4d5s, 5p and 5s5p, 5d, 6s, 6p configurations. From the comparison of the present data with available experimental and calculated ionization cross sections, the 5s + 4p 6 direct ionization cross section of rubidium atoms was determined with the maximum value of (7.2 ± 2.2) × 10 −16  cm 2 at 36 eV. It was also found that the 4p 6 excitation–autoionization is the dominant indirect ionization process contributing over 30% of the total single ionization of rubidium atoms by electron impact in the 15.3–50 eV energy range. (paper)

V0 production in p+A collisions at √(s)=41.6 GeV

International Nuclear Information System (INIS)

Abt, I.; Kisel, I.; Adams, M.; Cruse, C.; Ehret, K.; Funcke, M.; Schwenninger, B.; Wegener, D.; Agari, M.; Bauer, C.; Braeuer, M.; Hofmann, W.; Jagla, T.; Knoepfle, K.T.; Pleier, M.A.; Reeves, K.; Sanchez, F.; Schmelling, M.; Schwingenheuer, B.; Sciacca, F.; Albrecht, H.; Aplin, S.J.; Bagaturia, Y.; Egorytchev, V.; Emeliyanov, D.; Flammer, J.; Goloubkov, D.; Golubkov, Y.; Hohlmann, M.; Lewendel, B.; Lomonosov, B.; Masciocchi, S.; Medinnis, M.; Mevius, M.; Michetti, A.; Negodaev, M.; Noerenberg, M.; Nunez Pardo de Vera, M.T.; Padilla, C.; Ressing, D.; Riu, I.; Rybnikov, V.; Schmidt, B.; Schwarz, A.S.; Soezueer, L.; Somov, A.; Somov, S.; Spengler, J.; Wurth, R.; Aleksandrov, A.; Bohm, G.; Gellrich, A.; Hernandez, J.M.; Mankel, R.; Nowak, S.; Schreiner, A.; Schwanke, U.; Walter, M.; Amaral, V.; Amorim, A.; Bastos, J.; Batista, J.; Carvalho, J.; Silva, L.; Wolters, H.; Aushev, V.; Prystupa, S.; Pugatch, V.; Vassiliev, Yu.; Balagura, V.; Bobchenko, B.; Bogatyrev, A.; Danilov, M.; Essenov, S.; Fominykh, B.; Golutvin, A.; Gouchtchine, O.; Guilitsky, Yu.; Igonkina, O.; Khasanov, F.; Kvaratskheliia, T.; Matchikhilian, I.; Mikhailov, Yu.; Mizuk, R.; Popov, V.; Rostovtseva, I.; Tikhomirov, I.; Titov, M.; Zaitsev, Yu.; Zhelezov, A.; Bargiotti, M.; Bertin, A.; Bruschi, M.; De Castro, S.; Fabbri, L.; Faccioli, P.; Giacobbe, B.; Grimaldi, F.; Massa, I.; Piccinini, M.; Poli, M.; Semprini-Cesari, N.; Spighi, R.; Villa, M.; Vitale, A.; Zoccoli, A.; Barsukova, O.; Belkov, A.; Belkov, Ar.; Belotelov, I.; Golutvin, I.; Karpenko, N.; Kiryushin, Yu.; Lanyov, A.; Solunin, S.; Bauer, T.S.; Hulsbergen, W.; Sbrizzi, A.; Boecker, M.; Buchholz, P.; Husemann, U.; Keller, S.; Walenta, A.H.; Werthenbach, U.; Bruinsma, M.; Ouchrif, M.; Buran, T.; Danielsen, K.M.; Ould-Saada, F.; Pylypchenko, Y.; Conde, P.; Dam, M.; Groth-Jensen, J.; Hansen, J.D.; Klinkby, E.; Muresan, R.; Petersen, B.A.; Xella-Hansen, S.; Deppe, H.; Dreis, H.B.; Eisele, F.; Feuerstack-Raible, M.; Gradl, S.; Gradl, W.; Hott, T.; Kessler, J.; Krauss, C.; Rick, H.; Uwer, U.; Dong, X.; Jiang, C.; Zheng, Z.; Garrido, Ll.; Peralta, D.; Glaess, J.; Maenner, R.; Wurz, A.; Gorbounov, I.; Zeuner, T.; Gorisek, A.; Kupper, S.; Pestotnik, R.; Staric, M.; Zivko, T.; Goulart, D.C.; Schwartz, A.J.; Ispiryan, M.; Lau, K.; Pyrlik, J.; Subramania, H.S.; Kapitza, H.; Symalla, M.; Eldik, C. van; Karabekyan, S.; Pernack, R.; Schroeder, H.; Zimmermann, R.; Kolanoski, H.; Kruecker, D.; Lohse, T.; Medin, G.; Nedden, M. zur; Stegmann, C.; Korpar, S.; Kreuzer, P.; Krizan, P.; Stanovnik, A.; Pose, D.; Robmann, P.; Shuvalov, S.; Spiridonov, A.; Tsakov, I.; Vukotic, I.; Wahlberg, H.; Wang, J.J.; Zavertyaev, M.

2009-01-01

Inclusive doubly differential cross sections d 2 σ pA /dx F dp T 2 as a function of Feynman-x(x F ) and transverse momentum (p T ) for the production of K S 0 , Λ and anti Λ in proton-nucleus interactions at 920 GeV are presented. The measurements were performed by HERA-B in the negative x F range (-0.12 F T =1.6 GeV/c. Results for three target materials: carbon, titanium and tungsten are given. The ratios of production cross sections are presented and discussed. The Cronin effect is clearly observed for all three V 0 species. The atomic number dependence is parameterized as σ pA =σ pN .A α where σ pN is the proton-nucleon cross section. The measured values of α are all near one. The results are compared with EPOS 1.67 and PYTHIA 6.3. EPOS reproduces the data to within ∼20% except at very low transverse momentum. (orig.)

Infecção pulmonar por rhodococcus equi em doente vih+ - revisão baseada num caso clínico

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João Faria

2013-03-01

Full Text Available Apesar do aumento no número de casos de infecção humana por Rhodococcus equi (R. equi registado nas últimas décadas, sobretudo em indivíduos infectados pelo vírus da imunodeficiência humana (VIH, esse diagnóstico permanece uma raridade. Os autores apresentam um caso de infecção pulmonar por R. equi num homem de 36 anos com síndrome de imunodeficiência adquirida (SIDA. A doença manifestou-se de forma insidiosa, apresentando-se radiologicamente sob a forma de lesão pulmonar esquerda com cavitação e nível hidroaéreo, tendo sido inicialmente colocada a hipótese diagnóstica de tuberculose pulmonar (TP dada a pesquisa de bacilos álcool-ácido resistentes (BAAR na expectoração ter sido positiva. Posteriormente, foi possível isolar o R. equi nas culturas de expectoração e lavado broncoalveolar (LBA. O estudo anátomo-patológico confirmou a presença concomitante de malacoplaquia pulmonar. O doente cumpriu antibioterapia dirigida ao agente e teve boa evolução clínica, analítica e radiológica. A propósito deste caso, os autores fazem uma revisão teórica do tema à luz dos conhecimentos actuais.

Overexpression of Notch3 and pS6 Is Associated with Poor Prognosis in Human Ovarian Epithelial Cancer

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Zhaoxia Liu

2016-01-01

Full Text Available Notch3 and pS6 play important roles in tumor angiogenesis. To assess the expression of Notch3 and pS6 in Chinese ovarian epithelial cancer patients, a ten-year follow-up study was performed in ovarian epithelial cancer tissues from 120 specimens of human ovarian epithelial cancer, 30 specimens from benign ovarian tumors, and 30 samples from healthy ovaries by immunohistochemistry. The results indicate that the expression of Notch3 and pS6 was higher in ovarian epithelial cancer than in normal ovary tissues and in benign ovarian tumor tissues (p0.05 but positively associated with clinical stage, pathological grading, histologic type, lymph node metastasis, and ascites (p<0.05 or p<0.01. A follow-up survey of 64 patients with ovarian epithelial cancer showed that patients with high Notch3 and pS6 expression had a shorter survival time (p<0.01, in which the clinical stage (p<0.05 and Notch3 expression (p<0.01 played important roles. In conclusion, Notch3 and pS6 are significantly related to ovarian epithelial cancer development and prognosis, and their combination represents a potential biomarker and therapeutic target in ovarian tumor angiogenesis.

Glutamic Acid Residues in HIV-1 p6 Regulate Virus Budding and Membrane Association of Gag.

Science.gov (United States)

Friedrich, Melanie; Setz, Christian; Hahn, Friedrich; Matthaei, Alina; Fraedrich, Kirsten; Rauch, Pia; Henklein, Petra; Traxdorf, Maximilian; Fossen, Torgils; Schubert, Ulrich

2016-04-25

The HIV-1 Gag p6 protein regulates the final abscission step of nascent virions from the cell membrane by the action of its two late (L-) domains, which recruit Tsg101 and ALIX, components of the ESCRT system. Even though p6 consists of only 52 amino acids, it is encoded by one of the most polymorphic regions of the HIV-1 gag gene and undergoes various posttranslational modifications including sumoylation, ubiquitination, and phosphorylation. In addition, it mediates the incorporation of the HIV-1 accessory protein Vpr into budding virions. Despite its small size, p6 exhibits an unusually high charge density. In this study, we show that mutation of the conserved glutamic acids within p6 increases the membrane association of Pr55 Gag followed by enhanced polyubiquitination and MHC-I antigen presentation of Gag-derived epitopes, possibly due to prolonged exposure to membrane bound E3 ligases. The replication capacity of the total glutamic acid mutant E0A was almost completely impaired, which was accompanied by defective virus release that could not be rescued by ALIX overexpression. Altogether, our data indicate that the glutamic acids within p6 contribute to the late steps of viral replication and may contribute to the interaction of Gag with the plasma membrane.

High resolution emission Fourier transform infrared spectra of the 4p-5s and 5p-6s bands of ArH.

Science.gov (United States)

Baskakov, O I; Civis, S; Kawaguchi, K

2005-03-15

In the 2500-8500 cm(-1) region several strong emission bands of (40)ArH were observed by Fourier transform spectroscopy through a dc glow discharge in a mixture of argon and hydrogen. Rotational-electronic transitions of the two previously unstudied 4p-5s and 5p-6s,v = 0-0, bands of (40)ArH were measured and assigned in the 6060 and 3770 cm(-1) regions, respectively. A simultaneous fit of the emission transitions of the 4p-5s and 5p-6s bands and an extended set of transitions of the 6s-4p band observed by Dabrowski, Tokaryk, and Watson [J. Mol. Spectrosc. 189, 95 (1998)] and remeasured in the present work yielded consistent values of the spectroscopic parameters of the electronic states under investigation. In the branch of the 4p-5s band with transitions of type (Q)Q(f(3)e) we observed a narrowing in the linewidths with increasing rotational quantum number N. The rotational dependence of the linewidth is caused by predissociation of the 5s state by the repulsive ground 4s state through homogeneous coupling and changes in overlap integrals of the vibrational wave functions with the rotational level. Analysis was based on the Fermi's golden rule approximation model. In the 4p-5s band region a vibrational sequence ofv(')-v(")=1-1, 2-2, and 3-3 were recorded and a number of transitions belonging to the strongest (Q)Q(f(3)e) form branch of the 1-1 band were analyzed.

Study of the 6Li(p,π+)7Li reaction at 600 MeV

International Nuclear Information System (INIS)

Bauer, T.; Beurtey, R.; Boudard, A.; Bruge, G.; Chaumeaux, A.; Couvert, P.; Duhm, H.H.; Garreta, D.; Matoba, M.; Terrein, Y.; Aslanides, E.; Bertini, R.; Brochard, F.; Gorodetzky, Ph.; Hibou, F.; Bimbot, L.; Le Bornec, Y.; Tatischeff, B.; Dillig, M.

1977-01-01

The positive pion production through the 6 Li(p,π + ) reaction at 600 MeV has been studied using the high-resolution magnetic spectrometer SPES I. Differential cross sections have been measured from 5 0 to 35 0 sub(lab). The 6 Li(p,π + ) reaction feeds preferentially the 4.63 MeV 7/2 - level of 7 Li. The results of a calculation based on two- and three-nucleon diagrams with π and rho exchange between the projectile and a bound nucleon are also presented. (Auth.)

Extension of the chiral perturbation theory meson Lagrangian to order p{sup 6}

Energy Technology Data Exchange (ETDEWEB)

Fearing, H W; Scherer, S

1994-08-01

We have derived the most general chirally invariant Lagrangian L{sub 6} for the meson sector at order p{sup 6}. The result provides an extension of the standard Gasser-Leutwyler Lagrangian L{sub 4} to one higher order, including as well all the odd intrinsic parity terms in the Lagrangian. The most difficult part of the derivation was developing a systematic strategy so as to get all of the independent terms and eliminate the redundant ones in an efficient way. The equation of motion terms, which are redundant in the sense that they can be transformed away via field transformations, are separated out explicitly. The resulting Lagrangian has been separated into groupings of terms contributing to increasingly more complicated processes, so that one does not have to deal with the full result when calculating p{sup 6} contributions to simple processes. (author). 53 refs., 10 tabs.

Peculiarities of interaction of the p{sub z}-, π- electrons and the σ{sub p}-holes at the top 1–6 layers of HOPG

Energy Technology Data Exchange (ETDEWEB)

Dementjev, A.P., E-mail: demcarbon@yandex.ru; Ivanov, K.E.

2017-03-31

Graphical abstract: The formation of π-bands and σ{sub p}- holes as result of the p{sub z} → π transitions in 2–6 graphene layers HOPG. The valence band spectrum taken from Murday et al. (1981). - Abstract: The present work continues the analysis of results of Dementjev et al. (2015) in order to identify the interlayer interactions of the π-bands. Analysis of the N(E) C KVV Auger spectra of highly-ordered pyro-graphite showed the absence of the electron exchange between the π-bands in 1–6 layers. Since the π-bands are formed by the p{sub z} → π transitions, one can suggest that the π-band occupation at each graphene layer is formed by the p{sub z}-electrons of this layer. Since the p{sub z} electrons belong to the σ{sub p}-bands, the p{sub z} → π transitions in the σ{sub p}-bands in each of 2–6 graphene layers result in formation of holes H, whose concentration is equal to the concentration of electrons in the π-bands [H{sub i}] ≡ [π{sub i}]. This shows the origin of the ambipolar conductivity in graphene. The absence of the electronic interaction between the π-bands allows a suggestion that the interaction between top six graphene layers is due to the van der Waals electrostatic attractive forces. These forces promote the p{sub z} → π transitions in each of the 2–6 graphene layers and depend on the number of graphene layers above. The N(E) C KVV Auger spectra allow identification of number (1–6) of graphene layers and the π-band occupation at each of the layer. For the first time a specification of the van der Waals forces in HOPG was done.

Soroepidemiologia de Rhodococcus equi em equinos da região de Bagé, RS, pelo teste de inibição da hemólise sinérgica

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Lazzari Andrea

1997-01-01

Full Text Available Com o objetivo de avaliar a situação soroepidemiológica da infecção por Rhodococcus equi na região de Bagé, RS, foram testadas 290 amostras de soro sanguíneo de éguas e potros aparentemente sadios, obtidos de 6 haras com diferentes histórias de rodococose. Para relacionar o resultado sorológico com a presença deste agente bacteriano no trato intestinal destes animais, foram coletadas 123 amostras de fezes. O teste sorológico utilizado foi a inibição da hemólise sinérgica (IHS que detecta anticorpos neutralizantes contra o "fator equi". Um percentual de 87,93% (255/290 dos animais amostrados apresentaram estes anticorpos. O título médio geométrico (GMT destes anticorpos foi mais elevado nos potros do que nas éguas. A soropositividade destes equinos ao teste sorológico teve correlação com o isolamento do R. equi nas fezes dos respectivos animais. A maior taxa de isolamento de R. equi das fezes dos equinos e o maior GMT, ocorreu no único haras com casos clínicos recentes de enfermidade causada por esta bactéria. No entanto, todos os animais deste e dos demais haras, encontravam-se aparentemente sadios, sendo necessário, estabelecer em trabalho futuro, a possível relação entre títulos de anticorpos e sua importância na detecção da enfermidade.

The Equine Antimicrobial Peptide eCATH1 Is Effective against the Facultative Intracellular Pathogen Rhodococcus equi in Mice

Science.gov (United States)

Schlusselhuber, Margot; Torelli, Riccardo; Martini, Cecilia; Leippe, Matthias; Cattoir, Vincent; Leclercq, Roland; Laugier, Claire; Grötzinger, Joachim; Sanguinetti, Maurizio

2013-01-01

Rhodococcus equi, the causal agent of rhodococcosis, is a major pathogen of foals and is also responsible for severe infections in immunocompromised humans. Of great concern, strains resistant to currently used antibiotics have emerged. As the number of drugs that are efficient in vivo is limited because of the intracellular localization of the bacterium inside macrophages, new active but cell-permeant drugs will be needed in the near future. In the present study, we evaluated, by in vitro and ex vivo experiments, the ability of the alpha-helical equine antimicrobial peptide eCATH1 to kill intracellular bacterial cells. Moreover, the therapeutic potential of the peptide was assessed in experimental rhodococcosis induced in mice, while the in vivo toxicity was evaluated by behavioral and histopathological analysis. The study revealed that eCATH1 significantly reduced the number of bacteria inside macrophages. Furthermore, the bactericidal potential of the peptide was maintained in vivo at doses that appeared to have no visible deleterious effects for the mice even after 7 days of treatment. Indeed, daily subcutaneous injections of 1 mg/kg body weight of eCATH1 led to a significant reduction of the bacterial load in organs comparable to that obtained after treatment with 10 mg/kg body weight of rifampin. Interestingly, the combination of the peptide with rifampin showed a synergistic interaction in both ex vivo and in vivo experiments. These results emphasize the therapeutic potential that eCATH1 represents in the treatment of rhodococcosis. PMID:23817377

Two dimensional gel electrophoresis using narrow pH 3-5.6 immobilised pH gradient strips identifies potential novel disease biomarkers in plasma or serum

OpenAIRE

sprotocols

2015-01-01

Authors: Bevin Gangadharan & Nicole Zitzmann ### Abstract Two-dimensional gel electrophoresis (2-DE) is a protein separation technique often used to separate plasma or serum proteins in an attempt to identify novel biomarkers. This protocol describes how to run 2-DE gels using narrow pH 3-5.6 immobilised pH gradient strips to separate 2 mg of serum proteins. pH 3-6 ampholytes are used to enhance the solubility of proteins in this pH range before the serum proteins are separated in the...

IL-3 maintains activation of the P90S6K/RPS6 pathway and increases translation in human eosinophils1

Science.gov (United States)

Esnault, Stephane; Kelly, Elizabeth A.B.; Shen, Zhong-Jian; Johansson, Mats W.; Malter, James S.; Jarjour, Nizar N.

2015-01-01

IL-5 is a major therapeutic target to reduce eosinophilia. However, all of the eosinophil-activating cytokines IL-5, IL-3, and GM-CSF are typically present in atopic diseases including allergic asthma. Due to the functional redundancy of these 3 cytokines on eosinophils and the loss of IL-5 receptor on airway eosinophils, it is important to take IL-3 and GM-CSF into account to efficiently reduce tissue eosinophil functions. Moreover, these 3 cytokines signal through a common β-chain receptor, and yet differentially affect protein production in eosinophils. Notably, the increased ability of IL-3 to induce production of proteins such as semaphorin-7A without affecting mRNA level suggests a unique influence by IL-3 on translation. The purpose of this study is to identify the mechanisms by which IL-3 distinctively affects eosinophil function compared to IL-5 and GM-CSF, with a focus on protein translation. Peripheral blood eosinophils were used to study intracellular signaling and protein translation in cells activated with IL-3, GM-CSF or IL-5. We establish that, unlike GM-CSF or IL-5, IL-3 triggers prolonged signaling through activation of ribosomal protein (RP) S6 and the upstream kinase, p90S6K. Blockade of p90S6K activation inhibited phosphorylation of RPS6 and IL-3-enhanced semaphorin-7A translation. Furthermore, in an allergen-challenged environment, in vivo phosphorylation of RPS6 and p90S6K was enhanced in human airway compared to circulating eosinophils. Our findings provide new insights into the mechanisms underlying differential activation of eosinophils by IL-3, GM-CSF, and IL-5. These observations place IL-3 and its downstream intracellular signals as novel targets that should be considered to modulate eosinophil functions. PMID:26276876

Kinetics on the reaction of 6-chloroquinoline and p-substituted benzoylchlorides under high pressures

International Nuclear Information System (INIS)

Kim, Eung Ryul; Lim, Jong Wan; Kim, Se Kyong; Ko, Young Shin

2002-01-01

The reactions rates of substituted 6-chloroquinoline, with p-substituted benzoyichlorides (p-CH 3 , p-H, p-NO 2 ) have been measured by conductometry in acetonitrile, and the rate constants are determined at various temperatures (10,15,20,25 .deg. C) and pressures (1, 200, 500, 1000 bar). From the values of rate constants, the activation parameters (Ea, ΔV ≠ , ΔH ≠ , ΔS ≠ , and ΔG ≠ ) and the pressure dependence of Hammett ρ values were determined. The rate constants increased with increasing temperatures and pressures, and are further increased to introduction to the electron donor substituents in substrate (p-NO 2 ) with 6-chloroquinoline. When, the activation volume and the activation entropy are all negative. And the Hammett ρ values are positive for the substrate over the pressure and temperature range studied. The results of kinetic studies for pressure and substituent show that thease reactions proceed in typical S N 2 reaction mechanism and 'associative S N 2' in bond formation favored with increasing pressures

Extension of the chiral perturbation theory meson Lagrangian to order p6

International Nuclear Information System (INIS)

Fearing, H.W.; Scherer, S.

1996-01-01

We have constructed the most general chirally invariant Lagrangian scrL 6 for the meson sector at order p 6 . The result provides an extension of the standard Gasser-Leutwyler Lagrangian scrL 4 to one higher order, including as well all the odd intrinsic parity terms in the Lagrangian. The most difficult part of the construction was developing a systematic strategy so as to get all of the independent terms and eliminate the redundant ones in an efficient way. The claim to have obtained the most general Lagrangian relies on this systematic construction and on the elimination of redundant quantities using relations of which we are aware, rather than on a general formal proof of either completeness or independence. The open-quote open-quote equation-of-motion close-quote close-quote terms, which are redundant in the sense that they can be transformed away via field transformations, are separated out explicitly. The resulting Lagrangian has been separated into groupings of terms contributing to increasingly more complicated processes, so that one does not have to deal with the full result when calculating p 6 contributions to simple processes. copyright 1995 The American Physical Society

Molecular Mechanism of Enhanced Anticancer Effect of Nanoparticle Formulated LY2835219 via p16-CDK4/6-pRb Pathway in Colorectal Carcinoma Cell Line

Directory of Open Access Journals (Sweden)

Xu Tang

2016-01-01

Full Text Available LY2835219 is a dual inhibitor to CDK4 and CDK6. This study was to prepare LY2835219-loaded chitosan nanoparticles (CNP/LY and LY2835219-loaded hyaluronic acid-conjugated chitosan nanoparticles (HACNP/LY and revealed their anticancer effect and influence on p16-CDK4/6-pRb pathway against colon cell line. The nanoparticle sizes of CNP/LY and HACNP/LY were approximately 195±39.6 nm and 217±31.1 nm, respectively. The zeta potentials of CNP/LY and HACNP/LY were 37.3±1.5 mV and 30.3±2.2 mV, respectively. And the preparation process showed considerable drug encapsulation efficiency and loading efficiency. LY2835219, CNP/LY, and HACNP/LY inhibited HT29 cell proliferation with 0.68, 0.54, and 0.30 μM of IC50, respectively. G1 phase was arrested by LY2835219 and its formulations. Furthermore, inhibition of CDK4/6 by LY2835219 formulations induced CDK4, CDK6, cyclin D1, and pRb decrease and p16 increase at both protein and mRNA levels. Overall, nanoparticle formulated LY2835219 could enhance the cytotoxicity and cell cycle arrest, and HACNP/LY strengthened the trend furtherly compared to CNP/LY. It is the first time to demonstrate the anticancer effect and mechanism against HT29 by LY2835219 and its nanoparticles. The drug and its nanoparticle formulations delay the cell growth and arrest cell cycle through p16-CDK4/6-pRb pathway, while the nanoparticle formulated LY2835219 could strengthen the process.

Kinetics of hydrocarbon extraction from oil shale using biosurfactant producing bacteria

International Nuclear Information System (INIS)

Haddadin, Malik S.Y.; Abou Arqoub, Ansam A.; Abu Reesh, Ibrahim; Haddadin, Jamal

2009-01-01

This study was done to extract hydrocarbon compounds from El-Lajjun oil shale using biosurfactant produced from two strains Rhodococcus erythropolis and Rhodococcus ruber. The results have shown that, optimal biosurfactant production was found using naphthalene and diesel as a carbon source for R. erthropolis and R. ruber, respectively. Optimum nitrogen concentration was 9 g/l and 7 g/l for R. erthropolis and R. ruber, respectively. Optimum K 2 HPO 4 to KH 2 PO 4 ratio, temperature, pH, and agitation speeds were 2:1, 37 deg. C, 7 and 200 rpm. Under optimal conditions R. erthropolis and R. ruber produced 5.67 and 6.9 g/l biosurfactant, respectively. Maximum recovery of oil achieved with hydrogen peroxide pre-treatment was 25% and 26% at biosurfactant concentration of 8 g/l and 4 g/l for R. erthropolis and R. ruber, respectively. The extent desorption of hydrocarbons from the pre-treated oil shale by biosurfactant were inversely related to the concentration of high molecular weight hydrocarbons, asphaltenes compounds. Pre- treatment of oil shale with hydrogen peroxide produced better improvement in aromatic compounds extraction in comparison with improvement which resulted from demineralization of the oil shale

Structural basis for the substrate specificity and the absence of dehalogenation activity in 2-chloromuconate cycloisomerase from Rhodococcus opacus 1CP.

Science.gov (United States)

Kolomytseva, Marina; Ferraroni, Marta; Chernykh, Alexey; Golovleva, Ludmila; Scozzafava, Andrea

2014-09-01

2-Chloromuconate cycloisomerase from the Gram-positive bacterium Rhodococcus opacus 1CP (Rho-2-CMCI) is an enzyme of a modified ortho-pathway, in which 2-chlorophenol is degraded using 3-chlorocatechol as the central intermediate. In general, the chloromuconate cycloisomerases catalyze not only the cycloisomerization, but also the process of dehalogenation of the chloromuconate to dienelactone. However Rho-2-CMCI, unlike the homologous enzymes from the Gram-negative bacteria, is very specific for only one position of the chloride on the substrate chloromuconate. Furthermore, Rho-2-CMCI is not able to dehalogenate the 5-chloromuconolactone and therefore it cannot generate the dienelactone. The crystallographic structure of the homooctameric Rho-2-CMCI was solved by molecular replacement using the coordinates of the structure of chloromuconate cycloisomerase from Pseudomonas putida PRS2000. The structure was analyzed and compared to the other already known structures of (chloro)muconate cycloisomerases. In addition to this, molecular docking calculations were carried out, which allowed us to determine the residues responsible for the high substrate specificity and the lack of dehalogenation activity of Rho-2-CMCI. Our studies highlight that a histidine, located in a loop that closes the active site cavity upon the binding of the substrate, could be related to the dehalogenation inability of Rho-2-CMCI and in general of the muconate cycloisomerases. Copyright © 2014 Elsevier B.V. All rights reserved.

ADP-ribosylation factor arf6p may function as a molecular switch of new end take off in fission yeast

International Nuclear Information System (INIS)

Fujita, Atsushi

2008-01-01

Small GTPases act as molecular switches in a wide variety of cellular processes. In fission yeast Schizosaccharomyces pombe, the directions of cell growth change from a monopolar manner to a bipolar manner, which is known as 'New End Take Off' (NETO). Here I report the identification of a gene, arf6 + , encoding an ADP-ribosylation factor small GTPase, that may be essential for NETO. arf6Δ cells completely fail to undergo NETO. arf6p localizes at both cell ends and presumptive septa in a cell-cycle dependent manner. And its polarized localization is not dependent on microtubules, actin cytoskeletons and some NETO factors (bud6p, for3p, tea1p, tea3p, and tea4p). Notably, overexpression of a fast GDP/GTP-cycling mutant of arf6p can advance the timing of NETO. These findings suggest that arf6p functions as a molecular switch for the activation of NETO in fission yeast

Determination of the effective radiative lifetimes of the 6 3P1 atomic mercury level in low-pressure mercury discharges

International Nuclear Information System (INIS)

van de Weijer, P.; Cremers, R.M.M.

1984-01-01

Experiments are described in which low-pressure mercury, mercury-argon and mercury-krypton discharges were irradiated with a dye laser pulse at 365.5 nm, thus exciting mercury atoms from the metastable 6 3 P 2 level to the 6 3 D 2 level. The 6 3 D 2 level decays radiatively to the 6 P levels. By recording the time dependence of the overpopulation in the 6 3 P 1 and the 6 1 P 1 level at the fluorescence signals at 254 nm and 185 nm, respectively, the effective radiative lifetime of these levels were determined. The effective radiative lifetime of the 6 3 P 1 level was measured in the k 0 R regime 0.1-500. The 6 1 P 1 lifetime was determined for the following discharge conditions: tube diameter 10-36 mm, mercury density 7.10 18 -2.10 21 m -3 , and noble gas pressure 0, 130, 400 Pa

cap alpha. -transfer reactions /sup 27/Al(/sup 6/Li, d)/sup 31/P, /sup 29/Si(/sup 6/Li, d)/sup 33/S and /sup 31/P(/sup 6/Li, d)/sup 35/Cl at 36 MeV. [Angular distributions, EFR DWBA, spectroscopic strengths

Energy Technology Data Exchange (ETDEWEB)

Eswaran, M A; Gove, H E; Cook, R; Sikora, B [Rochester Univ., NY (USA). Nuclear Structure Research Lab.

1979-08-13

The ..cap alpha..-transfer reactions /sup 27/Al(/sup 6/Li,d)/sup 31/P,/sup 29/Si(/sup 6/Li,d) /sup 33/S and /sup 31/P(Li,d)/sup 35/Cl have been studied at a /sup 6/Li energy of 36 MeV. Absolute cross sections and angular distributions have been measured and an exact finite-range distorted-wave Born approximation analysis assuming a direct cluster transfer has been used to extract from the data ..cap alpha..-particle spectroscopic strengths for levels populated in /sup 31/P, /sup 33/S and /sup 35/Cl in three reactions respectively. The results show that in the case of most of the low-lying excited states of /sup 31/P a single value of L of the transferred ..cap alpha..-particle contributes, though a multiplicity of L-values are allowed by angular momentum selection rules. It is also found that the ..cap alpha..-particle spectroscopic strength of the ground state of /sup 31/P is a factor of 2 more than the strengths of the ground states of /sup 33/S and /sup 35/Cl. The ..cap alpha..-spectroscopic strengths of ground states of these, as well as other odd-A s-d shell nuclei, are compared with the presently available shell model calculations.

Majanduse üldolukord 2003. a septembris ja 6 kuu pärast

Index Scriptorium Estoniae

2003-01-01

Ilmunud ka: Konjunktuur : Quarterly Review of the Estonian Economy Sep nr. 3, lk. 4-11. Eesti Konjunktuuriinstituudi hinnangul püsib majanduse üldolukord soodne. Eelseisva 6 kuu suhtes domineerivad positiivsed ootused, tööstusettevõtted prognoosivad välisnõudluse suurenemist. Tabelid ja diagrammid

A Unique Failure Mechanism in the Nexus 6P Lithium-Ion Battery

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Saurabh Saxena

2018-04-01

Full Text Available Nexus 6P smartphones have been beset by battery drain issues, which have been causing premature shutdown of the phone even when the charge indicator displays a significant remaining runtime. To investigate the premature battery drain issue, two Nexus 6P smartphones (one new and one used were disassembled and their batteries were evaluated using computerized tomography (CT scan analysis, electrical performance (capacity, resistance, and impedance tests, and cycle life capacity fade tests. The “used” smartphone battery delivered only 20% of the rated capacity when tested in a first capacity cycle and then 15% of the rated capacity in a second cycle. The new smartphone battery exceeded the rated capacity when first taken out of the box, but exhibited an accelerated capacity fade under C/2 rate cycling and decreased to 10% of its initial capacity in just 50 cycles. The CT scan results revealed the presence of contaminant materials inside the used battery, raising questions about the quality of the manufacturing process.

TRAF6 promotes myogenic differentiation via the TAK1/p38 mitogen-activated protein kinase and Akt pathways.

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Fang Xiao

Full Text Available p38 mitogen-activated protein kinase (MAPK is an essential kinase involved in myogenic differentiation. Although many substrates of p38 MAPK have been identified, little is known about its upstream activators during myogenic differentiation. TRAF6 is known to function in cytokine signaling during inflammatory responses. However, not much is known about its role in myogenic differentiation and muscle regeneration. We showed here that TRAF6 and its intrinsic ubiquitin E3 ligase activity are required for myogenic differentiation. In mouse myoblasts, knockdown of TRAF6 compromised the p38 MAPK and Akt pathways, while deliberate activation of either pathway rescued the differentiation defect caused by TRAF6 knockdown. TAK1 acted as a key signal transducer downstream of TRAF6 in myogenic differentiation. In vivo, knockdown of TRAF6 in mouse muscles compromised the injury-induced muscle regeneration without impairing macrophage infiltration and myoblast proliferation. Collectively, we demonstrated that TRAF6 promotes myogenic differentiation and muscle regeneration via the TAK1/p38 MAPK and Akt pathways.

Reconstruction of structural evolution in the trnL intron P6b loop of symbiotic Nostoc (Cyanobacteria).

Science.gov (United States)

Olsson, Sanna; Kaasalainen, Ulla; Rikkinen, Jouko

2012-02-01

In this study we reconstruct the structural evolution of the hyper-variable P6b region of the group I trnLeu intron in a monophyletic group of lichen-symbiotic Nostoc strains and establish it as a useful marker in the phylogenetic analysis of these organisms. The studied cyanobacteria occur as photosynthetic and/or nitrogen-fixing symbionts in lichen species of the diverse Nephroma guild. Phylogenetic analyses and secondary structure reconstructions are used to improve the understanding of the replication mechanisms in the P6b stem-loop and to explain the observed distribution patterns of indels. The variants of the P6b region in the Nostoc clade studied consist of different combinations of five sequence modules. The distribution of indels together with the ancestral character reconstruction performed enables the interpretation of the evolution of each sequence module. Our results indicate that the indel events are usually associated with single nucleotide changes in the P6b region and have occurred several times independently. In spite of their homoplasy, they provide phylogenetic information for closely related taxa. Thus we recognize that features of the P6b region can be used as molecular markers for species identification and phylogenetic studies involving symbiotic Nostoc cyanobacteria.

Particle production in Si + A and p + A collisions at 14.6 A·GeV/c

International Nuclear Information System (INIS)

Miake, Y.

1990-01-01

Particle production (π ± , K ± , p) has been measured in both Si+A and p+A collisions at 14.6 A·GeV/c. Comparisons of m t and dn/dy distributions between p+Be, p+Au and central Si+Au collisions are discussed. 8 refs., 3 figs

Cytochrome P450IID6 recognized by LKM1 antibody is not exposed on the surface of hepatocytes.

Science.gov (United States)

Yamamoto, A M; Mura, C; De Lemos-Chiarandini, C; Krishnamoorthy, R; Alvarez, F

1993-06-01

LKM1 autoantibody, directed against P450IID6, is accepted as a marker of a particular type of autoimmune hepatitis, but its role in the pathogenesis of the disease is controversial. Localization of P450IID6 on the cell surface of rat hepatocytes was previously reported, suggesting that membrane-bound P450IID6 could be the target of LKM1 antibodies, thus allowing immune lysis of hepatocytes. The objective of the present study was to determine, using various methods, the cell localization of P450IID6 in human and rat hepatocytes. Incubation of rat and human hepatocytes with LKM1-positive serum showed slight, if any, cell membrane staining using immunofluorescence, immunoperoxidase and immunoelectron microscopic studies. No staining of the plasma membrane of human hepatocytes was observed when incubations were carried out with immunoaffinity-purified antibody directed against peptide 254-271, the main epitope of P450IID6 recognized by all LKM1 sera tested. Chinese hamster ovary cells, transfected with the complete P450IID6 cDNA and incubated with the supernatant from a B cell lymphoblastoid cell line prepared with the lymphocytes of a LKM1-positive patient, did not show any staining of the cell surface by immunofluorescence. Incubation of rat microsomal fraction vesicles with LKM1-positive serum, followed by protein A-gold immunoelectron microscopy, displayed a staining of almost all vesicles, confirming that P450IID6 is present on the cytoplasmic side of the microsomal membrane, which makes it unable to be expressed on the cell surface even if it were transported from the endoplasmic reticulum (ER). Sulpho NHS Biotin labelling of rat hepatocyte cell membranes did not show the presence of a 50-kD molecule that could have reacted with LKM1 antibody. DNA sequencing of exon 1 of the CYP2D6 gene of a patient positive for LKM1 antibody did not show any difference from that of the normal published sequence of the gene. This does not favour an alteration of the NH2 terminal

HSP90 inhibitors potentiate PGF2α-induced IL-6 synthesis via p38 MAP kinase in osteoblasts.

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Kazuhiko Fujita

Full Text Available Heat shock protein 90 (HSP90 that is ubiquitously expressed in various tissues, is recognized to be a major molecular chaperone. We have previously reported that prostaglandin F2α (PGF2α, a potent bone remodeling mediator, stimulates the synthesis of interleukin-6 (IL-6 through p44/p42 mitogen-activated protein (MAP kinase and p38 MAP kinase in osteoblast-like MC3T3-E1 cells, and that Rho-kinase acts at a point upstream of p38 MAP kinase. In the present study, we investigated the involvement of HSP90 in the PGF2α-stimulated IL-6 synthesis and the underlying mechanism in MC3T3-E1 cells. Geldanamycin, an inhibitor of HSP90, significantly amplified both the PGF2α-stimulated IL-6 release and the mRNA expression levels. In addition, other HSP90 inhibitors, 17-allylamino-17demethoxy-geldanamycin (17-AAG and 17-dimethylamino-ethylamino-17-demethoxy-geldanamycin (17-DMAG and onalespib, enhanced the PGF2α-stimulated IL-6 release. Geldanamycin, 17-AAG and onalespib markedly strengthened the PGF2α-induced phosphorylation of p38 MAP kinase. Geldanamycin and 17-AAG did not affect the PGF2α-induced phosphorylation of p44/p42 MAP kinase and myosin phosphatase targeting subunit (MYPT-1, a substrate of Rho-kinase, and the protein levels of RhoA and Rho-kinase. In addition, HSP90-siRNA enhanced the PGF2α-induced phosphorylation of p38 MAP kinase. Furthermore, SB203580, an inhibitor of p38 MAP kinase, significantly suppressed the amplification by geldanamycin, 17-AAG or 17-DMAG of the PGF2α-stimulated IL-6 release. Our results strongly suggest that HSP90 negatively regulates the PGF2α-stimulated IL-6 synthesis in osteoblasts, and that the effect of HSP90 is exerted through regulating p38 MAP kinase activation.

Expression of Bmi-1, P16, and CD44v6 in Uterine Cervical Carcinoma and Its Clinical Significance

International Nuclear Information System (INIS)

Weng, Mei-ying; Li, Lin; Feng, Shu-ying; Hong, Shun-jia

2012-01-01

Bmi-1, a putative proto-oncogene, is a core member of the polycomb gene family, which is expressed in many human tumors. The p16 protein negatively regulated cell proliferation, whereas CD44v6 is associated with proliferation as an important protein. Additionally, CD44v6 is an important nuclear antigen closely correlated to tumor metastasis. The present study aims to investigate the expression and significance of Bmi-1, p16, and CD44v6 in uterine cervical carcinoma (UCC). A total of 62 UCC, 30 cervical neoplasic, and 20 normal cervical mucosal tissues were used in the current study. The expression of Bmi-1, p16, and CD44v6 in these tissues was determined using immunohistochemical assay. The relationships among the expression of these indices, the clinicopathologic features of UCC, and the survival rate of UCC patients were also discussed. The correlation between Bmi-1 protein expression and p16 or CD44v6 protein in UCC was analyzed. The expression of Bmi-1, p16, and CD44v6 was significantly high in cervical carcinoma compared with that in the cervical neoplasia and normal colorectal mucosa (P<0.05). The over-expression of Bmi-1 protein in UCC was apparently related to the distant metastasis (P<0.01) and the tumor, nodes and metastasis-classification, i.e. the TNM staging, World Health Organization (P<0.05). Nevertheless, the positive expression of p16 protein in UCC was not significantly associated with the clinicopathologic features (P>0.05). The Kaplan–Meier survival analysis showed that the over-expression of Bmi-1 significantly decreased the survival rate of UCC patients (P<0.05). A strong correlation indicated that there was statistical significance between the expression of Bmi-1 and CD44V6 proteins in UCC (r=0.419, P=0.001). The over-expression of Bmi-1 and CD44v6 protein closely correlate to the tumorigenesis, metastasis, and prognosis of UCC. Bmi-1 and CD44v6 may be used to predict the prognosis of cervical carcinoma. Bmi-1 may indirectly regulate the

Immunohistochemical Analysis of the Activation Status of the Akt/mTOR/pS6 Signaling Pathway in Oral Lichen Planus

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Georgios Prodromidis

2013-01-01

Full Text Available Introduction. Aberrations of the Akt/mTOR/pS6 pathway have been linked to various types of human cancer, including oral squamous cell carcinoma (OSCC. The purpose of this study was to evaluate the activation status of Akt, mTOR, and pS6 in oral lichen planus (OLP in comparison with oral premalignant and malignant lesions and normal oral mucosa (NM. Materials and Methods. Immunohistochemistry for p-Akt, p-mTOR, and phospho-pS6 was performed in 40 OLP, 20 oral leukoplakias (OL, 10 OSCC, and 10 control samples of NM. Results. Nuclear p-Akt expression was detected in the vast majority of cases in all categories, being significantly higher in OL. Cytoplasmic p-Akt and p-mTOR staining was present only in a minority of OLP cases, being significantly lower compared to OL and OSCC. Phospho-pS6 showed cytoplasmic positivity in most OLP cases, which however was significantly lower compared to OL and OSCC. Conclusions. Overall, cytoplasmic p-Akt, p-mTOR, and phospho-pS6 levels appear to be significantly lower in OLP compared to OL and OSCC. However, the expression of these molecules in a subset of OLP cases suggests that activation of Akt/mTOR/pS6 may occur in the context of OLP, possibly contributing to the premalignant potential of individual cases.

Removal of polycyclic aromatic hydrocarbons in soil spiked with model mixtures of petroleum hydrocarbons and heterocycles using biosurfactants from Rhodococcus ruber IEGM 231.

Science.gov (United States)

Ivshina, Irina; Kostina, Ludmila; Krivoruchko, Anastasiya; Kuyukina, Maria; Peshkur, Tatyana; Anderson, Peter; Cunningham, Colin

2016-07-15

Removal of polycyclic aromatic hydrocarbons (PAHs) in soil using biosurfactants (BS) produced by Rhodococcus ruber IEGM 231 was studied in soil columns spiked with model mixtures of major petroleum constituents. A crystalline mixture of single PAHs (0.63g/kg), a crystalline mixture of PAHs (0.63g/kg) and polycyclic aromatic sulfur heterocycles (PASHs), and an artificially synthesized non-aqueous phase liquid (NAPL) containing PAHs (3.00g/kg) dissolved in alkanes C10-C19 were used for spiking. Percentage of PAH removal with BS varied from 16 to 69%. Washing activities of BS were 2.5 times greater than those of synthetic surfactant Tween 60 in NAPL-spiked soil and similar to Tween 60 in crystalline-spiked soil. At the same time, amounts of removed PAHs were equal and consisted of 0.3-0.5g/kg dry soil regardless the chemical pattern of a model mixture of petroleum hydrocarbons and heterocycles used for spiking. UV spectra for soil before and after BS treatment were obtained and their applicability for differentiated analysis of PAH and PASH concentration changes in remediated soil was shown. The ratios A254nm/A288nm revealed that BS increased biotreatability of PAH-contaminated soils. Copyright © 2016 Elsevier B.V. All rights reserved.

Epitaxial growth of matched metallic ErP0.6As0.4 layers on GaAs

International Nuclear Information System (INIS)

Guivarc'h, A.; Le Corre, A.; Gaulet, J.; Guenais, B.; Minier, M.; Ropars, G.; Badoz, P.A.; Duboz, J.Y.

1990-01-01

Successful growth of (001)ErP 0.6 As 0.4 single crystal film on (001) GaAs has been demonstrated. The epitaxial metallic layers reproducibly showed lattice mismatch below 5 10 -4 . This is, to the authors' knowledge, the first report of a stable, epitaxial and lattice-matched metal/compound semiconductor heterostructure. The ErP 0.6 As 0.4 /n-GaAs diodes yielded excellent I-V characteristics with an ideality factor of 1.1 and barrier height of 0.88 eV. For a 240 Angstrom- thick film, metallic behavior was observed with resistivities of 25 and 86 μΩcm at 1.5 K and room temperature, respectively. As the other Er compounds ErP, ErAs, ErSb and ErSi 2 , ErP 0.6 As 0.4 presents an abrupt drop in resistivity in the vicinity of the liquid helium temperature, due to a paramagnetic to antiferromagnetic phase transition

Clonal Evolution and Clinical Significance of Copy Number Neutral Loss of Heterozygosity of Chromosome Arm 6p in Acquired Aplastic Anemia

Science.gov (United States)

Betensky, Marisol; Babushok, Daria; Roth, Jacquelyn J.; Mason, Philip J; Biegel, Jaclyn A.; Busse, Tracy M; Li, Yimei; Lind, Curt; Papazoglou, Anna; Monos, Dimitri; Podsakoff, Gregory; Bessler, Monica; Olson, Timothy S.

2015-01-01

Acquired aplastic anemia (aAA) results from the T cell-mediated autoimmune destruction of hematopoietic stem cells. Factors predicting response to immune suppression therapy (IST) or development of myelodysplastic syndrome (MDS) are beginning to be elucidated. Our recent data suggest most patients with aAA treated with IST develop clonal somatic genetic alterations in hematopoietic cells. One frequent acquired abnormality is copy-number neutral loss of heterozygosity on chromosome 6p (6p CN-LOH) involving the human leukocyte antigen (HLA) locus. We hypothesized that because 6p CN-LOH clones may arise from selective pressure to escape immune surveillance through deletion of HLA alleles, the development of 6p CN-LOH may affect response to IST. We used single nucleotide polymorphism array genotyping and targeted next-generation sequencing of HLA alleles to assess frequency of 6p CN-LOH, identity of HLA alleles lost through 6p CN-LOH, and impact of 6p CN-LOH on response to IST. 6p CN-LOH clones were present in 11.3% of patients, remained stable over time, and were not associated with development of MDS-defining cytogenetic abnormalities. Notably, no patient with 6p CN-LOH treated with IST achieved a complete response. In summary, clonal 6p CN-LOH in aAA defines a unique subgroup of patients that may provide insights into hematopoietic clonal evolution. PMID:26702937

Increased p50/p50 NF-κB Activation in Human Papillomavirus Type 6- or Type 11-Induced Laryngeal Papilloma Tissue

Science.gov (United States)

Vancurova, Ivana; Wu, Rong; Miskolci, Veronika; Sun, Shishinn

2002-01-01

We have observed elevated NF-κB DNA-binding activity in nuclear extracts from human papillomavirus type 6- and 11-infected laryngeal papilloma tissues. The predominant DNA-binding species is the p50/p50 homodimer. The elevated NF-κB activity could be correlated with a reduced level of cytoplasmic IκBβ and could be associated with the overexpression of p21CIP1/WAF1 in papilloma cells. Increased NF-κB activity and cytoplasmic accumulation of p21CIP1/WAF1 might counteract death-promoting effects elicited by overexpressed PTEN and reduced activation of Akt and STAT3 previously noted in these tissues. PMID:11773428

The potential of species-specific tagatose-6-phosphate (T6P) pathway in Lactobacillus casei group for galactose reduction in fermented dairy foods.

Science.gov (United States)

Wu, Qinglong; Shah, Nagendra P

2017-04-01

Residual lactose and galactose in fermented dairy foods leads to several industrial and health concerns. There is very little information pertaining to manufacture of fermented dairy foods that are low in lactose and galactose. In the present study, comparative genomic survey demonstrated the constant presence of chromosome-encoded tagatose-6-phosphate (T6P) pathway in Lactobacillus casei group. Lactose/galactose utilization tests and β-galactosidase assay suggest that PTS Gal system, PTS Lac system and T6P pathway are major contributors for lactose/galactose catabolism in this group of organisms. In addition, it was found than lactose catabolism by Lb. casei group accumulated very limited galactose in the MRS-lactose medium and in reconstituted skim milk, whereas Streptococcus thermophilus and Lb. delbrueckii subsp. bulgaricus (Lb. bulgaricus) strains secreted high amount of galactose extracellularly. Moreover, co-culturing Lb. casei group with Str. thermophilus showed significant reduction in galactose content, while co-culturing Lb. casei group with Lb. bulgaricus showed significant reduction in lactose content but significant increase in galactose content in milk. Overall, the present study highlighted the potential of Lb. casei group for reducing galactose accumulation in fermented milks due to its species-specific T6P pathway. Copyright © 2016 Elsevier Ltd. All rights reserved.

ANISOTROPY EFFECTS IN ELECTRON-CAPTURE BY O6+ FROM ALIGNED NA-ASTERISK(3P)

NARCIS (Netherlands)

SCHLATMANN, AR; WIERSEMA, WP; HOEKSTRA, R; MORGENSTERN, R; OLSON, RE; PASCALE, J

1994-01-01

We report results of one electron capture by highly charged ions colliding with laser excited aligned Na*(3p). The 0 vi(10-->8) photon emission cross section after electron capture by the O6+ projectile is measured in the collision energy range 2-8 keV/amu. Effects of the Na*(3p) orbital alignment

p70S6 kinase signals cell survival as well as growth, inactivating the pro-apoptotic molecule BAD

DEFF Research Database (Denmark)

Harada, H; Andersen, Jens S.; Mann, M

2001-01-01

Cytokines often deliver simultaneous, yet distinct, cell growth and cell survival signals. The 70-kDa ribosomal protein S6 kinase (p70S6K) is known to regulate cell growth by inducing protein synthesis components. We purified membrane-based p70S6K as a kinase responsible for site-specific phospho...

Transportation research synthesis : state DOT experiences with Primavera P6 project management software.

Science.gov (United States)

2010-03-01

The eight agencies we interviewed all reported general satisfaction with Primavera P6 as a project management tool within their organizations, although they noted that a significant commitment to training is required. Most states have not implemented...

Phenotypic and molecular assessment of seven patients with 6p25 deletion syndrome: Relevance to ocular dysgenesis and hearing impairment

Directory of Open Access Journals (Sweden)

Ritch Robert

2004-06-01

Full Text Available Abstract Background Thirty-nine patients have been described with deletions involving chromosome 6p25. However, relatively few of these deletions have had molecular characterization. Common phenotypes of 6p25 deletion syndrome patients include hydrocephalus, hearing loss, and ocular, craniofacial, skeletal, cardiac, and renal malformations. Molecular characterization of deletions can identify genes that are responsible for these phenotypes. Methods We report the clinical phenotype of seven patients with terminal deletions of chromosome 6p25 and compare them to previously reported patients. Molecular characterization of the deletions was performed using polymorphic marker analysis to determine the extents of the deletions in these seven 6p25 deletion syndrome patients. Results Our results, and previous data, show that ocular dysgenesis and hearing impairment are the two most highly penetrant phenotypes of the 6p25 deletion syndrome. While deletion of the forkhead box C1 gene (FOXC1 probably underlies the ocular dysgenesis, no gene in this region is known to be involved in hearing impairment. Conclusions Ocular dysgenesis and hearing impairment are the two most common phenotypes of 6p25 deletion syndrome. We conclude that a locus for dominant hearing loss is present at 6p25 and that this locus is restricted to a region distal to D6S1617. Molecular characterization of more 6p25 deletion patients will aid in refinement of this locus and the identification of a gene involved in dominant hearing loss.

Biotransformation of benzonitrile herbicides via the nitrile hydratase-amidase pathway in rhodococci.

Science.gov (United States)

Veselá, Alicja B; Pelantová, Helena; Sulc, Miroslav; Macková, Martina; Lovecká, Petra; Thimová, Markéta; Pasquarelli, Fabrizia; Pičmanová, Martina; Pátek, Miroslav; Bhalla, Tek Chand; Martínková, Ludmila

2012-12-01

The aim of this work was to determine the ability of rhodococci to transform 3,5-dichloro-4-hydroxybenzonitrile (chloroxynil), 3,5-dibromo-4-hydroxybenzonitrile (bromoxynil), 3,5-diiodo-4-hydroxybenzonitrile (ioxynil) and 2,6-dichlorobenzonitrile (dichlobenil); to identify the products and determine their acute toxicities. Rhodococcus erythropolis A4 and Rhodococcus rhodochrous PA-34 converted benzonitrile herbicides into amides, but only the former strain was able to hydrolyze 2,6-dichlorobenzamide into 2,6-dichlorobenzoic acid, and produced also more of the carboxylic acids from the other herbicides compared to strain PA-34. Transformation of nitriles into amides decreased acute toxicities for chloroxynil and dichlobenil, but increased them for bromoxynil and ioxynil. The amides inhibited root growth in Lactuca sativa less than the nitriles but more than the acids. The conversion of the nitrile group may be the first step in the mineralization of benzonitrile herbicides but cannot be itself considered to be a detoxification.

Error-prone bypass of O6-methylguanine by DNA polymerase of Pseudomonas aeruginosa phage PaP1.

Science.gov (United States)

Gu, Shiling; Xiong, Jingyuan; Shi, Ying; You, Jia; Zou, Zhenyu; Liu, Xiaoying; Zhang, Huidong

2017-09-01

O 6 -Methylguanine (O 6 -MeG) is highly mutagenic and is commonly found in DNA exposed to methylating agents, generally leads to G:C to A:T mutagenesis. To study DNA replication encountering O 6 -MeG by the DNA polymerase (gp90) of P. aeruginosa phage PaP1, we analyzed steady-state and pre-steady-state kinetics of nucleotide incorporation opposite O 6 -MeG by gp90 exo - . O 6 -MeG partially inhibited full-length extension by gp90 exo - . O 6 -MeG greatly reduces dNTP incorporation efficiency, resulting in 67-fold preferential error-prone incorporation of dTTP than dCTP. Gp90 exo - extends beyond T:O 6 -MeG 2-fold more efficiently than C:O 6 -MeG. Incorporation of dCTP opposite G and incorporation of dCTP or dTTP opposite O 6 -MeG show fast burst phases. The pre-steady-state incorporation efficiency (k pol /K d,dNTP ) is decreased in the order of dCTP:G>dTTP:O 6 -MeG>dCTP:O 6 -MeG. The presence of O 6 -MeG at template does not affect the binding affinity of polymerase to DNA but it weakened their binding in the presence of dCTP and Mg 2+ . Misincorporation of dTTP opposite O 6 -MeG further weakens the binding affinity of polymerase to DNA. The priority of dTTP incorporation opposite O 6 -MeG is originated from the fact that dTTP can induce a faster conformational change step and a faster chemical step than dCTP. This study reveals that gp90 bypasses O 6 -MeG in an error-prone manner and provides further understanding in DNA replication encountering mutagenic alkylation DNA damage for P. aeruginosa phage PaP1. Copyright © 2017 Elsevier B.V. All rights reserved.

Sensitized fluorescence in thallium induced in collisions with Hg(6/sup 3/P/sub 1/) atoms

Energy Technology Data Exchange (ETDEWEB)

Wade, M K; Czajkowski, M; Krause, L [Windsor Univ., Ontario (Canada). Dept. of Physics

1978-07-01

The transfer of excitation from excited mercury atoms to ground-state thallium atoms was investigated using techniques of sensitized fluorescence. A Hg-Tl vapor mixture contained in a quartz cell was irradiated with Hg 2537 A resonance radiation which caused the mercury atoms to become excited to the 6/sup 3/P/sub 1/ state. Subsequent collisions between the Hg(6/sup 3/P/sub 1/) and Tl(6/sup 2/Psub(1/2)) atoms resulted in the population of the 8/sup 2/Ssub(1/2), 6/sup 2/D, and 7/sup 2/Ssub(1/2) thallium states, whose decay gave rise to sensitized fluorescence of wavelengths 3231, 3520, 3776, and 5352 A. Intensity measurements on the sensitized fluorescence and on the Hg 2537 A resonance fluorescence, observed at right angles to the direction of excitation, yielded cross sections of 3.0, 0.3, and 0.05 A/sup 2/ for collisional excitation transfer from Hg(6/sup 3/P/sub 1/) to the 8/sup 2/Ssub(1/2), 6/sup 2/D, and 7/sup 2/Ssub(1/2) states in thallium, respectively. The results are fully consistent with previously determined cross sections for excitation transfer in other binary metallic vapor systems.

Acupressure at acupoint P6 for prevention of postoperative nausea and vomiting: a randomised clinical trial

DEFF Research Database (Denmark)

Majholm, Birgitte; Møller, Ann M

2011-01-01

Postoperative nausea and vomiting causes discomfort in many patients despite both antiemetic prophylactics and improved anaesthetic techniques. Stimulation of acupoint P6 is described as an alternative method for prophylaxis of postoperative nausea and vomiting.In a randomised, double-blinded stu......, we aimed to investigate the effect of P6 acupoint stimulation on the incidence of postoperative nausea and vomiting within 24 h postoperatively with an acupressure wristband: Vital-Band....

Clonal evolution and clinical significance of copy number neutral loss of heterozygosity of chromosome arm 6p in acquired aplastic anemia.

Science.gov (United States)

Betensky, Marisol; Babushok, Daria; Roth, Jacquelyn J; Mason, Philip J; Biegel, Jaclyn A; Busse, Tracy M; Li, Yimei; Lind, Curt; Papazoglou, Anna; Monos, Dimitri; Podsakoff, Gregory; Bessler, Monica; Olson, Timothy S

2016-01-01

Acquired aplastic anemia (aAA) results from the T cell-mediated autoimmune destruction of hematopoietic stem cells. Factors predicting response to immune suppression therapy (IST) or development of myelodysplastic syndrome (MDS) are beginning to be elucidated. Our recent data suggest most patients with aAA treated with IST develop clonal somatic genetic alterations in hematopoietic cells. One frequent acquired abnormality is copy-number neutral loss of heterozygosity on chromosome 6p (6p CN-LOH) involving the human leukocyte antigen (HLA) locus. We hypothesized that because 6p CN-LOH clones may arise from selective pressure to escape immune surveillance through deletion of HLA alleles, the development of 6p CN-LOH may affect response to IST. We used single nucleotide polymorphism array genotyping and targeted next-generation sequencing of HLA alleles to assess frequency of 6p CN-LOH, identity of HLA alleles lost through 6p CN-LOH, and impact of 6p CN-LOH on response to IST. 6p CN-LOH clones were present in 11.3% of patients, remained stable over time, and were not associated with development of MDS-defining cytogenetic abnormalities. Notably, no patient with 6p CN-LOH treated with IST achieved a complete response. In summary, clonal 6p CN-LOH in aAA defines a unique subgroup of patients that may provide insights into hematopoietic clonal evolution. Copyright © 2016 Elsevier Inc. All rights reserved.

Contribution to the study of excited levels of {sup 7}Be obtained from the reaction {sup 6}Li (p, {alpha}); Contribution a l'etude des niveaux excites du {sup 7}Be obtenus par la reaction {sup 6}Li (p, {alpha})

Energy Technology Data Exchange (ETDEWEB)

Beaumevieille, H. [Commissariat a l' Energie Atomique, Grenoble (France). Centre d' Etudes Nucleaires

1964-05-01

The interpretation of the results of the reaction {sup 6}Li (p, {alpha}) in the energy range 100 keV to 3 MeV has been done with the next levels of {sup 7}Be : 3/2- (5,9 MeV), 3/2+ (6,2 MeV), 5/2- (7,18 MeV) and a level the characteristics of which may be 1/2+ or {sup 4}P (9,5 MeV). (author) [French] L'interpretation des resultats de la reaction {sup 6}Li (p, {alpha}) dans la gamme d'energie 100 keV a 3 MeV a ete faite avec les niveaux suivants du {sup 7}Be : 3/2- (5,9 MeV), 3/2 + (6,2 MeV), 5/2- (7,18 MeV) et un niveau dont les caracteristiques doivent etre 1/2+ ou {sup 4}P (9,5 MeV). (auteur)

Contribution to the study of excited levels of {sup 7}Be obtained from the reaction {sup 6}Li (p, {alpha}); Contribution a l'etude des niveaux excites du {sup 7}Be obtenus par la reaction {sup 6}Li (p, {alpha})

Energy Technology Data Exchange (ETDEWEB)

Beaumevieille, H [Commissariat a l' Energie Atomique, Grenoble (France). Centre d' Etudes Nucleaires

1964-05-01

The interpretation of the results of the reaction {sup 6}Li (p, {alpha}) in the energy range 100 keV to 3 MeV has been done with the next levels of {sup 7}Be : 3/2- (5,9 MeV), 3/2+ (6,2 MeV), 5/2- (7,18 MeV) and a level the characteristics of which may be 1/2+ or {sup 4}P (9,5 MeV). (author) [French] L'interpretation des resultats de la reaction {sup 6}Li (p, {alpha}) dans la gamme d'energie 100 keV a 3 MeV a ete faite avec les niveaux suivants du {sup 7}Be : 3/2- (5,9 MeV), 3/2 + (6,2 MeV), 5/2- (7,18 MeV) et un niveau dont les caracteristiques doivent etre 1/2+ ou {sup 4}P (9,5 MeV). (auteur)

Inhibitor of DNA binding 1 (Id1) induces differentiation and proliferation of mouse embryonic carcinoma P19CL6 cells

International Nuclear Information System (INIS)

Meng, Qingzhen; Jia, Zhuqing; Wang, Weiping; Li, Binhong; Ma, Kangtao; Zhou, Chunyan

2011-01-01

Highlights: → Id1 was upregulated during the cardiac differentiation process of P19CL6 cells. → Id1 upregulated expression of cardiac specific genes Gata4, α-MHC and ISL1. → Id1 promoted proliferation of P19CL6 cells. → Overexpression of Id1 increased activity of TOP flash. → Wnt3a or LiCl treatment promoted Id1 expression in P19CL6 cells. -- Abstract: The inhibitor of DNA binding (Id) family of genes encodes negative regulators of basic helix-loop-helix transcription factors and has been implicated in such diverse cellular processes as differentiation, proliferation, apoptosis and migration. Id knockout mouse embryos display multiple cardiac defects but the specific role of Id1 in cardiac differentiation is unclear. In the present study, we investigated the function of Id1 in DMSO-induced P19CL6 cells, a widely-accepted cell model of cardiac differentiation. We found that Id1 was upregulated during the cardiac differentiation of P19CL6 cells. The expression of cardiac specific marker genes, Gata4, α-MHC and ISL1, was upregulated in P19CL6 cells stably transfected with Id1 (P19CL6-Id1) during cardiac differentiation. The overexpression of Id1 reduced the number of cells in G1 phase and increased the cell population in G2, M and S phases, while knockdown of Id1 increased the number of cells in G1 phase from 48.6 ± 2.51% to 62.2 ± 1.52% at day 0 of cardiac induction, and from 52.5 ± 3.41% to 63.7 ± 1.02% at day 3 after cardiac induction, indicating that Id1 promoted proliferation of P19CL6 cells. Luciferase assays showed that the activity of TOP flash was higher in P19CL6-Id1 cells than wildtype P19CL6 cells, while Id1 expression was also upregulated in P19CL6 cells treated with Wnt3a or LiCl. This indicates that there may be positive feedback between Id1 and Wnt signaling which plays an important role in cardiac differentiation.

Systematics of proton absorption deduced from ( p,p) and ( p,n) cross sectionsfor 2.0- to 6.7-MeV protons on /sup 107,109/Ag and 115In

International Nuclear Information System (INIS)

Hershberger, R.L.; Flynn, D.S.; Gabbard, F.; Johnson, C.H.

1980-01-01

The ( p,p) and ( p,n) cross sections were measured to accuracies of +- 2% and +- 3%, respectively, for 2.0- to 6.7-MeV protons on /sup 107,109/Ag and 115 In. Hauser-Feshbach calculations, which included γ-ray emission channels, were used to convert the ( p,n) cross sections to proton absorption cross sections. Analysis of the ( p,p) and deduced proton absorption cross sections were made simultaneously using a conventional optical-model potential. The measured cross sections can be described using parameters extrapolated from the Sn region in a systematic way, except for a large increase required for the depth of the absorptive potential

Autocrine Regulation of UVA-Induced IL-6 Production via Release of ATP and Activation of P2Y Receptors

Science.gov (United States)

Kawano, Ayumi; Kadomatsu, Remi; Ono, Miyu; Kojima, Shuji; Tsukimoto, Mitsutoshi; Sakamoto, Hikaru

2015-01-01

Extracellular nucleotides, such as ATP, are released from cells in response to various stimuli and act as intercellular signaling molecules through activation of P2 receptors. Exposure to the ultraviolet radiation A (UVA) component of sunlight causes molecular and cellular damage, and in this study, we investigated the involvement of extracellular nucleotides and P2 receptors in the UVA-induced cellular response. Human keratinocyte-derived HaCaT cells were irradiated with a single dose of UVA (2.5 J/cm2), and ATP release and interleukin (IL)-6 production were measured. ATP was released from cells in response to UVA irradiation, and the release was blocked by pretreatment with inhibitors of gap junction hemichannels or P2X7 receptor antagonist. IL-6 production was increased after UVA irradiation, and this increase was inhibited by ecto-nucleotidase or by antagonists of P2Y11 or P2Y13 receptor. These results suggest that UVA-induced IL-6 production is mediated by release of ATP through hemichannels and P2X7 receptor, followed by activation of P2Y11 and P2Y13 receptors. Interestingly, P2Y11 and P2Y13 were associated with the same pattern of IL-6 production, though they trigger different intracellular signaling cascades: Ca2+-dependent and PI3K-dependent, respectively. Thus, IL-6 production in response to UVA-induced ATP release involves at least two distinct pathways, mediated by activation of P2Y11 and P2Y13 receptors. PMID:26030257

Neutron diffraction analysis of HRh[P(C6H5)3]4

International Nuclear Information System (INIS)

Bau, R.; Stevens, R.C.; McLean, M.; Koetzle, T.F.

1987-01-01

We have collected neutron diffraction data on a large single crystal of the title compound. The most surprising result is an extremely short Rh-H distance of 1.31(8) A, presumably caused by steric interactions involving the bulky triphenyl phosphine ligands. Crystallographic details: HRh[P(C 6 H 5 ) 3 ] 4 . 1 / 2 C 6 H 6 crystallizes in the space group Pa3, with a = b = c = 22.776(3) A, Z = 8. Data were collected at the Brookhaven High Flux Beam reactor at a temperature of -23 0 C, λ = 1.15882(7) A -1 . Least-squares refinement (in which the phenyl rings were treated as rigid groups) resulted in an R factor [based on data with f > 4σ(F)] of 0.12 for 914 reflections and 95 parameters. 10 refs

Structure and Function of p97 and Pex1/6 Type II AAA+ Complexes.

Science.gov (United States)

Saffert, Paul; Enenkel, Cordula; Wendler, Petra

2017-01-01

Protein complexes of the Type II AAA+ (ATPases associated with diverse cellular activities) family are typically hexamers of 80-150 kDa protomers that harbor two AAA+ ATPase domains. They form double ring assemblies flanked by associated domains, which can be N-terminal, intercalated or C-terminal to the ATPase domains. Most prominent members of this family include NSF (N-ethyl-maleimide sensitive factor), p97/VCP (valosin-containing protein), the Pex1/Pex6 complex and Hsp104 in eukaryotes and ClpB in bacteria. Tremendous efforts have been undertaken to understand the conformational dynamics of protein remodeling type II AAA+ complexes. A uniform mode of action has not been derived from these works. This review focuses on p97/VCP and the Pex1/6 complex, which both structurally remodel ubiquitinated substrate proteins. P97/VCP plays a role in many processes, including ER- associated protein degradation, and the Pex1/Pex6 complex dislocates and recycles the transport receptor Pex5 from the peroxisomal membrane during peroxisomal protein import. We give an introduction into existing knowledge about the biochemical and cellular activities of the complexes before discussing structural information. We particularly emphasize recent electron microscopy structures of the two AAA+ complexes and summarize their structural differences.

Genus beta human papillomavirus E6 proteins vary in their effects on the transactivation of p53 target genes.

Science.gov (United States)

White, Elizabeth A; Walther, Johanna; Javanbakht, Hassan; Howley, Peter M

2014-08-01

The genus beta human papillomaviruses (beta HPVs) cause cutaneous lesions and are thought to be involved in the initiation of some nonmelanoma skin cancers (NMSCs), particularly in patients with the genetic disorder epidermodysplasia verruciformis (EV). We have previously reported that at least two of the genus beta HPV E6 proteins bind to and/or increase the steady-state levels of p53 in squamous epithelial cells. This is in contrast to a well-characterized ability of the E6 proteins of cancer-associated HPVs of genus alpha HPV, which inactivate p53 by targeting its ubiquitin-mediated proteolysis. In this study, we have investigated the ability of genus beta E6 proteins from eight different HPV types to block the transactivation of p53 target genes following DNA damage. We find that the E6 proteins from diverse beta HPV species and types vary in their capacity to block the induction of MDM2, p21, and proapoptotic genes after genotoxic stress. We conclude that some genus beta HPV E6 proteins inhibit at least some p53 target genes, although perhaps not by the same mechanism or to the same degree as the high-risk genus alpha HPV E6 proteins. This study addresses the ability of various human papillomavirus E6 proteins to block the activation of p53-responsive cellular genes following DNA damage in human keratinocytes, the normal host cell for HPVs. The E6 proteins encoded by the high-risk, cancer-associated HPV types of genus alpha HPV have a well-established activity to target p53 degradation and thereby inhibit the response to DNA damage. In this study, we have investigated the ability of genus beta HPV E6 proteins from eight different HPV types to block the ability of p53 to transactivate downstream genes following DNA damage. We find that some, but not all, genus beta HPV E6 proteins can block the transactivation of some p53 target genes. This differential response to DNA damage furthers the understanding of cutaneous HPV biology and may help to explain the

Effect of microorganisms on the plutonium oxidation states

International Nuclear Information System (INIS)

Lukšienė, Benedikta; Druteikienė, Rūta; Pečiulytė, Dalia; Baltrūnas, Dalis; Remeikis, Vidmantas; Paškevičius, Algimantas

2012-01-01

Particular microbes from substrates at the low-level radioactive waste repository in the Ignalina NPP territory were exposed to 239 Pu (IV) at low pH under aerobic conditions. Pu(III) and Pu(IV) were separated and quantitatively evaluated using the modified anion exchange method and alpha spectrometry. Tested bacteria Bacillus mycoides and Serratia marcescens were more effective in Pu reduction than Rhodococcus fascians. Fungi Paecillomyces lilacinus and Absidia spinosa var. spinosa as well as bacterium Rhodococcus fascians did not alter the plutonium oxidation state. - Highlights: ► Particular microbes from low-level radioactive waste repository were exposed to Pu (IV). ► Some tested bacteria induced slight Pu (IV) reduction at low pH under aerobic conditions. ► Tested fungi did not show peculiarities to alter Pu oxidation state. ► The modified radiochemical method was applied to differentiate Pu oxidation states.

Structural, Mechanical, Anisotropic, and Thermal Properties of AlAs in oC12 and hP6 Phases under Pressure

Directory of Open Access Journals (Sweden)

Wei Zhang

2018-05-01

Full Text Available The structural, mechanical, anisotropic, and thermal properties of oC12-AlAs and hP6-AlAs under pressure have been investigated by employing first-principles calculations based on density functional theory. The elastic constants, bulk modulus, shear modulus, Young’s modulus, B/G ratio, and Poisson’s ratio for oC12-AlAs and hP6-AlAs have been systematically investigated. The results show that oC12-AlAs and hP6-AlAs are mechanically stable within the considered pressure. Through the study of lattice constants (a, b, and c with pressure, we find that the incompressibility of oC12-AlAs and hP6-AlAs is the largest along the c-axis. At 0 GPa, the bulk modulus B of oC12-AlAs, hP6-AlAs, and diamond-AlAs are 76 GPa, 75 GPa, and 74 Gpa, respectively, indicating that oC12-AlAs and hP6-AlAs have a better capability of resistance to volume than diamond-AlAs. The pressure of transition from brittleness to ductility for oC12-AlAs and hP6-AlAs are 1.21 GPa and 2.11 GPa, respectively. The anisotropy of Young’s modulus shows that oC12-AlAs and hP6-AlAs have greater isotropy than diamond-AlAs. To obtain the thermodynamic properties of oC12-AlAs and hP6-AlAs, the sound velocities, Debye temperature, and minimum thermal conductivity at considered pressure were investigated systematically. At ambient pressure, oC12-AlAs (463 K and hP6-AlAs (471 K have a higher Debye temperature than diamond-AlAs (433 K. At T = 300 K, hP6-AlAs (0.822 W/cm·K−1 has the best thermal conductivity of the three phases, and oC12-AlAs (0.809 W/cm·K−1 is much close to diamond-AlAs (0.813 W/cm·K−1.

Interleukin-6 stimulates Akt and p38 MAPK phosphorylation and fibroblast migration in non-diabetic but not diabetic mice.

Directory of Open Access Journals (Sweden)

Tsubame Nishikai-Yan Shen

Full Text Available Persistent inflammatory environment and abnormal macrophage activation are characteristics of chronic diabetic wounds. Here, we attempted to characterize the differences in macrophage activation and temporal variations in cytokine expression in diabetic and non-diabetic wounds, with a focus on interleukin (IL-6 mRNA expression and the p38 MAPK and PI3K/Akt signaling pathways. Cutaneous wound closure, CD68- and arginase-1 (Arg-1-expressing macrophages, and cytokine mRNA expression were examined in non-diabetic and streptozotocin-induced type 1 diabetic mice at different time points after injury. The effect of IL-6 on p38 MAPK and Akt phosphorylation was investigated, and an in vitro scratch assay was performed to determine the role of IL-6 in primary skin fibroblast migration. Before injury, mRNA expression levels of the inflammatory markers iNOS, IL-6, and TNF-α were higher in diabetic mice; however, IL-6 expression was significantly lower 6 h post injury in diabetic wounds than that in non-diabetic wounds. Non-diabetic wounds exhibited increased p38 MAPK and Akt phosphorylation; however, no such increase was found in diabetic wounds. In fibroblasts from non-diabetic mice, IL-6 increased the phosphorylation of p38 MAPK and levels of its downstream factor CREB, and also significantly increased Akt phosphorylation and levels of its upstream factor P13K. These effects of IL-6 were not detected in fibroblasts derived from the diabetic mice. In scratch assays, IL-6 stimulated the migration of primary cultured skin fibroblasts from the non-diabetic mice, and the inhibition of p38 MAPK was found to markedly suppress IL-6-stimulated fibroblast migration. These findings underscore the critical differences between diabetic and non-diabetic wounds in terms of macrophage activation, cytokine mRNA expression profile, and involvement of the IL-6-stimulated p38 MAPK-Akt signaling pathway. Aberrant macrophage activation and abnormalities in the cytokine m

Degradation/solubilization of Chinese lignite by Penicillium sp. P6

Energy Technology Data Exchange (ETDEWEB)

Yuan, H.L.; Yang, J.S.; Wang, F.Q.; Chen, W.X. [China Agricultural University, Beijing (China). Key Laboratory of Agro-Microbial Resource and Application, Ministry of Agrio, College of Biological Science

2006-01-15

Penicillium sp. P6, isolated from coal mine soil at the Qiantong colliery Liaoning Province, Northeast China, can degrade Chinese lignite in 36 h on a plate colony and in 48 h using a 4-day cultured cell-free filtrate. Results of elemental analysis and IR spectrometry indicated that solubilized products exhibited some alterations in comparison to the original lignite. The amount of fulvic acid extracted from the biodegraded lignite was high, and the molecular distribution of the humic acids from biodegraded lignite changed distinctively in comparison to which extracted from the control lignite, possibly due to the depolymerization associated with fungal biodegradation.

Association between PI3K/Akt/mTOR/p70S6K signaling pathway and hepatic fibrosis

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WU Changhui

2015-11-01

Full Text Available Phosphoinositide 3-kinase (PI3K/protein kinase-B (AkT/mammalian target of rapamycin (mTOR/70-kDa ribosomal protein S6 kinase (p70S6K, PI3K/Akt/mTOR/p70S6K, is an important signaling pathway in the life activities of cells, and it plays an important role in promoting the growth, proliferation, invasion, and anti-apoptosis of cells and promoting angiogenesis. It was clarified that the PI3K/Akt/mTOR/p70S6K signaling pathway is involved in regulating the activities of hepatic stellate cell(HSC, thus influencing the development and progression of hepatic fibrosis. Analysis demonstrated that blocking any target of the PI3K/Akt/mTOR/p70S6K signaling pathway can inhibit the activation and proliferation of HSC, promote the apoptosis of HSC, inhibit the extracellular matrix secretion from HSC, and delay the progression of hepatic fibrosis. Blocking the pathway is expected to be a treatment strategy for hepatic fibrosis.

Solution structure of the Equine Infectious Anemia Virus p9 protein: a rationalization of its different ALIX binding requirements compared to the analogous HIV-p6 protein

Directory of Open Access Journals (Sweden)

Henklein Peter

2009-12-01

Full Text Available Abstract Background The equine infection anemia virus (EIAV p9 Gag protein contains the late (L- domain required for efficient virus release of nascent virions from the cell membrane of infected cell. Results In the present study the p9 protein and N- and C-terminal fragments (residues 1-21 and 22-51, respectively were chemically synthesized and used for structural analyses. Circular dichroism and 1H-NMR spectroscopy provide the first molecular insight into the secondary structure and folding of this 51-amino acid protein under different solution conditions. Qualitative 1H-chemical shift and NOE data indicate that in a pure aqueous environment p9 favors an unstructured state. In its most structured state under hydrophobic conditions, p9 adopts a stable helical structure within the C-terminus. Quantitative NOE data further revealed that this α-helix extends from Ser-27 to Ser-48, while the N-terminal residues remain unstructured. The structural elements identified for p9 differ substantially from that of the functional homologous HIV-1 p6 protein. Conclusions These structural differences are discussed in the context of the different types of L-domains regulating distinct cellular pathways in virus budding. EIAV p9 mediates virus release by recruiting the ALG2-interacting protein X (ALIX via the YPDL-motif to the site of virus budding, the counterpart of the YPXnL-motif found in p6. However, p6 contains an additional PTAP L-domain that promotes HIV-1 release by binding to the tumor susceptibility gene 101 (Tsg101. The notion that structures found in p9 differ form that of p6 further support the idea that different mechanisms regulate binding of ALIX to primary versus secondary L-domains types.

Metabolism of bilirubin by human cytochrome P450 2A6

Energy Technology Data Exchange (ETDEWEB)

Abu-Bakar, A' edah, E-mail: a.abubakar@uq.edu.au [The University of Queensland, National Research Centre for Environmental Toxicology (Entox), 4072 Brisbane, Queensland (Australia); Arthur, Dionne M. [The University of Queensland, National Research Centre for Environmental Toxicology (Entox), 4072 Brisbane, Queensland (Australia); Cooperative Research Centre for Contamination Assessment and Remediation of the Environment, Adelaide (Australia); Wikman, Anna S. [The University of Queensland, National Research Centre for Environmental Toxicology (Entox), 4072 Brisbane, Queensland (Australia); Department of Pharmaceutical Biosciences, Uppsala University, SE-75123 Uppsala (Sweden); Rahnasto, Minna; Juvonen, Risto O.; Vepsäläinen, Jouko; Raunio, Hannu [School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, POB 1627, 70211 Kuopio (Finland); Ng, Jack C. [The University of Queensland, National Research Centre for Environmental Toxicology (Entox), 4072 Brisbane, Queensland (Australia); Cooperative Research Centre for Contamination Assessment and Remediation of the Environment, Adelaide (Australia); Lang, Matti A. [The University of Queensland, National Research Centre for Environmental Toxicology (Entox), 4072 Brisbane, Queensland (Australia)

2012-05-15

The mouse cytochrome P450 (CYP) 2A5 has recently been shown to function as hepatic “Bilirubin Oxidase” (Abu-Bakar, A., et al., 2011. Toxicol. Appl. Pharmacol. 257, 14–22). To date, no information is available on human CYP isoforms involvement in bilirubin metabolism. In this paper we provide novel evidence for human CYP2A6 metabolising the tetrapyrrole bilirubin. Incubation of bilirubin with recombinant yeast microsomes expressing the CYP2A6 showed that bilirubin inhibited CYP2A6-dependent coumarin 7-hydroxylase activity to almost 100% with an estimated K{sub i} of 2.23 μM. Metabolite screening by a high-performance liquid chromatography/electrospray ionisation mass spectrometry indicated that CYP2A6 oxidised bilirubin to biliverdin and to three other smaller products with m/z values of 301, 315 and 333. Molecular docking analyses indicated that bilirubin and its positively charged intermediate interacted with key amino acid residues at the enzyme's active site. They were stabilised at the site in a conformation favouring biliverdin formation. By contrast, the end product, biliverdin was less fitting to the active site with the critical central methylene bridge distanced from the CYP2A6 haem iron facilitating its release. Furthermore, bilirubin treatment of HepG2 cells increased the CYP2A6 protein and activity levels with no effect on the corresponding mRNA. Co-treatment with cycloheximide (CHX), a protein synthesis inhibitor, resulted in increased half-life of the CYP2A6 compared to cells treated only with CHX. Collectively, the observations indicate that the CYP2A6 may function as human “Bilirubin Oxidase” where bilirubin is potentially a substrate and a regulator of the enzyme. -- Highlights: ► Human CYP2A6 interacts with bilirubin with a high affinity. ► Bilirubin docking to the CYP2A6 active site is more stable than biliverdin docking. ► Recombinant CYP2A6 microsomes metabolised bilirubin to biliverdin. ► Bilirubin increased the hepatic

Metabolism of bilirubin by human cytochrome P450 2A6

International Nuclear Information System (INIS)

Abu-Bakar, A'edah; Arthur, Dionne M.; Wikman, Anna S.; Rahnasto, Minna; Juvonen, Risto O.; Vepsäläinen, Jouko; Raunio, Hannu; Ng, Jack C.; Lang, Matti A.

2012-01-01

The mouse cytochrome P450 (CYP) 2A5 has recently been shown to function as hepatic “Bilirubin Oxidase” (Abu-Bakar, A., et al., 2011. Toxicol. Appl. Pharmacol. 257, 14–22). To date, no information is available on human CYP isoforms involvement in bilirubin metabolism. In this paper we provide novel evidence for human CYP2A6 metabolising the tetrapyrrole bilirubin. Incubation of bilirubin with recombinant yeast microsomes expressing the CYP2A6 showed that bilirubin inhibited CYP2A6-dependent coumarin 7-hydroxylase activity to almost 100% with an estimated K i of 2.23 μM. Metabolite screening by a high-performance liquid chromatography/electrospray ionisation mass spectrometry indicated that CYP2A6 oxidised bilirubin to biliverdin and to three other smaller products with m/z values of 301, 315 and 333. Molecular docking analyses indicated that bilirubin and its positively charged intermediate interacted with key amino acid residues at the enzyme's active site. They were stabilised at the site in a conformation favouring biliverdin formation. By contrast, the end product, biliverdin was less fitting to the active site with the critical central methylene bridge distanced from the CYP2A6 haem iron facilitating its release. Furthermore, bilirubin treatment of HepG2 cells increased the CYP2A6 protein and activity levels with no effect on the corresponding mRNA. Co-treatment with cycloheximide (CHX), a protein synthesis inhibitor, resulted in increased half-life of the CYP2A6 compared to cells treated only with CHX. Collectively, the observations indicate that the CYP2A6 may function as human “Bilirubin Oxidase” where bilirubin is potentially a substrate and a regulator of the enzyme. -- Highlights: ► Human CYP2A6 interacts with bilirubin with a high affinity. ► Bilirubin docking to the CYP2A6 active site is more stable than biliverdin docking. ► Recombinant CYP2A6 microsomes metabolised bilirubin to biliverdin. ► Bilirubin increased the hepatic CYP2A6

^2H(^18F,p)^19F Study at 6 MeV/u

Science.gov (United States)

Kozub, R. L.; Nesaraja, C. D.; Moazen, B. H.; Scott, J. P.; Bardayan, D. W.; Blackmon, J. C.; Gross, C. J.; Shapira, D.; Smith, M. S.; Batchelder, J. C.; Brune, C. R.; Champagne, A. E.; Sahin, L.; Cizewski, J. A.; Thomas, J. S.; Davinson, T.; Woods, P. J.; Greife, U.; Jewett, C.; Livesay, R. J.; Ma, Z.; Parker, P. D.

2003-04-01

The degree to which the (p,α) and (p,γ) reactions destroy ^18F at temperatures ˜1-4 x 10^8 K is important for understanding the synthesis of nuclei in nova explosions and for using ^18F as a monitor of nova mechanisms in gamma ray astronomy. The reactions are dominated by low-lying proton resonances near the ^18F+p threshold (E_x=6.411 MeV excitation energy in ^19Ne). To gain further information about these resonances, we have used the inverse ^18F(d,p)^19F neutron transfer reaction at the Holifield Radioactive Ion Beam Facility to selectively populate corresponding mirror states in ^19F. Proton angular distributions were measured for states in ^19F in the excitation energy range 0-9 MeV. Results and implications for the ^18F+p reactions and nuclear structure will be presented. ^1Supported by DOE. ^2ORNL is managed by UT-Battelle, LLC, for the USDOE.

Metallicity of Ca{sub 2}Cu{sub 6}P{sub 5} with single and double copper-pnictide layers

Energy Technology Data Exchange (ETDEWEB)

Li, Li, E-mail: lil2@ornl.gov [Materials Science & Technology Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831 (United States); Parker, David [Materials Science & Technology Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831 (United States); Chi, Miaofang [Center for Nanophase Materials Sciences, Oak Ridge National Laboratory, Oak Ridge, TN 37831 (United States); Tsoi, Georgiy M.; Vohra, Yogesh K. [Department of Physics, University of Alabama at Birmingham, Birmingham, AL 35294 (United States); Sefat, Athena S., E-mail: sefata@ornl.gov [Materials Science & Technology Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831 (United States)

2016-06-25

We report thermodynamic and transport properties, and also theoretical calculations, for Cu-based compound Ca{sub 2}Cu{sub 6}P{sub 5} and compare with CaCu{sub 2-δ}P{sub 2}. Both materials have layers of edge-sharing copper pnictide tetrahedral CuP{sub 4}, similar to Fe–As and Fe–Se layers (with FeAs{sub 4}, FeSe{sub 4}) in the iron-based superconductors. Despite the presence of this similar transition-metal pnictide layer, we find that both Ca{sub 2}Cu{sub 6}P{sub 5} and CaCu{sub 2-δ}P{sub 2} have temperature-independent magnetic susceptibility and show metallic behavior with no evidence of either magnetic ordering or superconductivity down to 1.8 K CaCu{sub 2-δ}P{sub 2} is slightly off-stoichiometric, with δ = 0.14. Theoretical calculations suggest that unlike Fe 3d-based magnetic materials with a large density of states (DOS) at the Fermi surface, Cu have comparatively low DOS, with the majority of the 3d spectral weight located well below Fermi level. The room-temperature resistivity value of Ca{sub 2}Cu{sub 6}P{sub 5} is only 9 μΩ-cm, due to a substantial plasma frequency and an inferred electron-phonon coupling λ of 0.073 (significantly smaller than that of metallic Cu). Also, microscopy result shows that Cu–Cu distance along the c-axis within the double layers can be very short (2.5 Å), even shorter than metallic elemental copper bond (2.56 Å). The value of dρ/dT for CaCu{sub 2-δ}P{sub 2} at 300 K is approximately three times larger than in Ca{sub 2}Cu{sub 6}P{sub 5}, which suggests the likelihood of stronger electron-phonon coupling. This study shows that the details of Cu–P layers and bonding are important for their transport characteristics. In addition, it emphasizes the remarkable character of the DOS of ‘122’ iron-based materials, despite much structural similarities. - Highlights: • A comprehensive study on Cu-based compound Ca{sub 2}Cu{sub 6}P{sub 5} and compare with CaCu{sub 2-δ}P{sub 2}. • Both materials have layers of

The relevance of the M6P/IGF2R status for the tumorigenicity and invasiveness of liver and skin cancer cells

International Nuclear Information System (INIS)

Puxbaum, V.

2010-01-01

The mannose 6-phosphate/insulin-like growth factor 2 receptor (M6P/IGF2R), a multifunctional membrane-associated protein, plays a central role in targeting of lysosomal enzymes and controlling of the bioavailability of insulin-like growth factor II (IGF-II). In addition to intracellular sorting and endocytosis of M6P-containing ligands and IGF-II, the receptor also interacts with urokinase-type plasminogen activator receptor (uPAR) and plasminogen at the cell surface. M6P/IGF2R is considered a putative tumor suppressor, and its expression may modulate the invasiveness of cancer cells. Importantly, the M6P/IGF2R gene is frequently lost or mutated in a wide range of malignant tumors including hepatocellular and squamous cell carcinomas. However, the impact of the receptor on tumor invasion and metastasis is still poorly understood. FRL14 and SCC-VII cells are M6P/IGF2R deficient and thus secrete large amounts of lysosomal enzymes. Reconstitution of functional M6P/IGF2R expression restores transport of lysosomal enzymes to these compartments and drastically reduces the invasive potential of the cells. In addition, the presence of ectopic M6P/IGF2R compromises the growth of FRL14 and SCC-VII cells both in vitro and in vivo. Conversely, M6P/IGF2R knock-down in receptor-positive MIM-1-4 hepatocytes leads to increased lysosomal enzyme secretion and enhanced invasiveness. To assess the relevance of different ligand-binding sites for the biological activities of M6P/IGF2R, several mutant forms of the receptor were stably expressed in FRL14 and SCC-VII cells. Functional M6P-binding sites proved to be important for the anti-invasive potential of M6P/IGF2R, whereas the interactions of the receptor with IGF-II and uPAR/plasminogen were found to be dispensable for its biological activities. These results suggest that the M6P/IGF2R status influences the metastatic propensity of hepatocellular and squamous cell carcinomas, and that functional M6P-binding sites are crucial for the

Insights on the Effects of Heat Pretreatment, pH, and Calcium Salts on Isolation of Rare Actinobacteria from Karstic Caves

Science.gov (United States)

Fang, Bao-Zhu; Salam, Nimaichand; Han, Ming-Xian; Jiao, Jian-Yu; Cheng, Juan; Wei, Da-Qiao; Xiao, Min; Li, Wen-Jun

2017-01-01

The phylum Actinobacteria is one of the most ubiquitously present bacterial lineages on Earth. In the present study, we try to explore the diversity of cultivable rare Actinobacteria in Sigangli Cave, Yunnan, China by utilizing a combination of different sample pretreatments and under different culture conditions. Pretreating the samples under different conditions of heat, setting the isolation condition at different pHs, and supplementation of media with different calcium salts were found to be effective for isolation of diverse rare Actinobacteria. During our study, a total of 204 isolates affiliated to 30 genera of phylum Actinobacteria were cultured. Besides the dominant Streptomyces, rare Actinobacteria of the genera Actinocorallia, Actinomadura, Agromyces, Alloactinosynnema, Amycolatopsis, Beutenbergia, Cellulosimicrobium, Gordonia, Isoptericola, Jiangella, Knoellia, Kocuria, Krasilnikoviella, Kribbella, Microbacterium, Micromonospora, Mumia, Mycobacterium, Nocardia, Nocardioides, Nocardiopsis, Nonomuraea, Oerskovia, Pseudokineococcus, Pseudonocardia, Rhodococcus, Saccharothrix, Streptosporangium, and Tsukamurella were isolated from these cave samples. PMID:28848538

The 5s25p2 - (5s25p5d + 5s5p3 + 5s25p6s + 5s25p7s) transitions in Sb II and 5s25p - (5s5p2 + 5s2nl) transitions in Sb III

International Nuclear Information System (INIS)

Arcimowicz, B.; Joshi, Y.N.; Kaufman, V.

1989-01-01

The spectrum of antimony was photographed in the 575-2300 A region (1A 10 -10 m) using a hollow cathode and a triggered spark source. The analysis of the 5s 2 5p 2 - (5s 2 5p5d + 5s5p 3 + 5s 2 5p6s + 5s 2 5p7s) transitions in Sb II spectrum was revised and interpreted on the basis of multiconfiguration interaction calculations. Accurate wavelength measurements of Sb III lines lead to a revised ground-state 5s 2 5p 2 P interval value of 6574.5 cm -1 . (author). 15 refs., 9 tabs., 1 fig

Isolation of probes specific to human chromosomal region 6p21 from immunoselected irradiation-fusion gene transfer hybrids

International Nuclear Information System (INIS)

Ragoussis, J.; Jones, T.A.; Sheer, D.; Shrimpton, A.E.; Goodfellow, P.N.; Trowsdale, J.; Ziegler, A.

1991-01-01

A hybrid cell line (R21/B1) containing a truncated human chromosome 6 (6pter-6q21) and a human Y chromosome on a hamster background was irradiated and fused to A23 (TK-) or W3GH (HPRT-) hamster cells. Clones containing expressed HLA class I genes (4/40) were selected using monoclonal antibodies. These clones were recloned and analyzed with a panel of probes from the HLA region. One hybrid (4G6) contained the entire HLA complex. Two other hybrids (4J4 and 4H2) contained only the HLA class I region, while the fourth hybrid (5P9) contained HLA class I and III genes in addition to other genes located in the 6p21 chromosomal region. In situ hybridization showed that the hybrid cells contained more than one fragment of human DNA. Alu and LINE PCR products were derived from these cells and compared to each other as well as to products from two somatic cell hybrids having the 6p21 region in common. The PCR fragments were then screened on conventional Southern blots of the somatic cell hybrids to select a panel of novel probes encompassing the 6p21 region. In addition, the origin of the human DNA fragments in hybrid 4J4 was determined by regional mapping of PCR products

P6 Electroacupuncture Improved QTc Interval Prolongation by Upregulation of Connexin43 in Droperidol Treated Rats

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Feng Zhao

2014-01-01

Full Text Available Aim. This study investigated the effect of P6 EA on droperidol-induced QTc interval prolongation and Cx43 expression in ventricular muscle of rats. Methods. Twenty-four rats were randomly divided into control group (C, droperidol group (D, or EA group (E. C group rats were injected with normal saline. D group rats were injected with droperidol 0.13 mg/kg. E group rats were pretreated with EA at left P6 acupoint for 30 min and then injected with droperidol (0.13 mg/kg. QTc intervals were recorded at lead II in ECG within 120 min. Cx43 expression was measured by RT-PCR and western blotting. Result. Droperidol significantly prolonged QTc intervals compared with controls at 5 min, 10 min, 15 min, and 30 min (P0.05. Conclusion. P6 EA could improve QTc interval prolongation induced by droperidol, which may relate to upregulation of Cx43 mRNA and protein. Antiemetic dose of droperidol had minor effects on Cx43 mRNA and protein expression at 120 min.

A high-throughput cellular assay to quantify the p53-degradation activity of E6 from different human papillomavirus types.

Science.gov (United States)

Gagnon, David; Archambault, Jacques

2015-01-01

A subset of human papillomaviruses (HPVs), known as the high-risk types, are the causative agents of cervical cancer and other malignancies of the anogenital region and oral mucosa. The capacity of these viruses to induce cancer and to immortalize cells in culture relies in part on a critical function of their E6 oncoprotein, that of promoting the poly-ubiquitination of the cellular tumor suppressor protein p53 and its subsequent degradation by the proteasome. Here, we describe a cellular assay to measure the p53-degradation activity of E6 from different HPV types. This assay is based on a translational fusion of p53 to Renilla luciferase (Rluc-p53) that remains sensitive to degradation by high-risk E6 and whose steady-state levels can be accurately measured in standard luciferase assays. The p53-degradation activity of any E6 protein can be tested and quantified in transiently transfected cells by determining the amount of E6-expression vector required to reduce by half the levels of RLuc-p53 luciferase activity (50 % effective concentration [EC50]). The high-throughput and quantitative nature of this assay makes it particularly useful to compare the p53-degradation activities of E6 from several HPV types in parallel.

Aluminum triiodide induced complexation and structural rearrangement of the hexaphospha-pentaprismane cage, P6C4tBu4

International Nuclear Information System (INIS)

Al-Ktaifani, M. M.

2009-01-01

Treatment of P 6 C 4 t Bu 4 with the Lewis acid AlI 3 in toluene at room temperature unexpectedly results in a structural rearrangement affording the ionic complex [P 6 C 4 t Bu 4 H][AlI 4 ] which was fully characterized by multinuclear NMR spectroscopy and a single crystal X-ray diffraction study. Supportive evidence that the reaction might involve a simple intermediate Lewis acid-base adduct [AlI 3 (η 1 -P 6 C 4 t Bu 4 )] came from the isolation on one occasion of small number of crystals of the bis-Al adduct [(AlI)μ-(P 6 C 4 t Bu 4 )(AlI 3 )], whose molecular structure was also confirmed by a single crystal X-ray diffraction study. (author)

Structure of Rhodococcus equi virulence-associated protein B (VapB) reveals an eight-stranded antiparallel β-barrel consisting of two Greek-key motifs

International Nuclear Information System (INIS)

Geerds, Christina; Wohlmann, Jens; Haas, Albert; Niemann, Hartmut H.

2014-01-01

The structure of VapB, a member of the Vap protein family that is involved in virulence of the bacterial pathogen R. equi, was determined by SAD phasing and reveals an eight-stranded antiparallel β-barrel similar to avidin, suggestive of a binding function. Made up of two Greek-key motifs, the topology of VapB is unusual or even unique. Members of the virulence-associated protein (Vap) family from the pathogen Rhodococcus equi regulate virulence in an unknown manner. They do not share recognizable sequence homology with any protein of known structure. VapB and VapA are normally associated with isolates from pigs and horses, respectively. To contribute to a molecular understanding of Vap function, the crystal structure of a protease-resistant VapB fragment was determined at 1.4 Å resolution. The structure was solved by SAD phasing employing the anomalous signal of one endogenous S atom and two bound Co ions with low occupancy. VapB is an eight-stranded antiparallel β-barrel with a single helix. Structural similarity to avidins suggests a potential binding function. Unlike other eight- or ten-stranded β-barrels found in avidins, bacterial outer membrane proteins, fatty-acid-binding proteins and lysozyme inhibitors, Vaps do not have a next-neighbour arrangement but consist of two Greek-key motifs with strand order 41238567, suggesting an unusual or even unique topology

Structure of Rhodococcus equi virulence-associated protein B (VapB) reveals an eight-stranded antiparallel β-barrel consisting of two Greek-key motifs

Energy Technology Data Exchange (ETDEWEB)

Geerds, Christina [Bielefeld University, Universitaetsstrasse 25, 33615 Bielefeld (Germany); Wohlmann, Jens; Haas, Albert [University of Bonn, Ulrich-Haberland Strasse 61a, 53121 Bonn (Germany); Niemann, Hartmut H., E-mail: hartmut.niemann@uni-bielefeld.de [Bielefeld University, Universitaetsstrasse 25, 33615 Bielefeld (Germany)

2014-06-18

The structure of VapB, a member of the Vap protein family that is involved in virulence of the bacterial pathogen R. equi, was determined by SAD phasing and reveals an eight-stranded antiparallel β-barrel similar to avidin, suggestive of a binding function. Made up of two Greek-key motifs, the topology of VapB is unusual or even unique. Members of the virulence-associated protein (Vap) family from the pathogen Rhodococcus equi regulate virulence in an unknown manner. They do not share recognizable sequence homology with any protein of known structure. VapB and VapA are normally associated with isolates from pigs and horses, respectively. To contribute to a molecular understanding of Vap function, the crystal structure of a protease-resistant VapB fragment was determined at 1.4 Å resolution. The structure was solved by SAD phasing employing the anomalous signal of one endogenous S atom and two bound Co ions with low occupancy. VapB is an eight-stranded antiparallel β-barrel with a single helix. Structural similarity to avidins suggests a potential binding function. Unlike other eight- or ten-stranded β-barrels found in avidins, bacterial outer membrane proteins, fatty-acid-binding proteins and lysozyme inhibitors, Vaps do not have a next-neighbour arrangement but consist of two Greek-key motifs with strand order 41238567, suggesting an unusual or even unique topology.

Hypoxia-induced invadopodia formation involves activation of NHE-1 by the p90 ribosomal S6 kinase (p90RSK.

Directory of Open Access Journals (Sweden)

Fabrice Lucien

Full Text Available The hypoxic and acidic microenvironments in tumors are strongly associated with malignant progression and metastasis, and have thus become a central issue in tumor physiology and cancer treatment. Despite this, the molecular links between acidic pH- and hypoxia-mediated cell invasion/metastasis remain mostly unresolved. One of the mechanisms that tumor cells use for tissue invasion is the generation of invadopodia, which are actin-rich invasive plasma membrane protrusions that degrade the extracellular matrix. Here, we show that hypoxia stimulates the formation of invadopodia as well as the invasive ability of cancer cells. Inhibition or shRNA-based depletion of the Na(+/H(+ exchanger NHE-1, along with intracellular pH monitoring by live-cell imaging, revealed that invadopodia formation is associated with alterations in cellular pH homeostasis, an event that involves activation of the Na(+/H(+ exchange rate by NHE-1. Further characterization indicates that hypoxia triggered the activation of the p90 ribosomal S6 kinase (p90 RSK, which resulted in invadopodia formation and site-specific phosphorylation and activation of NHE-1. This study reveals an unsuspected role of p90RSK in tumor cell invasion and establishes p90RS kinase as a link between hypoxia and the acidic microenvironment of tumors.

Quasirelativistic calculation of 4s24p5, 4s24p44d and 4s4p6 configuration spectroscopic parameters for the W39+ ion

International Nuclear Information System (INIS)

Bogdanovich, P; Karpuškienė, R; Kisielius, R

2015-01-01

The ab initio quasirelativistic Hartree–Fock method developed specifically for the calculation of spectral parameters of heavy atoms and highly charged ions is used to derive spectral data for the 4s 2 4p 5 , 4s 2 4p 4 4d and 4s4p 6 configurations of the multicharged tungsten ion W 39+ . The relativistic effects are taken into account in the Breit–Pauli approximation for the quasirelativistic Hartree–Fock radial orbitals. The configuration interaction method is applied to include the electron correlation effects. Produced data are compared with existing experimental measurements and theoretical calculations. (paper)

MG132 plus apoptosis antigen-1 (APO-1) antibody cooperate to restore p53 activity inducing autophagy and p53-dependent apoptosis in HPV16 E6-expressing keratinocytes.

Science.gov (United States)

Lagunas-Martínez, Alfredo; García-Villa, Enrique; Arellano-Gaytán, Magaly; Contreras-Ochoa, Carla O; Dimas-González, Jisela; López-Arellano, María E; Madrid-Marina, Vicente; Gariglio, Patricio

2017-01-01

The E6 oncoprotein can interfere with the ability of infected cells to undergo programmed cell death through the proteolytic degradation of proapoptotic proteins such as p53, employing the proteasome pathway. Therefore, inactivation of the proteasome through MG132 should restore the activity of several proapoptotic proteins. We investigated whether in HPV16 E6-expressing keratinocytes (KE6 cells), the restoration of p53 levels mediated by MG132 and/or activation of the CD95 pathway through apoptosis antigen-1 (APO-1) antibody are responsible for the induction of apoptosis. We found that KE6 cells underwent apoptosis mainly after incubation for 24 h with MG132 alone or APO-1 plus MG132. Both treatments activated the extrinsic and intrinsic apoptosis pathways. Autophagy was also activated, principally by APO-1 plus MG132. Inhibition of E6-mediated p53 proteasomal degradation by MG132 resulted in the elevation of p53 protein levels and its phosphorylation in Ser46 and Ser20; the p53 protein was localized mainly at nucleus after treatment with MG132 or APO-1 plus MG132. In addition, induction of its transcriptional target genes such as p21, Bax and TP53INP was observed 3 and 6 h after treatment. Also, LC3 mRNA was induced after 3 and 6 h, which correlates with lipidation of LC3B protein and induction of autophagy. Finally, using pifithrin alpha we observed a decrease in apoptosis induced by MG132, and by APO-1 plus MG132, suggesting that restoration of APO-1 sensitivity occurs in part through an increase in both the levels and the activity of p53. The use of small molecules to inhibit the proteasome pathway might permit the activation of cell death, providing new opportunities for CC treatment.

Identification of the 15FRFG domain in HIV-1 Gag p6 essential for Vpr packaging into the virion

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Zhu Henghu

2004-09-01

Full Text Available Abstract The auxiliary regulatory protein Vpr of HIV-1 is packaged in the virion through interaction with the Gag C-terminal p6 domain. Virion packaging of Vpr is critical for Vpr to exert functions in the HIV-1 life cycle. Previous studies suggest that Vpr interacts with a (Lxx4 domain in p6 for virion packaging. In the present study, mutational analysis of HIV-1 Gag p6 domain was performed in the context of the HIV-1 genome to examine the effect on virion packaging of Vpr. Surprisingly, Ala substitutions for Leu44 and Phe45 in the (Lxx4 domain or deletion of the whole (Lxx4 domain (amino acid #35–52 of the Gag p6 domain did not affect Vpr virion packaging. Vpr virion packaging was normal when amino acid #1–23 of the Gag p6 domain was preserved. Most importantly, Ala substitutions for Phe15, Arg16 and Phe17 in the context of amino acid #1–23 of the Gag p6 domain abolished Vpr virion packaging. Single Ala substitutions for Phe15 and Phe17 also abolished Vpr virion packaging, whereas Ala substitution for Arg16 had no effect. Our studies have revealed a novel signal sequence for Vpr packaging into the HIV-1 virion. The 15FRFG domain in p6 resembles the FxFG repeat sequences commonly found in proteins of the nuclear pore complex. These results have provided novel insights into the process of virion packaging of Vpr and suggest for the first time that Vpr may recognize the FxFG domain for both virion packaging and association with nuclear pores.

Kinetics of hydrocarbon extraction from oil shale using biosurfactant producing bacteria

Energy Technology Data Exchange (ETDEWEB)

Haddadin, Malik S.Y.; Abou Arqoub, Ansam A.; Abu Reesh, Ibrahim [Faculty of Graduate Studies, Jordan University, Queen Rania Street, Amman, 11942 (Jordan); Haddadin, Jamal [Faculty of Agriculture, Mutah University, P.O. Box 59, Mutah 61710 (Jordan)

2009-04-15

This study was done to extract hydrocarbon compounds from El-Lajjun oil shale using biosurfactant produced from two strains Rhodococcus erythropolis and Rhodococcus ruber. The results have shown that, optimal biosurfactant production was found using naphthalene and diesel as a carbon source for R. erthropolis and R. ruber, respectively. Optimum nitrogen concentration was 9 g/l and 7 g/l for R. erthropolis and R. ruber, respectively. Optimum K{sub 2}HPO{sub 4} to KH{sub 2}PO{sub 4} ratio, temperature, pH, and agitation speeds were 2:1, 37 C, 7 and 200 rpm. Under optimal conditions R. erthropolis and R. ruber produced 5.67 and 6.9 g/l biosurfactant, respectively. Maximum recovery of oil achieved with hydrogen peroxide pre-treatment was 25% and 26% at biosurfactant concentration of 8 g/l and 4 g/l for R. erthropolis and R. ruber, respectively. The extent desorption of hydrocarbons from the pre-treated oil shale by biosurfactant were inversely related to the concentration of high molecular weight hydrocarbons, asphaltenes compounds. Pre-treatment of oil shale with hydrogen peroxide produced better improvement in aromatic compounds extraction in comparison with improvement which resulted from demineralization of the oil shale. (author)

Overexpression of cytochrome P450 CYP6BG1 may contribute to chlorantraniliprole resistance in Plutella xylostella (L.).

Science.gov (United States)

Li, Xiuxia; Li, Ran; Zhu, Bin; Gao, Xiwu; Liang, Pei

2018-06-01

The diamondback moth Plutella xylostella (L.) is the most widely distributed pest of cruciferous crops and has developed resistance to most commonly used insecticides, including chlorantraniliprole. Resistance to chlorantraniliprole is likely caused by mutations of the target, the ryanodine receptor, and/or mediated by an increase in detoxification enzyme activities. Although target-site resistance is documented in detail, resistance mediated by increased metabolism has rarely been reported. The activity of cytochrome P450 was significantly higher in two resistant P. xylostella populations than in a susceptible one. Among ten detected cytochrome P450 genes, CYP6BG1 was significantly overexpressed (over 80-fold) in a field-resistant population compared with expression in a susceptible one. Knockdown of CYP6BG1 by RNA interference dramatically reduced the 7-ethoxycoumarin-O-deethylase (7-ECOD) activity of P450 by 45.5% and increased the toxicity of chlorantraniliprole toward P. xylostella by 26.8% at 48 h postinjection of double-stranded RNA. By contrast, overexpression of CYP6BG1 in a transgenic Drosophila melanogaster line significantly decreased the toxicity of the insecticide to the transgenic flies. Overexpression of CYP6BG1 may contribute to chlorantraniliprole resistance in P. xylostella. Our findings will provide new insights into the mechanisms of resistance to diamide insecticides in other insect pests. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Effect of microorganisms on the plutonium oxidation states

Energy Technology Data Exchange (ETDEWEB)

Luksiene, Benedikta, E-mail: bena@ar.fi.lt [Center for Physical Sciences and Technology, Savanoriu ave 231, LT-02300 Vilnius (Lithuania); Druteikiene, Ruta [Center for Physical Sciences and Technology, Savanoriu ave 231, LT-02300 Vilnius (Lithuania); Peciulyte, Dalia [Nature Research Centre, Akademijos street 2, LT-08412 Vilnius (Lithuania); Baltrunas, Dalis; Remeikis, Vidmantas [Center for Physical Sciences and Technology, Savanoriu ave 231, LT-02300 Vilnius (Lithuania); Paskevicius, Algimantas [Nature Research Centre, Akademijos street 2, LT-08412 Vilnius (Lithuania)

2012-03-15

Particular microbes from substrates at the low-level radioactive waste repository in the Ignalina NPP territory were exposed to {sup 239}Pu (IV) at low pH under aerobic conditions. Pu(III) and Pu(IV) were separated and quantitatively evaluated using the modified anion exchange method and alpha spectrometry. Tested bacteria Bacillus mycoides and Serratia marcescens were more effective in Pu reduction than Rhodococcus fascians. Fungi Paecillomyces lilacinus and Absidia spinosa var. spinosa as well as bacterium Rhodococcus fascians did not alter the plutonium oxidation state. - Highlights: Black-Right-Pointing-Pointer Particular microbes from low-level radioactive waste repository were exposed to Pu (IV). Black-Right-Pointing-Pointer Some tested bacteria induced slight Pu (IV) reduction at low pH under aerobic conditions. Black-Right-Pointing-Pointer Tested fungi did not show peculiarities to alter Pu oxidation state. Black-Right-Pointing-Pointer The modified radiochemical method was applied to differentiate Pu oxidation states.

Complete Genome Sequence of vB_BveP-Goe6, a Virus Infecting Bacillus velezensis FZB42.

Science.gov (United States)

Schilling, Tobias; Hoppert, Michael; Daniel, Rolf; Hertel, Robert

2018-02-22

The new virus vB_BveP-Goe6 was isolated on the host organism Bacillus velezensis FZB42. The virus morphology indicated its association with the genus Phi29virus The genome of vB_BveP-Goe6 (19,105 bp) comprises a linear chromosome with a GC content of 39.99%. The genome harbors 26 putative protein-coding genes and a noncoding packaging RNA. Copyright © 2018 Schilling et al.

Design, synthesis and in vitro evaluation on glucosamine-6P synthase of aromatic analogs of 2-Aminohexitols-6P

Energy Technology Data Exchange (ETDEWEB)

Dias, Danielle F.; Alves, Ricardo J., E-mail: ricardodylan@farmacia.ufmg.b [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Faculdade de Farmacia; Roux, Celine; Durand, Philippe; Iorga, Bogdan; Badet-Denisot, Marie A.; Badet, Bernard [Centre National de la Recherche Scientifique (CNRS), Gif-sur-Yvette (France). Inst. de Chimie des Substances Naturelles

2010-07-01

The aminosugars are very important structural components of bacterial and fungi cell walls. Glucosamine-6-phosphate synthase (GlmS), which catalyses the first step of the aminosugar biosynthetic pathway i.e. the formation of D-glucosamine-6-phosphate from D-fructose-6-phosphate, is therefore an interesting target in the fight against microorganisms. In this work is described the synthesis of aromatic analogs of 2-amino-2-deoxy-D-glucitol-6-phosphate (ADGP) and its epimer 2-amino-2-deoxy-D-manitol-6-phosphate (ADMP), two important inhibitors of GlmS. The aromatic analogs displayed modest inhibitory activity against GlmS, with IC{sub 50} in the mmol L{sup -1} range. (author)

Accumulation of thorium and uranium by microbes. The effect of pH, concentration of metals, and time course on the accumulation of both elements using streptomyces levoris

International Nuclear Information System (INIS)

Tsuruta, Takehiko

2006-01-01

The accumulation of thorium and uranium by various microorganisms from a solution containing both metals at pH 3.5 was examined. Among the tested species, a high accumulation ability for thorium was exhibited by strains of gram-positive bacteria, such as Arthrobacter nicotianae, Bacillus megaterium, B. subtilis, Micrococcus luteus, Rhodococcus erythropolis, and Streptomyces levoris. Though uranium was accumulated in small amounts by most of microorganisms. A. nicotianae, S. flavoviridis, and S. levoris had relatively high uranium accumulation abilities. In these high performance thorium- and uranium-accumulating microorganisms, S. levoris, which accumulated the largest amount of uranium from the solution containing only uranium at pH 3.5, accumulated about 300 μmol thorium and 133 μmol uranium per gram dry weight of microbial cells from a solution containing both thorium and uranium at pH 3.5. The amount and time course of the thorium accumulation were almost unaffected by the co-existing uranium, while those of uranium were strongly affected by the co-existing thorium. The effects of pH, the thorium and uranium concentrations, and time course on both metal accumulations were also evaluated by numerical formulas. (author)

Relativistic and non-relativistic electronic molecular-structure calculations for dimers of 4p-, 5p-, and 6p-block elements.

Science.gov (United States)

Höfener, Sebastian; Ahlrichs, Reinhart; Knecht, Stefan; Visscher, Lucas

2012-12-07

We report results of non-relativistic and two-component relativistic single-reference coupled-cluster with single and double and perturbative triple excitations [CCSD(T)] treatments for the 4p-block dimers Ga(2) to Br(2) , the 5p-block dimers In(2) to I(2) , and their atoms. Extended basis sets up to pentuple zeta are employed and energies extrapolated to the complete basis-set limit. Relativistic and non-relativistic results for the dissociation energy D(e) are in close agreement with each other and previously published data, provided non-relativistic or scalar-relativistic results are corrected for spin-orbit contributions taken from the literature. An exception is Te(2) where theoretical results scatter by 0.085 eV. By virtue of this agreement it is unexpected that comparison with the experimental D(0) or D(e) dissociation energies (zero-point vibrational effects are negligible in this context) reveal errors larger than 0.1 eV for Ga(2), Ge(2), and Sb(2). Only relativistic treatments are presented for the 6p-block cases Tl(2) to At(2). Sufficient agreement with experimental data is found only for Pb(2) and Bi(2), the deviation of the computed and experimental D(0) values for Po(2) is again larger than 0.1 eV. Deviations of 0.1 eV between the computed and experimental D(0) values are a major reason for concern and call for additional investigations in both fields to clarify the situation. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

η → π0γγ to O(p6) in chiral perturbation theory

International Nuclear Information System (INIS)

Jetter, M.

1995-08-01

The decay η → π0γγ is discussed in the framework of SU(3) chiral perturbation theory. The process is dominated by the O(p 6 ) in the momentum expansion where tree-level amplitudes from the effective Lagrangian L 6 enter together with one-loop contributions from L 4 and two-loop contributions from L 2 . We estimate the 6 independent L 6 coupling constants by resonance saturation consistent with the pion production process γγ → π 0 π 0 and calculate the pion-loop part of the one-and two-loop amplitude. Predictions for the total rate and spectrum of η → π 0 γγ are given together with a discussion of the uncertainties involved. (author). 27 refs., 8 figs.,3 tabs

New, well-retained myocardial imaging agent: radioiodinated 15-(p-iodophenyl)-6-tellurapentadecanoic acid

International Nuclear Information System (INIS)

Goodman, M.M.; Knapp, F.F. Jr.; Callahan, A.P.; Ferren, L.A.

1982-01-01

A method involving the acid-catalyzed decomposition of a piperidyltriazene intermediate in the presence of radioiodide has been developed for the synthesis of radioiodinated 15-(p-iodophenyl)-6-tellurapentadecanoic acid. The iodine-125-labeled agent shows rapid, pronounced myocardial uptake in rats (5.30-6.45% injected dose/g after 5 min) and also exhibits the prolonged retention previously observed with 9-[/sup 123m/Te]telluraheptadecanoic acid (9-[/sup 123m/Te]HDA). After 6 hr, the heart uptake remained high (3.89-5.33% dose/g) and decreased only to 3.02-3.41% dose/g after 24 hr. Very low blood activity was detected (0.24-0.27% dose/g at 5 min; 0.29-0.32% dose/g at 6 hr) and the heart-to-blood ratios were high (22:1 at 5 min; 15:1 at 6 hr). Minimal deiodination was demonstrated by the low thyroid uptake (1.41-1.63% dose/g at 5 min; 5.33-7.08% dose/g at 6 hr). The rapid and pronounced uptake, prolonged myocardial retention, and low in vivo deiodination make this agent attractive for further evaluation

Highly conserved serine residue 40 in HIV-1 p6 regulates capsid processing and virus core assembly

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Solbak Sara MØ

2011-02-01

Full Text Available Abstract Background The HIV-1 p6 Gag protein regulates the final abscission step of nascent virions from the cell membrane by the action of two late assembly (L- domains. Although p6 is located within one of the most polymorphic regions of the HIV-1 gag gene, the 52 amino acid peptide binds at least to two cellular budding factors (Tsg101 and ALIX, is a substrate for phosphorylation, ubiquitination, and sumoylation, and mediates the incorporation of the HIV-1 accessory protein Vpr into viral particles. As expected, known functional domains mostly overlap with several conserved residues in p6. In this study, we investigated the importance of the highly conserved serine residue at position 40, which until now has not been assigned to any known function of p6. Results Consistently with previous data, we found that mutation of Ser-40 has no effect on ALIX mediated rescue of HIV-1 L-domain mutants. However, the only feasible S40F mutation that preserves the overlapping pol open reading frame (ORF reduces virus replication in T-cell lines and in human lymphocyte tissue cultivated ex vivo. Most intriguingly, L-domain mediated virus release is not dependent on the integrity of Ser-40. However, the S40F mutation significantly reduces the specific infectivity of released virions. Further, it was observed that mutation of Ser-40 selectively interferes with the cleavage between capsid (CA and the spacer peptide SP1 in Gag, without affecting cleavage of other Gag products. This deficiency in processing of CA, in consequence, led to an irregular morphology of the virus core and the formation of an electron dense extra core structure. Moreover, the defects induced by the S40F mutation in p6 can be rescued by the A1V mutation in SP1 that generally enhances processing of the CA-SP1 cleavage site. Conclusions Overall, these data support a so far unrecognized function of p6 mediated by Ser-40 that occurs independently of the L-domain function, but selectively

Common variation at 3q26.2, 6p21.33, 17p11.2 and 22q13.1 influences multiple myeloma risk

Science.gov (United States)

Broderick, Peter; Chen, Bowang; Johnson, David C; Försti, Asta; Vijayakrishnan, Jayaram; Migliorini, Gabriele; Dobbins, Sara E; Holroyd, Amy; Hose, Dirk; Walker, Brian A; Davies, Faith E; Gregory, Walter A; Jackson, Graham H; Irving, Julie A; Pratt, Guy; Fegan, Chris; Fenton, James AL; Neben, Kai; Hoffmann, Per; Nöthen, Markus M; Mühleisen, Thomas W; Eisele, Lewin; Ross, Fiona M; Straka, Christian; Einsele, Hermann; Langer, Christian; Dörner, Elisabeth; Allan, James M; Jauch, Anna; Morgan, Gareth J; Hemminki, Kari; Houlston, Richard S; Goldschmidt, Hartmut

2016-01-01

To identify variants for multiple myeloma risk, we conducted a genome-wide association study with validation in additional series totaling 4,692 cases and 10,990 controls. We identified four risk loci at 3q26.2 (rs10936599, P=8.70x10-14), 6p21.33 (rs2285803, PSORS1C2; P= 9.67x10-11), 17p11.2 (rs4273077, TNFRSF13B; P=7.67x10-9) and 22q13.1 (rs877529, CBX7; P=7.63x10-16). These data provide further evidence for genetic susceptibility to this B-cell hematological malignancy and insight into the biological basis of predisposition. PMID:23955597

Curcumin I mediates neuroprotective effect through attenuation of quinoprotein formation, p-p38 MAPK expression, and caspase-3 activation in 6-hydroxydopamine treated SH-SY5Y cells.