Effects of paraquat on Escherichia coli: Differences between B and K-12 strains

International Nuclear Information System (INIS)

Kitzler, J.W.; Minakami, H.; Fridovich, I.

1990-01-01

Escherichia coli B and K-12 are equally susceptible to the bacteriostatic effects of aerobic paraquat, but they differed strikingly when the lethality of paraquat was evaluated. E. coli B suffered an apparent loss of viability when briefly exposed to paraquat, whereas E. coli K-12 did not. This difference depended on the ability of the B-strain, but not the K-12 strain, to retain internalized paraquat; the B strain was killed on aerobic tryptic soy-yeast extract plates during the incubation which preceded the counting of colonies. This difference in retention of paraquat between strains was demonstrated by delayed loss of viability, by growth inhibition, and by cyanide-resistant respiration after brief exposure to paraquat, washing, and testing in fresh medium. This difference was also shown by using [ 14 C]paraquat. This previously unrecognized difference between E. coli B and K-12 has been the cause of apparently contradictory reports and should lead to some reevaluation of the pertinent literature

Evolution of Escherichia coli to 42 °C and Subsequent Genetic Engineering Reveals Adaptive Mechanisms and Novel Mutations

DEFF Research Database (Denmark)

Sandberg, Troy E.; Pedersen, Margit; LaCroix, Ryan A.

2014-01-01

Adaptive laboratory evolution (ALE) has emerged as a valuable method by which to investigate microbial adaptation to a desired environment. Here, we performed ALE to 42 °C of ten parallel populations of Escherichia coli K-12 MG1655 grown in glucose minimal media. Tightly controlled experimental c...... targets for additional ameliorating mutations. Overall, the results of this study provide insight into the adaptation process and yield lessons important for the future implementation of ALE as a tool for scientific research and engineering....

Draft Genome Sequence of Escherichia coli K-12 (ATCC 10798)

OpenAIRE

Dimitrova, Daniela; Engelbrecht, Kathleen C.; Putonti, Catherine; Koenig, David W.; Wolfe, Alan J.

2017-01-01

ABSTRACT Here, we present the draft genome sequence of Escherichia coli ATCC 10798. E.?coli ATCC 10798 is a K-12 strain, one of the most well-studied model microorganisms. The size of the genome was 4,685,496?bp, with a G+C content of 50.70%. This assembly consists of 62 contigs and the F plasmid.

The lon gene and photoprotection in Escherichia coli K-12

International Nuclear Information System (INIS)

Waksman, G.; Thomas, G.; Favre, A.

1984-01-01

Photoprotection, i.e. the increased resistance of the cells preilluminated with near ultraviolet light (300-380 nm) to the lethal action of 254nm radiations requires either an integrated prophage or a recA mutation in Escherichia coli K12 strains. Significant photoprotection occurs in an Escherichia coli K12 recA + cell containing the lon allele responsible for filamentous growth after 254nm irradiation. The Fil phenotype can be suppressed by the sfiA or sfiB suppressor genes. Since the E. coli K12 recA + lon sfiB strain exhibits no more photoprotection, it is concluded that in lon strains photoprotection is due to the abolition of the 254nm induced filamentation by the near ultraviolet treatment. In addition, near ultraviolet illumination of the cells leads to a severe restriction of the bulk protein synthesis. This effect is observed only in nuv + cells that contain 4-thiouridine the chromophore responsible for photoprotection. It is proposed that in lon (lysogenic strains) photoprotection is due to prevention of the SOS response. During the growth lag, the low residual level of protein synthesis does not allow the induction of the SOS response and accordingly prevents filamentation (the lytic cycle). (author)

Draft Genome Sequence of Escherichia coli K-12 (ATCC 10798).

Science.gov (United States)

Dimitrova, Daniela; Engelbrecht, Kathleen C; Putonti, Catherine; Koenig, David W; Wolfe, Alan J

2017-07-06

Here, we present the draft genome sequence of Escherichia coli ATCC 10798. E. coli ATCC 10798 is a K-12 strain, one of the most well-studied model microorganisms. The size of the genome was 4,685,496 bp, with a G+C content of 50.70%. This assembly consists of 62 contigs and the F plasmid. Copyright © 2017 Dimitrova et al.

Small noncoding RNA GcvB is a novel regulator of acid resistance in Escherichia coli

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Jin Ye

2009-04-01

Full Text Available Abstract Background The low pH environment of the human stomach is lethal for most microorganisms; but not Escherichia coli, which can tolerate extreme acid stress. Acid resistance in E. coli is hierarchically controlled by numerous regulators among which are small noncoding RNAs (sncRNA. Results In this study, we individually deleted seventy-nine sncRNA genes from the E. coli K12-MG1655 chromosome, and established a single-sncRNA gene knockout library. By systematically screening the sncRNA mutant library, we show that the sncRNA GcvB is a novel regulator of acid resistance in E. coli. We demonstrate that GcvB enhances the ability of E. coli to survive low pH by upregulating the levels of the alternate sigma factor RpoS. Conclusion GcvB positively regulates acid resistance by affecting RpoS expression. These data advance our understanding of the sncRNA regulatory network involved in modulating acid resistance in E. coli.

Escherichia coli K-12 pathogenicity in the pea aphid, Acyrthosiphon pisum, reveals reduced antibacterial defense in aphids.

Science.gov (United States)

Altincicek, Boran; Ter Braak, Bas; Laughton, Alice M; Udekwu, Klas I; Gerardo, Nicole M

2011-10-01

To better understand the molecular basis underlying aphid immune tolerance to beneficial bacteria and immune defense to pathogenic bacteria, we characterized how the pea aphid Acyrthosiphon pisum responds to Escherichia coli K-12 infections. E. coli bacteria, usually cleared in the hemolymph of other insect species, were capable of growing exponentially and killing aphids within a few days. Red fluorescence protein expressing E. coli K-12 laboratory strain multiplied in the aphid hemolymph as well as in the digestive tract, resulting in death of infected aphids. Selected gene deletion mutants of the E. coli K-12 predicted to have reduced virulence during systemic infections showed no difference in either replication or killing rate when compared to the wild type E. coli strain. Of note, however, the XL1-Blue E. coli K-12 strain exhibited a significant lag phase before multiplying and killing aphids. This bacterial strain has recently been shown to be more sensitive to oxidative stress than other E. coli K-12 strains, revealing a potential role for reactive oxygen species-mediated defenses in the otherwise reduced aphid immune system. Copyright © 2011 Elsevier Ltd. All rights reserved.

EchoBASE: an integrated post-genomic database for Escherichia coli.

Science.gov (United States)

Misra, Raju V; Horler, Richard S P; Reindl, Wolfgang; Goryanin, Igor I; Thomas, Gavin H

2005-01-01

EchoBASE (http://www.ecoli-york.org) is a relational database designed to contain and manipulate information from post-genomic experiments using the model bacterium Escherichia coli K-12. Its aim is to collate information from a wide range of sources to provide clues to the functions of the approximately 1500 gene products that have no confirmed cellular function. The database is built on an enhanced annotation of the updated genome sequence of strain MG1655 and the association of experimental data with the E.coli genes and their products. Experiments that can be held within EchoBASE include proteomics studies, microarray data, protein-protein interaction data, structural data and bioinformatics studies. EchoBASE also contains annotated information on 'orphan' enzyme activities from this microbe to aid characterization of the proteins that catalyse these elusive biochemical reactions.

Cloning and expression of the Escherichia coli K-12 sad gene.

OpenAIRE

Marek, L E; Henson, J M

1988-01-01

The Escherichia coli K-12 sad gene, which encodes an NAD-dependent succinic semialdehyde dehydrogenase, was cloned into a high-copy-number vector. Minicells carrying a sad+ plasmid produced a 55,000-dalton peptide, the probable sad gene product.

The role of Cra in regulating acetate excretion and osmotic tolerance in E. coli K-12 and E. coli B at high density growth.

Science.gov (United States)

Son, Young-Jin; Phue, Je-Nie; Trinh, Loc B; Lee, Sang Jun; Shiloach, Joseph

2011-06-30

E. coli B (BL21), unlike E.coli K-12 (JM109) is insensitive to glucose concentration and, therefore, grows faster and produces less acetate than E. coli K-12, especially when growing to high cell densities at high glucose concentration. By performing genomic analysis, it was demonstrated that the cause of this difference in sensitivity to the glucose concentration is the result of the differences in the central carbon metabolism activity. We hypothesized that the global transcription regulator Cra (FruR) is constitutively expressed in E. coli B and may be responsible for the different behaviour of the two strains. To investigate this possibility and better understand the function of Cra in the two strains, cra - negative E. coli B (BL21) and E. coli K-12 (JM109) were prepared and their growth behaviour and gene expression at high glucose were evaluated using microarray and real-time PCR. The deletion of the cra gene in E. coli B (BL21) minimally affected the growth and maximal acetate accumulation, while the deletion of the same gene in E.coli K-12 (JM109) caused the cells to stop growing as soon as acetate concentration reached 6.6 g/L and the media conductivity reached 21 mS/cm. ppsA (gluconeogenesis gene), aceBA (the glyoxylate shunt genes) and poxB (the acetate producing gene) were down-regulated in both strains, while acs (acetate uptake gene) was down-regulated only in E.coli B (BL21). These transcriptional differences had little effect on acetate and pyruvate production. Additionally, it was found that the lower growth of E. coli K-12 (JM109) strain was the result of transcription inhibition of the osmoprotectant producing bet operon (betABT). The transcriptional changes caused by the deletion of cra gene did not affect the activity of the central carbon metabolism, suggesting that Cra does not act alone; rather it interacts with other pleiotropic regulators to create a network of metabolic effects. An unexpected outcome of this work is the finding that cra

GenoBase: comprehensive resource database of Escherichia coli K-12.

Science.gov (United States)

Otsuka, Yuta; Muto, Ai; Takeuchi, Rikiya; Okada, Chihiro; Ishikawa, Motokazu; Nakamura, Koichiro; Yamamoto, Natsuko; Dose, Hitomi; Nakahigashi, Kenji; Tanishima, Shigeki; Suharnan, Sivasundaram; Nomura, Wataru; Nakayashiki, Toru; Aref, Walid G; Bochner, Barry R; Conway, Tyrrell; Gribskov, Michael; Kihara, Daisuke; Rudd, Kenneth E; Tohsato, Yukako; Wanner, Barry L; Mori, Hirotada

2015-01-01

Comprehensive experimental resources, such as ORFeome clone libraries and deletion mutant collections, are fundamental tools for elucidation of gene function. Data sets by omics analysis using these resources provide key information for functional analysis, modeling and simulation both in individual and systematic approaches. With the long-term goal of complete understanding of a cell, we have over the past decade created a variety of clone and mutant sets for functional genomics studies of Escherichia coli K-12. We have made these experimental resources freely available to the academic community worldwide. Accordingly, these resources have now been used in numerous investigations of a multitude of cell processes. Quality control is extremely important for evaluating results generated by these resources. Because the annotation has been changed since 2005, which we originally used for the construction, we have updated these genomic resources accordingly. Here, we describe GenoBase (http://ecoli.naist.jp/GB/), which contains key information about comprehensive experimental resources of E. coli K-12, their quality control and several omics data sets generated using these resources. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

Metabolic engineering of the Stevia rebaudiana ent-kaurene biosynthetic pathway in recombinant Escherichia coli.

Science.gov (United States)

Kong, Min Kyung; Kang, Hyun-Jun; Kim, Jin Ho; Oh, Soon Hwan; Lee, Pyung Cheon

2015-11-20

The ent-kaurene is a dedicated precursor pool and is responsible for synthesizing natural sweeteners such as steviol glycosides. In this study, to produce ent-kaurene in Escherichia coli, we modularly constructed and expressed two ent-kaurene genes encoding ent-copalyl diphosphate synthase (CPPS) and ent-kaurene synthase (KS) from Stevia rebaudiana known as a typical plant producing steviol glycoside. The CPPS and KS from S. rebaudiana were functionally expressed in a heterologous host E. coli. Furthermore, in order to enhance ent-kaurene production in E. coli, six geranylgeranyl diphosphate synthases (GGPPS) from various microorganisms and eight strains of E. coli as host were compared by measuring ent-kaurene production. The highest ent-kaurene production of approximately 41.1mg/L was demonstrated in E. coli strain MG1655 co-expressing synthetic CPPS-KS module and GGPPS from Rhodobacter sphaeroides. The ent-kaurene production was further increased up to 179.6 mg/L by overexpression of the three key enzymes for isoprenoid precursor, 1-deoxyxylulose-5-phosphate synthase (DXS), farnesyl diphosphate synthase (IspA) and isopentenyl diphosphate isomerase (IDI) from E. coli. Finally, the highest titer of ent-kaurene (578 mg/L) with a specific yield of ent-kaurene of 143.5mg/g dry cell weight was obtained by culturing E. coli strain MG1655 co-expressing the ent-kaurene module, DXS, IDI and IspA in 1L bioreactor containing 20 g/L glycerol. Copyright © 2015 Elsevier B.V. All rights reserved.

Role of SbmA in the Uptake of Peptide Nucleic Acid (PNA)-Peptide Conjugates in E. coli

DEFF Research Database (Denmark)

Ghosal, Anubrata; Vitali, Ally; Stach, James E M

2013-01-01

Antisense PNA oligomers targeting essential genes (acpP or ftsZ) and conjugated to the delivery peptide L((KFF)(3)K) show complete growth inhibition of wild type E. coli strain (MG1655) with submicromolar MIC. In this study we show that resistant mutants generated against such PNA......-peptide conjugates had disruptions in the region of sbmA, a gene encoding an inner membrane peptide transporter. The wild type sensitivity to the PNA conjugates was re-established in the resistance mutants by complementation with sbmA. Furthermore, deletion of sbmA in E. coli AS19, a strain that is sensitive...

4-thiouridine and photoprotection in Escherichia coli K12

International Nuclear Information System (INIS)

Thomas, Gilles; Favre, Alain

1977-01-01

A high level of protection is observed in the Escherichia coli K 12 strain AB 1157 rec A 1 nuv + whose transfer RNA contains 4-thiouridine. In contrast, the photoprotection level is low and observed at higher doses in a strain which differs from the former by a single mutation nuv - , (lack of 4-thiouridine). This nucleoside is therefore an important chromophore leading to photoprotection. This conclusion is corroborated by the similarity of the action spectra for 8-13 link formation in tRNA and for photoprotection [fr

Iodo-gen-catalysed iodination for identification of surface-exposed outer membrane proteins of Escherichia coli K12

International Nuclear Information System (INIS)

Ferreira, L.C.S.; Almeida, D.F. de

1987-01-01

Surface proteins of Escherichia coli K12 were identified by radiolabelling using 1,3,4,6 - tatrachloro, 3-alpha, 6-alpha - diphenylgycoluryl (Iodo-Gen) and 131 I. Labelled proteins were localized in the outer membrane of the cells. Using this technique it has been possible to observe technique it has been possible to observe that the eletrophoretic pattern of surface proteins changes according to the growth phases in culture. Radiolabelling of E.coli cells inculbated at 42 0 C showed that the syntheses of two surface proteins were temperature-inducible. At least one such protein may be involved in the process of cell division in E.coli K12. (author) [pt

Iodo-gen-catalysed iodination for identification of surface-exposed outer membrane proteins of Escherichia coli K12

Energy Technology Data Exchange (ETDEWEB)

Ferreira, L C.S.; Almeida, D.F. de

1987-12-01

Surface proteins of Escherichia coli K12 were identified by radiolabelling using 1,3,4,6 - tatrachloro, 3-alpha, 6-alpha - diphenylgycoluryl (Iodo-Gen) and /sup 131/I. Labelled proteins were localized in the outer membrane of the cells. Using this technique it has been possible to observe technique it has been possible to observe that the eletrophoretic pattern of surface proteins changes according to the growth phases in culture. Radiolabelling of E.coli cells inculbated at 42/sup 0/C showed that the syntheses of two surface proteins were temperature-inducible. At least one such protein may be involved in the process of cell division in E.coli K12.

Respiration shutoff in Escherichia coli K12 strains is induced by far ultraviolet radiations and by mitomycin C

International Nuclear Information System (INIS)

Swenson, P.A.; Norton, I.L.

1984-01-01

Near ultraviolet radiations (UV) cause respiration to shutoff in Escherichia coli B/r. It has been reported that E. coli K12 strains do not shut off respiration after UV. It is also reported that mitomycin C did not cause this 'SOS' response. In this paper it is reported that higher UV fluences than were previously used will cause respiration shutoff in K12 strain W3110 and that cyclic AMP increases the sensitivity of respiration shutoff of irradiated cell suspensions. Also mitomycin C shuts off respiration in this strain. Neither UV nor mitomycin C causes respiration shutoff in the recA56 derivative of W3110. Thus respiration shutoff is a recA dependent response to UV and mitomycin C in E. coli K12 strains. (Auth.)

Correlation of radiation sensitivity and nitrofurantoin sensitivity of Escherichia coli K-12

International Nuclear Information System (INIS)

Kloeck, K.

1981-01-01

The Uvr- and rec-mutants of E.coli K-12 have been tested with a view to their radiation- and nitrofuration sensitivity. The tests showed that all mutants tested were more radiation- and NF-sensitive than the wild type AB 1157. When the NF-sensitivity had been compared to the UV- and X-ray sensitivity it became obvious that the NF-sensitivity is correlated to the UV-sensitivity. Studies carried out with regard to the time dependence of the NF-effect on E.coli showed that the effect of NF on E. Coli became weaker after about 1 1/2 to 2 hours. That is possibly caused by the fact that the E. coli bacteria succeed in reducing the NF to an inactive form. By means of nitrosoguanidine mutants of E-coli AB 1157 had been induced and by means of the Replicite Plating Method, NF-sensible mutants had been isolated from the plutonium mixture. Among the mutants which had been isolated by this method, 74% had been more UV-sensitive than the wild type and 55% more X-ray sensitive. Thus NF-sensitive mutants have not necessarily to be considered as rec-mutants as there are also uvr-mutants in the mixture. (orig.) [de

Role of bolA and rpoS genes in biofilm formation and adherence pattern by Escherichia coli K-12 MG1655 on polypropylene, stainless steel, and silicone surfaces.

Science.gov (United States)

Adnan, Mohd; Sousa, Ana Margarida; Machado, Idalina; Pereira, Maria Olivia; Khan, Saif; Morton, Glyn; Hadi, Sibte

2017-06-01

Escherichia coli has developed sophisticated means to sense, respond, and adapt in stressed environment. It has served as a model organism for studies in molecular genetics and physiology since the 1960s. Stress response genes are induced whenever a cell needs to adapt and survive under unfavorable growth conditions. Two of the possible important genes are rpoS and bolA. The rpoS gene has been known as the alternative sigma (σ) factor, which controls the expression of a large number of genes, which are involved in responses to various stress factors as well as transition to stationary phase from exponential form of growth. Morphogene bolA response to stressed environment leads to round morphology of E. coli cells, but little is known about its involvement in biofilms and its development or maintenance. This study has been undertaken to address the adherence pattern and formation of biofilms by E. coli on stainless steel, polypropylene, and silicone surfaces after 24 h of growth at 37 °C. Scanning electron microscopy was used for direct examination of the cell attachment and biofilm formation on various surfaces and it was found that, in the presence of bolA, E. coli cells were able to attach to the stainless steel and silicone very well. By contrast, polypropylene surface was not found to be attractive for E. coli cells. This indicates that bolA responded and can play a major role in the presence and absence of rpoS in cell attachment.

Emergence of hyper-resistant Escherichia coli MG1655 derivative strains after applying sub-inhibitory doses of individual constituents of essential oils

Directory of Open Access Journals (Sweden)

Beatriz eChueca

2016-03-01

Full Text Available The improvement of food preservation by using essential oils (EOs and their individual constituents (ICs is attracting enormous interest worldwide. Until now, researchers considered that treatments with such antimicrobial compounds did not induce bacterial resistance via a phenotypic (i.e. transient response. Nevertheless, the emergence of genotypic (i.e. stable resistance after treatment with these compounds had not been previously tested. Our results confirm that growth of Escherichia coli MG1655 in presence of sub-inhibitory concentrations of the ICs carvacrol, citral, and (+-limonene oxide do not increase resistance to further treatments with either the same IC (direct resistance or with other preservation treatments (cross-resistance such as heat or pulsed electric fields (PEF. Bacterial mutation frequency was likewise lower when those IC’s were applied; however, after 10 days of re-culturing cells in presence of sub-inhibitory concentrations of the ICs, we were able to isolate several derivative strains (i.e. mutants displaying an increased minimum inhibitory concentration to those ICs. Furthermore, when compared to the wild type (WT strain, they also displayed direct resistance and cross-resistance. Derivative strains selected with carvacrol and citral also displayed morphological changes involving filamentation along with cell counts at late-stationary growth phase that were lower than the WT strain. In addition, co-cultures of each derivative strain with the WT strain resulted in a predominance of the original strain in absence of ICs, indicating that mutants would not out-compete WT cells under optimal growth conditions. Nevertheless, growth in the presence of ICs facilitated the selection of these resistant mutants. Thus, as a result, subsequent food preservation treatments of these bacterial cultures might be less effective than expected for WT cultures. In conclusion, this study recommends that treatment with ICs at sub

Emergence of Hyper-Resistant Escherichia coli MG1655 Derivative Strains after Applying Sub-Inhibitory Doses of Individual Constituents of Essential Oils.

Science.gov (United States)

Chueca, Beatriz; Berdejo, Daniel; Gomes-Neto, Nelson J; Pagán, Rafael; García-Gonzalo, Diego

2016-01-01

The improvement of food preservation by using essential oils (EOs) and their individual constituents (ICs) is attracting enormous interest worldwide. Until now, researchers considered that treatments with such antimicrobial compounds did not induce bacterial resistance via a phenotypic (i.e., transient) response. Nevertheless, the emergence of genotypic (i.e., stable) resistance after treatment with these compounds had not been previously tested. Our results confirm that growth of Escherichia coli MG1655 in presence of sub-inhibitory concentrations of the ICs carvacrol, citral, and (+)-limonene oxide do not increase resistance to further treatments with either the same IC (direct resistance) or with other preservation treatments (cross-resistance) such as heat or pulsed electric fields (PEF). Bacterial mutation frequency was likewise lower when those IC's were applied; however, after 10 days of re-culturing cells in presence of sub-inhibitory concentrations of the ICs, we were able to isolate several derivative strains (i.e., mutants) displaying an increased minimum inhibitory concentration to those ICs. Furthermore, when compared to the wild type (WT) strain, they also displayed direct resistance and cross-resistance. Derivative strains selected with carvacrol and citral also displayed morphological changes involving filamentation along with cell counts at late-stationary growth phase that were lower than the WT strain. In addition, co-cultures of each derivative strain with the WT strain resulted in a predominance of the original strain in absence of ICs, indicating that mutants would not out-compete WT cells under optimal growth conditions. Nevertheless, growth in the presence of ICs facilitated the selection of these resistant mutants. Thus, as a result, subsequent food preservation treatments of these bacterial cultures might be less effective than expected for WT cultures. In conclusion, this study recommends that treatment with ICs at sub

Phleomycin-induced lethality and DNA degradation in Escherichia coli K12

Energy Technology Data Exchange (ETDEWEB)

Nakayama, H

1975-01-01

The cell lethality and DNA fragmentation caused by phleomycin (PM) were studied in E. coli K12 strains with special reference to the effects of repair or recombination deficiencies and metabolic inhibitors. Unlike excision-defective derivatives of E. coli B, uvrA, uvrB, and uvrC mutants of strain K12 showed no peculiarities compared with wild type in regard to cell survival. Likewise, mutant alleles at uvrD and polA loci had no effect. In contrast, rec mutants were more sensitive to PM-killing than were rec/sup +/ strains. PM-induced strand breakage in DNA was observed in all strains tested including the above-mentioned mutants. There was no significant distinction between the uvr mutants and the wild type strain, indicating that the uvr-endonuclease was not responsible for the strand breaks. Involvement of endonuclease I was also ruled out. At least some of the PM-induced strand breaks were repairable. PM-induced lethality and strand breakage were totally dependent on energy supply. Inhibition of protein synthesis resulted in a partial and parallel suppression of the two effects. Our results suggest that the lethality is due to DNA strand breakage and the repair of such damage is postulated to be controlled by rec genes.

Radiation inactivation of Salmonella panama and Escherichia coli K 12 present on deep-frozen broiler carcasses

International Nuclear Information System (INIS)

Mulder, R.W.A.W.

1976-01-01

Low doses of ionizing radiation have been used to extend the shelf life of refrigerated poultry carcasses and to reduce the numbers of Salmonellae present. This report gives results of experiments on irradiation of deep-frozen poultry carcasses which were, before freezing, artificially contaminated with Salmonella panama and with a nalidixic acid-resistant Escherichia coli K 12. The D-values (decimal reduction) obtained with the inoculated carcasses were compared with D-values obtained with carcasses which were slaughtered in the normal way. The D-values for S.panama and for E.coli K 12 were 64.9 krad and 55.9 krad in the dripwater. Under commercial conditions approximately 100 krad were required for one decimal reduction of the Enterobacteriaceae present. The D-values estimated on the skin were higher for S.panama than for E.coli K 12 (128.6 krad vs 57.6 krad). If it is assumed that 1 positive carcass in 10,000 is allowed, the deep-frozen carcasses should be irradiated with doses of at least 700 krad to be sure of the absence of the tested S.panama strain. (orig.) [de

Radiation inactivation of Salmonella panama and Escherichia coli K 12 present on deep-frozen broiler carcasses

Energy Technology Data Exchange (ETDEWEB)

Mulder, R W.A.W. [Spelderholt Inst. for Poultry Research, Beekbergen (Netherlands). Processing Dept.

1976-01-01

Low doses of ionizing radiation have been used to extend the shelf life of refrigerated poultry carcasses and to reduce the numbers of Salmonellae present. This report gives results of experiments on irradiation of deep-frozen poultry carcasses which were, before freezing, artificially contaminated with Salmonella panama and with a nalidixic acid-resistant Escherichia coli K 12. The D-values (decimal reduction) obtained with the inoculated carcasses were compared with D-values obtained with carcasses which were slaughtered in the normal way. The D-values for S.panama and for E.coli K 12 were 64.9 krad and 55.9 krad in the dripwater. Under commercial conditions approximately 100 krad were required for one decimal reduction of the Enterobacteriaceae present. The D-values estimated on the skin were higher for S.panama than for E.coli K 12 (128.6 krad vs 57.6 krad). If it is assumed that 1 positive carcass in 10,000 is allowed, the deep-frozen carcasses should be irradiated with doses of at least 700 krad to be sure of the absence of the tested S.panama strain.

The EcoCyc database: reflecting new knowledge about Escherichia coli K-12.

Science.gov (United States)

Keseler, Ingrid M; Mackie, Amanda; Santos-Zavaleta, Alberto; Billington, Richard; Bonavides-Martínez, César; Caspi, Ron; Fulcher, Carol; Gama-Castro, Socorro; Kothari, Anamika; Krummenacker, Markus; Latendresse, Mario; Muñiz-Rascado, Luis; Ong, Quang; Paley, Suzanne; Peralta-Gil, Martin; Subhraveti, Pallavi; Velázquez-Ramírez, David A; Weaver, Daniel; Collado-Vides, Julio; Paulsen, Ian; Karp, Peter D

2017-01-04

EcoCyc (EcoCyc.org) is a freely accessible, comprehensive database that collects and summarizes experimental data for Escherichia coli K-12, the best-studied bacterial model organism. New experimental discoveries about gene products, their function and regulation, new metabolic pathways, enzymes and cofactors are regularly added to EcoCyc. New SmartTable tools allow users to browse collections of related EcoCyc content. SmartTables can also serve as repositories for user- or curator-generated lists. EcoCyc now supports running and modifying E. coli metabolic models directly on the EcoCyc website. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

Restriction alleviation of phage λ in Escherichia Coli K-12 cells after γ-irradiation

International Nuclear Information System (INIS)

Rabinkova, E.V.; Torosyan, M.V.; Fradkin, G.E.

1987-01-01

In γ-irradiated cells of Escherichia coli K-12 restriction allevation of an unmodified phage λ is only observed in AB1157 strain. No restriction allevation by γ-rays is registered in AB1157 mutants (rec A and ssb-1)

Environmental and genetic factors affecting mutability to aminoglycoside antibiotics among Escherichia coli K12 strains

Directory of Open Access Journals (Sweden)

Monteiro A.C.M.

2003-01-01

Full Text Available Environmental and genetic factors affecting the in vitro spontaneous mutation frequencies to aminoglycoside resistance in Escherichia coli K12 were investigated. Spontaneous mutation frequencies to kanamycin resistance were at least 100 fold higher on modified Luria agar (L2 plates, when compared to results obtained in experiments carried out with Nutrient agar (NA plates. In contrast to rifampincin, the increased mutability to kanamycin resistance could not be attributed to a mutator phenotype expressed by DNA repair defective strains. Kanamycin mutant selection windows and mutant preventive concentrations on L2 plates were at least fourfold higher than on NA plates, further demonstrating the role of growth medium composition on the mutability to aminoglycosides. Mutability to kanamycin resistance was increased following addition of sorbitol, suggesting that osmolarity is involved on the spontaneous mutability of E. coli K12 strains to aminoglycosides. The spontaneous mutation rates to kanamycin resistance on both L2 and NA plates were strictly associated with the selective antibiotic concentrations. Moreover, mutants selected at different antibiotic concentrations expressed heterogeneous resistance levels to kanamycin and most of them expressing multiple resistance to all tested aminoglycoside antibiotics (gentamicin, neomycin, amykacin and tobramycin. These results will contribute to a better understanding of the complex nature of aminoglycoside resistance and the emergence of spontaneous resistant mutants among E. coli K12 strains.

Global impact of mature biofilm lifestyle on Escherichia coli K-12 gene expression

DEFF Research Database (Denmark)

Beloin, C.; Valle, J.; Latour-Lambert, P.

2004-01-01

The formation of biofilm results in a major lifestyle switch that is thought to affect the expression of multiple genes and operons. We used DNA arrays to study the global effect of biofilm formation on gene expression in mature Escherichia coli K-12 biofilm. We show that, when biofilm is compared...... that 20 of these genes are required for the formation of mature biofilm. This group includes 11 genes of previously unknown function. These results constitute a comprehensive analysis of the global transcriptional response triggered in mature E. coli biofilms and provide insights into its physiological...

Ultraviolet radiation-induced mutability of isogenic uvrA and uvrB strains of Escherichia coli K-12 W3110

International Nuclear Information System (INIS)

Barfknecht, T.R.; Smith, K.C.

1977-01-01

E. coli K-12 W3110 uvrB5 strain has been shown to have a higher UV induced reversion frequency than its wild-type parent when plotted on the basis of mutation frequency versus survival. However for the E. coli B/r WP2s uvrA strain this higher mutability has been observed only at survival levels of 80-100%. A study was undertaken to determine if these differences in UV mutability were due primarily to the uvrA and uvrB mutations, or to other genetic background differences. Isogenic strains of E. coli K-12 W3110 carrying uvrA6, uvrB5, uvrA6 and uvrB5, and the uvrA allele from E.coli B/r WP2s were used. Results indicate that the enrichment of minimal medium with a small amount of nutrient broth is sufficient to inhibit minimal medium recovery (MMR) and to enhance leu + reversion of the leu B missense mutation in these uvr - strains. This suggests that there may be a relationship between MMR and error-free postreplication repair. Further research is in progress to clarify the relationship between MMR and broth enhancement of UV-induced mutagenesis in uvr - strains of E. Coli K-12 W3110. (author)

Ultraviolet radiation-induced mutability of isogenic uvrA and uvrB strains of Escherichia coli K-12 W3110

Energy Technology Data Exchange (ETDEWEB)

Barfknecht, T R; Smith, K C [Stanford Univ., Calif. (USA). Dept. of Radiology

1977-12-01

Escherichia coli K-12 W3110 uvrB5 strain has been shown to have a higher uv-induced reversion frequency than its wild-type parent when plotted on the basis of mutation frequency versus survival. However for the E. coli B/r WP2s uvrA strain this higher mutability has been observed only at survival levels of 80 to 100%. A study was undertaken to determine if ly to the uvrA and uvrB mutations, or to other genetic background differences. Isogenic strains of E. coli K-12 W3110 carrying uvrA6, uvrB5, uvrA6, and uvrB5, and the uvrA allele from E.coli B/r WP2s were used. Results indicate that the enrichment of minimal medium with a small amount of nutrient broth is sufficient to inhibit minimal medium recovery (MMR) and to enhance leu/sup +/ reversion of the leu B missense mutation in these uvr/sup -/ strains. This suggests that there may be a relationship between MMR and error-free postreplication repair. Further research is in progress to clarify the relationship between MMR and broth enhancement of uv-induced mutagenesis in uvr/sup -/ strains of E. Coli K-12 W3110.

Fulltext PDF

Indian Academy of Sciences (India)

2014-07-01

Jul 1, 2014 ... RpoS is a stationary-phase sigma factor which is required for the expression of several ... that happened in laboratory strains over the last six decades in retrospect. Representatives of the two E. coli strains K12 and B ... W3110 but not MG1655 has six nonfunctional genes, namely, rpoS, dcuA, rcsC, gatA, ...

Graphene-Based FET Detector for E. coli K12 Real-Time Monitoring and Its Theoretical Analysis

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Jieyi Zhu

2016-01-01

Full Text Available This paper presents a theoretical analysis for a graphene-based FET real-time detector of the target bacteria E. coli K12. The motivation for this study is to design a sensor device for detection of bacteria in food and water in order to guarantee food safety. Graphene is chosen as our material for sensor design, which has outstanding electrical, physical, and optical performance. In our sensor structure, graphene-based solution gate field effect transistor (FET is the device model; fabrication and functionalization protocol are presented together in this paper. What is more, a real-time signal display system is the accompanied equipment for our designed biosensor device. In this system, the sensor bias current signal Ids would change obviously when the target bacteria are attached to the sensor surface. And the bias current Ids increases when the E. coli concentration increases. In the latter part, a theoretical interpretation of the sensor signal is to explain the bias current Ids increasing after the E. coli K12 attachment.

Associations of Escherichia coli K-12 OmpF trimers with rough and smooth lipopolysaccharides

International Nuclear Information System (INIS)

Diedrich, D.L.; Stein, M.A.; Schnaitman, C.A.

1990-01-01

The associations of both rough and smooth lipopolysaccharides (LPS) with the OmpF porin of Escherichia coli K-12 were examined in galE strains deleted for ompC. Transformation with pSS37 and growth with galactose conferred the ability to assemble a Shigella dysenteriae O antigen onto the core oligosaccharide of E. coli K-12 LPS. The association of LPS with OmpF trimers was assessed by staining, autoradiography of LPS specifically labeled with [1-14C]galactose, and Western immunoblotting with a monoclonal antibody specific for OmpF trimers. These techniques revealed that the migration distances and multiple banding patterns of OmpF porin trimers in sodium dodecyl sulfate-polyacrylamide gels were dictated by the chemotype of associated LPS. Expression of smooth LPS caused almost all of the trimeric OmpF to run in gels with a slower mobility than trimers from rough strains. The LPS associated with trimers from a smooth strain differed from the bulk-phase LPS by consisting almost exclusively of molecules with O antigen

Regulatory role of XynR (YagI) in catabolism of xylonate in Escherichia coli K-12.

Science.gov (United States)

Shimada, Tomohiro; Momiyama, Eri; Yamanaka, Yuki; Watanabe, Hiroki; Yamamoto, Kaneyoshi; Ishihama, Akira

2017-12-01

The genome of Escherichia coli K-12 contains ten cryptic phages, altogether constituting about 3.6% of the genome in sequence. Among more than 200 predicted genes in these cryptic phages, 14 putative transcription factor (TF) genes exist, but their regulatory functions remain unidentified. As an initial attempt to make a breakthrough for understanding the regulatory roles of cryptic phage-encoded TFs, we tried to identify the regulatory function of CP4-6 cryptic prophage-encoded YagI with unknown function. After SELEX screening, YagI was found to bind mainly at a single site within the spacer of bidirectional transcription units, yagA (encoding another uncharacterized TF) and yagEF (encoding 2-keto-3-deoxy gluconate aldolase, and dehydratase, respectively) within this prophage region. YagEF enzymes are involved in the catabolism of xylose downstream from xylonate. We then designated YagI as XynR (regulator of xylonate catabolism), one of the rare single-target TFs. In agreement with this predicted regulatory function, the activity of XynR was suggested to be controlled by xylonate. Even though low-affinity binding sites of XynR were identified in the E. coli K-12 genome, they all were inside open reading frames, implying that the regulation network of XynR is still fixed within the CR4-6 prophage without significant influence over the host E. coli K-12. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection.

Science.gov (United States)

Baba, Tomoya; Ara, Takeshi; Hasegawa, Miki; Takai, Yuki; Okumura, Yoshiko; Baba, Miki; Datsenko, Kirill A; Tomita, Masaru; Wanner, Barry L; Mori, Hirotada

2006-01-01

We have systematically made a set of precisely defined, single-gene deletions of all nonessential genes in Escherichia coli K-12. Open-reading frame coding regions were replaced with a kanamycin cassette flanked by FLP recognition target sites by using a one-step method for inactivation of chromosomal genes and primers designed to create in-frame deletions upon excision of the resistance cassette. Of 4288 genes targeted, mutants were obtained for 3985. To alleviate problems encountered in high-throughput studies, two independent mutants were saved for every deleted gene. These mutants-the 'Keio collection'-provide a new resource not only for systematic analyses of unknown gene functions and gene regulatory networks but also for genome-wide testing of mutational effects in a common strain background, E. coli K-12 BW25113. We were unable to disrupt 303 genes, including 37 of unknown function, which are candidates for essential genes. Distribution is being handled via GenoBase (http://ecoli.aist-nara.ac.jp/).

Physiological Function of Rac Prophage During Biofilm Formation and Regulation of Rac Excision in Escherichia coli K-12

Science.gov (United States)

including Escherichia coli, Salmonella spp. and Shigellaspp. Here, we found that rac excision is induced during biofilm formation, and the isogenic...stain without rac is more motile and forms more biofilms in nutrient-rich medium at early stages in E.coli K-12. Additionally, the presence of rac...genes increases cell lysis during biofilm development. In most E. coli strains, rac is integrated into the ttcA gene which encodes a tRNA-thioltransferase

Tif-stimulated deoxyribonucleic acid repair in Escherichia coli K-12

International Nuclear Information System (INIS)

Castellazzi, M.; Jacques, M.; George, J.

1980-01-01

Bacterial survival is significantly increased after ultraviolet irradiation in tif sfi cells, provided that the thermosensitive tif mutation has been expressed at 41 0 C before irradiation. This tif-mediated reactivation of ultraviolet irradiated bacteria needs de novo protein synthesis, as is the case for the tif-mediated reactivation of ultraviolet-irradiated phage lambda. However, in striking contrast to the phage reactivation process, this tif-mediated reactivation is no longer associated with mutagenesis. It also requires the presence of the uvrA + excision function. These results strongly suggest the existence in Escherichia coli K-12 of a repair pathway acting on bacterial deoxyribonucleic acid which is inducible, error free, and uvr dependent

Attachment behaviour of Escherichia coli K12 and Salmonella Typhimurium P6on food contact surfaces for food transportation

DEFF Research Database (Denmark)

Abban, Stephen; Jakobsen, Mogens; Jespersen, Lene

2012-01-01

The role of cargo container lining materials aluminium, a fibre reinforced plastic (FRP) and stainless steel in bacterial cross contamination during transport was assessed. For this, attachment and detachment of Escherichia coli K12 and Salmonella Typhimurium P6 on the three surfaces in the absence....... Typhimurium P6 respectively. Correlation with roughness average was poor; r = -0.425 and -0.413 respectively for E. coli K12 and S. Typhimurium P6. Presence of residue caused significant reduction (p ... material sections of the same surfaces. We report these observations for the first time for aluminium and the FRP material and in part for stainless steel. The S. Typhimurium P6 strain also had significantly higher level of attachment than the E. coli K12 strain. Our findings show that food residue...

Polyamine stress at high pH in Escherichia coli K-12

Directory of Open Access Journals (Sweden)

Tate Daniel P

2005-10-01

Full Text Available Abstract Background Polyamines such as spermine and spermidine are required for growth of Escherichia coli; they interact with nucleic acids, and they bind to ribosomes. Polyamines block porins and decrease membrane permeability, activities that may protect cells in acid. At high concentrations, however, polyamines impair growth. They impair growth more severely at high pH, probably due to their increased uptake as membrane-permeant weak bases. The role of pH is critical in understanding polyamine stress. Results The effect of polyamines was tested on survival of Escherichia coli K-12 W3110 in extreme acid or base (pH conditions outside the growth range. At pH 2, 10 mM spermine increased survival by 2-fold, and putrescine increased survival by 30%. At pH 9.8, however, E. coli survival was decreased 100-fold by 10 mM spermine, putrescine, cadaverine, or spermidine. At pH 8.5, spermine decreased the growth rate substantially, whereas little effect was seen at pH 5.5. Spermidine required ten-fold higher concentrations to impair growth. On proteomic 2-D gels, spermine and spermidine caused differential expression of 31 different proteins. During log-phase growth at pH 7.0, 1 mM spermine induced eight proteins, including PykF, GlpK, SerS, DeaD, OmpC and OmpF. Proteins repressed included acetate-inducible enzymes (YfiD, Pta, Lpd as well as RapA (HepA, and FabB. At pH 8.5, spermine induced additional proteins: TnaA, OmpA, YrdA and NanA (YhcJ and also repressed 17 proteins. Four of the proteins that spermine induced (GlpK, OmpA, OmpF, TnaA and five that were repressed (Lpd, Pta, SucB, TpiA, YfiD show similar induction or repression, respectively, in base compared to acid. Most of these base stress proteins were also regulated by spermidine, but only at ten-fold higher concentration (10 mM at high pH (pH 8.5. Conclusion Polyamines increase survival in extreme acid, but decrease E. coli survival in extreme base. Growth inhibition by spermine and

5 CFR 1655.14 - Loan payments.

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2010-01-01

... 5 Administrative Personnel 3 2010-01-01 2010-01-01 false Loan payments. 1655.14 Section 1655.14 Administrative Personnel FEDERAL RETIREMENT THRIFT INVESTMENT BOARD LOAN PROGRAM § 1655.14 Loan payments. (a) Loan payments must be made through payroll deduction in accordance with the loan agreement. Once loan...

Multiplex polymerase chain reaction for identification of Escherichia coli, Escherichia albertii and Escherichia fergusonii.

Science.gov (United States)

Lindsey, Rebecca L; Garcia-Toledo, L; Fasulo, D; Gladney, L M; Strockbine, N

2017-09-01

Escherichia coli, Escherichia albertii, and Escherichia fergusonii are closely related bacteria that can cause illness in humans, such as bacteremia, urinary tract infections and diarrhea. Current identification strategies for these three species vary in complexity and typically rely on the use of multiple phenotypic and genetic tests. To facilitate their rapid identification, we developed a multiplex PCR assay targeting conserved, species-specific genes. We used the Daydreamer™ (Pattern Genomics, USA) software platform to concurrently analyze whole genome sequence assemblies (WGS) from 150 Enterobacteriaceae genomes (107 E. coli, 5 Shigella spp., 21 E. albertii, 12 E. fergusonii and 5 other species) and design primers for the following species-specific regions: a 212bp region of the cyclic di-GMP regulator gene (cdgR, AW869_22935 from genome K-12 MG1655, CP014225) for E. coli/Shigella; a 393bp region of the DNA-binding transcriptional activator of cysteine biosynthesis gene (EAKF1_ch4033 from genome KF1, CP007025) for E. albertii; and a 575bp region of the palmitoleoyl-acyl carrier protein (ACP)-dependent acyltransferase (EFER_0790 from genome ATCC 35469, CU928158) for E. fergusonii. We incorporated the species-specific primers into a conventional multiplex PCR assay and assessed its performance with a collection of 97 Enterobacteriaceae strains. The assay was 100% sensitive and specific for detecting the expected species and offers a quick and accurate strategy for identifying E. coli, E. albertii, and E. fergusonii in either a single reaction or by in silico PCR with sequence assemblies. Published by Elsevier B.V.

5 CFR 1655.7 - Interest rate.

Science.gov (United States)

2010-01-01

... 5 Administrative Personnel 3 2010-01-01 2010-01-01 false Interest rate. 1655.7 Section 1655.7 Administrative Personnel FEDERAL RETIREMENT THRIFT INVESTMENT BOARD LOAN PROGRAM § 1655.7 Interest rate. (a... interest rate established by the Department of the Treasury in effect on the date the TSP record keeper...

The absence of the luxS gene increases swimming motility and flagella synthesis in Escherichia coli K12

International Nuclear Information System (INIS)

Ling, Hua; Kang, Aram; Tan, Mui Hua; Qi, Xiaobao; Chang, Matthew Wook

2010-01-01

Research highlights: → This paper provides the first evidence that luxS deletion enhances swimming motility and flagella synthesis in Escherichia coli K12 based on motility, transcriptome, and scanning electron microscopy analyses. → A conceptual genetic regulatory network underlying the increased flagella synthesis was constructed based on the transcriptome and network component analyses, and previously known regulatory relations. → The genetic regulatory network suggests that the increased flagella synthesis and motility might be contributed to by increased flhDC transcription level and/or decreased c-di-GMP concentration in luxS-deficient E. coli. -- Abstract: Despite the significant role of S-ribosylhomocysteinase (LuxS) in the activated methyl cycle pathway and quorum sensing, the connectivity between luxS and other cellular functions remains incomplete. Herein, we show that luxS deletion significantly increases swimming motility and flagella synthesis in Escherichia coli K12 using motility, transcriptome, and scanning electron microscopy assays. Further, based on the transcriptome and network component analyses, and known regulatory relations, we propose a conceptual genetic regulatory network underlying the increased flagella synthesis in response to luxS deletion.

The absence of the luxS gene increases swimming motility and flagella synthesis in Escherichia coli K12

Energy Technology Data Exchange (ETDEWEB)

Ling, Hua; Kang, Aram; Tan, Mui Hua; Qi, Xiaobao [School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 637459 (Singapore); Chang, Matthew Wook, E-mail: Matthewchang@ntu.edu.sg [School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 637459 (Singapore)

2010-10-29

Research highlights: {yields} This paper provides the first evidence that luxS deletion enhances swimming motility and flagella synthesis in Escherichia coli K12 based on motility, transcriptome, and scanning electron microscopy analyses. {yields} A conceptual genetic regulatory network underlying the increased flagella synthesis was constructed based on the transcriptome and network component analyses, and previously known regulatory relations. {yields} The genetic regulatory network suggests that the increased flagella synthesis and motility might be contributed to by increased flhDC transcription level and/or decreased c-di-GMP concentration in luxS-deficient E. coli. -- Abstract: Despite the significant role of S-ribosylhomocysteinase (LuxS) in the activated methyl cycle pathway and quorum sensing, the connectivity between luxS and other cellular functions remains incomplete. Herein, we show that luxS deletion significantly increases swimming motility and flagella synthesis in Escherichia coli K12 using motility, transcriptome, and scanning electron microscopy assays. Further, based on the transcriptome and network component analyses, and known regulatory relations, we propose a conceptual genetic regulatory network underlying the increased flagella synthesis in response to luxS deletion.

5 CFR 1655.21 - Loan fee.

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2010-01-01

... 5 Administrative Personnel 3 2010-01-01 2010-01-01 false Loan fee. 1655.21 Section 1655.21 Administrative Personnel FEDERAL RETIREMENT THRIFT INVESTMENT BOARD LOAN PROGRAM § 1655.21 Loan fee. The TSP will charge a participant a $50.00 loan fee when it disburses the loan and will deduct the fee from the...

A model‐driven quantitative metabolomics analysis of aerobic and anaerobic metabolism in E. coli K‐12 MG1655 that is biochemically and thermodynamically consistent

DEFF Research Database (Denmark)

McCloskey, Douglas; Gangoiti, Jon A.; King, Zachary A.

2014-01-01

in metabolomes between anaerobic and aerobic growth of Escherichia coli. Constraint‐based modeling was utilized to deduce a target list of compounds for downstream method development. An analytical and experimental methodology was developed and tailored to the compound chemistry and growth conditions of interest....... This included the construction of a rapid sampling apparatus for use with anaerobic cultures. The resulting genome‐scale data sets for anaerobic and aerobic growth were validated by comparison to previous small‐scale studies comparing growth of E. coli under the same conditions. The metabolomics data were......‐oxidation pathway for synthesis of fatty acids. This analysis also identified enzyme promiscuity for the pykA gene, that is critical for anaerobic growth, and which has not been previously incorporated into metabolic models of E coli. Biotechnol....

Nonthermal inactivation of Escherichia coli K12 in buffered peptone water using a pilot-plant scale supercritical carbon dioxide system with gas-liquid porous metal contractor

Science.gov (United States)

This study evaluated the effectiveness of a supercritical carbon dioxide (SCCO2) system, with a gas-liquid CO2 contactor, for reducing Escherichia coli K12 in diluted buffered peptone water. 0.1% (w/v) buffered peptone water inoculated with E. coli K12 was processed using the SCCO2 system at CO2 con...

DNA synthesis and uv resistance in Escherichia coli K12 cells

Energy Technology Data Exchange (ETDEWEB)

Slezarikova, V [Slovenska Akademia Vied, Bratislava (Czechoslovakia). Vyskumny Ustav Onkologicky

1976-01-01

The influence was studied of preirradiation inhibition of proteosynthesis by amino acids starvation on survival and DNA synthesis in E. coli K 12 cells, which differ by their genetic features with regard to a certain type of repair. The surviving fraction was studied by appropriate dilution of cell suspension and spreading on agar plates. DNA synthesis was investigated by the incorporation of thymine-2-/sup 14/C. In our conditions a correlation was found between cell survival and the resistance of DNA replication to UV radiation in cells proficient in excision and post-replication repair. This correlation was not found in the excision deficient strain. It is concluded that enhanced resistance of DNA replication is not a sufficient condition for enhanced cell resistance.

In Vitro Assembly of the Outer Core of the Lipopolysaccharide from Escherichia coli K-12 and Salmonella typhimurium

Science.gov (United States)

2015-01-01

There are five distinct core structures in the lipopolysaccharides of Escherichia coli and at least two in Salmonella isolates, which vary principally in the outer core oligosaccharide. Six outer core glycosyltransferases, E. coli K-12 WaaG, WaaB, and WaaO and Salmonella typhimurium WaaI, WaaJ, and WaaK, were cloned, overexpressed, and purified. A novel substrate for WaaG was isolated from ΔwaaG E. coli overexpressing the lipid A phosphatase lpxE and the lipid A late acyltransferase lpxM. The action of lpxE and lpxM in the ΔwaaG background yielded heptose2-1-dephospho Kdo2-lipid A, a 1-dephosphorylated hexa-acylated lipid A with the inner core sugars that is easily isolated by organic extraction. Using this structurally defined acceptor and commercially available sugar nucleotides, each outer core glycosyltransferases was assayed in vitro. We show that WaaG and WaaB add a glucose and galactose sequentially to heptose2-1-dephospho Kdo2-lipid A. E. coli K-12 WaaO and S. typhimurium WaaI add a galactose to the WaaG/WaaB product but can also add a galactose to the WaaG product directly without the branched core sugar added by WaaB. Both WaaI and WaaO require divalent metal ions for optimal activity; however, WaaO, unlike WaaI, can add several glucose residues to its lipid acceptor. Using the product of WaaG, WaaB, and WaaI, we show that S. typhimurium WaaJ and WaaK transfer a glucose and N-acetylglucosamine, respectively, to yield the full outer core. This is the first demonstration of the in vitro assembly of the outer core of the lipopolysaccharide using defined lipid A-oligosaccharide acceptors and sugar donors. PMID:24479701

Induction of Shiga toxin-converting prophage in Escherichia coli by high hydrostatic pressure.

Science.gov (United States)

Aertsen, Abram; Faster, David; Michiels, Chris W

2005-03-01

Since high hydrostatic pressure is becoming increasingly important in modern food preservation, its potential effects on microorganisms need to be thoroughly investigated. In this context, mild pressures (pressures. In this report, we extend this observation to lambdoid Shiga toxin (Stx)-converting bacteriophages in MG1655, which constitute an important virulence trait in Stx-producing E. coli strains (STEC). The window of pressures capable of inducing Stx phages correlated well with the window of bacterial survival. When pressure treatments were conducted in whole milk, which is known to promote bacterial survival, Stx phage induction could be observed at up to 250 MPa in E. coli MG1655 and at up to 300 MPa in a pressure-resistant mutant of this strain. In addition, we found that the intrinsic pressure resistance of two types of Stx phages was very different, with one type surviving relatively well treatments of up to 400 MPa for 15 min at 20 degrees C. Interestingly, and in contrast to UV irradiation or mitomycin C treatment, pressure was not able to induce Stx prophage or an SOS response in several natural Stx-producing STEC isolates.

Increased riboflavin production by knockout of 6-phosphofructokinase I and blocking the Entner-Doudoroff pathway in Escherichia coli.

Science.gov (United States)

Liu, Shuang; Kang, Pei; Cui, Zhenzhen; Wang, Zhiwen; Chen, Tao

2016-08-01

To construct an Escherichia coli strain capable of producing riboflavin with high titer and yield. A low copy number plasmid pLS01 containing a riboflavin operon under the control of a constitutive promoter was constructed and introduced into Escherichia coli MG1655. Subsequently, the pfkA, edd and ead genes were disrupted, and the resulting strain LS02T produced 667 mg riboflavin/l in MSY medium supplied with 10 g glucose/l in flask cultivation. In a fed-batch process, riboflavin production of the strain reached 10.4 g/l with a yield of 56.8 mg riboflavin/g glucose. To our knowledge, this is the first report of engineered E. coli strains that can produce more than 10 g riboflavin/l in fed-batch cultivation, indicating that E. coli has potential for riboflavin production.

Genetic basis of growth adaptation of Escherichia coli after deletion of pgi, a major metabolic gene.

Directory of Open Access Journals (Sweden)

Pep Charusanti

2010-11-01

Full Text Available Bacterial survival requires adaptation to different environmental perturbations such as exposure to antibiotics, changes in temperature or oxygen levels, DNA damage, and alternative nutrient sources. During adaptation, bacteria often develop beneficial mutations that confer increased fitness in the new environment. Adaptation to the loss of a major non-essential gene product that cripples growth, however, has not been studied at the whole-genome level. We investigated the ability of Escherichia coli K-12 MG1655 to overcome the loss of phosphoglucose isomerase (pgi by adaptively evolving ten replicates of E. coli lacking pgi for 50 days in glucose M9 minimal medium and by characterizing endpoint clones through whole-genome re-sequencing and phenotype profiling. We found that 1 the growth rates for all ten endpoint clones increased approximately 3-fold over the 50-day period; 2 two to five mutations arose during adaptation, most frequently in the NADH/NADPH transhydrogenases udhA and pntAB and in the stress-associated sigma factor rpoS; and 3 despite similar growth rates, at least three distinct endpoint phenotypes developed as defined by different rates of acetate and formate secretion. These results demonstrate that E. coli can adapt to the loss of a major metabolic gene product with only a handful of mutations and that adaptation can result in multiple, alternative phenotypes.

Comparison of hydrogen-production capability of four different Enterobacteriaceae strains under growing and non-growing conditions

Energy Technology Data Exchange (ETDEWEB)

Seol, Eunhee; Kim, Seohyoung; Raj, S. Mohan; Park, Sunghoon [Department of Chemical and Biochemical Engineering, Pusan National University, Busan 609-735 (Korea)

2008-10-15

Non-growing cells can function as whole-cell biocatalysts for hydrogen (H{sub 2}) production, a process that has recently drawn much attention. In order to evaluate their potential as whole-cell biocatalysts, we compared the H{sub 2}-production capability of four Enterobacteriaceae strains (Citrobacter amalonaticus Y19, Escherichia coli K-12 MG1655, Escherichia coli DJT135, and Enterobacter aerogenes) under growing and non-growing conditions. We evaluated their H{sub 2}-production activity at varying temperatures (25-45 C) and pH conditions (6.0-8.0) using glucose or formate as the carbon source. Under growing conditions with 10 mM glucose as a substrate, E. aerogenes exhibited the highest H{sub 2}-production activity (17.0 {+-} 0.2 {mu}mol H{sub 2} mg cell{sup -1} h{sup -1}) among the four strains, but the final H{sub 2} yield was similar (1.7-1.8 mol H{sub 2} mol{sup -1} glucose) in all four strains. H{sub 2} production in the four strains proceeded through a formate-dependent pathway that involved the formate hydrogen lyase (FHL) complex. Under non-growing conditions with 20 mM formate as a substrate, we obtained high H{sub 2}-production activities, in the range of 95.5-195.2 {mu}mol H{sub 2} mg cell{sup -1} h{sup -1}, with E. coli DJT135 exhibiting the highest activity (195.2 {mu}mol H{sub 2} mg{sup -1} h{sup -1}) at pH 6.0 and 45 C. In contrast, using glucose as the carbon substrate in non-growing cell experiments greatly reduced the H{sub 2}-production activity to 6.1-7.7 {mu}mol H{sub 2} mg cell{sup -1} h{sup -1}. This study indicated that formate is a better substrate than glucose for H{sub 2} production by non-growing cells, and that the H{sub 2}-production performance among the strains did not vary significantly, with the exception of E. coli K-12 MG1655. (author)

Uvm mutants of Escherichia coli K 12 deficient in UV mutagenesis. Pt. 1

International Nuclear Information System (INIS)

Steinborn, G.

1978-01-01

Selection for defective reversion induction, after UV treatment of E. coli K 12, yielded uvm mutants. These mutants exhibited highly reduced or no UV mutability for all loci tested although they were moderately and normally mutable by X-rays and EMS, respectively. Uvm mutations confer only a slight sensitivity to killing by UV and X-rays and no clear sensitivity to the lethal effect of HN2, EMS or MMS. Growth and viability of untreated uvm cells were normal. The properties of uvm mutants are discussed in relation to those of other relevant mutant types and to some actual problems of induced mutagenesis. (orig.) 891 AJ [de

Mutants of Escherichia coli K-12 with enhanced resistance to ionizing radiation

International Nuclear Information System (INIS)

Verbenko, V.N.; Akhmedov, A.T.; Kalinin, V.L.

1986-01-01

By means of one-dimensional electrophoresis, it is shown that in radiation-resistant Gam 444 ad Gam 445 mutants of Escherichia coli K-12 high-molecular weight heat shock proteins are hyperproduced at 32-37 deg C and are induced more intensively during heat shock (in comparison to the parental) wild-tupe strain AB parallel 57). When the missense htp R15 mutation of the positive regulatory htpR gene for heat shock proteins was introduced by transduction into genome of the Gam 444 mutant, its enhanced radiation-resistance disappeared but could not be restored upon introduction of pKV3 plasmid bearing the htpR, gene. These data show that heat shock Protens are participating in the enhanced radioresistance of Gam mutants

Heterologous co-expression of accA, fabD, and thioesterase genes for improving long-chain fatty acid production in Pseudomonas aeruginosa and Escherichia coli.

Science.gov (United States)

Lee, Sunhee; Jeon, Eunyoung; Jung, Yeontae; Lee, Jinwon

2012-05-01

The goal of the present study was to increase the content of intracellular long-chain fatty acids in two bacterial strains, Pseudomonas aeruginosa PA14 and Escherichia coli K-12 MG1655, by co-overexpressing essential enzymes that are involved in the fatty acid synthesis metabolic pathway. Recently, microbial fatty acids and their derivatives have been receiving increasing attention as an alternative source of fuel. By introducing two genes (accA and fabD) of P. aeruginosa into the two bacterial strains and by co-expressing with them the fatty acyl-acyl carrier protein thioesterase gene of Streptococcus pyogenes (strain MGAS10270), we have engineered recombinant strains that are efficient producers of long-chain fatty acids (C16 and C18). The recombinant strains exhibit a 1.3-1.7-fold increase in the production of long-chain fatty acids over the wild-type strains. To enhance the production of total long-chain fatty acids, we researched the carbon sources for optimized culture conditions and results were used for post-culture incubation period. E. coli SGJS17 (containing the accA, fabD, and thioesterase genes) produced the highest content of intracellular total fatty acids; in particular, the unsaturated fatty acid content was about 20-fold higher than that in the wild-type E. coli.

Relevance of DNA repair pathways on ascorbic acid effects on Echerichia Coli K-12 cells

International Nuclear Information System (INIS)

Slyus, M.A. van; Oliveira, R.L.B. da C.; Felzenszwalb, I.; Gomes, R.A.; Menck, C.F.

1985-01-01

Inactivation kinetics were performed with repair proficient and deficient Escherichia coli K-12 cells treated with oxidized solutions of ascorbic acid. The repair pathways controlled by the recA and uvrA gene products are essential for cell survival to the treatment. However, SOS chromotest result indicates that the SOS functions are only induced at high and toxic concentrations of the drug. Moreover, single strand breaks in DNA from treated cells are detected, demonstrating genome damage promoted by oxidized solutions of ascorbate. (M.A.C.) [pt

Escherichia coli yjjPB genes encode a succinate transporter important for succinate production.

Science.gov (United States)

Fukui, Keita; Nanatani, Kei; Hara, Yoshihiko; Yamakami, Suguru; Yahagi, Daiki; Chinen, Akito; Tokura, Mitsunori; Abe, Keietsu

2017-09-01

Under anaerobic conditions, Escherichia coli produces succinate from glucose via the reductive tricarboxylic acid cycle. To date, however, no genes encoding succinate exporters have been established in E. coli. Therefore, we attempted to identify genes encoding succinate exporters by screening an E. coli MG1655 genome library. We identified the yjjPB genes as candidates encoding a succinate transporter, which enhanced succinate production in Pantoea ananatis under aerobic conditions. A complementation assay conducted in Corynebacterium glutamicum strain AJ110655ΔsucE1 demonstrated that both YjjP and YjjB are required for the restoration of succinate production. Furthermore, deletion of yjjPB decreased succinate production in E. coli by 70% under anaerobic conditions. Taken together, these results suggest that YjjPB constitutes a succinate transporter in E. coli and that the products of both genes are required for succinate export.

Induction of the lambda bacteriophage synthesis in Escherichia coli K 12 by polonium alpha rays; Induction de la synthese du bacteriophage lambda chez Escherichia coli K 12 par les rayons alpha du polonium

Energy Technology Data Exchange (ETDEWEB)

Devoret, Raymond

1958-06-15

This research thesis reports the study of the inducing action of polonium alpha radiations in Escherichia Coli K 12 by using an external irradiation device. This work comprised the development of a method to spread bacteria in layer with a thickness less than 20 microns, and the measurement of the number of α particles falling on the irradiated surface. This measurement has been performed by using a nuclear emulsion and a simple photographic film. It appears that alpha radiations have an inducing action, and that at most 15 per cent of bacteria can be induced. The comparison of the induction curve with the survival curves of lysogen and sensitive stains shows that there is no abortive induction. Thus, it appears that this inducing action is not due to an indirect effect of the irradiated medium [French] Dans ce travail on a etudie l'action inductrice des rayons alphas du polonium chez Escherichia Coli K 12 par un diapoaitif d'irradiation externe. Son utilisation necessitait: - une methode d'etalement des bacteries en couche de moins de 20 microns d'epaisseur; - une mesure du nombre des particules alpha tombant sur la surface etendue irradiee. Les mesures ont ete faites a l'aide d'une emulsion nucleaire et d'un film photographique ordinaire. 1) Les rayons alphas ont une action inductrice. Au plus 15 pc des bacteries peuvent etre induites. 2) La oomparaison de la courbe d'induction et des courbes de survie des souches lysogene et sensible montre qu'il n'y a pas d'inductions abortives. 3) Cette action inductrice n'est pas due a un effet indirect du milieu irradie. (auteur)

Molecular structure of the pyruvate dehydrogenase complex from Escherichia coli K-12.

Science.gov (United States)

Vogel, O; Hoehn, B; Henning, U

1972-06-01

The pyruvate dehydrogenase core complex from E. coli K-12, defined as the multienzyme complex that can be obtained with a unique polypeptide chain composition, has a molecular weight of 3.75 x 10(6). All results obtained agree with the following numerology. The core complex consists of 48 polypeptide chains. There are 16 chains (molecular weight = 100,000) of the pyruvate dehydrogenase component, 16 chains (molecular weight = 80,000) of the dihydrolipoamide dehydrogenase component, and 16 chains (molecular weight = 56,000) of the dihydrolipoamide dehydrogenase component. Usually, but not always, pyruvate dehydrogenase complex is produced in vivo containing at least 2-3 mol more of dimers of the pyruvate dehydrogenase component than the stoichiometric ratio with respect to the core complex. This "excess" component is bound differently than are the eight dimers in the core complex.

Deficiency in L-serine deaminase interferes with one-carbon metabolism and cell wall synthesis in Escherichia coli K-12.

Science.gov (United States)

Zhang, Xiao; El-Hajj, Ziad W; Newman, Elaine

2010-10-01

Escherichia coli K-12 provided with glucose and a mixture of amino acids depletes L-serine more quickly than any other amino acid even in the presence of ammonium sulfate. A mutant without three 4Fe4S L-serine deaminases (SdaA, SdaB, and TdcG) of E. coli K-12 is unable to do this. The high level of L-serine that accumulates when such a mutant is exposed to amino acid mixtures starves the cells for C(1) units and interferes with cell wall synthesis. We suggest that at high concentrations, L-serine decreases synthesis of UDP-N-acetylmuramate-L-alanine by the murC-encoded ligase, weakening the cell wall and producing misshapen cells and lysis. The inhibition by high L-serine is overcome in several ways: by a large concentration of L-alanine, by overproducing MurC together with a low concentration of L-alanine, and by overproducing FtsW, thus promoting septal assembly and also by overexpression of the glycine cleavage operon. S-Adenosylmethionine reduces lysis and allows an extensive increase in biomass without improving cell division. This suggests that E. coli has a metabolic trigger for cell division. Without that reaction, if no other inhibition occurs, other metabolic functions can continue and cells can elongate and replicate their DNA, reaching at least 180 times their usual length, but cannot divide.

Competitive accumulation of betaines by Escherichia coli K-12 and derivative strains lacking betaine porters.

Science.gov (United States)

Randall, K; Lever, M; Peddie, B A; Chambers, S T

1995-08-17

Escherichia coli was grown in hyperosmotic media containing both glycine betaine and one other betaine. E. coli K-12 derivative WG439 (putP- proP- proU-) did not accumulate any of 15 betaines. Strains WG445 (putP- proP- proU+), WG443 (putP- proP+ proU-) and the control strains all accumulated less betaine, (CH3)3N(+)-(CH2)n-COO-, when n was greater than 1. Accumulation was not detectable when n = 5. Both L- and D-isomers of alpha-substituted betaines were accumulated by both strains WG443 and WG445, the D-isomers more slowly. Hydroxylated alpha-substituted betaines were accumulated relatively more through the osmoregulated transport protein ProU than through ProP. In actively growing cultures glycine betaine appeared to be the preferred substrate for accumulation, but the proportion of the second accumulated betaine increased as cultures approached stationary phase.

Crystallization and preliminary X-ray diffraction analysis of ybfF, a new esterase from Escherichia coli K12

Energy Technology Data Exchange (ETDEWEB)

Park, Suk-Youl; Lee, Sang-Hak; Lee, Jieun; Jung, Che-Hun; Kim, Jeong-Sun, E-mail: jsunkim@chonnam.ac.kr [Department of Chemistry and Institute of Basic Sciences, Chonnam National University, Gwangju 500-757 (Korea, Republic of)

2007-12-01

The crystallization of ybfF, a new esterase from E. coli, and the collection of diffraction data to 1.1 Å resolution are reported. The product of the recently discovered ybfF gene, which belongs to the esterase family, does not show high sequence similarity to other esterases. To provide the molecular background to the enzymatic mechanism of the ybfF esterase, the ybfF protein from Escherichia coli K12 (Ec-ybfF) was cloned, expressed and purified. The Ec-ybfF protein was crystallized from 60% Tacsimate and 0.1 M bis-Tris propane buffer pH 7.0. Diffraction data were collected to 1.10 Å resolution using synchrotron radiation. The crystal belongs to the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 66.09, b = 90.71, c = 92.88 Å. With two Ec-ybfF molecules in the asymmetric unit, the crystal volume per unit protein weight is 2.17 Å{sup 3} Da{sup −1}, corresponding to a solvent content of 42%.

Deficiency in l-Serine Deaminase Interferes with One-Carbon Metabolism and Cell Wall Synthesis in Escherichia coli K-12▿

Science.gov (United States)

Zhang, Xiao; El-Hajj, Ziad W.; Newman, Elaine

2010-01-01

Escherichia coli K-12 provided with glucose and a mixture of amino acids depletes l-serine more quickly than any other amino acid even in the presence of ammonium sulfate. A mutant without three 4Fe4S l-serine deaminases (SdaA, SdaB, and TdcG) of E. coli K-12 is unable to do this. The high level of l-serine that accumulates when such a mutant is exposed to amino acid mixtures starves the cells for C1 units and interferes with cell wall synthesis. We suggest that at high concentrations, l-serine decreases synthesis of UDP-N-acetylmuramate-l-alanine by the murC-encoded ligase, weakening the cell wall and producing misshapen cells and lysis. The inhibition by high l-serine is overcome in several ways: by a large concentration of l-alanine, by overproducing MurC together with a low concentration of l-alanine, and by overproducing FtsW, thus promoting septal assembly and also by overexpression of the glycine cleavage operon. S-Adenosylmethionine reduces lysis and allows an extensive increase in biomass without improving cell division. This suggests that E. coli has a metabolic trigger for cell division. Without that reaction, if no other inhibition occurs, other metabolic functions can continue and cells can elongate and replicate their DNA, reaching at least 180 times their usual length, but cannot divide. PMID:20729359

Development of an Escherichia coli K12-specific quantitative polymerase chain reaction assay and DNA isolation suited to biofilms associated with iron drinking water pipe corrosion products

Science.gov (United States)

Escherichia coli is one of the most commonly used fecal indicator organisms for drinking water and groundwater systems. In order to understand various biogeochemical and biophysical factors affecting its interactions with biofilms, E. coli K12 was chosen as a model organism. A Ta...

Control of utilization of L-arginine, L-ornithine, agmatine, and putrescine as nitrogen sources in Escherichia coli K-12.

OpenAIRE

Shaibe, E; Metzer, E; Halpern, Y S

1985-01-01

The regulation of the synthesis of the enzymes involved in the utilization of L-arginine, L-ornithine, agmatine, and putrescine as a sole nitrogen source in Escherichia coli K-12 was examined. The synthesis of agmatine ureohydrolase, putrescine aminotransferase, and pyrroline dehydrogenase is dually controlled by catabolite repression and nitrogen availability. Catabolite repression of agmatine ureohydrolase, but not that of putrescine aminotransferase or pyrroline dehydrogenase, is relieved ...

DnaB gene product-independence of DNA polymerase III-directed repair synthesis in Escherichia coli K-12

International Nuclear Information System (INIS)

Billen, D.; Hellermann, G.R.

1977-01-01

An investigation has been carried out into the role of dnaB gene product in X-ray-induced repair synthesis carried out by DNA polymerase III in toluene-treated Escherichia coli K-12. A polAl polBlOO dnaB mutant deficient in both DNA polymerase I and II activities was used, and it was shown that the level of X-ray-induced, ATP-dependent, non-conservative DNA synthesis was, unlike semi-conservative DNA synthesis, unaffected by a temperature shift from 30 0 to 42 0 C. The dnaB gene product was not therefore necessary for DNA polymerase III-directed repair synthesis, which occurred in the absence of replicative synthesis. (U.K.)

Search for 12 C+ 12 C clustering in 24 Mg ground state

Indian Academy of Sciences (India)

Home; Journals; Pramana – Journal of Physics; Volume 88; Issue 2. Search for 12C+12C clustering in 24Mg ground state. B N JOSHI ARUN K JAIN D C BISWAS B V JOHN Y K GUPTA L S DANU R P VIND G K PRAJAPATI S MUKHOPADHYAY A SAXENA. Regular Volume 88 Issue 2 February 2017 Article ID 29 ...

The steady-state kinetics of the NADH-dependent nitrite reductase from Escherichia coli K 12. Nitrite and hydroxylamine reduction.

OpenAIRE

Jackson, R H; Cole, J A; Cornish-Bowden, A

1981-01-01

The reduction of both NO2- and hydroxylamine by the NADH-dependent nitrite reductase of Escherichia coli K 12 (EC 1.6.6.4) appears to follow Michaelis-Menten kinetics over a wide range of NADH concentrations. Substrate inhibition can, however, be detected at low concentrations of the product NAD+. In addition, NAD+ displays mixed product inhibition with respect to NADH and mixed or uncompetitive inhibition with respect to hydroxylamine. These inhibition characteristics are consistent with a m...

Genetical and functional investigation of fliC genes encoding flagellar serotype H4 in wildtype strains of Escherichia coli and in a laboratory E. coli K-12 strain expressing flagellar antigen type H48

Directory of Open Access Journals (Sweden)

Schaudinn Christoph

2005-01-01

Full Text Available Abstract Background Serotyping of O-(lipopolysaccharide and H-(flagellar antigens is a wideley used method for identification of pathogenic strains and clones of Escherichia coli. At present, 176 O- and 53 H-antigens are described for E. coli which occur in different combinations in the strains. The flagellar antigen H4 is widely present in E. coli strains of different O-serotypes and pathotypes and we have investigated the genetic relationship between H4 encoding fliC genes by PCR, nucleotide sequencing and expression studies. Results The complete nucleotide sequence of fliC genes present in E. coli reference strains U9-41 (O2:K1:H4 and P12b (O15:H17 was determined and both were found 99.3% (1043 of 1050 nucleotides identical in their coding sequence. A PCR/RFLP protocol was developed for typing of fliC-H4 strains and 88 E. coli strains reacting with H4 antiserum were investigated. Nucleotide sequencing of complete fliC genes of six E. coli strains which were selected based on serum agglutination titers, fliC-PCR genotyping and reference data revealed 96.6 to 100% identity on the amino acid level. The functional expression of flagellin encoded by fliC-H4 from strain U9-41 and from our strain P12b which is an H4 expressing variant type was investigated in the E. coli K-12 strain JM109 which encodes flagellar type H48. The fliC recombinant plasmid carrying JM109 strains reacted with both H4 and H48 specific antisera whereas JM109 reacted only with the H48 antiserum. By immunoelectron microscopy, we could show that the flagella made by the fliC-H4 recombinant plasmid carrying strain are constituted of H48 and H4 flagellins which are co-assembled into functional flagella. Conclusion The flagellar serotype H4 is encoded by closely related fliC genes present in serologically different types of E. coli strainswhich were isolated at different time periods and geographical locations. Our expression studies show for the first time, that flagellins of

In silico genomic analyses reveal three distinct lineages of Escherichia coli O157:H7, one of which is associated with hyper-virulence.

Science.gov (United States)

Laing, Chad R; Buchanan, Cody; Taboada, Eduardo N; Zhang, Yongxiang; Karmali, Mohamed A; Thomas, James E; Gannon, Victor Pj

2009-06-29

Many approaches have been used to study the evolution, population structure and genetic diversity of Escherichia coli O157:H7; however, observations made with different genotyping systems are not easily relatable to each other. Three genetic lineages of E. coli O157:H7 designated I, II and I/II have been identified using octamer-based genome scanning and microarray comparative genomic hybridization (mCGH). Each lineage contains significant phenotypic differences, with lineage I strains being the most commonly associated with human infections. Similarly, a clade of hyper-virulent O157:H7 strains implicated in the 2006 spinach and lettuce outbreaks has been defined using single-nucleotide polymorphism (SNP) typing. In this study an in silico comparison of six different genotyping approaches was performed on 19 E. coli genome sequences from 17 O157:H7 strains and single O145:NM and K12 MG1655 strains to provide an overall picture of diversity of the E. coli O157:H7 population, and to compare genotyping methods for O157:H7 strains. In silico determination of lineage, Shiga-toxin bacteriophage integration site, comparative genomic fingerprint, mCGH profile, novel region distribution profile, SNP type and multi-locus variable number tandem repeat analysis type was performed and a supernetwork based on the combination of these methods was produced. This supernetwork showed three distinct clusters of strains that were O157:H7 lineage-specific, with the SNP-based hyper-virulent clade 8 synonymous with O157:H7 lineage I/II. Lineage I/II/clade 8 strains clustered closest on the supernetwork to E. coli K12 and E. coli O55:H7, O145:NM and sorbitol-fermenting O157 strains. The results of this study highlight the similarities in relationships derived from multi-locus genome sampling methods and suggest a "common genotyping language" may be devised for population genetics and epidemiological studies. Future genotyping methods should provide data that can be stored centrally and

In silico genomic analyses reveal three distinct lineages of Escherichia coli O157:H7, one of which is associated with hyper-virulence

Directory of Open Access Journals (Sweden)

Karmali Mohamed A

2009-06-01

Full Text Available Abstract Background Many approaches have been used to study the evolution, population structure and genetic diversity of Escherichia coli O157:H7; however, observations made with different genotyping systems are not easily relatable to each other. Three genetic lineages of E. coli O157:H7 designated I, II and I/II have been identified using octamer-based genome scanning and microarray comparative genomic hybridization (mCGH. Each lineage contains significant phenotypic differences, with lineage I strains being the most commonly associated with human infections. Similarly, a clade of hyper-virulent O157:H7 strains implicated in the 2006 spinach and lettuce outbreaks has been defined using single-nucleotide polymorphism (SNP typing. In this study an in silico comparison of six different genotyping approaches was performed on 19 E. coli genome sequences from 17 O157:H7 strains and single O145:NM and K12 MG1655 strains to provide an overall picture of diversity of the E. coli O157:H7 population, and to compare genotyping methods for O157:H7 strains. Results In silico determination of lineage, Shiga-toxin bacteriophage integration site, comparative genomic fingerprint, mCGH profile, novel region distribution profile, SNP type and multi-locus variable number tandem repeat analysis type was performed and a supernetwork based on the combination of these methods was produced. This supernetwork showed three distinct clusters of strains that were O157:H7 lineage-specific, with the SNP-based hyper-virulent clade 8 synonymous with O157:H7 lineage I/II. Lineage I/II/clade 8 strains clustered closest on the supernetwork to E. coli K12 and E. coli O55:H7, O145:NM and sorbitol-fermenting O157 strains. Conclusion The results of this study highlight the similarities in relationships derived from multi-locus genome sampling methods and suggest a "common genotyping language" may be devised for population genetics and epidemiological studies. Future genotyping

Genes and proteins of Escherichia coli K-12.

Science.gov (United States)

Riley, M

1998-01-01

GenProtEC is a database of Escherichia coli genes and their gene products, classified by type of function and physiological role and with citations to the literature for each. Also present are data on sequence similarities among E.coli proteins, representing groups of paralogous genes, with PAM values, percent identity of amino acids, length of alignment and percent aligned. GenProtEC can be accessed at the URL http://www.mbl.edu/html/ecoli.html

A Negative Thermal Expansion Material of ZrMgMo3O12

International Nuclear Information System (INIS)

Song Wen-Bo; Liang Er-Jun; Liu Xian-Sheng; Li Zhi-Yuan; Yuan Bao-He; Wang Jun-Qiao

2013-01-01

A material with the formula ZrMgMo 3 O 12 having negative thermal expansion is presented and characterized. It is shown that ZrMgMo 3 O 12 crystallizes in an orthorhombic symmetry with space group Pnma(62) or Pna2 1 (33) and exhibits negative thermal expansion in a large temperature range (α l = −3.8 × 10 −6 K −1 from 300K to 1000K by x-ray diffraction and α l = −3.73 × 10 −6 K −1 from 295K to 775K by dilatometer). ZrMgMo 3 O 12 remains the orthorhombic structure without phase transition or decomposition at least from 123K to 1200K and is not hygroscopic. These properties make it an excellent material with negative thermal expansion for a variety of applications

Expression of ultraviolet-induced restriction alleviation in Escherichia coli K-12

International Nuclear Information System (INIS)

Thoms, B.; Wackernagel, W.

1983-01-01

Ultraviolet-induced restriction alleviation is an SOS function which partially relieves the K-12-specific DNA restriction in Escherichia coli. Restriction alleviation is determined by observing elevated survival of unmodified phage lambda in cells irradiated with ultraviolet prior to infection. The authors demonstrate that restriction of lambda is also relieved when log-phase cells are irradiated as late as 50 min after adsorption of lambda. At this time more than 60% of the lambda DNA is already released as acid-soluble material from the cells. Experiments involving reextraction of lambda DNA from infected cells and a mild detergent treatment removing adsorbed phages from the cellular surface showed that only a small specific fraction of all lambda infections is destined to escape restriction due to restriction alleviation. This fraction (10-20%) has a retarded mode of DNA injection (60 min or longer) after adsorption which allows the expression of the restriction alleviation function before the phage DNA is exposed to restriction endonucleases. This behaviour of a fraction of lambda phages explains why the SOS function restriction alleviation could initially be discovered. The authors show that the retarded mode of DNA injection is not required for another SOS function acting on lambda DNA, the increased repair of ultraviolet-irradiated DNA (Weigle reactivation). (Auth.)

Characterization of Escherichia coli MG1655 grown in a low-shear modeled microgravity environment

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Pierson Duane L

2007-03-01

Full Text Available Abstract Background Extra-cellular shear force is an important environmental parameter that is significant both medically and in the space environment. Escherichia coli cells grown in a low-shear modeled microgravity (LSMMG environment produced in a high aspect rotating vessel (HARV were subjected to transcriptional and physiological analysis. Results Aerobic LSMMG cultures were grown in rich (LB and minimal (MOPS + glucose medium with a normal gravity vector HARV control. Reproducible changes in transcription were seen, but no specific LSMMG responsive genes were identified. Instead, absence of shear and a randomized gravity vector appears to cause local extra-cellular environmental changes, which elicit reproducible cellular responses. In minimal media, the majority of the significantly up- or down-regulated genes of known function were associated with the cell envelope. In rich medium, most LSMMG down-regulated genes were involved in translation. No observable changes in post-culture stress responses and antibiotic sensitivity were seen in cells immediately after exposure to LSMMG. Comparison with earlier studies of Salmonella enterica serovar Typhimurium conducted under similar growth conditions, revealed essentially no similarity in the genes that were significantly up- or down-regulated. Conclusion Comparison of these results to previous studies suggests that different organisms may dramatically differ in their responses to medically significant low-shear and space environments. Depending on their specific response, some organisms, such as Salmonella, may become preadapted in a manner that predisposes them to increased virulence.

The oxygen effect in E.coli K-12 cells of various repair genotypes exposed to neutrons and gamma rays

International Nuclear Information System (INIS)

Komova, O.V.; Golovacheva, E.V.

1988-01-01

The oxygen enchancement ratio, as estimated after the effect of 137 Cs-γ-quanta, depends on the repair genotype of E. coli K-12 cells and increases in the studied strains in the following order: recA - uvrA - →recA - →wild type→polA - . These variations are levelled with the effect of fast neutrons of divison spectrum (0.75 MeV); the oxygen enhancement ratio for the strains under study decrease, while the oxygen effect is virtually absent in recA - uvrA - -mutant

The mechanism of uncoupling by picrate in Escherichia coli K-12 membrane systems.

Science.gov (United States)

Michels, M; Bakker, E P

1981-06-01

The mechanism of action of the uncoupler picrate on intact cells and everted membrane vesicles of Escherichia coli K-12 was investigated. Like in mitochondria [Hanstein, W. G. and Hatefi, Y. (1974) Proc. Natl Acad. Sci. USA, 71, 288-292], it was observed that picrate uncoupled energy-linked functions only in everted, but not in intact membrane systems. In the vesicles picrate also decreased the magnitude of the transmembrane proton-motive force at concentrations similar to those at which it caused uncoupling. Experiments with 14C-labelled picrate showed that this compound bound both to deenergized intact cells and everted vesicles. However, upon energization of the membrane, picrate was extruded from the intact cell and taken up to a larger extent by the vesicles. These energy-dependent changes in picrate uptake correlated with the magnitude of the transmembrane electrical potential, delta psi. It is therefore proposed that picrate is a permeant uncoupler, that delta psi is the driving force for picrate movement across biological membranes, and that the uncoupling activity of picrate in everted membrane systems is due to its protonophoric action.

Biogenic synthesis and characterization of gold nanoparticles by Escherichia coli K12 and its heterogeneous catalysis in degradation of 4-nitrophenol

Science.gov (United States)

Srivastava, Sarvesh Kumar; Yamada, Ryosuke; Ogino, Chiaki; Kondo, Akihiko

2013-02-01

Room-temperature extracellular biosynthesis of gold nanoparticles (Au NPs) was achieved using Escherichia coli K12 cells without the addition of growth media, pH adjustments or inclusion of electron donors/stabilizing agents. The resulting nanoparticles were analysed by ultraviolet-visible (UV-vis) spectrophotometry, atomic force microscopy, transmission electron microscopy and X-ray diffraction. Highly dispersed gold nanoplates were achieved in the order of around 50 nm. Further, the underlying mechanism was found to be controlled by certain extracellular membrane-bound proteins, which was confirmed by Fourier transformation-infrared spectroscopy and sodium dodecyl sulfate polyacrylamide gel electrophoresis. We observed that certain membrane-bound peptides are responsible for reduction and subsequent stabilization of Au NPs (confirmed by zeta potential analysis). Upon de-activation of these proteins, no nanoparticle formation was observed. Also, we prepared a novel biocatalyst with Au NPs attached to the membrane-bound fraction of E. coli K12 cells serving as an efficient heterogeneous catalyst in complete reduction of 4-nitrophenol in the presence of NaBH4 which was studied with UV-vis spectroscopy. This is the first report on bacterial membrane-Au NP nanobiocomposite serving as an efficient heterogeneous catalyst in complete reduction of nitroaromatic pollutant in water.

Oxygen effect of E.coli K-12 with different repair genotype at the bombardment by neutrons and γ-rays

International Nuclear Information System (INIS)

Komova, O.V.; Golovacheva, E.V.

1986-01-01

It is shown that the value of oxygen enhancement ratio (OER) depends essentually on repair possibilities of cells E.coli K-12 at 137 Cs - γ-irradiation. It increases in a range of investigated strains rec A - uvr A - → rec A - → wild type → pol A - . These differences disappear under action of fast neutron fission spectra with 0.75 MeV mean energy. OER values for all strains have been reduced in this case, and double mutant rec A - uvr A - practically has not any oxygen effects

W-reactivation of phage lambda in X-irradiated mutants of Escherichia coli K-12

Energy Technology Data Exchange (ETDEWEB)

Martignoni, K D; Haselbacher, I [Muenchen Univ. (Germany, F.R.). Strahlenbiologisches Inst.

1980-07-01

The survival of UV irradiated phage lambda was increased on X-irradiated E.coli K-12 host cells over that on unirradiated cells. The frequency of c mutants among the surviving phages was increased to a similar extent by the X-ray exposure of the host cells as by UV light. This W-reactivation of phage lambda occurred in uvrA, polA, and recB mutants besides the wild type at about equal X-ray doses, but at a reduced reactivation efficiency compared with the wild type. W-reactivation was undetectable in recA mutants. While maximal UV induced W-reactivation occured 30 min after irradiation, the maximal X-ray induced reactivation was found immediately after irradiation. Chloramphenicol (100 ..mu..g/ml) and nitrofurantoin (50 ..mu..g/ml) inhibited W-reactivation of phage lambda if added before irradiation of the host cells, indicating the necessity of protein synthesis for W-reactivation.

Induction of the lambda bacteriophage synthesis in Escherichia coli K 12 by polonium alpha rays

International Nuclear Information System (INIS)

Devoret, Raymond

1958-06-01

This research thesis reports the study of the inducing action of polonium alpha radiations in Escherichia Coli K 12 by using an external irradiation device. This work comprised the development of a method to spread bacteria in layer with a thickness less than 20 microns, and the measurement of the number of α particles falling on the irradiated surface. This measurement has been performed by using a nuclear emulsion and a simple photographic film. It appears that alpha radiations have an inducing action, and that at most 15 per cent of bacteria can be induced. The comparison of the induction curve with the survival curves of lysogen and sensitive stains shows that there is no abortive induction. Thus, it appears that this inducing action is not due to an indirect effect of the irradiated medium [fr

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

Science.gov (United States)

Miki, Takeyoshi; Yamamoto, Yoshihiro; Matsuda, Hideo

2008-01-01

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

Characterization of new radiation-sensitive mutant, Escherichia coli K-12 radC102

International Nuclear Information System (INIS)

Felzenszwalb, I.; Sargentini, N.J.; Smith, K.C.

1984-01-01

A new radiation-sensitive mutant, radC, has been isolated. The radC gene is located at 81.0 min on the Escherichia coli K-12 linkage map. The radC mutation sensitized cells to uv radiation, but unlike most DNA repair mutations, sensitization to X rays was observed only for rich medium-grown cells. For cells grown in rich medium, the radC mutant was normal for γ radiation mutagenesis, but showed less uv-radiation mutagenesis than the wild-type strain; it showed normal amount of X- and uv-radiation-induced DNA degradation, and it wasapprox. =60% deficient in recombination ability. The radC strain was normal for host cell reactivation of γ and uv-irradiated bacteriophage the radC mutation did not sensitize a recA strain, but did sensitize a radA and a polA strain to X and uv radiation and a uvrA strain to uv radiation. Therefore, it is suggested that the radC gene product plays a role in the growth medium-dependent, recA gene-dependent repair of DNA single-strand breaks after X irradiation, and in postreplication repair after uv irradiation

Two proline porters in Escherichia coli K-12.

Science.gov (United States)

Stalmach, M E; Grothe, S; Wood, J M

1983-11-01

Escherichia coli mutants defective at putP and putA lack proline transport via proline porter I and proline dehydrogenase activity, respectively. They retain a proline uptake system (proline porter II) that is induced during tryptophan-limited growth and are sensitive to the toxic L-proline analog, 3,4-dehydroproline. 3,4-Dehydroproline-resistant mutants derived from a putP putA mutant lack proline porter II. Auxotrophic derivatives derived from putP+ or putP bacteria can grow if provided with proline at low concentration (25 microM); those derived from the 3,4-dehydroproline-resistant mutants require high proline for growth (2.5 mM). We conclude that E. coli, like Salmonella typhimurium, possesses a second proline porter that is inactivated by mutations at the proP locus.

Adherence to abiotic surface induces SOS response in Escherichia coli K-12 strains under aerobic and anaerobic conditions.

Science.gov (United States)

Costa, Suelen B; Campos, Ana Carolina C; Pereira, Ana Claudia M; de Mattos-Guaraldi, Ana Luiza; Júnior, Raphael Hirata; Rosa, Ana Cláudia P; Asad, Lídia M B O

2014-09-01

During the colonization of surfaces, Escherichia coli bacteria often encounter DNA-damaging agents and these agents can induce several defence mechanisms. Base excision repair (BER) is dedicated to the repair of oxidative DNA damage caused by reactive oxygen species (ROS) generated by chemical and physical agents or by metabolism. In this work, we have evaluated whether the interaction with an abiotic surface by mutants derived from E. coli K-12 deficient in some enzymes that are part of BER causes DNA damage and associated filamentation. Moreover, we studied the role of endonuclease V (nfi gene; 1506 mutant strain) in biofilm formation. Endonuclease V is an enzyme that is involved in DNA repair of nitrosative lesions. We verified that endonuclease V is involved in biofilm formation. Our results showed more filamentation in the xthA mutant (BW9091) and triple xthA nfo nth mutant (BW535) than in the wild-type strain (AB1157). By contrast, the mutant nfi did not present filamentation in biofilm, although its wild-type strain (1466) showed rare filaments in biofilm. The filamentation of bacterial cells attaching to a surface was a consequence of SOS induction measured by the SOS chromotest. However, biofilm formation depended on the ability of the bacteria to induce the SOS response since the mutant lexA Ind(-) did not induce the SOS response and did not form any biofilm. Oxygen tension was an important factor for the interaction of the BER mutants, since these mutants exhibited decreased quantitative adherence under anaerobic conditions. However, our results showed that the presence or absence of oxygen did not affect the viability of BW9091 and BW535 strains. The nfi mutant and its wild-type did not exhibit decreased biofilm formation under anaerobic conditions. Scanning electron microscopy was also performed on the E. coli K-12 strains that had adhered to the glass, and we observed the presence of a structure similar to an extracellular matrix that depended on the

Characterization of WbiQ: An α1,2-fucosyltransferase from Escherichia coli O127:K63(B8), and synthesis of H-type 3 blood group antigen

International Nuclear Information System (INIS)

Pettit, Nicholas; Styslinger, Thomas; Mei, Zhen; Han, Weiqing; Zhao, Guohui; Wang, Peng George

2010-01-01

Research highlights: → WbiQ is an α1,2-fucosyltransferase from Escherichia coli O127. → WbiQ demonstrates strict substrate specificity for the Gal-β1,3-GalNAc acceptor. → WbiQ was used to synthesize milligram scale of the H-type 3 blood group antigen. -- Abstract: Escherichia coli O127:K63(B8) possesses high human blood group H (O) activity due to its O-antigen repeating unit structure. In this work, the wbiQ gene from E. coli O127:K63(B8) was expressed in E. coli BL21 (DE3) and purified as a fusion protein containing an N-terminal GST affinity tag. Using the GST-WbiQ fusion protein, the wbiQ gene was identified to encode an α1,2-fucosyltransferase using a radioactivity based assay, thin-layer chromatography assay, as well confirming product formation by using mass spectrometry and NMR spectroscopy. The fused enzyme (GST-WbiQ) has an optimal pH range from 6.5 to 7.5 and does not require the presence of a divalent metal to be enzymatically active. WbiQ displays strict substrate specificity, displaying activity only towards acceptors that contain Gal-β1,3-GalNAc-α-OR linkages; indicating that both the Gal and GalNAc residues are vital for enzymatic activity. In addition, WbiQ was used to prepare the H-type 3 blood group antigen, Fuc-α1,2-Gal-β1,3-GalNAc-α-OMe, on a milligram scale.

The SOS response is permitted in Escherichia coli strains deficient in the expression of the mazEF pathway.

Science.gov (United States)

Kalderon, Ziva; Kumar, Sathish; Engelberg-Kulka, Hanna

2014-01-01

The Escherichia coli (E. coli) SOS response is the largest, most complex, and best characterized bacterial network induced by DNA damage. It is controlled by a complex network involving the RecA and LexA proteins. We have previously shown that the SOS response to DNA damage is inhibited by various elements involved in the expression of the E. coli toxin-antitoxin mazEF pathway. Since the mazEF module is present on the chromosomes of most E. coli strains, here we asked: Why is the SOS response found in so many E. coli strains? Is the mazEF module present but inactive in those strains? We examined three E. coli strains used for studies of the SOS response, strains AB1932, BW25113, and MG1655. We found that each of these strains is either missing or inhibiting one of several elements involved in the expression of the mazEF-mediated death pathway. Thus, the SOS response only takes place in E. coli cells in which one or more elements of the E. coli toxin-antitoxin module mazEF or its downstream pathway is not functioning.

32 CFR 165.5 - Responsibilities.

Science.gov (United States)

2010-07-01

... NONRECURRING COSTS ON SALES OF U.S. ITEMS § 165.5 Responsibilities. (a) The Comptroller of the Department of... of this part. (2) Review and approve nonrecurring cost recoupment charges and nonrecurring cost... military sales. (3) Ensure publication of a listing of items developed for or by the Department of Defense...

Transcriptome of E. coli K1 bound to human brain microvascular endothelial cells

OpenAIRE

Xie, Yi; Parthasarathy, Geetha; Di Cello, Francescopaolo; Teng, Ching-Hao; Paul-Satyaseela, Maneesh; Kim, Kwang Sik

2007-01-01

Escherichia coli K1 is the most common Gram-negative organism causing neonatal meningitis. Binding to human brain microvascdular endothelial cells (HBMEC) is an essential step for E. coli K1 traversal of the blood-brain barrier. In this study, we examined expression profiles of E. coli K1 strain RS218 during its binding to HBMEC. Comparison of HBMEC-bound E. coli K1 with collagen-bound E. coli revealed more than one hundred genes whose expression patterns were significantly changed in HBMEC-b...

21 CFR 184.1655 - Propane.

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2010-04-01

... CONSUMPTION (CONTINUED) DIRECT FOOD SUBSTANCES AFFIRMED AS GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS § 184.1655 Propane. (a) Propane (empirical formula C3H8, CAS Reg. No. 74-98-6) is also known as dimethylmethane or propyl hydrid. It is a colorless, odorless, flammable gas at normal...

Probiotic Escherichia coli Nissle 1917 inhibits leaky gut by enhancing mucosal integrity.

Directory of Open Access Journals (Sweden)

Sya N Ukena

2007-12-01

Full Text Available Probiotics are proposed to positively modulate the intestinal epithelial barrier formed by intestinal epithelial cells (IECs and intercellular junctions. Disruption of this border alters paracellular permeability and is a key mechanism for the development of enteric infections and inflammatory bowel diseases (IBDs.To study the in vivo effect of probiotic Escherichia coli Nissle 1917 (EcN on the stabilization of the intestinal barrier under healthy conditions, germfree mice were colonized with EcN or K12 E. coli strain MG1655. IECs were isolated and analyzed for gene and protein expression of the tight junction molecules ZO-1 and ZO-2. Then, in order to analyze beneficial effects of EcN under inflammatory conditions, the probiotic was orally administered to BALB/c mice with acute dextran sodium sulfate (DSS induced colitis. Colonization of gnotobiotic mice with EcN resulted in an up-regulation of ZO-1 in IECs at both mRNA and protein levels. EcN administration to DSS-treated mice reduced the loss of body weight and colon shortening. In addition, infiltration of the colon with leukocytes was ameliorated in EcN inoculated mice. Acute DSS colitis did not result in an anion secretory defect, but abrogated the sodium absorptive function of the mucosa. Additionally, intestinal barrier function was severely affected as evidenced by a strong increase in the mucosal uptake of Evans blue in vivo. Concomitant administration of EcN to DSS treated animals resulted in a significant protection against intestinal barrier dysfunction and IECs isolated from these mice exhibited a more pronounced expression of ZO-1.This study convincingly demonstrates that probiotic EcN is able to mediate up-regulation of ZO-1 expression in murine IECs and confer protection from the DSS colitis-associated increase in mucosal permeability to luminal substances.

Correlation between survival, ability to rejoin DNA and stability of DNA after preirradiation inhibition of protein synthesis in a rec- mutant of Escherichia coli K12

International Nuclear Information System (INIS)

Pirsel, M.; Slezarikova, V.

1977-01-01

A 90 min inhibition of protein synthesis induced by starvation for amino acids (AA - ) or by chloramphenicol (CAP) treatment prior to UV irradiation (2.5 J m -2 ) increased more than tenfold the resistance of the strain Escherichia coli K12 SR19 to UV radiation. Under these conditions, cultures in which protein synthesis was inhibited before the UV irradiation rejoin short regions of DNA synthesized after the irradiation to a normal-size molecule, whereas an exponentially growing culture does not rejoin DNA synthesized after UV irradiation to a molecule of a normal size. In the exponentially growing culture both the parental and the newly synthesized DNA are unstable after the irradiation. In cultures with inhibited protein synthesis only the parental DNA is somewhat unstable. In Escherichia coli K12 SR19 where protein synthesis was inhibited before the irradiation, a correlation between the survival of cells, the ability to rejoin short regions of DNA synthesized after UV irradiation, and a higher stability of both parental and newly synthesized DNAs could be demonstrated. (author)

Supercritical CO2 induces marked changes in membrane phospholipids composition in Escherichia coli K12.

Science.gov (United States)

Tamburini, Sabrina; Anesi, Andrea; Ferrentino, Giovanna; Spilimbergo, Sara; Guella, Graziano; Jousson, Olivier

2014-06-01

Supercritical carbon dioxide (SC-CO2) treatment is one of the most promising alternative techniques for pasteurization of both liquid and solid food products. The inhibitory effect of SC-CO2 on bacterial growth has been investigated in different species, but the precise mechanism of action remains unknown. Membrane permeabilization has been proposed to be the first event in SC-CO2-mediated inactivation. Flow cytometry, high performance liquid chromatography–electrospray ionization–mass spectrometry and NMR analyses were performed to investigate the effect of SC-CO2 treatment on membrane lipid profile and membrane permeability in Escherichia coli K12. After 15 min of SC-CO2 treatment at 120 bar and 35 °C, the majority of bacterial cells dissipated their membrane potential (95 %) and lost membrane integrity, as 81 % become partially permeabilized and 18 % fully permeabilized. Membrane permeabilization was associated with a 20 % decrease in bacterial biovolume and to a strong (>50 %) reduction in phosphatidylglycerol (PG) membrane lipids, without altering the fatty acid composition and the degree of unsaturation of acyl chains. PGs are thought to play an important role in membrane stability, by reducing motion of phosphatidylethanolamine (PE) along the membrane bilayer, therefore promoting the formation of inter-lipid hydrogen bonds. In addition, the decrease in intracellular pH induced by SC-CO2 likely alters the chemical properties of phospholipids and the PE/PG ratio. Biophysical effects of SC-CO2 thus cause a strong perturbation of membrane architecture in E. coli, and such alterations are likely associated with its strong inactivation effect.

40 CFR 60.1655 - Who must complete the plant-specific training course?

Science.gov (United States)

2010-07-01

...-specific training course? All employees with responsibilities that affect how a municipal waste combustion... 40 Protection of Environment 6 2010-07-01 2010-07-01 false Who must complete the plant-specific training course? 60.1655 Section 60.1655 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY...

Effects of starvation on the transport of Escherichia coli K12 in saturated porous media are dependent on pH and ionic strength

Science.gov (United States)

Xu, S.; Walczak, J. J.; Wang, L.; Bardy, S. L.; Li, J.

2010-12-01

In this research, we investigate the effects of starvation on the transport of E. coli K12 in saturated porous media. Particularly, we examine the relationship between such effects and the pH and ionic strength of the electrolyte solutions that were used to suspend bacterial cells. E. coli K12 (ATCC 10798) cells were cultured using either Luria-Bertani Miller (LB-Miller) broth (10 g trypton, 5 g yeast extract and 10 g NaCl in 1 L of deionized water) or LB-Luria broth (10 g tryptone, 5 g yeast extract and 0.5 g NaCl in 1 L of deionized water). Both broths had similar pH (~7.1) but differed in ionic strength (LB-Miller: ~170 mM, LB-Luria: ~ 8 mM). The bacterial cells were then harvested and suspended using one of the following electrolyte solutions: phosphate buffered saline (PBS) (pH ~7.2; ionic strength ~170 mM), 168 mM NaCl (pH ~5.7), 5% of PBS (pH ~ 7.2; ionic strength ~ 8 mM) and 8 mM NaCl (pH ~ 5.7). Column transport experiments were performed at 0, 21 and 48 hours following cell harvesting to evaluate the change in cell mobility over time under “starvation” conditions. Our results showed that 1) starvation increased the mobility of E. coli K12 cells; 2) the most significant change in mobility occurred when bacterial cells were suspended in an electrolyte solution that had different pH and ionic strength (i.e., LB-Miller culture suspended in 8 mM NaCl and LB-Luria culture suspended in 168 mM Nacl); and 3) the change in cell mobility primarily occurred within the first 21 hours. The size of the bacterial cells was measured and the surface properties (e.g., zeta potential, hydrophobicity, cell-bound protein, LPS sugar content, outer membrane protein profiles) of the bacterial cells were characterized. We found that the measured cell surface properties could not fully explain the observed changes in cell mobility caused by starvation.

Iron-hydroxamate transport in Escherichia coli K12

International Nuclear Information System (INIS)

Prody, C.A.

1984-01-01

FhuB mutants, which are deficient in ferrichrome transport, were isolated and characterized. They were found to be deficient in the utilization of all hydroxamate-type siderophores. They were, however, able to transport enterobactin. A number of analogs of hydroxamate-type siderophores were tested for biological activity in E. coli, and about half of these were active. In addition, two rhodotorulic acid analogs were able to supply iron to fhuB mutants. A search for the fhuB gene product, using one and two-dimensional polyacrylamide gels of proteins from fhuB and wild type strains proved fruitless, and it appeared that the fhuB gene product is expressed at a very low level. Therefore, the fhuB gene was subcloned from a plasmid in the Carbon bank onto plasmid vectors containing the E. coli lac UV-5 and tacI promoters as a device to amplify the fhuB gene. One of these recombinant plasmids carried an 8Kb insert which contained both the tonA and fhuB genes. This plasmid synthesized five proteins of molecular weights 78,000, 40,000, 30,000, 24,000, and 13,700 in maxicell strain CSR603. By use of deletions, the approximate order of the genes for these proteins was determined. Although 3 He-ferrichrome is transported into E. coli cells and vesicles, 3 He-ferric rhodotorulate is not, and so the mechanism of transport for these two siderophores must be different. To examine this further, mutants were obtained that could transport ferrichrome but not rhodotorulic acid. These map in the region between tonA and fhuB, and most are able to transport aerobactin, when carrying the ColV plasmid, but not schizokinen

Probiotic Mixture Golden Bifido Prevents Neonatal Escherichia coli K1 Translocation via Enhancing Intestinal Defense

Directory of Open Access Journals (Sweden)

Qing Zeng

2017-09-01

Full Text Available Escherichia coli (E. coli K1 sepsis and meningitis is a severe infection characterized by high mortality in neonates. Successful colonization and translocation across the intestinal mucosa have been regarded as the critical steps for E. coli K1 sepsis and meningitis. We recently reported that the probiotic mixture, Golden Bifido (containing live Lactobacillus bulgaricus, Bifidobacterium, and Streptococcus thermophilus, LBS has a preventive role against neonatal E. coli K1 bacteremia and meningitis. However, the interaction between the neonatal gut barrier, probiotics and E. coli K1 is still not elucidated. The present study aims to investigate how LBS exerts its protective effects on neonatal gut barrier during E. coli K1 infection. The beneficial effects of LBS were explored in vitro and in vivo using human colon carcinoma cell lines HT-29 and rat model of neonatal E. coli K1 infection, respectively. Our results showed that stimulation with E. coli K1 was able to cause intestinal barrier dysfunction, which were reflected by E. coli K1-induced intestinal damage and apoptosis of intestinal epithelial cells, reduction of mucin, immunoglobulin A (IgA and tight junction proteins expression, as well as increase in intestinal permeability, all these changes facilitate E. coli K1 intestinal translocation. However, these changes were alleviated when HT-29 cells were treated with LBS before E. coli K1 infection. Furthermore, we found that LBS-treated neonatal rats (without E. coli K1 infection have showed higher production of mucin, ZO-1, IgA, Ki67 in intestinal mucosa as well as lower intestinal permeability than that of non-treated rats, indicating that LBS could accelerate the development of neonatal intestinal defense. Taken together, our results suggest that enhancement of the neonatal intestinal defense to fight against E. coli K1 translocation could be the potential mechanism to elucidate how LBS confers a protective effect against neonatal E

Ionizing radiation damage to the folded chromosome of Escherichia coli K-12: repair of double-strand breaks in deoxyribonucleic acid

International Nuclear Information System (INIS)

Ulmer, M.K.; Gomez, R.F.; Sinskevy, A.J.

1979-01-01

The extremely gentle lysis and unfolding procedures that have been developed for the isolation of nucleoid deoxyribonucleic acid yield undamaged, replicating genomes, thus permitting direct measurement of the formation and repair of DNA double-strand breaks at biologically significant doses of ionizing radiation. Repair of ionizing radiation damage to folded chromosomes of Escherichia coli K-12 strain AB2497 was observed within 2 to 3 h of post-irradiation incubation in growth medium. Such behavior was not observed after post-irradiation incubation in growth medium of a recA13 strain (strain AB2487). A model based on recombinational repair is proposed to explain the formation of 2,200 to 2,300S material during early stages of incubation and to explain subsequent changes in the gradient profiles. Association of unrepaired DNA with the plasma membrane is proposed to explain the formation of a peak of rapidly sedimenting material (greater than 3,100S) during the later stage of repair. Direct evidence of repair of double-strand breaks during post-irradiation incubation in growth medium was obtained from gradient profiles of DNA from ribonuclease-digested chromosomes. The sedimentation coefficient of broken molecules was restored to the value of unirradiated DNA after 2 to 3 h of incubation, and the fraction of the DNA repaired in this fashion was equal to the fraction of cells that survived at the same dose. An average of 2.7 double-strand breaks per genome per lethal event was observed, suggesting that one to two double-strand breaks per genome are repairable in E. coli K-12 strain AB2497

Impact of metal ion homeostasis of genetically modified Escherichia coli Nissle 1917 and K12 (W3110) strains on colonization properties in the murine intestinal tract.

Science.gov (United States)

Kupz, Andreas; Fischer, André; Nies, Dietrich H; Grass, Gregor; Göbel, Ulf B; Bereswill, Stefan; Heimesaat, Markus M

2013-09-01

Metal ions are integral parts of pro- as well as eukaryotic cell homeostasis. Escherichia coli proved a valuable in vitro model organism to elucidate essential mechanisms involved in uptake, storage, and export of metal ions. Given that E. coli Nissle 1917 is able to overcome murine colonization resistance, we generated several E. coli Nissle 1917 mutants with defects in zinc, iron, copper, nickel, manganese homeostasis and performed a comprehensive survey of the impact of metal ion transport and homeostasis for E. coli colonization capacities within the murine intestinal tract. Seven days following peroral infection of conventional mice with E. coli Nissle 1917 strains exhibiting defined defects in zinc or iron uptake, the respective mutant and parental strains could be cultured at comparable, but low levels from the colonic lumen. We next reassociated gnotobiotic mice in which the microbiota responsible for colonization resistance was abrogated by broad-spectrum antibiotics with six different E. coli K12 (W3110) mutants. Seven days following peroral challenge, each mutant and parental strain stably colonized duodenum, ileum, and colon at comparable levels. Taken together, defects in zinc, iron, copper, nickel, and manganese homeostasis do not compromise colonization capacities of E. coli in the murine intestinal tract.

Development and maturation of Escherichia coli K-12 biofilms

DEFF Research Database (Denmark)

Reisner, A.; Haagensen, J.A.J.; Schembri, Mark

2003-01-01

The development and maturation of E. coli biofilms in flow-chambers was investigated. We found that the presence of transfer constitutive IncF plasmids induced biofilm development forming structures resembling those reported for Pseudomonas aeruginosa . The development occurred in a step...... occurred in conjugation pilus proficient plasmid-carrying strains. The final shapes of the expanding structures in the mature biofilm seem to be determined by the pilus configuration, as various mutants affected in the processing and activity of the transfer pili displayed differently structured biofilms....... We further provide evidence that flagella, type 1 fimbriae, curli and Ag43 are all dispensable for the observed biofilm maturation. In addition, our results indicate that cell-to-cell signalling mediated by autoinducer 2 (AI-2) is not required for differentiation of E. coli within a biofilm community...

Conjugation in Escherichia coli

Science.gov (United States)

Boyer, Herbert

1966-01-01

Boyer, Herbert (Yale University, New Haven, Conn.). Conjugation in Escherichia coli. J. Bacteriol. 91:1767–1772. 1966.—The sex factor of Escherichia coli K-12 was introduced into an E. coli B/r strain by circumventing the host-controlled modification and restriction incompatibilities known to exist between these closely related strains. The sexual properties of the constructed F+ B strain and its Hfr derivatives were examined. These studies showed that the E. coli strain B/r F+ and Hfr derivatives are similar to the E. coli strain K-12 F+ and Hfr derivatives. However, the site of sex factor integration was found to be dependent on the host genome. PMID:5327905

Protection against Escherichia coli K1 infection in newborn rats by antibody to K1 capsular polysaccharide antigen.

OpenAIRE

Bortolussi, R; Ferrier, P

1980-01-01

The protective value of antibody to the K1 capsular polysaccharide antigen of Escherichia coli was investigated in a newborn rat model of E. coli K1 infection. Pregnant rats were immunized intravenously with E. coli, and the agglutinating titer to meningococcal group B polysaccharide, which is identical to K1 polysaccharide, was measured in the serum of rats and their offspring. Convalescent serum from rat mothers showed an increased antibody titer in animals injected twice but not once with ...

Interaction of Escherichia coli K1 and K5 with Acanthamoeba castellanii Trophozoites and Cysts

OpenAIRE

Matin, Abdul; Jung, Suk-Yul

2011-01-01

The existence of symbiotic relationships between Acanthamoeba and a variety of bacteria is well-documented. However, the ability of Acanthamoeba interacting with host bacterial pathogens has gained particular attention. Here, to understand the interactions of Escherichia coli K1 and E. coli K5 strains with Acanthamoeba castellanii trophozoites and cysts, association assay, invasion assay, survival assay, and the measurement of bacterial numbers from cysts were performed, and nonpathogenic E. ...

The inclusions of Mg-B (MgB12?) as potential pinning centres in high-pressure-high-temperature-synthesized or sintered magnesium diboride

International Nuclear Information System (INIS)

Prikhna, T A; Gawalek, W; Savchuk, Ya M; Habisreuther, T; Wendt, M; Sergienko, N V; Moshchil, V E; Nagorny, P; Schmidt, Ch; Dellith, J; Dittrich, U; Litzkendorf, D; Melnikov, V S; Sverdun, V B

2007-01-01

A systematic study of the structure and superconductive characteristics of high-pressure-high-temperature (2 GPa, 700-1000 deg. C )-synthesized and sintered MgB 2 without additions from different initial powders was performed. Among various secondary phases Mg-B inclusions with a stoichiometry close to MgB 12 were identified. With an increasing amount of these inclusions the critical current density increased. So these inclusions can be feasible pinning centres in MgB 2 . The highest j c values in zero field were 1300 kA cm -2 at 10 K, 780 kA cm -2 at 20 K and 62 kA cm -2 at 35 K and in 1 T field were 1200 kA cm -2 at 10 K, 515 kA cm -2 at 20 K and 0.1 kA cm -2 at 35 K for high-pressure-synthesized magnesium diboride and the field of irreversibility at 20 K reached 8 T. The average grain sizes calculated from x-ray examinations in materials having high j c were 15-37 nm

The organization of the fuc regulon specifying L-fucose dissimilation in Escherichia coli K12 as determined by gene cloning.

Science.gov (United States)

Chen, Y M; Zhu, Y; Lin, E C

1987-12-01

In Escherichia coli the six known genes specifying the utilization of L-fucose as carbon and energy source cluster at 60.2 min and constitute a regulon. These genes include fucP (encoding L-fucose permease), fucI (encoding L-fucose isomerase), fucK (encoding L-fuculose kinase), fucA (encoding L-fuculose 1-phosphate aldolase), fucO (encoding L-1,2-propanediol oxidoreductase), and fucR (encoding the regulatory protein). In this study the fuc genes were cloned and their positions on the chromosome were established by restriction endonuclease and complementation analyses. Clockwise, the gene order is: fucO-fucA-fucP-fucI-fucK-fucR. The operons comprising the structural genes and the direction of transcription were determined by complementation analysis and Southern blot hybridization. The fucPIK and fucA operons are transcribed clockwise. The fucO operon is transcribed counterclockwise. The fucR gene product activates the three structural operons in trans.

Binding of a nitroxyl to radiation-induced DNA transients in repair and repair deficient of E. coli K-12

Energy Technology Data Exchange (ETDEWEB)

Wold, E; Brustad, T [Norsk Hydros Institutt for Kreftforskning, Oslo

1975-01-01

Binding of tritiated 2,2,6,6-tetramethyl-4-piperidone-N-oxyl (/sup 3/H-TAN) to radiation-induced DNA-transients in E. coli K-12 strains AB 1157 and JO 307 rec A uvr A has been studied under in vivo conditions. After irradiation the cells were washed and resuspended in growth medium and left overnight at 37 deg C. Within an uncertainty of about 10 %, no effect of repair could be detected on the yield of TAN bound to DNA for any of the strains. During the period after resuspension TAN or fragments of TAN leaked out of the irradiated cell samples. This leakage may be attributed to semi-permanent association between TAN and radiation-induced radicals within the cell. The relevance of different interactions between TAN and transients in DNA is discussed.

Plasmid (pKM101)-mediated enhancement of repair and mutagenesis: dependence on chromosomal genes in 'Escherichia coli' K-12

International Nuclear Information System (INIS)

Walker, G.C.

1977-01-01

The drug resistance plasmid pKM101 plays a major role in the Ames Salmonella/microsome carcinogen detecting system by enhancing chemical mutagenesis. It is shown that in Escherichia coli K-12 the plasmid pKM101 enhances both spontaneous and methyl methanesulfonate-caused reversion of an ochre mutation, bacterial survival after ultaviolet irradiation, and reactivation of ultraviolet-irradiated lambda in unirradiated cells. All these effects are shown to be dependent on the recA + lexA + genotype but not on the recB + recC + or recF + genotypes. The recA lexA-dependence of the plasmid-mediated repair and mutagenesis suggests an interaction with the cell's inducible error-prone repair system. The presence of pKM101 is shown to cause an additional increase in methyl methanesulfonate mutagenesis in a tif mutant beyond that caused by growth at 42 0 . The presence of the plasmid raises the level of the Weigle-reactivation curve for the reactivation of ultraviolet-irradiated lambda in E. coli and causes a shift of the maximum to a higher UV fluence. These observations suggest that pKM101 does not exert its effects by altering the regulation of the cell's error-prone repair system but rather by supplying a mechanistic component or components. (orig.) [de

Ultraviolet radiation-induced mutability of uvrD3 strains of Escherichia coli B/r and K-12: a problem in analyzing mutagenesis data

International Nuclear Information System (INIS)

Smith, K.C.

1976-01-01

The involvement of the uvrD gene product in UV-induced mutagenesis in Escherichia coli was studied by comparing wild-type and uvrA or uvrB strains with their uvrD derivatives in B/r and K-12(W3110) backgrounds. Mutations per survivor (reversions to prototrophy) were compared as a function of surviving fraction and of UV fluence. While recognizing that both methods are not without problems, arguments are presented for favoring the former rather than the latter method of presenting the data when survival is less than 100%. When UV-induced mutation frequencies were plotted as a function of surviving fraction, the uvrD derivatives were less mutable than the corresponding parent strains. The B/r strains exhibited higher mutation frequencies than did the K-12(W3110) strains. A uvrB mutation increased the mutation frequency of its parental K-12 strain, but a uvrA mutation only increased the mutation frequency of its parental B/r strain at UV survivals greater than approximately 80%. Both the uvrA and uvrB mutations increased the mutation frequencies of the uvrD strains in the B/r and K-12 backgrounds, respectively. Rather different conclusions would be drawn if mutagenesis were considered as a function of UV fluence rather than of survival, a situation that calls for further work and discussion. Ideally mutation efficiencies should be compared as a function of the number of repair events per survivor, a number that is currently unobtainable. (author)

Enzymatic induction of DNA double-strand breaks in γ-irradiated Escherichia coli K-12

International Nuclear Information System (INIS)

Bonura, T.; Smith, K.C.; Kaplan, H.S.

1975-01-01

The polA1 mutation increases the sensitivity of E. coli K-12 to killing by γ-irradiation in air by a factor of 2.9 and increases the yield of DNA double-strand breaks by a factor of 2.5. These additional DNA double-strand breaks appear to be due to the action of nucleases in the polA1 strain rather than to the rejoining of radiation-induced double-strand breaks in the pol + strain. This conclusion is based upon the observation that γ-irradiation at 3 0 did not affect the yield of DNA double-strand breaks in the pol + strain, but decreased the yield in the polA1 strain by a factor of 2.2. Irradiation of the polA1 strain at 3 0 followed by incubation at 3 0 for 20 min before plating resulted in approximately a 1.5-fold increase in the D 0 . The yield of DNA double-strand breaks was reduced by a factor of 1.5. The pol + strain, however, did not show the protective effect of the low temperature incubation upon either survival or DNA double-strand breakage. We suggest that the increased yield of DNA double-strand breaks in the polA 1 strain may be the result of the unsuccessful excision repair of ionizing radiation-induced dna base damage

Genome-wide Reconstruction of OxyR and SoxRS Transcriptional Regulatory Networks under Oxidative Stress in Escherichia coli K-12 MG1655

DEFF Research Database (Denmark)

Seo, Sang Woo; Kim, Donghyuk; Szubin, Richard

2015-01-01

Three transcription factors (TFs), OxyR, SoxR, and SoxS, play a critical role in transcriptional regulation of the defense system for oxidative stress in bacteria. However, their full genome-wide regulatory potential is unknown. Here, we perform a genome-scale reconstruction of the OxyR, SoxR, an...

Revealing genome-scale transcriptional regulatory landscape of OmpR highlights its expanded regulatory roles under osmotic stress in Escherichia coli K-12 MG1655

DEFF Research Database (Denmark)

Seo, Sang Woo; Gao, Ye; Kim, Donghyuk

2017-01-01

) belong to OmpR regulon. Among them, 26 genes show more than two-fold changes in expression level in an OmpR knock-out strain. Specifically, we find that: 1) OmpR regulates mostly membrane-located gene products involved in diverse fundamental biological processes, such as narU (encoding nitrate/nitrite...

Probiotic Mixture Golden Bifido Prevents Neonatal Escherichia coli K1 Translocation via Enhancing Intestinal Defense

OpenAIRE

Qing Zeng; Xiaolong He; Santhosh Puthiyakunnon; Hansen Xiao; Zelong Gong; Swapna Boddu; Lecheng Chen; Huiwen Tian; Huiwen Tian; Sheng-He Huang; Sheng-He Huang; Hong Cao

2017-01-01

Escherichia coli (E. coli) K1 sepsis and meningitis is a severe infection characterized by high mortality in neonates. Successful colonization and translocation across the intestinal mucosa have been regarded as the critical steps for E. coli K1 sepsis and meningitis. We recently reported that the probiotic mixture, Golden Bifido (containing live Lactobacillus bulgaricus, Bifidobacterium, and Streptococcus thermophilus, LBS) has a preventive role against neonatal E. coli K1 bacteremia and men...

Effects of casein glycomacropeptide supplementation on growth performance, intestinal morphology, intestinal barrier permeability and inflammatory responses in Escherichia coli K88 challenged piglets

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Yili Rong

2015-06-01

Full Text Available Casein glycomacropeptide (CGMP is a bioactive peptide derived from milk with multiple functions. This study was aimed at evaluating the effects of CGMP as a potential feed additive on growth performance, intestinal morphology, intestinal barrier permeability and inflammatory responses of Escherichia coli K88 (E. coli K88 challenged piglets. Eighteen weaning piglets were randomly assigned to three groups. Control group and K88 challenged group received a basal diet, and CGMP treated group received the basal diet supplemented with 1% of CGMP powder. The trail lasted for 12 days, K88 was orally administered to the piglets of K88 challenged group and CGMP treated group on days 8–10. The results showed that the diet containing 1% CGMP significantly alleviated the decrease in average daily gain (P  0.05 and barrier permeability damage (P < 0.05, and acute inflammatory response (P < 0.05 induced by E. coli K88 infection. In conclusion, CGMP supplementation in the diet protected the weaning piglets against E. coli K88 infection.

Activation of the Glutamic Acid-Dependent Acid Resistance System in Escherichia coli BL21(DE3) Leads to Increase of the Fatty Acid Biotransformation Activity.

Science.gov (United States)

Woo, Ji-Min; Kim, Ji-Won; Song, Ji-Won; Blank, Lars M; Park, Jin-Byung

The biosynthesis of carboxylic acids including fatty acids from biomass is central in envisaged biorefinery concepts. The productivities are often, however, low due to product toxicity that hamper whole-cell biocatalyst performance. Here, we have investigated factors that influence the tolerance of Escherichia coli to medium chain carboxylic acid (i.e., n-heptanoic acid)-induced stress. The metabolic and genomic responses of E. coli BL21(DE3) and MG1655 grown in the presence of n-heptanoic acid indicated that the GadA/B-based glutamic acid-dependent acid resistance (GDAR) system might be critical for cellular tolerance. The GDAR system, which is responsible for scavenging intracellular protons by catalyzing decarboxylation of glutamic acid, was inactive in E. coli BL21(DE3). Activation of the GDAR system in this strain by overexpressing the rcsB and dsrA genes, of which the gene products are involved in the activation of GadE and RpoS, respectively, resulted in acid tolerance not only to HCl but also to n-heptanoic acid. Furthermore, activation of the GDAR system allowed the recombinant E. coli BL21(DE3) expressing the alcohol dehydrogenase of Micrococcus luteus and the Baeyer-Villiger monooxygenase of Pseudomonas putida to reach 60% greater product concentration in the biotransformation of ricinoleic acid (i.e., 12-hydroxyoctadec-9-enoic acid (1)) into n-heptanoic acid (5) and 11-hydroxyundec-9-enoic acid (4). This study may contribute to engineering E. coli-based biocatalysts for the production of carboxylic acids from renewable biomass.

Activation of the Glutamic Acid-Dependent Acid Resistance System in Escherichia coli BL21(DE3 Leads to Increase of the Fatty Acid Biotransformation Activity.

Directory of Open Access Journals (Sweden)

Ji-Min Woo

Full Text Available The biosynthesis of carboxylic acids including fatty acids from biomass is central in envisaged biorefinery concepts. The productivities are often, however, low due to product toxicity that hamper whole-cell biocatalyst performance. Here, we have investigated factors that influence the tolerance of Escherichia coli to medium chain carboxylic acid (i.e., n-heptanoic acid-induced stress. The metabolic and genomic responses of E. coli BL21(DE3 and MG1655 grown in the presence of n-heptanoic acid indicated that the GadA/B-based glutamic acid-dependent acid resistance (GDAR system might be critical for cellular tolerance. The GDAR system, which is responsible for scavenging intracellular protons by catalyzing decarboxylation of glutamic acid, was inactive in E. coli BL21(DE3. Activation of the GDAR system in this strain by overexpressing the rcsB and dsrA genes, of which the gene products are involved in the activation of GadE and RpoS, respectively, resulted in acid tolerance not only to HCl but also to n-heptanoic acid. Furthermore, activation of the GDAR system allowed the recombinant E. coli BL21(DE3 expressing the alcohol dehydrogenase of Micrococcus luteus and the Baeyer-Villiger monooxygenase of Pseudomonas putida to reach 60% greater product concentration in the biotransformation of ricinoleic acid (i.e., 12-hydroxyoctadec-9-enoic acid (1 into n-heptanoic acid (5 and 11-hydroxyundec-9-enoic acid (4. This study may contribute to engineering E. coli-based biocatalysts for the production of carboxylic acids from renewable biomass.

Transcriptional and Physiological Characterizations of Escherichia coli MG1655 that have been grown under Low Shear Stress Environment for 1000 Generations

Science.gov (United States)

Karouia, Fathi; Tirumalai, Madhan R.; Nelman-Gonzalez, Mayra A.; Sams, Clarence F.; Ott, Mark C.; Pierson, Duane L.; Fofanov, Yuriy; Willson, Richard C.; Fox, George E.

Human space travelers experience a unique environment that affects homeostasis and physio-logic adaptation. One of the important regulatory biology interactions affected by space flight is the alteration of the immune response. As such, the impairment of the immune system may lead to higher risk of bacterial and/or viral infection during human space flight missions. Mi-crobiological contaminants have been a source of concern over the years for NASA and there is evidence to suggest that microbes in space do not behave like they do on Earth. Previ-ous studies have examined the physiological response of bacteria when exposed to short-term microgravity either during spaceflight or in a Low Shear Modeled Microgravity (LSMMG) en-vironment. Exposure to these environments has been found to induce increased resistance to stresses and antibiotics, and in one case increase of virulence. As NASA increases the duration of space flight missions and is starting to envision human presence on the lunar surface and Mars, it becomes legitimate to question the long-term effects of microgravity on bacteria. The effect of long-term exposure to LSMMG on microbial gene expression and physiology in Escherichia coli (E. coli) is being examined using functional genomics, and molecular tech-niques. In previous E. coli short term studies, reproducible changes in transcription were seen but no direct responses to changes in the gravity vector were identified. Instead, absence of shear and a randomized gravity vector appeared to cause local extra-cellular environmental changes, which elicited cellular responses. In order to evaluate the long-term effects of micro-gravity on bacteria, E. coli was grown under simulated microgravity for 1000 generations and gene expression patterns and cellular physiology were analyzed in comparison with short-term exposure. The analysis revealed that the long-term response differed significantly from the short-term exposure and 357 genes were expressed

NCBI nr-aa BLAST: CBRC-STRI-01-1655 [SEVENS

Lifescience Database Archive (English)

Full Text Available CBRC-STRI-01-1655 ref|NP_997055.1| G protein-coupled receptor for asthma susceptibi...rotein coupled receptor 154; AltName: Full=G-protein coupled receptor for asthma susceptibility; AltName: Fu

Effect of the uvr D3 mutation on ultraviolet radiation-induced DNA-repair replication in Escherichia coli K12

International Nuclear Information System (INIS)

Carlson, K.M.; Smith, K.C.

1981-01-01

Ultraviolet-radiation-induced DNA-repair replication was measured in wild-type, polA1, uvrD3, and polA1 uvrD3 strains of Escherichia coli K 12. A large stimulation of repair replication was observed in the uvrD3 strain, compared to the wild-type and polA1 strains. This enhanced repair replication was reduced in the polA1 uvrD3 strain. Therefore, a uvrD3 mutation appears to affect the amount of repair replication performed by DNA polymerase I. In the polA1 strain, there also appears to be an effect of the uvrD3 mutation on the amount of repair replication performed by DNA polymerase III (and/or II). The enhanced repair replication observed for the uvrD3 strains appears to be in response to the enhanced DNA degradation observed for these strains. (orig.)

Cytolysin a expressing E. coli a promising candidate for imageable therapeutic probe

International Nuclear Information System (INIS)

Nguyen, Vu Hong; Phan, Thuy Xuan; Hong, Yeoung Jin; Min, Jung Joon

2007-01-01

Using bacteria for cancer treatment has a long history. Discovery of optical reporter genes consisting of fluorescent and luminescent protein facilitates the monitor of bacteria in vivo, non-invasively and repeatedly. E. coli, the natural enteric bacteria possessing capacity of tumor-targeting ability, seems to be suitable candidate for cancer treatment. In this study, we established the strain light-emitting E. coli for diagnostic purpose and Cytolysin A (Cly A) expressing E. coli for therapeutic purpose. E. coli (MG1655, wild type strain) was transformed plasmid pUC19 carrying lux gene to create the light expressing bacteria and test the tumor targeting-capacity by injecting the bacteria into CT26-tumor bearing mice via tail vein. On the other hand, for therapeutic purpose, plasmid containing Cly A gene, which is encoded for a pore-forming protein toxin, was introduced into E. coli. The toxicity of Cly A was evaluated in vitro by inoculating the bacteria with various cultured cancer cell lines. On the other hand, to test the therapeutic effect, the bacteria were injected intratumorally and intravenously into s.c.CT26-bearing as well as CT26-lung metastasized Balb/c mice. In vivo imaging data showed that the E. coli strains selectively located in the tumor. The in vitro result showed that the number of death cells were significantly higher in the samples containing E. coli expressing Cly A (E. coli Cly A) compared with the samples containing wild type strain. The growth of tumors was repressed in mice injected with either E. coli Cly A (significantly) or wild type E. coli (mildly), while tumors in no treatment group still grew fast. Furthermore, the tumors inoculated with E. coli cly A were necrotized but not with wild type E. coli. In the CT26-lung metastasized mouse model, the life span of mice was elongated when inject E. coli and longer in the group injected with E. coli cly A. Cly A expressing E. coli can become an effective candidate for imageable

Oxygen limitation modulates pH regulation of catabolism and hydrogenases, multidrug transporters, and envelope composition in Escherichia coli K-12

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Radmacher Michael D

2006-10-01

Full Text Available Abstract Background In Escherichia coli, pH regulates genes for amino-acid and sugar catabolism, electron transport, oxidative stress, periplasmic and envelope proteins. Many pH-dependent genes are co-regulated by anaerobiosis, but the overall intersection of pH stress and oxygen limitation has not been investigated. Results The pH dependence of gene expression was analyzed in oxygen-limited cultures of E. coli K-12 strain W3110. E. coli K-12 strain W3110 was cultured in closed tubes containing LBK broth buffered at pH 5.7, pH 7.0, and pH 8.5. Affymetrix array hybridization revealed pH-dependent expression of 1,384 genes and 610 intergenic regions. A core group of 251 genes showed pH responses similar to those in a previous study of cultures grown with aeration. The highly acid-induced gene yagU was shown to be required for extreme-acid resistance (survival at pH 2. Acid also up-regulated fimbriae (fimAC, periplasmic chaperones (hdeAB, cyclopropane fatty acid synthase (cfa, and the "constitutive" Na+/H+ antiporter (nhaB. Base up-regulated core genes for maltodextrin transport (lamB, mal, ATP synthase (atp, and DNA repair (recA, mutL. Other genes showed opposite pH responses with or without aeration, for example ETS components (cyo,nuo, sdh and hydrogenases (hya, hyb, hyc, hyf, hyp. A hypF strain lacking all hydrogenase activity showed loss of extreme-acid resistance. Under oxygen limitation only, acid down-regulated ribosome synthesis (rpl,rpm, rps. Acid up-regulated the catabolism of sugar derivatives whose fermentation minimized acid production (gnd, gnt, srl, and also a cluster of 13 genes in the gadA region. Acid up-regulated drug transporters (mdtEF, mdtL, but down-regulated penicillin-binding proteins (dacACD, mreBC. Intergenic regions containing regulatory sRNAs were up-regulated by acid (ryeA, csrB, gadY, rybC. Conclusion pH regulates a core set of genes independently of oxygen, including yagU, fimbriae, periplasmic chaperones, and nha

The detection of K88, K99 fimbrial antigen and enterotoxin genes of Escherichia coli isolated from piglets and calves with diarrhoea in Indonesia

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Supar

1996-03-01

Full Text Available Enterotoxigenic Escherichia coli (ETEC strains cause diarrhoeal disease in piglets and calves in Indonesia. These strains possess two virulence factors namely attachment and enterotoxin antigens . These factors could be detected phenotypically and genetically. Haemolytic Escherichia coli (E coli isolates possessing K88 fimbrial antigen associated with 0-group 108 and 149. They were positive for K88 gene and demonstrated their ability to produce heat labile enterotoxin (LT and genetically were all positive for LT gene . Seventeen isolates ofE coli K88 which associated with 0-group 149 were positive forSTb gene, other O-serotypes were negative . Ten isolates of Ecoli K88 which associated with 0-group 108 possessed K88, K99, LT and STa genes, but negative for STb gene . However, phenotypically the K99 antigen and STa toxin were not expressed under laboratory conditions, the reason was not well understood . E. coli K99 strains isolated from calves wit h diarrhoea were all associated with 0-group 9 and produced STa toxin when tested by suckling mousse bioassay. The E. coli K99 calf isolates were all hybridized with K99 and STa gene only . It is likely that K99 gene is associated with STa gene . The DNA hybridization technique is more convenience to be used for confirmation diagnosis of colibacillosis, however, not all veterinary laboratories could perform these tests .

The Capsule Supports Survival but Not Traversal of Escherichia coli K1 across the Blood-Brain Barrier

OpenAIRE

Hoffman, Jill A.; Wass, Carol; Stins, Monique F.; Kim, Kwang Sik

1999-01-01

The vast majority of cases of gram-negative meningitis in neonates are caused by K1-encapsulated Escherichia coli. The role of the K1 capsule in the pathogenesis of E. coli meningitis was examined with an in vivo model of experimental hematogenous E. coli K1 meningitis and an in vitro model of the blood-brain barrier. Bacteremia was induced in neonatal rats with the E. coli K1 strain C5 (O18:K1) or its K1− derivative, C5ME. Subsequently, blood and cerebrospinal fluid (CSF) were obtained for c...

Expression and purification of functional human mu opioid receptor from E.coli.

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Yanbin Ma

Full Text Available N-terminally his-tagged human mu opioid receptor, a G protein-coupled receptor was produced in E.coli employing synthetic codon-usage optimized constructs. The receptor was expressed in inclusion bodies and membrane-inserted in different E.coli strains. By optimizing the expression conditions the expression level for the membrane-integrated receptor was raised to 0.3-0.5 mg per liter of culture. Milligram quantities of receptor could be enriched by affinity chromatography from IPTG induced cultures grown at 18°C. By size exclusion chromatography the protein fraction with the fraction of alpha-helical secondary structure expected for a 7-TM receptor was isolated, by CD-spectroscopy an alpha-helical content of ca. 45% was found for protein solubilised in the detergent Fos-12. Receptor in Fos-12 micelles was shown to bind endomorphin-1 with a K(D of 61 nM. A final yield of 0.17 mg functional protein per liter of culture was obtained.

Genomic Comparison of Escherichia coli K1 Strains Isolated from the Cerebrospinal Fluid of Patients with Meningitis †

OpenAIRE

Yao, Yufeng; Xie, Yi; Kim, Kwang Sik

2006-01-01

Escherichia coli is a major cause of enteric/diarrheal diseases, urinary tract infections, and sepsis. E. coli K1 is the leading gram-negative organism causing neonatal meningitis, but the microbial basis of E. coli K1 meningitis is incompletely understood. Here we employed comparative genomic hybridization to investigate 11 strains of E. coli K1 isolated from the cerebrospinal fluid (CSF) of patients with meningitis. These 11 strains cover the majority of common O serotypes in E. coli K1 iso...

Genetic control of near-UV (300-400 nm) sensitivity independent of the recA gene in strains of Escherichia coli K12

International Nuclear Information System (INIS)

Tuveson, R.W.; Jonas, R.B.

1979-01-01

Stationary cells of isogenic pairs of Escherichia coli K12 strains presumably differing only in the recA function, were inactivated with near-UV (300-400 nm) radiation. Based on near-UV inactivation kinetics, the strains can be divided into two discrete categories in which near-UV sensitivity does not necessarily correlate with far-UV sensitivity conferred by two different recA alleles. Lack of overlap between near-UV and far-UV (recA) sensitivity can be explained by assuming that a different chromosomal gene (nur) controls near-UV sensitivity. Support for this hypothesis came from a mating experiment in which four selected recombinants, isogenic with respect to auxotrophic markers, were identified exhibiting all four possible combinations of far-UV (recA1 vs recA + ) and near-UV sensitivity (nur vs nur + ). Transduction with phase P1 showed that introduction of the recA1 allele into a recA + recipient did not affect the near-UV sensitivity of the recipient. Additional matings together with transduction experiments suggested that the nur gene is located at a position on the E. coli linkage map clearly separable from recA (minute 58). (author)

Monitoring bacteriolytic therapy of salmonella typhimurium with optical imaging system

International Nuclear Information System (INIS)

Kim, Sun A; Min, Jung Joon; Moon, Sung Min; Kim, Hyun Ju; Kim, Sung Mi; Song, Ho Cheon; Choy, Hyon E.; Bom, Hee Seung

2005-01-01

Systemically administrated Salmonella has been studied for targeting tumor and developed as an anticancer agent. In Salmonella, because msbB gene plays role in the terminal myristoylation of lipid A and induces tumor necrosis factor a (TNF-a) -mediated septic shock, Salmonella msbB mutant strain is safe and useful for tumor-targeting therapy. Here we report that Salmonella msbB mutant strain induce onco lysis after intravenous injection in tumor bearing mice. The CT26 mouse colon cancer cells were stably transfected with firefly luciferase gene and subcutaneously implantated in Balb/C mice. After establishing subcutaneous tumor mass, we intravenously injected 1x108 cfu Salmonella msbB mutant strain or MG1655 E coli strain. Not only tumor size but also total photon flux from the tumor mass were monitored. everyday and compared among experimental groups (No treatment, Salmonella treatment, E. coli MG1655 treatment group). After intraperitoneal injection of D-Iuciferin (3 mg/animal), in vivo optical imaging for firefly luciferase was performed using cooled CCD camera. Imaging signal from Salmonella injected group were significantly lower than that of no treatment or E. coli treatment group on day 2 after injection. On day 4 after injection, imaging signal of salmonella-injected group was 43.8 or 20.7 times lower than that of no treatment or E. coli treatment group, respectively (no treatment: 2.78E+07 p/s/cm 2 /sr, Salmonella treatment: 6.35E+05 p/s/cm 2 /sr, E. coli treatment: 1.29E+07 p/s/cm 2 /sr, P<0.05). However. when we injected E. coli MG1655 into tumor bearing mice, the intensity of imaging signal was not different from no treatment group. These findings suggest that Salmonella msbB mutant strain retains its tumor-targeting properties and have therapeutical effect. Bioluminescent tumor bearing animal model was useful for assessing tumor viability after bacteriolytic therapy using Salmonella

Modular Engineering of l-Tyrosine Production in Escherichia coli

Science.gov (United States)

Juminaga, Darmawi; Baidoo, Edward E. K.; Redding-Johanson, Alyssa M.; Batth, Tanveer S.; Burd, Helcio; Mukhopadhyay, Aindrila; Petzold, Christopher J.

2012-01-01

Efficient biosynthesis of l-tyrosine from glucose is necessary to make biological production economically viable. To this end, we designed and constructed a modular biosynthetic pathway for l-tyrosine production in E. coli MG1655 by encoding the enzymes for converting erythrose-4-phosphate (E4P) and phosphoenolpyruvate (PEP) to l-tyrosine on two plasmids. Rational engineering to improve l-tyrosine production and to identify pathway bottlenecks was directed by targeted proteomics and metabolite profiling. The bottlenecks in the pathway were relieved by modifications in plasmid copy numbers, promoter strength, gene codon usage, and the placement of genes in operons. One major bottleneck was due to the bifunctional activities of quinate/shikimate dehydrogenase (YdiB), which caused accumulation of the intermediates dehydroquinate (DHQ) and dehydroshikimate (DHS) and the side product quinate; this bottleneck was relieved by replacing YdiB with its paralog AroE, resulting in the production of over 700 mg/liter of shikimate. Another bottleneck in shikimate production, due to low expression of the dehydroquinate synthase (AroB), was alleviated by optimizing the first 15 codons of the gene. Shikimate conversion to l-tyrosine was improved by replacing the shikimate kinase AroK with its isozyme, AroL, which effectively consumed all intermediates formed in the first half of the pathway. Guided by the protein and metabolite measurements, the best producer, consisting of two medium-copy-number, dual-operon plasmids, was optimized to produce >2 g/liter l-tyrosine at 80% of the theoretical yield. This work demonstrates the utility of targeted proteomics and metabolite profiling in pathway construction and optimization, which should be applicable to other metabolic pathways. PMID:22020510

The Escherichia coli argW-dsdCXA genetic island is highly variable, and E. coli K1 strains commonly possess two copies of dsdCXA.

Science.gov (United States)

Moritz, Rebecca L; Welch, Rodney A

2006-11-01

The genome sequences of Escherichia coli pathotypes reveal extensive genetic variability in the argW-dsdCXA island. Interestingly, the archetype E. coli K1 neonatal meningitis strain, strain RS218, has two copies of the dsdCXA genes for d-serine utilization at the argW and leuX islands. Because the human brain contains d-serine, an epidemiological study emphasizing K1 isolates surveyed the dsdCXA copy number and function. Forty of 41 (97.5%) independent E. coli K1 isolates could utilize d-serine. Southern blot hybridization revealed physical variability within the argW-dsdC region, even among 22 E. coli O18:K1:H7 isolates. In addition, 30 of 41 K1 strains, including 21 of 22 O18:K1:H7 isolates, had two dsdCXA loci. Mutational analysis indicated that each of the dsdA genes is functional in a rifampin-resistant mutant of RS218, mutant E44. The high percentage of K1 strains that can use d-serine is in striking contrast to our previous observation that only 4 of 74 (5%) isolates in the diarrheagenic E. coli (DEC) collection have this activity. The genome sequence of diarrheagenic E. coli isolates indicates that the csrRAKB genes for sucrose utilization are often substituted for dsdC and a portion of dsdX present at the argW-dsdCXA island of extraintestinal isolates. Among DEC isolates there is a reciprocal pattern of sucrose fermentation versus d-serine utilization. The ability to use d-serine is a trait strongly selected for among E. coli K1 strains, which have the ability to infect a wide range of extraintestinal sites. Conversely, diarrheagenic E. coli pathotypes appear to have substituted sucrose for d-serine as a potential nutrient.

Crystallization Behavior of A Bulk Amorphous Mg62Cu26Y12 Alloy

Science.gov (United States)

Wu, Shyue-Sheng; Chin, Tsung-Shune; Su, Kuo-Chang

1994-07-01

The crystallization temperature, the associated activation energy and the crystallized structure of a bulk amorphous Mg62Cu26Y12 alloy with a diameter of 2.5 mm were studied. It possesses a one-step crystallization behavior. The crystallization reaction was found to be represented by: AM(MG62Cu26Y12)→Mg2Cu+MgY+CuY+Mg, ( Tx=188°C, Eac=134 kJ/mol) where AM represents the amorphous state, T x the crystallization temperature at an infinitesimal heating rate, and E ac the associated activation energy. The amount of crystalline phases were found to be Mg2Cu:MgY:CuY=76:17:7. The Mg phase is identifiable only by high resolution electron microscopy, not by X-ray diffraction. The crystallization leads to a sharp rise in electrical resistivity which is reversed to those of iron-based amorphous alloys.

Cell survival, UV-reactivation and induction of prophage lambda in Escherichia coli K12 overproducing RecA protein

International Nuclear Information System (INIS)

Quillardet, P.; Moreau, P.L.; Devoret, R.; Ginsburg, H.; Mount, D.W.

1982-01-01

The effect of the cellular level of RecA protein on the ability of E. coli K12 bacteria to (I) survive UV-irradiation (II) promote UV-reactivation of UV-damaged phage lambda (III) induce prophage lambda was determined in bacterial mutants with discrete increasing levels of RecA protein. The various levels of RecA protein were obtained by combining lexA and recA alleles. Except for the double mutant lexA3 recAo98, whose repair ability was 25% less than that observed in wild type bacteria, bacterial survival was proportional to the level of ReCA protein measured after 90 min of incubation. In lexA3 recAo98 bacteria, RecA protein, at a constitutive high basal level, failed to compensate totally for the lack of LexA repressor cleavage; UV-reactivation of UV-damaged phage lambda was not restored; yet, prophage lambda was induced with 35% efficiency. Efficient UV-induction of prophage lambda is linked to the induction of lexA-controlled host processes that repair the UV-damaged prophage. (orig.)

Genes Required for Growth at High Hydrostatic Pressure in Escherichia coli K-12 Identified by Genome-Wide Screening

Science.gov (United States)

Black, S. Lucas; Dawson, Angela; Ward, F. Bruce; Allen, Rosalind J.

2013-01-01

Despite the fact that much of the global microbial biosphere is believed to exist in high pressure environments, the effects of hydrostatic pressure on microbial physiology remain poorly understood. We use a genome-wide screening approach, combined with a novel high-throughput high-pressure cell culture method, to investigate the effects of hydrostatic pressure on microbial physiology in vivo. The Keio collection of single-gene deletion mutants in Escherichia coli K-12 was screened for growth at a range of pressures from 0.1 MPa to 60 MPa. This led to the identification of 6 genes, rodZ, holC, priA, dnaT, dedD and tatC, whose products were required for growth at 30 MPa and a further 3 genes, tolB, rffT and iscS, whose products were required for growth at 40 MPa. Our results support the view that the effects of pressure on cell physiology are pleiotropic, with DNA replication, cell division, the cytoskeleton and cell envelope physiology all being potential failure points for cell physiology during growth at elevated pressure. PMID:24040140

Use of Adaptive Laboratory Evolution To Discover Key Mutations Enabling Rapid Growth of Escherichia coli K-12 MG1655 on Glucose Minimal Medium

DEFF Research Database (Denmark)

LaCroix, Ryan A.; Sandberg, Troy E.; O'Brien, Edward J.

2015-01-01

Adaptive laboratory evolution (ALE) has emerged as an effective tool for scientific discovery and addressing biotechnological needs. Much of ALE's utility is derived from reproducibly obtained fitness increases. Identifying causal genetic changes and their combinatorial effects is challenging...

Deletion mutants of the Escherichia coli K-12 mannitol permease: dissection of transport-phosphorylation, phospho-exchange, and mannitol-binding activities.

Science.gov (United States)

Grisafi, P L; Scholle, A; Sugiyama, J; Briggs, C; Jacobson, G R; Lengeler, J W

1989-05-01

We have constructed a series of deletion mutations of the cloned Escherichia coli K-12 mtlA gene, which encodes the mannitol-specific enzyme II of the phosphoenolpyruvate (PEP)-dependent carbohydrate phosphotransferase system. This membrane-bound permease consists of 637 amino acid residues and is responsible for the concomitant transport and phosphorylation of D-mannitol in E. coli. Deletions into the 3' end of mtlA were constructed by exonuclease III digestion. Restriction mapping of the resultant plasmids identified several classes of deletions that lacked approximately 5% to more than 75% of the gene. Immunoblotting experiments revealed that many of these plasmids expressed proteins within the size range predicted by the restriction analyses, and all of these proteins were membrane localized, which demonstrated that none of the C-terminal half of the permease is required for membrane insertion. Functional analyses of the deletion proteins, expressed in an E. coli strain deleted for the chromosomal copy of mtlA, showed that all but one of the strains containing confirmed deletions were inactive in transport and PEP-dependent phosphorylation of mannitol, but deletions removing up to at least 117 amino acid residues from the C terminus of the permease were still active in catalyzing phospho exchange between mannitol 1-phosphate and mannitol. A deletion protein that lacked 240 residues from the C terminus of the permease was inactive in phospho exchange but still bound mannitol with high affinity. These experiments localize sites important for transport and PEP-dependent phosphorylation to the extreme C terminus of the mannitol permease, sites important for phospho exchange to between residues 377 and 519, and sites necessary for mannitol binding to the N-terminal 60% of the molecule. The results are discussed with respect to the fact that the mannitol permease consists of structurally independent N- and C-terminal domains.

Research on relation of HRS/IRR and anti-oxidases in E.coli implanted by 30 keV N+

International Nuclear Information System (INIS)

Yang Tianyou; Wang Tiegu; Tian Jing; Chang Jingling; Qing Guangyong

2010-01-01

The inducement of hyperradiosensitivity/increased radioresistance (HRS/IRR) and activities of anti-oxidases in Escherichia coli implanted by 30 keV N + and relationship between them were investigated. The results showed the change trend of the survival rate and the activity of superoxide dismutase (SOD) and Catalas(CAT) of E. coli K12 was consistent. The activity of SOD and CAT of E.coli were lower, less than 10 and 8 U/mg, when the HRS was induced at 0 - 5 x 10 14 N + /cm 2 doses. And the activity of SOD and CAT of E. coli increased and reached highest rate, were 58 and 26 U/mg respectively, when the IRR was induced at 5 x 10 14 - 10 x 10 14 N + /cm 2 doses. It could be concluded that the change of SOD and CAT activity was related with the HRS/IRR. (authors)

Excited states in 22Mg via the 12C(12C,2n)22Mg reaction

International Nuclear Information System (INIS)

Jewett, Cybele; Baktash, Cyrus; Bardayan, Daniel W.; Blackmon, Jeff C.; Chipps, K.; Galindo-Uribarri, Alfredo; Greife, U.; Gross, Carl J.; Jones, K. L.; Liang, Junjien; Livesay, Jake; Kozub, R. L.; Nesaraja, Caroline D; Radford, David C.; Sarazin, F.; Smith, Michael Scott; Thomas, J. S.; Yu, Chang-Hong

2007-01-01

The 12C(12C, 2n)22Mg reaction was measured with the CLARION array and the RMS separator at the Holifield Facility of Oak Ridge National Laboratory. This experiment was performed to gather more information on the excited states in 22Mg, which might be of relevance to recent radioactive ion beam measurements of the astrophysically important 21Na(p,γ)22Mg reaction. The results are compared to direct measurements, transfer experiments and a competing experiment performed with Gammasphere

A survey about prophage induction ability in Escherichia coli K-12(λ by ethnic medicinal plants of Kohgiluyeh va Boyerahmad, Iran

Directory of Open Access Journals (Sweden)

M. Hamzeloo-Moghadam

2014-10-01

Full Text Available Background and objectives: There is a growing trend towards investigating natural products as sources of compounds with biological effects and many researches have been carried out in order to find effective medications against many diseases. Cancer is no exception and studies focusing on evaluating the effects of different materials on DNA, give valuable information in cancer researches and carcinogenicity studies; thus the present study was focused on evaluating the impact of medicinal plants from  Kohgiluyeh va Boyerahmad province, Iran on DNA. Methods: Thirty five plant species collected have been investigated for prophage induction ability in Escherichia coli K-12(λthroughinductest. Results:The assay demonstrated that 8 plants were able to affect DNA. Conclusion: The results confirm the role of natural resources for biologic effects and what’s more, potential drug candidates in new drug discovery.

An Accurate Mass Determination for Kepler-1655b, a Moderately Irradiated World with a Significant Volatile Envelope

Science.gov (United States)

Haywood, Raphaëlle D.; Vanderburg, Andrew; Mortier, Annelies; Giles, Helen A. C.; López-Morales, Mercedes; Lopez, Eric D.; Malavolta, Luca; Charbonneau, David; Collier Cameron, Andrew; Coughlin, Jeffrey L.; Dressing, Courtney D.; Nava, Chantanelle; Latham, David W.; Dumusque, Xavier; Lovis, Christophe; Molinari, Emilio; Pepe, Francesco; Sozzetti, Alessandro; Udry, Stéphane; Bouchy, François; Johnson, John A.; Mayor, Michel; Micela, Giusi; Phillips, David; Piotto, Giampaolo; Rice, Ken; Sasselov, Dimitar; Ségransan, Damien; Watson, Chris; Affer, Laura; Bonomo, Aldo S.; Buchhave, Lars A.; Ciardi, David R.; Fiorenzano, Aldo F.; Harutyunyan, Avet

2018-05-01

We present the confirmation of a small, moderately irradiated (F = 155 ± 7 F ⊕) Neptune with a substantial gas envelope in a P = 11.8728787 ± 0.0000085 day orbit about a quiet, Sun-like G0V star Kepler-1655. Based on our analysis of the Kepler light curve, we determined Kepler-1655b’s radius to be 2.213 ± 0.082 R ⊕. We acquired 95 high-resolution spectra with Telescopio Nazionale Galileo/HARPS-N, enabling us to characterize the host star and determine an accurate mass for Kepler-1655b of 5.0{+/- }2.83.1 {M}\\oplus via Gaussian-process regression. Our mass determination excludes an Earth-like composition with 98% confidence. Kepler-1655b falls on the upper edge of the evaporation valley, in the relatively sparsely occupied transition region between rocky and gas-rich planets. It is therefore part of a population of planets that we should actively seek to characterize further.

The K1 capsule is the critical determinant in the development of Escherichia coli meningitis in the rat.

OpenAIRE

Kim, K S; Itabashi, H; Gemski, P; Sadoff, J; Warren, R L; Cross, A S

1992-01-01

Although Escherichia coli strains possessing the K1 capsule are predominant among isolates from neonatal E. coli meningitis and most of these K1 isolates are associated with a limited number of 0 lipopolysaccharide (LPS) types, the basis of this association of K1 and certain 0 antigens with neonatal E. coli meningitis is not clear. The present study examined in experimental E. coli bacteremia and meningitis in newborn and adult rats whether or not the K1 capsule and/or O-LPS antigen are criti...

Inhibition of factor-dependent transcription termination in ...

Indian Academy of Sciences (India)

Inhibition of factor-dependent transcription termination in Escherichia coli might relieve xenogene silencing by abrogating. H-NS-DNA interactions in vivo. DEEPTI CHANDRAPRAKASH and ASWIN SAI NARAIN SESHASAYEE. Chromatin immunoprecipitation. MG1655 hns::3xFLAG cells were grown in liquid LB me-.

Chromosomal features of Escherichia coli serotype O2:K2, an avian pathogenic E. coli.

Science.gov (United States)

Jørgensen, Steffen L; Kudirkiene, Egle; Li, Lili; Christensen, Jens P; Olsen, John E; Nolan, Lisa; Olsen, Rikke H

2017-01-01

Escherichia coli causing infection outside the gastrointestinal system are referred to as extra-intestinal pathogenic E. coli. Avian pathogenic E. coli is a subgroup of extra-intestinal pathogenic E. coli and infections due to avian pathogenic E. coli have major impact on poultry production economy and welfare worldwide. An almost defining characteristic of avian pathogenic E. coli is the carriage of plasmids, which may encode virulence factors and antibiotic resistance determinates. For the same reason, plasmids of avian pathogenic E. coli have been intensively studied. However, genes encoded by the chromosome may also be important for disease manifestation and antimicrobial resistance. For the E. coli strain APEC_O2 the plasmids have been sequenced and analyzed in several studies, and E. coli APEC_O2 may therefore serve as a reference strain in future studies. Here we describe the chromosomal features of E. coli APEC_O2. E. coli APEC_O2 is a sequence type ST135, has a chromosome of 4,908,820 bp (plasmid removed), comprising 4672 protein-coding genes, 110 RNA genes, and 156 pseudogenes, with an average G + C content of 50.69%. We identified 82 insertion sequences as well as 4672 protein coding sequences, 12 predicated genomic islands, three prophage-related sequences, and two clustered regularly interspaced short palindromic repeats regions on the chromosome, suggesting the possible occurrence of horizontal gene transfer in this strain. The wildtype strain of E. coli APEC_O2 is resistant towards multiple antimicrobials, however, no (complete) antibiotic resistance genes were present on the chromosome, but a number of genes associated with extra-intestinal disease were identified. Together, the information provided here on E. coli APEC_O2 will assist in future studies of avian pathogenic E. coli strains, in particular regarding strain of E. coli APEC_O2, and aid in the general understanding of the pathogenesis of avian pathogenic E. coli .

E. coli O124 K72 alters the intestinal barrier and the tight junctions proteins of guinea pig intestine.

Science.gov (United States)

Ren, Xiaomeng; Zhu, Yanyan; Gamallat, Yaser; Ma, Shenhao; Chiwala, Gift; Meyiah, Abdo; Xin, Yi

2017-10-01

Our research group previously isolated and identified a strain of pathogenic Escherichia coli from clinical samples called E. coli O124 K72. The present study was aimed at determining the potential effects of E. coli O124 K72 on intestinal barrier functions and structural proteins integrity in guinea pig. Guinea pigs were grouped into three groups; control (CG); E. coli O124 K72 (E. coli); and probiotics Lactobacillus rhamnosus (LGG). Initially, we create intestinal dysbiosis by giving all animals Levofloxacin for 10days, but the control group (CG) received the same volume of saline. Then, the animals received either E. coli O124 K72 (E. coli) or Lactobacillus rhamnosus (LGG) according to their assigned group. E. coli O124 K72 treatment significantly affected colon morphology and distorted intestinal barrier function by up-regulating Claudin2 and down-regulating Occludin. In addition, E. coli upregulated the mRNA expression of MUC1, MUC2, MUC13 and MUC15. Furthermore, suspected tumor was found in the E. coli treated animals. Our results suggested that E. coli O124 K72 strain has adverse effects on intestinal barrier functions and is capable of altering integrity of structural proteins in guinea pig model while at same time it may have a role in colon carcinogenesis. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

DNA synthesis and degradation in UV-irradiated toluene treated cells of E. coli K12: the role of polynucleotide ligase

International Nuclear Information System (INIS)

Strike, P.

1977-01-01

Toluene treated cells have been used to study the processes of DNA synthesis and DNA degradation in ultra-violet irradiated Escherichia coli K12. Synthesis and degradation are both shown to occur extensively if polynucleotide ligase is inhibited, and to occur to a much lesser extent if ligase activity is optimal. Extensive UV-induced DNA synthesis in toluene-treated cells requires ATP for the initial incision step, and DNA polymerase I. Extensive degradation also depends on the early ATP-dependent incision step, and the subsequent degradation shows a partial requirement for ATP. Curtailment of degradation by ligase requires DNA polymerase activity, but is not dependent upon DNA polymerase I. Apparently this process can be carried out with equal facility by either DNA polymerase II or polymerase III. These observations suggest that extensive DNA polymerase I-dependent repair synthesis and extensive DNA degradation are facets of two divergent pathways of excision repair, both of which depend upon the early uvrABC determined ATP-dependent incision step. (orig.) [de

GENETIC CONTROL OF RESTRICTION AND MODIFICATION IN ESCHERICHIA COLI1

Science.gov (United States)

Boyer, Herbert

1964-01-01

Boyer, Herbert (Yale University, New Haven, Conn.). Genetic control of restriction and modification in Escherichia coli. J. Bacteriol. 88:1652–1660. 1964.—Bacterial crosses with K-12 strains of Escherichia coli as Hfr donors (Hfr Hayes, Hfr Cavalli, and Hfr P4X-6) and B/r strains of E. coli as F− recipients were found to differ from crosses between K-12 Hfr donors and K-12 F− recipients in two ways: (i) recombinants (leu, pro, lac, and gal) did not appear at discrete time intervals but did appear simultaneously 30 min after matings were initiated, and (ii) the linkage of unselected markers to selected markers was reduced. Integration of a genetic region linked to the threonine locus of K-12 into the B/r genome resulted in a hybrid which no longer gave anomalous results in conjugation experiments. A similar region of the B strain was introduced into the K-12 strain, which then behaved as a typical B F− recipient. These observations are interpreted as the manifestation of host-controlled modification and restriction on the E. coli chromosome. This was verified by experiments on the restriction and modification of the bacteriophage lambda, F-lac, F-gal, and sex-factor, F1. It was found that the genetic region that controlled the mating responses of the K-12 and B/r strains also controlled the modification and restriction properties of these two strains. The genes responsible for the restricting and modifying properties of the K-12 and B strains of E. coli were found to be allelic, linked to each other, and linked to the threonine locus. PMID:14240953

Synthesis and crystal structure of MgB12

International Nuclear Information System (INIS)

Adasch, Volker; Hess, Kai-Uwe; Ludwig, Thilo; Vojteer, Natascha; Hillebrecht, Harald

2006-01-01

Single crystals of MgB 12 were synthesized from the elements in a Mg/Cu melt at 1600deg. C. MgB 12 crystallizes orthorhombic in space group Pnma with a=16.632(3)A, b=17.803(4)A and c=10.396(2)A. The crystal structure (Z=30, 5796 reflections, 510 variables, R 1 (F)=0.049, wR 2 (I)=0.134) consists of a three dimensional net of B 12 icosahedra and B 21 units in a ratio 2:1. The B 21 units are observed for the first time in a solid compound. Mg is on positions with partial occupation. The summation reveals the composition MgB 12.35 or Mg 0.97 B 12 , respectively. This is in good agreement with the value of MgB 11.25 as expected by electronic reasons to stabilize the boron polyhedra B 12 2- and B 21 4-

Escherichia coli K-12 survives anaerobic exposure at pH 2 without RpoS, Gad, or hydrogenases, but shows sensitivity to autoclaved broth products.

Directory of Open Access Journals (Sweden)

Daniel P Riggins

Full Text Available Escherichia coli and other enteric bacteria survive exposure to extreme acid (pH 2 or lower in gastric fluid. Aerated cultures survive via regulons expressing glutamate decarboxylase (Gad, activated by RpoS, cyclopropane fatty acid synthase (Cfa and others. But extreme-acid survival is rarely tested under low oxygen, a condition found in the stomach and the intestinal tract. We observed survival of E. coli K-12 W3110 at pH 1.2-pH 2.0, conducting all manipulations (overnight culture at pH 5.5, extreme-acid exposure, dilution and plating in a glove box excluding oxygen (10% H2, 5% CO2, balance N2. With dissolved O2 concentrations maintained below 6 µM, survival at pH 2 required Cfa but did not require GadC, RpoS, or hydrogenases. Extreme-acid survival in broth (containing tryptone and yeast extract was diminished in media that had been autoclaved compared to media that had been filtered. The effect of autoclaved media on extreme-acid survival was most pronounced when oxygen was excluded. Exposure to H2O2 during extreme-acid treatment increased the death rate slightly for W3110 and to a greater extent for the rpoS deletion strain. Survival at pH 2 was increased in strains lacking the anaerobic regulator fnr. During anaerobic growth at pH 5.5, strains deleted for fnr showed enhanced transcription of acid-survival genes gadB, cfa, and hdeA, as well as catalase (katE. We show that E. coli cultured under oxygen exclusion (<6 µM O2 requires mechanisms different from those of aerated cultures. Extreme acid survival is more sensitive to autoclave products under oxygen exclusion.

Acid Evolution of Escherichia coli K-12 Eliminates Amino Acid Decarboxylases and Reregulates Catabolism.

Science.gov (United States)

He, Amanda; Penix, Stephanie R; Basting, Preston J; Griffith, Jessie M; Creamer, Kaitlin E; Camperchioli, Dominic; Clark, Michelle W; Gonzales, Alexandra S; Chávez Erazo, Jorge Sebastian; George, Nadja S; Bhagwat, Arvind A; Slonczewski, Joan L

2017-06-15

Acid-adapted strains of Escherichia coli K-12 W3110 were obtained by serial culture in medium buffered at pH 4.6 (M. M. Harden, A. He, K. Creamer, M. W. Clark, I. Hamdallah, K. A. Martinez, R. L. Kresslein, S. P. Bush, and J. L. Slonczewski, Appl Environ Microbiol 81:1932-1941, 2015, https://doi.org/10.1128/AEM.03494-14). Revised genomic analysis of these strains revealed insertion sequence (IS)-driven insertions and deletions that knocked out regulators CadC (acid induction of lysine decarboxylase), GadX (acid induction of glutamate decarboxylase), and FNR (anaerobic regulator). Each acid-evolved strain showed loss of one or more amino acid decarboxylase systems, which normally help neutralize external acid (pH 5 to 6) and increase survival in extreme acid (pH 2). Strains from populations B11, H9, and F11 had an IS 5 insertion or IS-mediated deletion in cadC , while population B11 had a point mutation affecting the arginine activator adiY The cadC and adiY mutants failed to neutralize acid in the presence of exogenous lysine or arginine. In strain B11-1, reversion of an rpoC (RNA polymerase) mutation partly restored arginine-dependent neutralization. All eight strains showed deletion or downregulation of the Gad acid fitness island. Strains with the Gad deletion lost the ability to produce GABA (gamma-aminobutyric acid) and failed to survive extreme acid. Transcriptome sequencing (RNA-seq) of strain B11-1 showed upregulated genes for catabolism of diverse substrates but downregulated acid stress genes (the biofilm regulator ariR , yhiM , and Gad). Other strains showed downregulation of H 2 consumption mediated by hydrogenases ( hya and hyb ) which release acid. Strains F9-2 and F9-3 had a deletion of fnr and showed downregulation of FNR-dependent genes ( dmsABC , frdABCD , hybABO , nikABCDE , and nrfAC ). Overall, strains that had evolved in buffered acid showed loss or downregulation of systems that neutralize unbuffered acid and showed altered regulation of

Three-decade epidemiological analysis of Escherichia coli O15:K52:H1

DEFF Research Database (Denmark)

Olesen, Bente; Scheutz, Flemming; Menard, Megan

2009-01-01

The successful Escherichia coli O15:K52:H1 clonal group provides a case study for the emergence of multiresistant clonal groups of Enterobacteriaceae generally. Accordingly, we tested the hypotheses that, over time, the O15:K52:H1 clonal group has become increasingly (i) virulent and (ii) resistant...... to antibiotics. One hundred archived international E. coli O15:K52:[H1] clinical isolates from 100 unique patients (1975 to 2006) were characterized for diverse phenotypic and molecular traits. All 100 isolates derived from phylogenetic group D and, presumptively, sequence type ST393. They uniformly carried...

[Changes of biological behavioral of E. coli K1 after ppk1 gene deletion].

Science.gov (United States)

Peng, Liang; Pan, Jiayun; Luo, Su; Yang, Zhenghui; Huang, Mufang; Cao, Hong

2014-06-01

To study the changes in biological behaviors of meningitis E. coli K1 strain E44 after deletion of polyphosphate kinase 1 (ppk1) gene and explore the role of ppk1 in the pathogenesis of E. coli K1-induced meningitis. The wild-type strain E. coli K1 and ppk1 deletion mutant were exposed to heat at 56 degrees celsius; for 6 min, and their survival rates were determined. The adhesion and invasion of the bacteria to human brain microvascular endothelial cells (HBMECs) were observed using electron microscopy and quantitative tests. HBMECs were co-incubated with wild-type strain or ppk1 deletion mutant, and the cytoskeleton rearrangement was observed under laser scanning confocal microscope. The survival rate of the ppk1 deletion mutant was significantly lower than that of the wild-type strain after heat exposure. The ppk1 deletion mutant also showed lowered cell adhesion and invasion abilities and weakened ability to induce cytoskeleton rearrangement in HBMECs. ppk1 gene is important for E.coli K1 for heat resistance, cell adhesion and invasion, and for inducing cytoskeletal rearrangement in HBMECs.

An Investigative Study on the Effect of Silver Nanoparticles on E.Coli K12 in Various Sodium Chloride Concentrations

Science.gov (United States)

Levard, C.; Mitra, S.; Badireddy, A.; Jew, A. D.; Brown, G. E.

2011-12-01

Engineered nanomaterials have had an increasing presence in consumer products. Consequently, their release in wastewater systems is believed to pose a viable threat to the environment. NPs are used for drug delivery devices, imaging agents, and consumer products like sunscreens, paints, and cosmetics. Among the major types of manufactured nanoparticles, silver nanoparticles (Ag-NPs) are currently the most widely used in the nanotechnology industry. These particles have unique antibacterial, antiviral, and antifungal properties and as a result, there is a growing concern about the environmental impact of released Ag nanoparticles, particularly their unintended impact on organisms and ecosystems. Even though the toxicity of Ag-NPs has been extensively studied, the environmental transformations that the Ag-NPs may experience once released in the environment have not been considered. These transformations can readily impact their properties and therefore their behavior in terms of reactivity and toxicity. For example, it is known that silver strongly react with Chloride (Cl), which is ubiquitous in natural waters. At a low Cl/Ag ratio, Cl may precipitate on the surface and partly inhibit dissolution. On the contrary, for a high Cl/Ag ratio, chloride may enhance dissolution and therefore toxicity since soluble Ag species are a main source of toxicity. In this context, the focus of this study is on understanding the toxicity of coated Ag-NPs at various concentrations (1ppb-100ppm) on E.Coli (K12) in deionized water and various sodium chloride concentrations that mimic natural conditions (.5, .1 and .01 M NaCl). Ag+ ions (100 ppm-1ppb) were also tested in these salt concentrations as a control. Samples were inoculated in bacteria and incubated for 24 hours. Based on this test, we inferred that increasing concentrations of Ag+ ions/ AgNps played a role in the inhibition of growth of E.Coli K12. A live-dead staining test has shown the correlation between inhibition of

Effects of chloramphenicol and caffeine on postreplication repair in uvrA-umuC- and uvrA-recF- strains of Escherichia coli K-12

International Nuclear Information System (INIS)

Kato, T.

1977-01-01

Postreplication repair and its inhibition by chloramphenicol and caffeine, as seen in alkaline sucrose gradients, were compared between a UV nonmutable strain uvrA - umuC - and normally mutable strains uvrA - recF - and uvrA - umu + rec + of Escherichia coli K-12. The uvrA - umuC - strain performed postreplication repair as efficiently as the parental strain, while the repair in uvrA - recF - strain was dependent on UV dose. Both chloramphenicol and caffeine inhibited postreplication repair to an equal extent of about 25%, and 10%, respectively, in all three uvrA strains of umuC36, recF and umu + rec + . These observations suggest that postreplication repair is largely not responsible for UV mutagenesis. (orig.) [de

Increased Frequency of ColV Plasmids and Mannose-Resistant Hemagglutinating Activity in an Escherichia coli K1 Population

OpenAIRE

1984-01-01

The expression of traits linked to pathogenicity was studied in a population of Escherichia coli K1 strains. It was found that E. coli K1 strains isolated from extraintestinal infection harbor the ColV plasmid and express mannose-resistant hemagglutinating activity type VI with a high frequency. The presence of these properties may play a role in the ability of some E. coli K1 serogroups to invade.

Alignment of Escherichia coli K12 DNA sequences to a genomic restriction map.

Science.gov (United States)

Rudd, K E; Miller, W; Ostell, J; Benson, D A

1990-01-25

We use the extensive published information describing the genome of Escherichia coli and new restriction map alignment software to align DNA sequence, genetic, and physical maps. Restriction map alignment software is used which considers restriction maps as strings analogous to DNA or protein sequences except that two values, enzyme name and DNA base address, are associated with each position on the string. The resulting alignments reveal a nearly linear relationship between the physical and genetic maps of the E. coli chromosome. Physical map comparisons with the 1976, 1980, and 1983 genetic maps demonstrate a better fit with the more recent maps. The results of these alignments are genomic kilobase coordinates, orientation and rank of the alignment that best fits the genetic data. A statistical measure based on extreme value distribution is applied to the alignments. Additional computer analyses allow us to estimate the accuracy of the published E. coli genomic restriction map, simulate rearrangements of the bacterial chromosome, and search for repetitive DNA. The procedures we used are general enough to be applicable to other genome mapping projects.

K-12 Education

Science.gov (United States)

products laboratories publications nisee b.i.p. members education FAQs links education Education Program Internships K-12 Education Contact the PEER Education Program PEER's Educational Affiliates Student Design Competition Student Leadership Council Classes and Other Educational Activities Site Map Search K-12 Education

Patterning bacterial communities on epithelial cells.

Directory of Open Access Journals (Sweden)

Mohammed Dwidar

Full Text Available Micropatterning of bacteria using aqueous two phase system (ATPS enables the localized culture and formation of physically separated bacterial communities on human epithelial cell sheets. This method was used to compare the effects of Escherichia coli strain MG1655 and an isogenic invasive counterpart that expresses the invasin (inv gene from Yersinia pseudotuberculosis on the underlying epithelial cell layer. Large portions of the cell layer beneath the invasive strain were killed or detached while the non-invasive E. coli had no apparent effect on the epithelial cell layer over a 24 h observation period. In addition, simultaneous testing of the localized effects of three different bacterial species; E. coli MG1655, Shigella boydii KACC 10792 and Pseudomonas sp DSM 50906 on an epithelial cell layer is also demonstrated. The paper further shows the ability to use a bacterial predator, Bdellovibriobacteriovorus HD 100, to selectively remove the E. coli, S. boydii and P. sp communities from this bacteria-patterned epithelial cell layer. Importantly, predation and removal of the P. Sp was critical for maintaining viability of the underlying epithelial cells. Although this paper focuses on a few specific cell types, the technique should be broadly applicable to understand a variety of bacteria-epithelial cell interactions.

Genetic Control of the Secondary Modification of Deoxyribonucleic Acid in Escherichia coli1

Science.gov (United States)

Mamelak, Linda; Boyer, Herbert W.

1970-01-01

The wild-type restriction and modification alleles of Escherichia coli K-12 and B were found to have no measurable effect on the patterns of methylated bases in the deoxyribonucleic acid (DNA) of these strains. The genetic region controlling the methylation of cytosine in E. coli K-12 was mapped close to his, and the presence or absence of this gene in E. coli B or E. coli K had no effect on the restriction and modification properties of these strains. Thus, only a few of the methylated bases in the DNA of these strains are involved in host modification, and the biological role of the remainder remains obscure. PMID:4919756

Structure-function relationships of Na+, K+, ATP, or Mg2+ binding and energy transduction in Na,K-ATPase

DEFF Research Database (Denmark)

Jorgensen, Peter L.; Pedersen, Per Amstrup

2000-01-01

Na,K-ATPase; Mutagenesis; Na+ binding; K+ binding; Tl+ binding; Mg2+ binding; ATP binding; Cation binding site; Energy transduction......Na,K-ATPase; Mutagenesis; Na+ binding; K+ binding; Tl+ binding; Mg2+ binding; ATP binding; Cation binding site; Energy transduction...

Search for 12 C+ 12 C clustering in 24 Mg ground state

Indian Academy of Sciences (India)

In the backdrop of many models, the heavy cluster structure of the ground state of 24 Mg has been probed experimentally for the first time using the heavy cluster knockout reaction 24 Mg( 12 C, 212 C) 12 C in thequasifree scattering kinematic domain. In the ( 12 C, 212 C) reaction, the direct 12 C-knockout cross-section was ...

A structural study of the capsular antigens of Escherichia coli K36 and Klebsiella K68

International Nuclear Information System (INIS)

Stanley, S.M.R.

1986-01-01

This thesis discusses the structural study of the capsular antigens of the bacteria, Escherichia coli K36 and Klebsiella K68. In the elucidation of bacterial polysaccharides chemically based analytical procedures and instrumental analytical techniques were used. Nuclear magnetic resonance is a powerful tool for obtaining structural information on poly-, oligo- and di-saccharides and a detailed discussion of this technique, and the results obtained in this study, is given

Genes and proteins of Escherichia coli K-12 (GenProtEC).

Science.gov (United States)

Riley, M

1997-01-01

GenProtEC is a database of Escherichia coli genes and their gene products, classified by type of function and physiological role and with citations to the literature for each. Also present are data on sequence similarities amongE.coliproteins with PAM values, percent identity of amino acids, length of alignment and percent aligned. GenProtEC can also be accessed through the World Wide Web at URL http://mbl.edu/html/ecoli.html .

Single-strand breaks in the DNA of the uvrA and uvrB strains of Escherichia coli K-12 after ultraviolet irradiation

Energy Technology Data Exchange (ETDEWEB)

Youngs, D A; Smith, K C [Stanford Univ., Calif. (USA). Dept. of Radiology

1976-12-01

DNA single-strand breaks were produced in uvrA and uvrB strains of E.coli K-12 after UV (254 nm) irradiation. These breaks appeared to be produced both directly by photochemical events, and by a temperature-dependent process. Cyclobutane-type pyrimidine dimers are probably not the photoproducts that lead to the temperature-dependent breaks, since photoreactivation had no detectable effect on the final yield of breaks. The DNA strand breaks appeared to be repairable by a process that requires DNA polymerase I and polynucleotide ligase, but not the recA, recB, recF, lexA101 or uvrD gene products. It is hypothesized that these temperature-dependent breaks occur either as a result of breakdown of a thermolabile photoproduct, or as the initial endonucleolytic event of a uvrA, uvrB-independent excision repair process that acts on a UV photoproduct other than the cyclobutane-type pyrimidine dimer.

Inactivation of Escherichia coli inoculated onto fresh-cut chopped cabbage using electron-beam processing.

Science.gov (United States)

Grasso, Elizabeth M; Uribe-Rendon, Roberto M; Lee, Ken

2011-01-01

During the past decade there were more than 50 reported outbreaks involving leafy green vegetables contaminated with foodborne pathogens. Leafy greens, including cabbage, are fresh foods rarely heated before consumption, which enables foodborne illness. The need for improved safety of fresh food drives the demand for nonthermal food processes to decrease the risk of pathogens while maintaining fresh quality. This study examines the efficacy of electron-beam (e-beam) irradiation in decreasing indigenous microflora on fresh-cut cabbage and determines the optimal dosage to pasteurize fresh-cut cabbage inoculated with Escherichia coli K-12. Fresh-cut cabbage (100 g) was inoculated with ∼8 log E. coli K-12 and e-beam irradiated at doses of 0, 1.0, 2.3, or 4.0 kGy. At 2.3 kGy there was 7-log reduction of E. coli K-12 in the fresh-cut cabbage. The D(10)-value for E. coli K-12 in fresh-cut cabbage was 0.564 kGy. E-beam irradiation is thus a viable nonthermal treatment that extends the shelf life and increases the safety of fresh cabbage by reducing or eliminating indigenous microflora and unwanted pathogens.

A High-Throughput Oxidative Stress Biosensor Based on Escherichia coli roGFP2 Cells Immobilized in a k-Carrageenan Matrix

Directory of Open Access Journals (Sweden)

Lia Ooi

2015-01-01

Full Text Available Biosensors fabricated with whole-cell bacteria appear to be suitable for detecting bioavailability and toxicity effects of the chemical(s of concern, but they are usually reported to have drawbacks like long response times (ranging from hours to days, narrow dynamic range and instability during long term storage. Our aim is to fabricate a sensitive whole-cell oxidative stress biosensor which has improved properties that address the mentioned weaknesses. In this paper, we report a novel high-throughput whole-cell biosensor fabricated by immobilizing roGFP2 expressing Escherichia coli cells in a k-carrageenan matrix, for the detection of oxidative stress challenged by metalloid compounds. The E. coli roGFP2 oxidative stress biosensor shows high sensitivity towards arsenite and selenite, with wide linear range and low detection limit (arsenite: 1.0 × 10−3–1.0 × 101 mg·L−1, LOD: 2.0 × 10−4 mg·L−1; selenite: 1.0 × 10−5–1.0 × 102 mg·L−1, LOD: 5.8 × 10−6 mg·L−1, short response times (0–9 min, high stability and reproducibility. This research is expected to provide a new direction in performing high-throughput environmental toxicity screening with living bacterial cells which is capable of measuring the bioavailability and toxicity of environmental stressors in a friction of a second.

Influence of some bacterial and host factors on colonization and invasiveness of Escherichia coli K1 in neonatal rats.

OpenAIRE

Wullenweber, M; Beutin, L; Zimmermann, S; Jonas, C

1993-01-01

Of 209 healthy infants examined, 44 (21.1%) carried Escherichia coli K1 in their feces. Of these 44 isolates, 36 (81.8%) were attributed to 10 different known clonal groups of E. coli K1 and 4 isolates represented unknown types. The influence of mannose-resistant (MR) adhesins, aerobactin production, and resistance to serum on colonization and invasiveness of E. coli K1 in orally infected inbred LEW baby rats was investigated. Strains expressing MR adhesins had significantly higher colonizati...

Regulation of Toll-like receptor 2 interaction with Ecgp96 controls Escherichia coli K1 invasion of brain endothelial cells

OpenAIRE

Krishnan, Subramanian; Chen, Shuang; Turcatel, Gianluca; Arditi, Moshe; Prasadarao, Nemani V.

2012-01-01

The interaction of outer membrane protein A (OmpA) with its receptor, Ecgp96 (a homologue of Hsp90β) is critical for the pathogenesis of E. coli K1 meningitis. Since Hsp90 chaperones Toll-like receptors (TLRs), we examined the role of TLRs in E. coli K1 infection. Herein, we show that newborn TLR2−/− mice are resistant to E. coli K1 meningitis, while TLR4−/− mice succumb to infection sooner. In vitro, OmpA+ E. coli infection selectively upregulates Ecgp96 and TLR2 in human brain microvascular...

Comparative genomic analysis shows that avian pathogenic Escherichia coli isolate IMT5155 (O2:K1:H5; ST complex 95, ST140 shares close relationship with ST95 APEC O1:K1 and human ExPEC O18:K1 strains.

Directory of Open Access Journals (Sweden)

Xiangkai Zhu Ge

Full Text Available Avian pathogenic E. coli and human extraintestinal pathogenic E. coli serotypes O1, O2 and O18 strains isolated from different hosts are generally located in phylogroup B2 and ST complex 95, and they share similar genetic characteristics and pathogenicity, with no or minimal host specificity. They are popular objects for the study of ExPEC genetic characteristics and pathogenesis in recent years. Here, we investigated the evolution and genetic blueprint of APEC pathotype by performing phylogenetic and comparative genome analysis of avian pathogenic E. coli strain IMT5155 (O2:K1:H5; ST complex 95, ST140 with other E. coli pathotypes. Phylogeny analyses indicated that IMT5155 has closest evolutionary relationship with APEC O1, IHE3034, and UTI89. Comparative genomic analysis showed that IMT5155 and APEC O1 shared significant genetic overlap/similarities with human ExPEC dominant O18:K1 strains (IHE3034 and UTI89. Furthermore, the unique PAI I5155 (GI-12 was identified and found to be conserved in APEC O2 serotype isolates. GI-7 and GI-16 encoding two typical T6SSs in IMT5155 might be useful markers for the identification of ExPEC dominant serotypes (O1, O2, and O18 strains. IMT5155 contained a ColV plasmid p1ColV5155, which defined the APEC pathotype. The distribution analysis of 10 sequenced ExPEC pan-genome virulence factors among 47 sequenced E. coli strains provided meaningful information for B2 APEC/ExPEC-specific virulence factors, including several adhesins, invasins, toxins, iron acquisition systems, and so on. The pathogenicity tests of IMT5155 and other APEC O1:K1 and O2:K1 serotypes strains (isolated in China through four animal models showed that they were highly virulent for avian colisepticemia and able to cause septicemia and meningitis in neonatal rats, suggesting zoonotic potential of these APEC O1:K1 and O2:K1 isolates.

The ternary system K2SO4MgSO4CaSO4

Science.gov (United States)

Rowe, J.J.; Morey, G.W.; Silber, C.C.

1967-01-01

Melting and subsolidus relations in the system K2SO4MgSO4CaSO4 were studied using heating-cooling curves, differential thermal analysis, optics, X-ray diffraction at room and high temperatures and by quenching techniques. Previous investigators were unable to study the binary MgSO4CaSO4 system and the adjacent area in the ternary system because of the decomposition of MgSO4 and CaSO4 at high temperatures. This problem was partly overcome by a novel sealed-tube quenching method, by hydrothermal synthesis, and by long-time heating in the solidus. As a result of this study, we found: (1) a new compound, CaSO4??3MgSO4 (m.p. 1201??C) with a field extending into the ternary system; (2) a high temperature form of MgSO4 with a sluggishly reversible inversion. An X-ray diffraction pattern for this polymorphic form is given; (3) the inversion of ??-CaSO4 (anhydrite) to ??-CaSO4 at 1195??C, in agreement with grahmann; (1) (4) the melting point of MgSO4 is 1136??C and that of CaSO4 is 1462??C (using sealed tube methods to prevent decomposition of the sulphates); (5) calcium langbeinite (K2SO4??2CaSO4) is the only compound in the K2SO4CaSO4 binary system. This resolved discrepancies in the results of previous investigators; (6) a continuous solid solution series between congruently melting K2SOP4??2MgSO4 (langbeinite) and incongruently melting K2SO4??2CaSO4 (calcium langbeinite); (7) the liquidus in the ternary system consists of primary phase fields of K2SO4, MgSO4, CaSO4, langbeinite-calcium langbeinite solid solution, and CaSO4??3MgSO4. The CaSO4 field extends over a large portion of the system. Previously reported fields for the compounds (K2SO4??MgSO4??nCaSO4), K2SO4??3CaSO4 and K2SO4??CaSO4 were not found; (8) a minimum in the ternary system at: 740??C, 25% MgSO4, 6% CaSO4, 69% K2SO4; and ternary eutectics at 882??C, 49% MgSO4, 19% CaSO4, 32% K2SO4; and 880??, 67??5% MgSO4, 5% CaSO4, 27??5% K2SO4. ?? 1967.

Structure in the 12C+ 12C and 8Be+16O fission width distribution of 24Mg observed via the reaction 12C(16O,α)24Mg(→X+Y)

International Nuclear Information System (INIS)

Lazzarini, A.J.; Steadman, S.G.; Ledoux, R.J.; Sperduto, A.; Young, G.R.; Van Bibber, K.; Cosman, E.R.

1983-01-01

Prominent gross and intermediate width structures are observed in the 12 C+ 12 C, 12 C+ 12 C((2 + ), 1 C((2 + )+ 12 C((2 + ), 8 Be+ 16 O, and 8 Be+ 16 O + (3 - , O + ) decay channels following 24 Mg* population via the 12 C( 16 O,α) 24 Mg reaction at E/sub c.m./ = 33 MeV. Evidence that the 12 C( 16 O,α) 24 Mg reaction populates states in 24 Mg which are associated with 12 C+ 12 C resonances is presented in the form of correlation analyses between the α+ 12 C+ 12 C three-body spectra and previously measured 12 C+ 12 C elastic and inelastic excitation functions. Direct determination of 12 C+ 12 C widths from these measurements is obscured by a background of other strong transitions which appear to be present in the 12 C( 16 O,α) 24 Mg singles spectrum

R.b.e. of 50 kVp X-rays and 660 keV γ-rays (137Cs) with respect to the production of DNA damage, repair and cell-killing in Escherichia coli K-12

International Nuclear Information System (INIS)

Bonura, T.; Youngs, D.A.; Smith, K.C.

1975-01-01

A comparison has been made of the efficiency of cell-killing, DNA single-strand breakage and double-strand breakage in an Escherichia coli K-12 wild-type strain after irradiation with soft X-rays (50 kVp) and hard γ-rays (660 keV) under aerobic conditions. Irradiation with 50 kVp X-rays resulted in 1.47 times more cell-killing than was observed with 137 Cs γ-rays based on a comparison of D 0 values evaluated from the survival curves. DNA sedimentation studies showed that, although 50 kVp X-rays were 1.93 times more effective than 137 Cs γ-rays in producing DNA double-strand breaks, there was no significant difference between the two qualities of radiation with respect to the initial number of single-strand breaks produced. When the cells were irradiated and allowed to repair maximally in minimal medium, 1.57 times more unrepaired DNA single-strand breaks remained per krad after irradiation with 50 kVp X-rays than with 137 Cs γ-rays. The increased yield of DNA double-strand breaks resulting from 50 kVp X-irradiation may account for most of these additional unrepaired single-strand breaks, since single- and double-strand breaks are indistinguishable on alkaline sucrose gradients. These results suggest that the greater r.b.e. of 50 kVp X-rays may be related to an increased effectiveness for producing DNA double-strand breaks compared with the higher energy 137 Cs γ-rays. (author)

5 CFR 1655.3 - Information concerning the cost of a loan.

Science.gov (United States)

2010-01-01

... 5 Administrative Personnel 3 2010-01-01 2010-01-01 false Information concerning the cost of a loan... § 1655.3 Information concerning the cost of a loan. Information concerning the cost of a loan is provided... office or service, or from the TSP record keeper). From this information, a participant can determine the...

A study on preparation and hydriding of β-Mg2Al3 and γ-Mg17Al12

International Nuclear Information System (INIS)

Hadi Suwarno

2009-01-01

The mechanism of the synthetic formation of β-Mg 2 Al 3 and γ-Mg 17 Al 12 has been studied. Mechanical alloying of Mg and Al powders with the atomic ratio of Mg:Al = 2:3 in toluene solution yields β-Mg 2 Al 3 compound after milling for 30 h. The γ-Mg 17 Al 12 can be formed by heating the β-Mg 2 Al 3 at 430°C under high vacuum. The measured hydrogen capacities of β-Mg 2 Al 3 and γ-Mg 17 Al 12 as hydride at 300°C are 3.2 and 4.9 wt%, respectively. Microstructure of the Mg-Al specimen shows that on hydriding at 300°C the polygonal shape of the γ-Mg 17 Al 12 changes into irregular shapes which are composed of γ-MgH 2 and Al. (author)

Inhibition of X-ray-induced protection of Escherichia coli K-12 cells against the lethal effects of ultra-violet light by nitrofurantoin

Energy Technology Data Exchange (ETDEWEB)

Martignoni, K D [Muenchen Univ. (Germany, F.R.). Strahlenbiologisches Inst.

1978-06-01

Wild-type cells of E.coli K-12 showed increasing U.V. resistance if they were X-irradiated and incubated at 37/sup 0/C in growth medium before the U.V. exposure. Development of higher U.V. resistance could be inhibited by incubating the X-irradiated cells either at temperatures below 15/sup 0/C, or in the presence of 0.01 M KCN. Nitrofurantoin (NF), which was recently found specifically to inhibit inducible enzyme synthesis, had only a transient inhibitory effect on X-ray-induced U.V. resistance. Cells grown in glucose medium showed less inhibition by NF of X-radiation-induced resistance to U.V.-radiation than did cells grown in glycerol, or in glucose medium with added cyclic AMP. It is suggested that X-ray-induced U.V. resistance requires active cellular metabolism, but it is not subject to catabolite repression. The following hypothesis is offered to explain the action of NF : Under de-repressed conditions (without catabolite repression by glucose) nitrofurantoin could counteract the radiation-induced inhibition of a repair inhibitor (such as post-irradiation DNA degradation).

Prediction of transcriptional regulatory sites in the complete genome sequence of Escherichia coli K-12.

Science.gov (United States)

Thieffry, D; Salgado, H; Huerta, A M; Collado-Vides, J

1998-06-01

As one of the best-characterized free-living organisms, Escherichia coli and its recently completed genomic sequence offer a special opportunity to exploit systematically the variety of regulatory data available in the literature in order to make a comprehensive set of regulatory predictions in the whole genome. The complete genome sequence of E.coli was analyzed for the binding of transcriptional regulators upstream of coding sequences. The biological information contained in RegulonDB (Huerta, A.M. et al., Nucleic Acids Res.,26,55-60, 1998) for 56 different transcriptional proteins was the support to implement a stringent strategy combining string search and weight matrices. We estimate that our search included representatives of 15-25% of the total number of regulatory binding proteins in E.coli. This search was performed on the set of 4288 putative regulatory regions, each 450 bp long. Within the regions with predicted sites, 89% are regulated by one protein and 81% involve only one site. These numbers are reasonably consistent with the distribution of experimental regulatory sites. Regulatory sites are found in 603 regions corresponding to 16% of operon regions and 10% of intra-operonic regions. Additional evidence gives stronger support to some of these predictions, including the position of the site, biological consistency with the function of the downstream gene, as well as genetic evidence for the regulatory interaction. The predictions described here were incorporated into the map presented in the paper describing the complete E.coli genome (Blattner,F.R. et al., Science, 277, 1453-1461, 1997). The complete set of predictions in GenBank format is available at the url: http://www. cifn.unam.mx/Computational_Biology/E.coli-predictions ecoli-reg@cifn.unam.mx, collado@cifn.unam.mx

Metabolic flux balance analysis and the in silico analysis of Escherichia coli K-12 gene deletions

Directory of Open Access Journals (Sweden)

Edwards Jeremy S

2000-07-01

Full Text Available Abstract Background Genome sequencing and bioinformatics are producing detailed lists of the molecular components contained in many prokaryotic organisms. From this 'parts catalogue' of a microbial cell, in silico representations of integrated metabolic functions can be constructed and analyzed using flux balance analysis (FBA. FBA is particularly well-suited to study metabolic networks based on genomic, biochemical, and strain specific information. Results Herein, we have utilized FBA to interpret and analyze the metabolic capabilities of Escherichia coli. We have computationally mapped the metabolic capabilities of E. coli using FBA and examined the optimal utilization of the E. coli metabolic pathways as a function of environmental variables. We have used an in silico analysis to identify seven gene products of central metabolism (glycolysis, pentose phosphate pathway, TCA cycle, electron transport system essential for aerobic growth of E. coli on glucose minimal media, and 15 gene products essential for anaerobic growth on glucose minimal media. The in silico tpi-, zwf, and pta- mutant strains were examined in more detail by mapping the capabilities of these in silico isogenic strains. Conclusions We found that computational models of E. coli metabolism based on physicochemical constraints can be used to interpret mutant behavior. These in silica results lead to a further understanding of the complex genotype-phenotype relation. Supplementary information: http://gcrg.ucsd.edu/supplementary_data/DeletionAnalysis/main.htm

Polyelectrolyte-Functionalized Nanofiber Mats Control the Collection and Inactivation of Escherichia coli

Directory of Open Access Journals (Sweden)

Katrina A. Rieger

2016-04-01

Full Text Available Quantifying the effect that nanofiber mat chemistry and hydrophilicity have on microorganism collection and inactivation is critical in biomedical applications. In this study, the collection and inactivation of Escherichia coli K12 was examined using cellulose nanofiber mats that were surface-functionalized using three polyelectrolytes: poly (acrylic acid (PAA, chitosan (CS, and polydiallyldimethylammonium chloride (pDADMAC. The polyelectrolyte functionalized nanofiber mats retained the cylindrical morphology and average fiber diameter (~0.84 µm of the underlying cellulose nanofibers. X-ray photoelectron spectroscopy (XPS and contact angle measurements confirmed the presence of polycations or polyanions on the surface of the nanofiber mats. Both the control cellulose and pDADMAC-functionalized nanofiber mats exhibited a high collection of E. coli K12, which suggests that mat hydrophilicity may play a larger role than surface charge on cell collection. While the minimum concentration of polycations needed to inhibit E. coli K12 was 800 µg/mL for both CS and pDADMAC, once immobilized, pDADMAC-functionalized nanofiber mats exhibited a higher inactivation of E. coli K12, (~97%. Here, we demonstrate that the collection and inactivation of microorganisms by electrospun cellulose nanofiber mats can be tailored through a facile polyelectrolyte functionalization process.

Polyelectrolyte-Functionalized Nanofiber Mats Control the Collection and Inactivation of Escherichia coli

Science.gov (United States)

Rieger, Katrina A.; Porter, Michael; Schiffman, Jessica D.

2016-01-01

Quantifying the effect that nanofiber mat chemistry and hydrophilicity have on microorganism collection and inactivation is critical in biomedical applications. In this study, the collection and inactivation of Escherichia coli K12 was examined using cellulose nanofiber mats that were surface-functionalized using three polyelectrolytes: poly (acrylic acid) (PAA), chitosan (CS), and polydiallyldimethylammonium chloride (pDADMAC). The polyelectrolyte functionalized nanofiber mats retained the cylindrical morphology and average fiber diameter (~0.84 µm) of the underlying cellulose nanofibers. X-ray photoelectron spectroscopy (XPS) and contact angle measurements confirmed the presence of polycations or polyanions on the surface of the nanofiber mats. Both the control cellulose and pDADMAC-functionalized nanofiber mats exhibited a high collection of E. coli K12, which suggests that mat hydrophilicity may play a larger role than surface charge on cell collection. While the minimum concentration of polycations needed to inhibit E. coli K12 was 800 µg/mL for both CS and pDADMAC, once immobilized, pDADMAC-functionalized nanofiber mats exhibited a higher inactivation of E. coli K12, (~97%). Here, we demonstrate that the collection and inactivation of microorganisms by electrospun cellulose nanofiber mats can be tailored through a facile polyelectrolyte functionalization process. PMID:28773422

Mapping Stress-Induced Changes in Autoinducer AI-2 Production in Chemostat-Cultivated Escherichia coli K-12

Science.gov (United States)

DeLisa, Matthew P.; Valdes, James J.; Bentley, William E.

2001-01-01

Numerous gram-negative bacteria employ a cell-to-cell signaling mechanism, termed quorum sensing, for controlling gene expression in response to population density. Recently, this phenomenon has been discovered in Escherichia coli, and while pathogenic E. coli utilize quorum sensing to regulate pathogenesis (i.e., expression of virulence genes), the role of quorum sensing in nonpathogenic E. coli is less clear, and in particular, there is no information regarding the role of quorum sensing during the overexpression of recombinant proteins. The production of autoinducer AI-2, a signaling molecule employed by E. coli for intercellular communication, was studied in E. coli W3110 chemostat cultures using a Vibrio harveyi AI-2 reporter assay (M. G. Surrette and B. L. Bassler, Proc. Natl. Acad. Sci. USA 95:7046–7050, 1998). Chemostat cultures enabled a study of AI-2 regulation through steady-state and transient responses to a variety of environmental stimuli. Results demonstrated that AI-2 levels increased with the steady-state culture growth rate. In addition, AI-2 increased following pulsed addition of glucose, Fe(III), NaCl, and dithiothreitol and decreased following aerobiosis, amino acid starvation, and isopropyl-β-d-thiogalactopyranoside-induced expression of human interleukin-2 (hIL-2). In general, the AI-2 responses to several perturbations were indicative of a shift in metabolic activity or state of the cells induced by the individual stress. Because of our interest in the expression of heterologous proteins in E. coli, the transcription of four quorum-regulated genes and 20 stress genes was mapped during the transient response to induced expression of hIL-2. Significant regulatory overlap was revealed among several stress and starvation genes and known quorum-sensing genes. PMID:11292813

Establishing Streptomycin Epidemiological Cut-Off Values for Salmonella and Escherichia coli

DEFF Research Database (Denmark)

Migura, Lourdes Garcia; Sunde, Marianne; Karlsmose, Susanne

2011-01-01

This study was conducted to elucidate the accuracy of the current streptomycin epidemiological cut-off value (ECOFF) for Escherichia coli and Salmonella spp. A total of 236 Salmonella enterica and 208 E. coli isolates exhibiting MICs between 4 and 32 mg/L were selected from 12 countries. Isolates...

Inhibition of Inducible Nitric Oxide Controls Pathogen Load and Brain Damage by Enhancing Phagocytosis of Escherichia coli K1 in Neonatal Meningitis

OpenAIRE

Mittal, Rahul; Gonzalez-Gomez, Ignacio; Goth, Kerstin A.; Prasadarao, Nemani V.

2010-01-01

Escherichia coli K1 is a leading cause of neonatal meningitis in humans. In this study, we sought to determine the pathophysiologic relevance of inducible nitric oxide (iNOS) in experimental E. coli K1 meningitis. By using a newborn mouse model of meningitis, we demonstrate that E. coli infection triggered the expression of iNOS in the brains of mice. Additionally, iNOS−/− mice were resistant to E. coli K1 infection, displaying normal brain histology, no bacteremia, no disruption of the blood...

Molecular resonances in sub-Coulomb energy region (12C-12C, 12C-24Mg, 12C-9Be systems)

International Nuclear Information System (INIS)

Takimoto, Kiyohiko; Shimomura, Susumu; Tanaka, Makoto; Murakami, Tetsuya; Fukada, Mamoru; Sakaguchi, Atsushi

1982-01-01

Molecular resonance in sub-Coulomb energy region was studied on 12 C- 12 C, 12 C- 24 Mg and 12 C- 9 Be systems. The excitation functions and the angular distributions were measured on the reactions 12 C( 12 C, 8 Besub(g,s,)) 16 Osub(g,s,), 24 Mg( 12 C, α) 32 S and 9 Be ( 12 C, 8 Besub(g,s,)) 13 Csub(g,s,). Sub-Coulomb resonances were observed in all systems and the contribution of the 12 Csub(2nd)*(0 + , 7.65 MeV) state is proposed. (author)

Membrane engineering via trans unsaturated fatty acids production improves Escherichia coli robustness and production of biorenewables.

Science.gov (United States)

Tan, Zaigao; Yoon, Jong Moon; Nielsen, David R; Shanks, Jacqueline V; Jarboe, Laura R

2016-05-01

Constructing microbial biocatalysts that produce biorenewables at economically viable yields and titers is often hampered by product toxicity. For production of short chain fatty acids, membrane damage is considered the primary mechanism of toxicity, particularly in regards to membrane integrity. Previous engineering efforts in Escherichia coli to increase membrane integrity, with the goal of increasing fatty acid tolerance and production, have had mixed results. Herein, a novel approach was used to reconstruct the E. coli membrane by enabling production of a novel membrane component. Specifically, trans unsaturated fatty acids (TUFA) were produced and incorporated into the membrane of E. coli MG1655 by expression of cis-trans isomerase (Cti) from Pseudomonas aeruginosa. While the engineered strain was found to have no increase in membrane integrity, a significant decrease in membrane fluidity was observed, meaning that membrane polarization and rigidity were increased by TUFA incorporation. As a result, tolerance to exogenously added octanoic acid and production of octanoic acid were both increased relative to the wild-type strain. This membrane engineering strategy to improve octanoic acid tolerance was found to require fine-tuning of TUFA abundance. Besides improving tolerance and production of carboxylic acids, TUFA production also enabled increased tolerance in E. coli to other bio-products, e.g. alcohols, organic acids, aromatic compounds, a variety of adverse industrial conditions, e.g. low pH, high temperature, and also elevated styrene production, another versatile bio-chemical product. TUFA permitted enhanced growth due to alleviation of bio-product toxicity, demonstrating the general effectiveness of this membrane engineering strategy towards improving strain robustness. Copyright © 2016 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

Structural studies of some bacterial capsular polysaccharides from the family enterobacteriaceae: Klebsiella serotypes K79 and K35 and Escherichia coli serotype K44

Energy Technology Data Exchange (ETDEWEB)

Lim, A.V.S.

1986-01-01

The techniques of sugar analysis, methylation, chromic acid oxidation, deamination, base-catalyzed uronic acid degradation, Smith degradation and partial hydrolysis were used in the structural elucidation of bacterial capsular polysaccharides. Methods such as gas-liquid chromatography, gas chromatography-mass spectrometry, gel permeation, ion-exchange and paper-chromatography were used to isolate and characterize oligosaccharides obtained from degradative procedures. N.m.r. spectroscopy (/sup 1/H and /sup 13/C) was widely used in the characterization of the polysaccharides and of derived poly- and oligo-saccharides. In a few instances, n.m.r. spectroscopy and mass spectrometry were used to delineate the sequence of the sugars in the structure of the poly- and oligo-saccharides. The bacteriophage-induced depolymerizations of the capsular polysaccharides of Klebsiella K79 and E. coli K44 are also reported in this thesis. The sum of these experiments demonstrated that the endoglycanase associated with Klebsiella Phi 79 had ..beta..-galactosidase activity. The endoglycanase from E. coli Phi 44 exhibited ..beta..-N-acetyl-galactosaminidase activity which is novel, in that it is the first reported action of this nature in the bacteriophages isolated for the species E. coli.

Detection and molecular characterization of Escherichia coli CTX-M-15 and Klebsiella pneumoniae SHV-12 β-lactamases from bovine mastitis isolates in the United Kingdom.

Science.gov (United States)

Timofte, Dorina; Maciuca, Iuliana E; Evans, Nicholas J; Williams, Helen; Wattret, Andrew; Fick, Jenny C; Williams, Nicola J

2014-01-01

Recent reports raised concerns about the role that farm stock may play in the dissemination of extended-spectrum β-lactamase (ESBL)-producing bacteria. This study characterized the ESBLs in two Escherichia coli and three Klebsiella pneumoniae subsp. pneumoniae isolates from cases of clinical bovine mastitis in the United Kingdom. Bacterial culture and sensitivity testing of bovine mastitic milk samples identified Gram-negative cefpodoxime-resistant isolates, which were assessed for their ESBL phenotypes. Conjugation experiments and PCR-based replicon typing (PBRT) were used for characterization of transferable plasmids. E. coli isolates belonged to sequence type 88 (ST88; determined by multilocus sequence typing) and carried blaCTX-M-15 and blaTEM-1, while K. pneumoniae subsp. pneumoniae isolates carried blaSHV-12 and blaTEM-1. Conjugation experiments demonstrated that blaCTX-M-15 and blaTEM-1 were carried on a conjugative plasmid in E. coli, and PBRT identified this to be an IncI1 plasmid. The resistance genes were nontransferable in K. pneumoniae subsp. pneumoniae isolates. Moreover, in the E. coli isolates, an association of ISEcp1 and IS26 with blaCTX-M-15 was found where the IS26 element was inserted upstream of both ISEcp1 and the blaCTX-M promoter, a genetic arrangement highly similar to that described in some United Kingdom human isolates. We report the first cases in Europe of bovine mastitis due to E. coli CTX-M-15 and also of bovine mastitis due to K. pneumoniae subsp. pneumoniae SHV-12 β-lactamases in the United Kingdom. We also describe the genetic environment of blaCTX-M-15 and highlight the role that IncI1 plasmids may play in the spread and dissemination of ESBL genes, which have been described in both human and cattle isolates.

ECMDB: The E. coli Metabolome Database

OpenAIRE

Guo, An Chi; Jewison, Timothy; Wilson, Michael; Liu, Yifeng; Knox, Craig; Djoumbou, Yannick; Lo, Patrick; Mandal, Rupasri; Krishnamurthy, Ram; Wishart, David S.

2012-01-01

The Escherichia coli Metabolome Database (ECMDB, http://www.ecmdb.ca) is a comprehensively annotated metabolomic database containing detailed information about the metabolome of E. coli (K-12). Modelled closely on the Human and Yeast Metabolome Databases, the ECMDB contains >2600 metabolites with links to ?1500 different genes and proteins, including enzymes and transporters. The information in the ECMDB has been collected from dozens of textbooks, journal articles and electronic databases. E...

Mechanical properties and fracture mechanism of as-cast Mg77TM12Zn5Y6 (TM = Cu, Ni) bulk amorphous matrix composites

International Nuclear Information System (INIS)

Qiu, K.Q.; Hu, N.N.; Zhang, H.B.; Jiang, W.H.; Ren, Y.L.; Liaw, P.K.

2009-01-01

Comparative investigations on the microstructures, thermal stability and mechanical properties of Mg 77 Cu 12 Zn 5 Y 6 and Mg 77 Ni 12 Zn 5 Y 6 bulk metallic glass matrix composites were carried out by using scanning electron microscopy (SEM), DSC and compressive tester. The results show that the microstructure of as-cast samples with 3 mm in diameter for Cu-containing alloy is consisted of Mg flakes and dotted Mg 2 Cu phase in the amorphous matrix, while the as-cast Ni-containing alloy with the same diameter is mainly consisted of Mg flakes in the amorphous matrix. The glass transition temperature and supercooled liquid region are 413 K and 27 K for the Cu-containing, 443 K and 32 K for the Ni-containing amorphous matrix composites, respectively. The fracture strength, yield strength and plastic strain are 532 MPa, 390 MPa and 2.4% for the Cu-containing alloy, 667 MPa, 412 MPa and 7% for the Ni-containing alloy, respectively. Furthermore, the fracture mechanism for the amorphous matrix composites was discussed according to both the fracture surfaces and the stress-strain curves.

Detection of 12 micron Mg I and OH lines in stellar spectra

Science.gov (United States)

Jennings, D. E.; Deming, D.; Wiedemann, G. R.; Keady, J. J.

1986-01-01

Infrared lines of Mg I and OH have been detected in stellar spectra near 12.3 microns. The Mg I 7i-6h transition was seen in Alpha Ori and Alpha Tau, and the R2e(23.5) and R1f(24.5) transitions of OH were seen in Alpha Ori. All lines appear in absorption, in contrast to the solar spectrum where the Mg I line shows a prominent emission core. The lack of emission in these low surface gravity stars is due to a greatly reduced volume recombination rate for the high-n states of Mg I, which is not fully compensated by the increased chromospheric scale height. The OH equivalent widths are sensitive to the temperature structure of the upper photosphere of Alpha Ori, and they indicate that the photosphere near tau 5000 of about 10 to the -5th is approximately 100 K hotter than is given by flux constant models. The OH measurements agree more closely with the 1981 semiemprical model of Basri, Linsky, and Eriksson (1981), which is based on Ca II and Mg II ultraviolet features.

Transport proteins promoting Escherichia coli pathogenesis

Science.gov (United States)

Tang, Fengyi; Saier, Milton H.

2014-01-01

Escherichia coli is a genetically diverse species infecting hundreds of millions of people worldwide annually. We examined seven well-characterized E. coli pathogens causing urinary tract infections, gastroenteritis, pyelonephritis and haemorrhagic colitis. Their transport proteins were identified and compared with each other and a non-pathogenic E. coli K12 strain to identify transport proteins related to pathogenesis. Each pathogen possesses a unique set of protein secretion systems for export to the cell surface or for injecting effector proteins into host cells. Pathogens have increased numbers of iron siderophore receptors and ABC iron uptake transporters, but the numbers and types of low-affinity secondary iron carriers were uniform in all strains. The presence of outer membrane iron complex receptors and high-affinity ABC iron uptake systems correlated, suggesting co-evolution. Each pathovar encodes a different set of pore-forming toxins and virulence-related outer membrane proteins lacking in K12. Intracellular pathogens proved to have a characteristically distinctive set of nutrient uptake porters, different from those of extracellular pathogens. The results presented in this report provide information about transport systems relevant to various types of E. coli pathogenesis that can be exploited in future basic and applied studies. PMID:24747185

Transport proteins promoting Escherichia coli pathogenesis.

Science.gov (United States)

Tang, Fengyi; Saier, Milton H

2014-01-01

Escherichia coli is a genetically diverse species infecting hundreds of millions of people worldwide annually. We examined seven well-characterized E. coli pathogens causing urinary tract infections, gastroenteritis, pyelonephritis and haemorrhagic colitis. Their transport proteins were identified and compared with each other and a non-pathogenic E. coli K12 strain to identify transport proteins related to pathogenesis. Each pathogen possesses a unique set of protein secretion systems for export to the cell surface or for injecting effector proteins into host cells. Pathogens have increased numbers of iron siderophore receptors and ABC iron uptake transporters, but the numbers and types of low-affinity secondary iron carriers were uniform in all strains. The presence of outer membrane iron complex receptors and high-affinity ABC iron uptake systems correlated, suggesting co-evolution. Each pathovar encodes a different set of pore-forming toxins and virulence-related outer membrane proteins lacking in K12. Intracellular pathogens proved to have a characteristically distinctive set of nutrient uptake porters, different from those of extracellular pathogens. The results presented in this report provide information about transport systems relevant to various types of E. coli pathogenesis that can be exploited in future basic and applied studies. Copyright © 2014 Elsevier Ltd. All rights reserved.

Lactobacillus rhamnosus GG supernatant enhance neonatal resistance to systemic Escherichia coli K1 infection by accelerating development of intestinal defense

OpenAIRE

Xiaolong He; Qing Zeng; Santhosh Puthiyakunnon; Zhijie Zeng; Weijun Yang; Jiawen Qiu; Lei Du; Swapna Boddu; Tongwei Wu; Danxian Cai; Sheng-He Huang; Hong Cao

2017-01-01

The objective of this study was to determine whether Lactobacillus rhamnosus GG culture supernatant (LCS) has a preventive effect against gut-derived systemic neonatal Escherichia coli (E. coli) K1 infection. The preventive effects were evaluated in human colonic carcinoma cell line Caco-2 and neonatal rat models. Our in vitro results showed that LCS could block adhesion, invasion and translocation of E. coli K1 to Caco-2 monolayer via up-regulating mucin production and maintaining intestinal...

Activation of the NLRP3 Inflammasome Pathway by Uropathogenic Escherichia coli Is Virulence Factor-Dependent and Influences Colonization of Bladder Epithelial Cells

Directory of Open Access Journals (Sweden)

Isak Demirel

2018-03-01

Full Text Available The NLRP3 inflammasome and IL-1β release have recently been suggested to be important for the progression of urinary tract infection (UTI. However, much is still unknown regarding the interaction of UPEC and the NLRP3 inflammasome. The purpose of this study was to elucidate what virulence factors uropathogenic Escherichia coli (UPEC use to modulate NLRP3 inflammasome activation and subsequent IL-1β release and the role of NLRP3 for UPEC colonization of bladder epithelial cells. The bladder epithelial cell line 5637, CRISPR/Cas9 generated NLRP3, caspase-1 and mesotrypsin deficient cell lines and transformed primary bladder epithelial cells (HBLAK were stimulated with UPEC isolates and the non-pathogenic MG1655 strain. We found that the UPEC strain CFT073, but not MG1655, induced an increased caspase-1 activity and IL-1β release from bladder epithelial cells. The increase was shown to be mediated by α-hemolysin activation of the NLRP3 inflammasome in an NF-κB-independent manner. The effect of α-hemolysin on IL-1β release was biphasic, initially suppressive, later inductive. Furthermore, the phase-locked type-1-fimbrial ON variant of CFT073 inhibited caspase-1 activation and IL-1β release. In addition, the ability of CFT073 to adhere to and invade NLRP3 deficient cells was significantly reduced compare to wild-type cells. The reduced colonization of NLRP3-deficient cells was type-1 fimbriae dependent. In conclusion, we found that the NLRP3 inflammasome was important for type-1 fimbriae-dependent colonization of bladder epithelial cells and that both type-1 fimbriae and α-hemolysin can modulate the activity of the NLRP3 inflammasome.

Effects of limestone, N, K and Mg fertilizers on Mg absorption by oats and barley

Energy Technology Data Exchange (ETDEWEB)

Alston, A M

1966-01-01

Oats grown in a pot experiment on two sandy loams were sampled at four stages of growth. Neither KCl nor MgSO/sub 4/. 7H/sub 2/O had any effect on yield but % Mg and total Mg uptake were consistently decreased by applying K and increased by applying Mg. Ca(NO/sub 3/)/sub 2/ increased % Mg more than did (NH/sub 4/)/sub 2/SO/sub 4/, but yield and total Mg uptake were higher where (NH/sub 4/)/sub 2/SO/sub 4/ was applied. The effects of fertilizers were similar on both soils. The effects of applying (NH/sub 4/)/sub 2/SO/sub 4/ and NaNO/sub 2/ to the soil on % Mg in barley were compared in a field experiment on an acid loam to which several rates of limestone had been applied. Treatments had no effect on the % Mg in grain or straw at maturity. At four earlier stages of growth (NH/sub 4/)/sub 2/SO)/sub 4/ increased % Mg in the plants more than did NaNO/sub 2/. Limestone slightly increased % Mg. Nitrification of NH/sub 4/ in the soil was rapid.

The in vitro biocompatibility and macrophage phagocytosis of Mg17Al12 phase in Mg-Al-Zn alloys.

Science.gov (United States)

Liu, Chen; He, Peng; Wan, Peng; Li, Mei; Wang, Kehong; Tan, Lili; Zhang, Yu; Yang, Ke

2015-07-01

Mg alloys are gaining interest for applications as biodegradable medical implant, including Mg-Al-Zn series alloys with good combination of mechanical properties and reasonable corrosion resistance. However, whether the existence of second phase particles in the alloys exerts influence on the biocompatibility is still not clear. A deeper understanding of how the particles regulate specific biological responses is becoming a crucial requirement for their subsequent biomedical application. In this work, the in vitro biocompatibility of Mg17Al12 as a common second phase in biodegradable Mg-Al-Zn alloys was investigated via hemolysis, cytotoxicity, cell proliferation, and cell adhesion tests. Moreover, osteogenic differentiation was evaluated by the extracellular matrix mineralization assay. The Mg17Al12 particles were also prepared to simulate the real situation of second phase in the in vivo environment in order to estimate the cellular response in macrophages to the Mg17Al12 particles. The experimental results indicated that no hemolysis was found and an excellent cytocompatibility was also proved for the Mg17Al12 second phase when co-cultured with L929 cells, MC3T3-E1 cells and BMSCs. Macrophage phagocytosis co-culture test revealed that Mg17Al12 particles exerted no harmful effect on RAW264.7 macrophages and could be phagocytized by the RAW264.7 cells. Furthermore, the possible inflammatory reaction and metabolic way for Mg17Al12 phase were also discussed in detail. © 2014 Wiley Periodicals, Inc.

Solubilization of Na,K-ATPase from rabbit kidney outer medulla using only C12E8

Directory of Open Access Journals (Sweden)

H.L. Santos

2002-03-01

Full Text Available SDS, C12E8, CHAPS or CHAPSO or a combination of two of these detergents is generally used for the solubilization of Na,K-ATPase and other ATPases. Our method using only C12E8 has the advantage of considerable reduction of the time for enzyme purification, with rapid solubilization and purification in a single chromatographic step. Na,K-ATPase-rich membrane fragments of rabbit kidney outer medulla were obtained without adding SDS. Optimum conditions for solubilization were obtained at 4ºC after rapid mixing of 1 mg of membrane Na,K-ATPase with 1 mg of C12E8/ml, yielding 98% recovery of the activity. The solubilized enzyme was purified by gel filtration on a Sepharose 6B column at 4ºC. Non-denaturing PAGE revealed a single protein band with phosphomonohydrolase activity. The molecular mass of the purified enzyme estimated by gel filtration chromatography was 320 kDa. The optimum apparent pH obtained for the purified enzyme was 7.5 for both PNPP and ATP. The dependence of ATPase activity on ATP concentration showed high (K0.5 = 4.0 µM and low (K0.5 = 1.4 mM affinity sites for ATP, with negative cooperativity. Ouabain (5 mM, oligomycin (1 µg/ml and sodium vanadate (3 µM inhibited the ATPase activity of C12E8-solubilized and purified Na,K-ATPase by 99, 81 and 98.5%, respectively. We have shown that Na,K-ATPase solubilized only with C12E8 can be purified and retains its activity. The activity is consistent with the form of (alphaß2 association.

Small atomic displacements in the molybdenophosphates AMo2P3O12 (A = K, Rb, Tl)

International Nuclear Information System (INIS)

Leclaire, A.; Raveau, B.

1988-01-01

KMo 2 P 3 O 12 , M r = 515.90, orthorhombic, Pbnm, a = 17.6398(14), b = 9.1761(4), c = 12.3000(8) A, V = 1990.9(4) A 3 , Z=8, D x = 3.44 Mg m -3 , λ(Mo Kα) = 0.71069 A, μ = 3.42 mm -1 , F(000) = 1952. T = 294 K, R = 0.028 for 2123 reflections. RbMo 2 P 3 O 12 , M r = 562.26, orthorhombic, Pbcm, a = 8.8314(8), b = 9.2368(7), c = 12.3051(9) A, V = 1003.8(4) A 3 , Z=4, D x = 3.72 Mg m -3 , λ(Mo Kα) = 0.71069 A, μ = 8.08 mm -1 , F(000) = 1048, T = 294 K, R = 0.044 for 2073 reflections. The Mo 2 P 3 O 12 frameworks of the K, Rb and Tl compounds are almost the same. The main difference is in the position of the alkaline-earth ions in the tunnels, which induces, in the potassium compound, a superstructure along a. The alkaline-earth ions are slightly displaced as their size decreases in order that the A-O distances may agree with the sum of the ionic radii. (orig.)

Evaluación de la tolerancia a la crioconservación de dos cepas de Escherichia coli K12 de uso frecuente en biotecnología

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Diliana Celeste Pérez-Reytor*, Angela Estela Sosa Espinosa

2010-08-01

Full Text Available Uno de los métodos más recomendados para la conservación de E. coli es la congelación entre -20ºC y -70ºC, donde comúnmente se utiliza glicerol o dimetilsulfóxido como criopreservante. Los bancos de E. coli que se conservan por esta vía pueden mantener altas viabilidades por más de 10 años. A pesar de que es una práctica común están poco documentadas las diferencias que existen entre las cepas de interés biotecnológico en cuanto a la resistencia a la congelación. En este artículo se comparan dos cepas mutantes de E. coli K12: la cepa RRI y la cepa HB101, con el objetivo de evaluar el comportamiento de ambas a dos temperaturas de almacenamiento y diferentes concentraciones de glicerol, después de ocho ciclos sucesivos de congelación-descongelación. Las cepas estudiadas sólo difieren en su capacidad de reparación al daño sobre el ADN al presentar la HB101 una variante mutada del gen recA. Cuando se utilizó una temperatura de almacenamiento de -70 ºC y diferentes concentraciones de criopreservante las cepas tuvieron un comportamiento similar de supervivencia, después de los ocho ciclos. Sin embargo, el comportamiento de la supervivencia difiere cuando la temperatura de congelación en los ciclos es de -20 ºC. Además de la supervivencia y la estabilidad de los marcadores genéticos estudiados, observamos que el efecto de los ciclos de congelación y descongelación retardó el tiempo de aparición de las colonias en medio sólido.

Comparative Study on Different Expression Hosts for Alkaline Phytase Engineered in Escherichia coli.

Science.gov (United States)

Chen, Weiwei; Yu, Hongwei; Ye, Lidan

2016-07-01

The application of alkaline phytase as a feed additive is restricted by the poor specific activity. Escherichia coli is a frequently used host for directed evolution of proteins including alkaline phytase towards improved activity. However, it is not suitable for production of food-grade products due to potential pathogenicity. To combine the advantages of different expression systems, mutants of the alkaline phytase originated from Bacillus subtilis 168 (phy168) were first generated via directed evolution in E. coli and then transformed to food-grade hosts B. subtilis and Pichia pastoris for secretory expression. In order to investigate the suitability of different expression systems, the phy168 mutants expressed in different hosts were characterized and compared in terms of specific activity, pH profile, pH stability, temperature profile, and thermostability. The specific activity of B. subtilis-expressed D24G/K70R/K111E/N121S mutant at pH 7.0 and 60 °C was 30.4 U/mg, obviously higher than those in P. pastoris (22.7 U/mg) and E. coli (19.7 U/mg). Moreover, after 10 min incubation at 80 °C, the B. subtilis-expressed D24G/K70R/K111E/N121S retained about 70 % of the activity at pH 7.0 and 37 °C, whereas the values were only about 25 and 50 % when expressed in P. pastoris and E. coli, respectively. These results suggested B. subtilis as an appropriate host for expression of phy168 mutants and that the strategy of creating mutants in one host and expressing them in another might be a new solution to industrial production of proteins with desired properties.

Evaluation of Colicin Effect on the Induction of Treated Mice in Prevention of Infection Caused by Escherichia coli K99

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Yahya Tahamtan

2016-11-01

Full Text Available Background. Colicin produce by colicinogenic E. coli (CEC arenarrow limited spectrum antimicrobial agents that are able to kill or prevent close related strains. Objective. The objective of this study was to evaluation effect of Colicin to induce immunized mice to prevent infection caused by E. coliK99. Patient and Methods. The experiment was conducted into two mice groups (30 in each group with two weeks old. All mice were administered by streptomycin sulfate prior to treatment to eliminate resident E. coli. Group one was orally inoculated with PBS as control and the second was immunized by Colicin solution as immunize group. Both control and immunized group were challenged by 3 LD 50 of E. coli K99 and follow a week. Results. Immunized mice group were not showed severe clinical signs. While diarrhea with different sings of colibaccillosis was established in control group and infected mice was died. Conclusion. Overuse antibiotics developed serious new types of multi drug resistance in human medicine and therefore has limited their use in farm animals. The study indicates the use of Colicin and biotherapy instead of antibiotic is more safe and efficient for control of E. coliK99 infection. Immunized mice by Colicin solution protected E. coli K99 colonization and reduce fecal shedding. Investigation in livestock for applying Colicin in farm animal is recommended.

Rapid, large-scale purification and characterization of Ada protein (O sup 6 methylguanine-DNA methyltransferase) of E. coli

Energy Technology Data Exchange (ETDEWEB)

Bhattacharyya, D.; Tano, K.; Bunick, G.J.; Uberbacher, E.C.; Mitra, S. (Oak Ridge National Laboratory, TN (USA)); Behnke, W.D. (Univ. of Cincinnati College of Medicine, OH (USA))

1988-07-25

The E. coli Ada protein (O{sup 6}-methylguanine-DNA methyltransferase) has been purified using a high-level expression vector with a yield of about 3 mg per liter of E. coli culture. The 39-kDa protein has an extinction coefficient (E{sup 280nm}{sub 1%}) of 5.3. Its isoelectric point of 7.1 is lower than that predicted from the amino acid content. The homogeneous Ada protein is fully active as a methyl acceptor from O{sup 6}-methylguanine in DNA. Its reaction with O{sup 6}-methylguanine in a synthetic DNA has a second-order rate constant of 1.1 {times} 10{sup 9} M{sup {minus}1} min{sup {minus}1} at 0{degree}C. Both the native form and the protein methylated at Cys-69 are monomeric. The CD spectrum suggests a low {alpha}-helical content and the radius of gyration of 23 {angstrom} indicates a compact, globular shape. The middle region of the protein is sensitive to a variety of proteases, including an endogenous activity in E. coli, suggesting that the protein is composed of N-terminal and C-terminal domains connected by a hinge region. E. coli B has a higher level of this protease than does K12.

Limits on the quiescent radio emission from the black hole binaries GRO J1655−40 and XTE J1550−564

NARCIS (Netherlands)

Calvelo, D.E.; Fender, R.P.; Russell, D.M.; Gallo, E.; Corbel, S.; Tzioumis, A.K.; Bell, M.E.; Lewis, F.; Maccarone, T.J.

2010-01-01

We present the results of radio observations of the black hole binaries GRO J1655−40 and XTE J1550−564 in quiescence, with the upgraded Australia Telescope Compact Array. Neither system was detected. Radio flux density upper limits (3σ) of 26 μJy (at 5.5 GHz), 47 μJy (at 9 GHz) for GRO J1655−40 and

Norwegian patients and retail chicken meat share cephalosporin-resistant Escherichia coli and IncK/blaCMY-2 resistance plasmids.

Science.gov (United States)

Berg, E S; Wester, A L; Ahrenfeldt, J; Mo, S S; Slettemeås, J S; Steinbakk, M; Samuelsen, Ø; Grude, N; Simonsen, G S; Løhr, I H; Jørgensen, S B; Tofteland, S; Lund, O; Dahle, U R; Sunde, M

2017-06-01

In 2012 and 2014 the Norwegian monitoring programme for antimicrobial resistance in the veterinary and food production sectors (NORM-VET) showed that 124 of a total of 406 samples (31%) of Norwegian retail chicken meat were contaminated with extended-spectrum cephalosporin-resistant Escherichia coli. The aim of this study was to compare selected cephalosporin-resistant E. coli from humans and poultry to determine their genetic relatedness based on whole genome sequencing (WGS). Escherichia coli representing three prevalent cephalosporin-resistant multi-locus sequence types (STs) isolated from poultry (n=17) were selected from the NORM-VET strain collections. All strains carried an IncK plasmid with a bla CMY-2 gene. Clinical E. coli isolates (n=284) with AmpC-mediated resistance were collected at Norwegian microbiology laboratories from 2010 to 2014. PCR screening showed that 29 of the clinical isolates harboured both IncK and bla CMY-2 . All IncK/bla CMY-2 -positive isolates were analysed with WGS-based bioinformatics tools. Analysis of single nucleotide polymorphisms (SNP) in 2.5 Mbp of shared genome sequences showed close relationship, with fewer than 15 SNP differences between five clinical isolates from urinary tract infections (UTIs) and the ST38 isolates from poultry. Furthermore, all of the 29 clinical isolates harboured IncK/bla CMY-2 plasmid variants highly similar to the IncK/bla CMY-2 plasmid present in the poultry isolates. Our results provide support for the hypothesis that clonal transfer of cephalosporin-resistant E. coli from chicken meat to humans may occur, and may cause difficult-to-treat infections. Furthermore, these E. coli can be a source of AmpC-resistance plasmids for opportunistic pathogens in the human microbiota. Copyright © 2017 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Alkylation of acetohydroxyacid synthase I from Escherichia coli K-12 by 3-bromopyruvate: evidence for a single active site catalyzing acetolactate and acetohydroxybutyrate synthesis.

Science.gov (United States)

Silverman, P M; Eoyang, L

1987-01-01

Acetohydroxyacid synthase I (AHAS I) purified from Escherichia coli K-12 was irreversibly inactivated by incubation with 3-bromopyruvate. Inactivation was specific, insofar as bromoacetate and iodoacetate were much less effective than bromopyruvate. Inactivation was accompanied by incorporation of radioactivity from 3-bromo[2-14C]pyruvate into acid-insoluble material. More than 95% of the incorporated radioactivity coelectrophoresed with the 60-kilodalton IlvB subunit of the enzyme through a sodium dodecyl sulfate-polyacrylamide gel; less than 5% coelectrophoresed with the 11.2-kilodalton IlvN subunit. The stoichiometry of incorporation at nearly complete inactivation was 1 mol of 14C per mol of IlvB polypeptide. These data indicate that bromopyruvate inactivates AHAS I by alkylating an amino acid at or near a single active site located in the IlvB subunit of the enzyme. We confirmed that this alkylation inactivated both AHAS reactions normally catalyzed by AHAS I. These results provide the first direct evidence that AHAS I catalyzes both acetohydroxybutyrate and acetolactate synthesis from the same active site. Images PMID:3294793

Alkylation of acetohydroxyacid synthase I from Escherichia coli K-12 by 3-bromopyruvate: evidence for a single active site catalyzing acetolactate and acetohydroxybutyrate synthesis

International Nuclear Information System (INIS)

Silverman, P.M.; Eoyang, L.

1987-01-01

Acetohyroxyacid synthease I (AHAS I) purified from Escherichia coli K-12 was irreversibly inactivated by incubation with 3-bromopyruvate. Inactivation was specific, insofar as bromoacetate and iodoacetate were much less effective than bromopyruvate. Inactivation was accompanied by incorporation of radioactivity from 3-bromo[2- 14 C]pyruvate into acid-insoluble material. More than 95% of the incorporated radioactivity coelectrophoresed with the 60-kilodalton IlvB subunit of the enzyme through a sodium dodecyl sulfate-polyacrylamide gel; less than 5% coelectrophoresed with the 11.2-kilodalton IlvN subunit. The stoichiometry of incorporation at nearly complete inactivation was 1 mol of 14 C per mol of IlvB polypeptide. These data indicate that bromopyruvate inactivates AHAS I by alkylating an amino acid at or near a single active site located in the IlvB subunit of the enzyme. The authors confirmed that this alkylation inactivated both AHAS reactions normally catalyzed by AHAS I. These results provide the first direct evidence that AHAS I catalyzes both acetohydroxybutyrate and acetolactate synthesis from the same active site

The type III secretion system is involved in the invasion and intracellular survival of Escherichia coli K1 in human brain microvascular endothelial cells

OpenAIRE

Yao, Yufeng; Xie, Yi; Perace, Donna; Zhong, Yi; Lu, Jie; Tao, Jing; Guo, Xiaokui; Kim, Kwang Sik

2009-01-01

Type III secretion systems have been documented in many Gram-negative bacteria, including enterohemorrhagic Escherichia coli. We have previously shown the existence of a putative type III secretion system in meningitis-causing E. coli K1 strains, referred to as E. coli type III secretion 2 (ETT2). The sequence of ETT2 in meningitis-causing E. coli K1 strain EC10 (O7:K1) revealed that ETT2 comprises the epr, epa and eiv genes, but bears mutations, deletions and insertions. We constructed the E...

Pharmacokinetics, bioavailability and dose assessment of Cefquinome against Escherichia coli in black swans (Cygnus atratus).

Science.gov (United States)

Zhao, Dong-Hao; Wang, Xu-Feng; Wang, Qiang; Li, Liu-Dong

2017-07-28

The objective of this study is to investigate pharmacokinetics and dose regimens of cefquinome in black swans following intravenous (IV) and intramuscular (IM) administration at a single dose of 2 mg/kg. The MICs of cefquinome against 49 Escherichia coli isolates from black swans were determined. Monte Carlo simulation was applied to conduct the dose regimen assessment and optimization of cefquinome against E. coli in black swans, and a pharmacokinetic/pharmacodynamic (PK/PD) cutoff was established for E. coli isolates obtained in this study. The PK parameters of T 1/2α (0.31 h), T 1/2β (1.69 h) and Cl B (0.13 L/kg·h) indicated a rapid distribution and elimination of cefquinome in black swans after IV administration. After IM injection, the corresponding PK parameters of T 1/2Ka , T 1/2Ke , T max , C max , and F were 0.12 h, 1.62 h, 0.39 h, 5.71 μg/mL and 74.2%, respectively. The MICs of cefquinome against black swans E. coli ranged from 0.03 to 8 μg/mL, with MIC 50 and MIC 90 of 0.06 and 0.5 μg/mL, respectively. The PK/PD cutoff of cefquinome against E. coli was determined to be 0.2 μg/mL. Monte Carlo simulation showed that the nominal dose regimen (2 mg/kg/24 h) could not achieve a satisfactory probability of target attainment (PTA) for %T MIC  ≥ 50%, indicating a risk of treatment failure and the development of potential drug resistance. The current daily dosage of cefquinome when divided into 12-h interval (1 mg/kg/12 h) may be effective for the treatment of E. coli infections with an MIC ≤0.5 μg/mL.

Fcγ receptor I alpha chain (CD64) expression in macrophages is critical for the onset of meningitis by Escherichia coli K1.

Science.gov (United States)

Mittal, Rahul; Sukumaran, Sunil K; Selvaraj, Suresh K; Wooster, David G; Babu, M Madan; Schreiber, Alan D; Verbeek, J Sjef; Prasadarao, Nemani V

2010-11-18

Neonatal meningitis due to Escherichia coli K1 is a serious illness with unchanged morbidity and mortality rates for the last few decades. The lack of a comprehensive understanding of the mechanisms involved in the development of meningitis contributes to this poor outcome. Here, we demonstrate that depletion of macrophages in newborn mice renders the animals resistant to E. coli K1 induced meningitis. The entry of E. coli K1 into macrophages requires the interaction of outer membrane protein A (OmpA) of E. coli K1 with the alpha chain of Fcγ receptor I (FcγRIa, CD64) for which IgG opsonization is not necessary. Overexpression of full-length but not C-terminal truncated FcγRIa in COS-1 cells permits E. coli K1 to enter the cells. Moreover, OmpA binding to FcγRIa prevents the recruitment of the γ-chain and induces a different pattern of tyrosine phosphorylation of macrophage proteins compared to IgG2a induced phosphorylation. Of note, FcγRIa(-/-) mice are resistant to E. coli infection due to accelerated clearance of bacteria from circulation, which in turn was the result of increased expression of CR3 on macrophages. Reintroduction of human FcγRIa in mouse FcγRIa(-/-) macrophages in vitro increased bacterial survival by suppressing the expression of CR3. Adoptive transfer of wild type macrophages into FcγRIa(-/-) mice restored susceptibility to E. coli infection. Together, these results show that the interaction of FcγRI alpha chain with OmpA plays a key role in the development of neonatal meningitis by E. coli K1.

Isopiestic Investigation of the Osmotic and Activity Coefficients of {yMgCl2 + (1 - y)MgSO4}(aq) and the Osmotic Coefficients of Na2SO4.MgSO4(aq) at 298.15 K

Energy Technology Data Exchange (ETDEWEB)

Miladinovic, J; Ninkovic, R; Todorovic, M; Rard, J A

2007-06-06

Isopiestic vapor pressure measurements were made for {l_brace}yMgCl{sub 2} + (1-y)MgSO{sub 4}{r_brace}(aq) solutions with MgCl{sub 2} ionic strength fractions of y = 0, 0.1997, 0.3989, 0.5992, 0.8008, and (1) at the temperature 298.15 K, using KCl(aq) as the reference standard. These measurements for the mixtures cover the ionic strength range I = 0.9794 to 9.4318 mol {center_dot} kg{sup -1}. In addition, isopiestic measurements were made with NaCl(aq) as reference standard for mixtures of {l_brace}xNa{sub 2}SO{sub 4} + (1-x)MgSO{sub 4}{r_brace}(aq) with the molality fraction x = 0.50000 that correspond to solutions of the evaporite mineral bloedite (astrakanite), Na{sub 2}Mg(SO{sub 4}){sub 2} {center_dot} 4H{sub 2}O(cr). The total molalities, m{sub T} = m(Na{sub 2}SO{sub 4}) + m(MgSO{sub 4}), range from m{sub T} = 1.4479 to 4.4312 mol {center_dot} kg{sup -1} (I = 5.0677 to 15.509 mol {center_dot} kg{sup -1}), where the uppermost concentration is the highest oversaturation molality that could be achieved by isothermal evaporation of the solvent at 298.15 K. The parameters of an extended ion-interaction (Pitzer) model for MgCl2(aq) at 298.15 K, which were required for an analysis of the {l_brace}yMgCl{sub 2} + (1-y)MgSO{sub 4}{r_brace}(aq) mixture results, were evaluated up to I = 12.025 mol {center_dot} kg{sup -1} from published isopiestic data together with the six new osmotic coefficients obtained in this study. Osmotic coefficients of {l_brace}yMgCl{sub 2} + (1-y)MgSO{sub 4}{r_brace}(aq) solutions from the present study, along with critically-assessed values from previous studies, were used to evaluate the mixing parameters of the extended ion-interaction model.

Experimental validation of the predicted binding site of Escherichia coli K1 outer membrane protein A to human brain microvascular endothelial cells: identification of critical mutations that prevent E. coli meningitis.

Science.gov (United States)

Pascal, Tod A; Abrol, Ravinder; Mittal, Rahul; Wang, Ying; Prasadarao, Nemani V; Goddard, William A

2010-11-26

Escherichia coli K1, the most common cause of meningitis in neonates, has been shown to interact with GlcNAc1-4GlcNAc epitopes of Ecgp96 on human brain microvascular endothelial cells (HBMECs) via OmpA (outer membrane protein A). However, the precise domains of extracellular loops of OmpA interacting with the chitobiose epitopes have not been elucidated. We report the loop-barrel model of these OmpA interactions with the carbohydrate moieties of Ecgp96 predicted from molecular modeling. To test this model experimentally, we generated E. coli K1 strains expressing OmpA with mutations of residues predicted to be critical for interaction with the HBMEC and tested E. coli invasion efficiency. For these same mutations, we predicted the interaction free energies (including explicit calculation of the entropy) from molecular dynamics (MD), finding excellent correlation (R(2) = 90%) with experimental invasion efficiency. Particularly important is that mutating specific residues in loops 1, 2, and 4 to alanines resulted in significant inhibition of E. coli K1 invasion in HBMECs, which is consistent with the complete lack of binding found in the MD simulations for these two cases. These studies suggest that inhibition of the interactions of these residues of Loop 1, 2, and 4 with Ecgp96 could provide a therapeutic strategy to prevent neonatal meningitis due to E. coli K1.

Role of leader peptide synthesis in tryptophanase operon expression in Escherichia coli K-12.

OpenAIRE

Stewart, V; Yanofsky, C

1986-01-01

We used site-directed mutagenesis to replace the Escherichia coli tryptophanase (tna) operon leader peptide start codon with AUC. This change greatly decreased the uninduced rate of tna operon expression, and it also lowered the response to inducer. We conclude that leader peptide synthesis plays an essential role in tna operon expression.

Phylogenetische und funktionelle Analysen zur Kapsel O-Acetyltransferase NeuO von Escherichia coli K1

OpenAIRE

Mordhorst, Ines Louise

2010-01-01

Escherichia coli ist ein Kommensale des menschlichen und tierischen Gastrointestinaltraktes. Einige E. coli-Stämme sind in der Lage, extraintestinale Erkrankungen beim Menschen wie Harnwegsinfekte, Neugeborenen-Meningitis und Sepsis, sowie beim Tier aviäre Coliseptikämien, hervorzurufen. Ein wichtiger Virulenzfaktor des Bakteriums ist dabei die aus α-2,8-verknüpften Sialinsäuremonomeren aufgebaute K1-Kapsel, die phasenvariabel mit einer hohen Frequenz O-acetyliert werden kann. Im Jahr 20...

Chromosomal features of Escherichia coli serotype O2:K2, an avian pathogenic E. coli

DEFF Research Database (Denmark)

Jørgensen, Steffen L; Kudirkiene, Egle; Li, Lili

2017-01-01

Escherichia coli causing infection outside the gastrointestinal system are referred to as extra-intestinal pathogenic E. coli. Avian pathogenic E. coli is a subgroup of extra-intestinal pathogenic E. coli and infections due to avian pathogenic E. coli have major impact on poultry production econo...

Reversible Activation of Halophilic β-lactamase from Methanol-Induced Inactive Form: Contrast to Irreversible Inactivation of Non-Halophilic Counterpart.

Science.gov (United States)

Tokunaga, Hiroko; Maeda, Junpei; Arakawa, Tsutomu; Tokunaga, Masao

2017-06-01

Effects of a water-miscible organic solvent, methanol, on the structure and activity of halophilic β-lactamase derived from Chromohalobacter sp.560 (HaBla), were investigated by means of circular dichroism (CD) measurement and enzymatic activity determination. Beta-lactamase activity was enhanced about 1.2-fold in the presence of 10-20% methanol. CD measurement of HaBla revealed different structures depending on the methanol concentration: native-like active form (Form I) in 10-20% methanol and methanol-induced inactive form at higher concentration (Form II in 40-60% and Form III in 75-80% methanol). Incubation of HaBla with 40% methanol led to the complete loss of activity within ~80 min accompanied by the formation of Form II, whose activity was recovered promptly up to ~80% of full activity upon dilution of the methanol concentration to 10%. In addition, when the protein concentration was sufficiently high (e.g., 0.7 mg/ml), HaBla activity of Form III in 75% methanol could be recovered in the same way (with slightly slower recovery rate), upon dilution of the methanol concentration. In contrast, non-halophilic β-lactamase from Escherichia coli K12 strain MG1655 (EcBla) was irreversibly denatured in the presence of 40% methanol. HaBla showed remarkable ability to renature from the methanol-induced inactive states.

Effects of Saponins against Clinical E. coli Strains and Eukaryotic Cell Line

Science.gov (United States)

Arabski, Michał; Węgierek-Ciuk, Aneta; Czerwonka, Grzegorz; Lankoff, Anna; Kaca, Wiesław

2012-01-01

Saponins are detergent-like substances showing antibacterial as well as anticancer potential. In this study, the effects of saponins from Quillaja saponaria were analyzed against prokaryotic and eukaryotic cells. Multidrug-resistant clinical E. coli strains were isolated from human urine. As eukaryotic cells, the CHO-K1 cell lines were applied. Antibacterial effect of ampicillin, streptomycin, and ciprofloxacin in the presence of saponins was measured by cultivation methods. Properties of saponins against CHO-K1 cells were measured by the MTT test, hemolysis assay and flow cytometry. Saponin from Quillaja saponaria has a cytotoxic effect at concentrations higher than 25 μg/mL and in the range of 12–50 μg/mL significantly increases the level of early apoptotic cells. Saponin at dose of 12 μg/mL enhances the six E. coli strains growth. We postulate that saponins increase the influx of nutrients from the medium into E. coli cells. Saponins do not have synergetic effects on antibacterial action of tested antibiotics. In contrary, in the presence of saponins and antibiotics, more CFU/mL E. coli cells were observed. This effect was similar to saponins action alone towards E. coli cells. In conclusion, saponins was cytotoxic against CHO-K1 cells, whereas against E. coli cells this effect was not observed. PMID:22500084

Effects of Saponins against Clinical E. coli Strains and Eukaryotic Cell Line

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Michał Arabski

2012-01-01

Full Text Available Saponins are detergent-like substances showing antibacterial as well as anticancer potential. In this study, the effects of saponins from Quillaja saponaria were analyzed against prokaryotic and eukaryotic cells. Multidrug-resistant clinical E. coli strains were isolated from human urine. As eukaryotic cells, the CHO-K1 cell lines were applied. Antibacterial effect of ampicillin, streptomycin, and ciprofloxacin in the presence of saponins was measured by cultivation methods. Properties of saponins against CHO-K1 cells were measured by the MTT test, hemolysis assay and flow cytometry. Saponin from Quillaja saponaria has a cytotoxic effect at concentrations higher than 25 μg/mL and in the range of 12–50 μg/mL significantly increases the level of early apoptotic cells. Saponin at dose of 12 μg/mL enhances the six E. coli strains growth. We postulate that saponins increase the influx of nutrients from the medium into E. coli cells. Saponins do not have synergetic effects on antibacterial action of tested antibiotics. In contrary, in the presence of saponins and antibiotics, more CFU/mL E. coli cells were observed. This effect was similar to saponins action alone towards E. coli cells. In conclusion, saponins was cytotoxic against CHO-K1 cells, whereas against E. coli cells this effect was not observed.

X-ray-induced mutations in Escherichia coli K-12 strains with altered DNA polymerase I activities

International Nuclear Information System (INIS)

Nagata, Yuki; Kawata, Masakado; Komura, Jun-ichiro; Ono, Tetsuya; Yamamoto, Kazuo

2003-01-01

Spectra of ionizing radiation mutagenesis were determined by sequencing X-ray-induced endogenous tonB gene mutations in Escherichia coli polA strains. We used two polA alleles, the polA1 mutation, defective for Klenow domain, and the polA107 mutation, defective for flap domain. We demonstrated that irradiation of 75 and 50 Gy X-rays could induce 3.8- and 2.6-fold more of tonB mutation in polA1 and polA107 strains, respectively, than spontaneous level. The radiation induced spectrum of 51 tonB mutations in polA1 and 51 in polA107 indicated that minus frameshift, A:T→T:A transversion and G:C→T:A transversion were the types of mutations increased. Previously, we have reported essentially the same X-ray-induced tonB mutation spectra in the wild-type strain. These results indicate that (1) X-rays can induce minus frameshift, A:T→T:A transversion and G:C→T:A transversion in E. coli and (2) presence or absence of polymerase I (PolI) of E. coli does not have any effects on the process of X-ray mutagenesis

The Age-Precipitations Structure Of Al-Mg-Ge Alloy Aged At 473K

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Kawai A.

2015-06-01

Full Text Available The Al-Mg-Ge alloy is one of the age-hardening aluminum alloy after solution heat treatment. It has been proposed that the age-precipitation behavior of Al-Mg-Ge alloy is different from that of Al-Mg-Si alloy according to our previous works about the microstructure on Al-Mg-Ge alloy over-aged at 523K. For example, The hardness of peak aged Al-1.0mass%Mg2Ge alloy is higher than that of Al-1.0mass%Mg2Si alloy. The precipitates in the over-aged samples have been classified as some metastable phases, such as the β’-phase and Type-A precipitates and equilibrium phase of β-Mg2Ge by TEM observation. There a few reports about microstructure on Al-Mg-Ge alloys observed by TEM for different aging times. The age-precipitations structure of Al-Mg-Ge alloy has not been became clear. In this work, TEM observation was investigated the microstructure on Al-1.0mass%Mg2Ge alloy for difference aging times aged at 473K.

Photoreactivation and dark repair of environmental E. coli strains following 24 kHz continuous ultrasound and UV-C irradiation.

Science.gov (United States)

Kaur, Jasjeet; Karthikeyan, Raghupathy; Pillai, Suresh D

2016-07-02

In this study, effects of 24 kHz continuous ultrasound and UV-C on inactivation and potential repair of environmental E. coli strains were studied through a culture based method and a metabolic activity assay. Three environmental E. coli strains isolated from fecal samples of feral hog and deer and treated wastewater effluent were studied and compared with a laboratory E. coli strain (ATCC® 10798). Metabolic activity of E. coli cells during the inactivation and repair period was assessed using the AlamarBlue® assay. Transmission electron microscopy assays were also performed to evaluate morphological damage of bacterial cell wall. After 24 h of photoreactivation period, laboratory E. coli strain (ATCC® 10798) reactivated by 30% and 42% in contrast to E. coli isolate from treated wastewater effluent, which reactivated by 53% and 82% after ultrasound and UV-C treatment, respectively. Possible shearing and reduction in cell size of E. coli strains exposed to ultrasound was revealed by transmission electron micrographs. Metabolic activity of E. coli strains was greatly reduced due to morphological damage to cell membrane caused by 24 kHz continuous ultrasound. Based upon experimental data and TEM micrographs, it could be concluded that ultrasound irradiation has potential in advanced water treatment and water reuse applications.

CCQM-K11.2 determination of glucose in human serum and CCQM-K12.2 determination of creatinine in human serum

Science.gov (United States)

Wise, Stephen A.; Phinney, Karen W.; Duewer, David L.; Sniegoski, Lorna T.; Welch, Michael J.; Pritchett, Jeanita; Pabello, Guiomar; Avila Calderon, Marco A.; Balderas, Miryan; Qinde, Liu; Kooi, Lee Tong; Rego, Eliane; Garrido, Bruno; Allegri, Gabriella; de La Cruz, Marcia; Barrabin, Juliana; Monteiro, Tânia; Lee, Hwashim; Kim, Byungjoo; Delatour, Vincent; Peignaux, Maryline; Kawaguchi, Migaku; Bei, Xu; Can, Quan; Nammoonnoy, Jintana; Schild, Katrin; Ohlendorf, Rüdiger; Henrion, Andre; Ceyhan Gören, Ahmet; Yılmaz, Hasibe; Bilsel, Mine; Konopelko, L.; Krylov, A.; Lopushanskaya, E.

2018-01-01

Glucose and creatinine are two of the most frequently measured substances in human blood/serum for assessing the health status of individuals. Because of their clinical significance, CCQM-K11 glucose in human serum and CCQM-K12 creatinine in human serum were the fourth and fifth key comparisons (KCs) performed by the Organic Analysis Working Group (OAWG). These KCs were conducted in parallel and were completed in 2001. The initial subsequent KCs for glucose, CCQM-K11.1, and creatinine, CCQM-K12.1, were completed in 2005. Measurements for the next KCs for these two measurands, CCQM-K11.2 and CCQM-K12.2, were completed in 2013. While designed as subsequent KCs, systematic discordances between the participants' and the anchor institution's results in both comparisons lead the OAWG to request reference results from two experienced laboratories that had participated in the 2001 comparisons. Based on the totality of the available information, the OAWG converted both CCQM-K11.2 and CCQM-K12.2 to 'Track C' KCs where the key comparison reference value is estimated by consensus. These comparisons highlighted that carrying out comparisons for complex chemical measurements and expecting to be able to treat them under the approaches used for formal CIPM subsequent comparisons is not an appropriate strategy. The approach used here is a compromise to gain the best value from the comparison; it is not an approach that will be used in the future. Instead, the OAWG will focus on Track A and Track C comparisons that are treated as stand-alone entities. Participation in CCQM-K11.2 demonstrates a laboratory's capabilities to measure a polar (pKow > 2), low molecular mass (100 g/mol to 500 g/mol) metabolite in human serum at relatively high concentrations (0.1 mg/g to 10 mg/g). Participation in CCQM-K12.2 demonstrates capabilities to measure similar classes of metabolites at relatively low concentrations (1 μg/g to 30 μg/g). The capabilities required for the analysis of complex

Solid-liquid stable phase equilibria of the ternary systems MgCl2 + MgB6O10+ H2O AND MgSO4 + MgB6O10 + H2O at 308.15 K

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Lingzong Meng

2014-03-01

Full Text Available The solubilities and the relevant physicochemical properties of the ternary systems MgCl2 + MgB6O10 + H2O and MgSO4 + MgB6O10 + H2O at 308.15 K were investigated using an isothermal dissolution method. It was found that there is one invariant point, two univariant curves, and two crystallization regions of the systems. The systems belong to a simple co-saturated type, and neither double salts nor solid solutions were found. Based on the extended HW model and its temperature-dependent equations, the single-salt Pitzer parameters β(0, β(1, β(2 and CØ for MgCl2, MgSO4, and Mg(B6O7(OH6, the mixed ion-interaction parameters θCl,B6O10, θSO4,B6O10, ΨMg,Cl,B6O10, ΨMg,SO4,B6O10 of the systems at 308.15 K were fitted, In addition, the average equilibrium constants of the stable equilibrium solids at 308.15 K were obtained by a method using the activity product constant. Then the solubilities of the ternary systems are calculated. The calculated solubilities agree well with the experimental values.

A Raman spectroscopic study of the structural aspects of K2MgCl4 and Cs2MgCl4 as solid single crystals and molten salts

International Nuclear Information System (INIS)

Brooker, M.H.

1975-01-01

Polarized Raman spectra have been obtained for oriented single crystals of K 2 MgCl 4 and Cs 2 MgCl 4 at 77 and 298 K. The data are in excellent agreement with factor group analyses based on the space groups I 4 /mmm (D 17 4 /subh/) and Pnma (D 16 2 /subh/) for the K 2 MgCl 4 and Cs 2 MgCl 4 crystals. In K 2 MgCl 4 the magnesium is surrounded by six chloride ions in a distorted octahedral arrangement with a network structure such that neighboring octahedra share corners. In Cs 2 MgCl 4 a discrete tetrahedral MgCl 4 2- species is present. The 35 Cl-- 37 Cl isotopic splitting of the symmetric stetching mode of the tetrahedral MgCl 4 2- species has been resolved at 77 K and is similar to that observed for CCl 4 . Raman spectra for the high temperature solids and molten salts suggest that the coordination number of magnesium changes from six in solid K 2 MgCl 4 to four in the melt, whereas Cs 2 MgCl 4 melts with retention of the MgCl 4 2- tetrahedral complex. Additional evidence is presented to support previous reports that the MgCl 4 2- tetrahedral species is the principal complex ion in the melts, although a fraction of the magnesium appears to be present in a polynuclear complex, perhaps Mg 2 Cl 6 2-

Enhanced hydrogen storage properties of MgH2 co-catalyzed with K2NiF6 and CNTs.

Science.gov (United States)

Sulaiman, N N; Ismail, M

2016-12-06

The composite of MgH 2 /K 2 NiF 6 /carbon nanotubes (CNTs) is prepared by ball milling, and its hydrogenation properties are studied for the first time. MgH 2 co-catalyzed with K 2 NiF 6 and CNTs exhibited an improvement in the onset dehydrogenation temperature and isothermal de/rehydrogenation kinetics compared with the MgH 2 -K 2 NiF 6 composite. The onset dehydrogenation temperature of MgH 2 doped with 10 wt% K 2 NiF 6 and 5 wt% CNTs is 245 °C, which demonstrated a reduction of 25 °C compared with the MgH 2 + 10 wt% K 2 NiF 6 composite. In terms of rehydrogenation kinetics, MgH 2 doped with 10 wt% K 2 NiF 6 and 5 wt% CNTs samples absorbed 3.4 wt% of hydrogen in 1 min at 320 °C, whereas the MgH 2 + 10 wt% K 2 NiF 6 sample absorbed 2.6 wt% of hydrogen under the same conditions. For dehydrogenation kinetics at 320 °C, the MgH 2 + 10 wt% K 2 NiF 6 + 5 wt% CNTs sample released 3.3 wt% hydrogen after 5 min of dehydrogenation. By contrast, MgH 2 doped with 10 wt% K 2 NiF 6 released 3.0 wt% hydrogen in the same time period. The apparent activation energy, E a , for the dehydrogenation of MgH 2 doped with 10 wt% K 2 NiF 6 reduced from 100.0 kJ mol -1 to 70.0 kJ mol -1 after MgH 2 was co-doped with 10 wt% K 2 NiF 6 and 5 wt% CNTs. Based on the experimental results, the hydrogen storage properties of the MgH 2 /K 2 NiF 6 /CNTs composite is enhanced because of the catalytic effects of the active species of KF, KH and Mg 2 Ni that are formed in situ during dehydrogenation, as well as the unique structure of CNTs.

Biological effects of an aqueous extract of Salix alba on the survival of Escherichia coli AB1157 cultures submitted to the action of stannous chloride

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RAPHAEL S S SOUZA

2009-01-01

Full Text Available Stannous chloride (SnC12 is used in nuclear medicine as a reducing agent to obtain technetium-99m-radiopharmaceuticals. It have been reported that natural products might reduce the genotoxic and cytotoxic effects related to SnC12. This work evaluated the biological effects of an aqueous extract of Salix alba on the survival of Escherichia coli (E. coli AB1157 (wild type cultures submitted to the action of SnC12. E. coli AB1157 cultures (exponential growth phase were collected by centrifugation, washed and resuspended in 0.9%NaCl. Samples were incubated in water bath shaker with: (a SnC12 (25mg/ml, (bSalix alba extract(11.6mg/ml and (cSnC12(25mg/ml + Salix alba extract (11.6mg/ml. Incubation with 0.9% NaCl was also carried out (control. At 60 min intervals, aliquots were withdrawn, diluted, spread onto Petri dishes with solid LB medium and incubated overnight. The colonies formed were counted and the survival fractions calculated. The extract was not able to protect the E. coli cultures against the lesive action of SnC12. The extract also did not interfere with the survival of the cultures. It suggested that the substances present in the Salix alba aqueous extract did not interfere strongly with cellular metabolism and did not alter the survival fractions of E. coli AB 1157. It is speculated that this extract cannot interfere with the generation of free radicals, the possible main agent responsible for SnC12 lesive action.

APPLICATION OF KATG::LUX GENE CONSTRUCT FOR CYTOTOXICITY AND GENOTOXICITY MONITORING OF METOPROLOL IN ENVIRONMENT

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Eliza Hawrylik

2017-06-01

Full Text Available The aim of the study was the evaluation of usefulness of Escherichia coli K-12 RFM 443 katG::lux for cytotoxicity and genotoxicity monitoring of metoprolol in the environment. Metoprolol is one of the most popular cardiac drug which belongs to the group of β – blockers. The drug was applied at concentrations ranging from 10-1 mg/cm3 to 10-5 mg/cm3. Obtained data indicated the influence of metoprolol on lux gene expression and katG promotor activity in E.coli K-12. The results indicato the possibility of using of Escherichia coli K-12 RFM 443 strain with katG::lux gene construct in the monitoring of cytotoxicity and genotoxicity cardiac drug residues in the environment.

Building a complete image of genome regulation in the model organism Escherichia coli.

Science.gov (United States)

Ishihama, Akira

2018-01-15

The model organism, Escherichia coli, contains a total of more than 4,500 genes, but the total number of RNA polymerase (RNAP) core enzyme or the transcriptase is only about 2,000 molecules per genome. The regulatory targets of RNAP are, however, modulated by changing its promoter selectivity through two-steps of protein-protein interplay with 7 species of the sigma factor in the first step, and then 300 species of the transcription factor (TF) in the second step. Scientists working in the field of prokaryotic transcription in Japan have made considerable contributions to the elucidation of genetic frameworks and regulatory modes of the genome transcription in E. coli K-12. This review summarizes the findings by this group, first focusing on three sigma factors, the stationary-phase sigma RpoS, the heat-shock sigma RpoH, and the flagellar-chemotaxis sigma RpoF, as examples. It also presents an overview of the current state of the systematic research being carried out to identify the regulatory functions of all TFs from a single and the same bacterium E. coli K-12, using the genomic SELEX and PS-TF screening systems. All these studies have been undertaken with the aim of understanding the genome regulation in E. coli K-12 as a whole.

The steady-state kinetics of the NADH-dependent nitrite reductase from Escherichia coli K 12. Nitrite and hydroxylamine reduction.

Science.gov (United States)

Jackson, R H; Cole, J A; Cornish-Bowden, A

1981-01-01

The reduction of both NO2- and hydroxylamine by the NADH-dependent nitrite reductase of Escherichia coli K 12 (EC 1.6.6.4) appears to follow Michaelis-Menten kinetics over a wide range of NADH concentrations. Substrate inhibition can, however, be detected at low concentrations of the product NAD+. In addition, NAD+ displays mixed product inhibition with respect to NADH and mixed or uncompetitive inhibition with respect to hydroxylamine. These inhibition characteristics are consistent with a mechanism in which hydroxylamine binds during catalysis to a different enzyme form from that generated when NAD+ is released. The apparent maximum velocity with NADH as varied substrate increases as the NAD+ concentration increases from 0.05 to 0.7 mM with 1 mM-NO2- or 100 mM-hydroxylamine as oxidized substrate. This increase is more marked for hydroxylamine reduction than for NO2- reduction. Models incorporating only one binding site for NAD can account for the variation in the Michaelis-Menten parameters for both NADH and hydroxylamine with [NAD+] for hydroxylamine reduction. According to these models, activation of the reaction occurs by reversal of an over-reduction of the enzyme by NADH. If the observed activation of the enzyme by NAD+ derives both from activation of the generation of the enzyme-hydroxylamine complex from the enzyme-NO2- complex during NO2- reduction and from activation of the reduction of the enzyme-hydroxylamine complex to form NH4+, then the variation of Vapp. for NO2- or hydroxylamine with [NAD+] is consistent with the occurrence of the same enzyme-hydroxylamine complex as an intermediate in both reactions. PMID:6279095

Non-stoichiometry in the KMo2P3O12-tunnel structure: The oxide K0.75MoNbP3O12

International Nuclear Information System (INIS)

Leclaire, A.; Borel, M.M.; Grandin, A.; Raveau, B.

1990-01-01

K 0.75 MoNbP 3 O 12 , M r =503.009, orthorhombic, Pbcm, a=8.8518 (5), b=9.1453 (11), c=12.5174 (11) A, V=1013.3 (3) A 3 , Z=4, D x =3.300 Mg m -3 , λ(Mo Kα)=0.71073 A, μ=3.13 mm -1 , F(000)=953, T=294 K, R=0.029, wR=0.033 for 1235 observed reflections. This compound is isostructural with KMo 2 P 3 O 12 -type oxides. Its framework is built up from MoO 6 octahedra and PO 4 tetrahedra which delimit tunnels running along b. Different from KMo 2 P 3 O 12 , the tunnels are partly occupied by the potassium ions which are distributed at random. (orig.)

Converting hcp Mg-Al-Zn alloy into bcc Mg-Li-Al-Zn alloy by electrolytic deposition and diffusion of reduced lithium atoms in a molten salt electrolyte LiCl-KCl

International Nuclear Information System (INIS)

Lin, M.C.; Tsai, C.Y.; Uan, J.Y.

2007-01-01

A body-centered cubic (bcc) Mg-12Li-9Al-1Zn (wt.%) alloy was fabricated in air by electrolysis from LiCl-KCl molten salt at 500 deg. C. Electrolytic deposition of Li atoms on cathode (Mg-Al-Zn alloy) and diffusion of the Li atoms formed the bcc Mg-Li-Al-Zn alloy with 12 wt.% Li and only 0.264 wt.% K. Low K concentration in the bcc Mg alloy strip after the electrolysis process resulted from 47% atomic size misfit between K and Mg atoms and low solubility of K in Mg matrix

Synergistic effects in mixed Escherichia coli biofilms

DEFF Research Database (Denmark)

Reisner, A.; Holler, B.M.; Molin, Søren

2006-01-01

Bacterial biofilms, often composed of multiple species and genetically distinct strains, develop under complex influences of cell-cell interactions. Although detailed knowledge about the mechanisms underlying formation of single-species laboratory biofilms has emerged, little is known about...... the pathways governing development of more complex heterogeneous communities. In this study, we established a laboratory model where biofilm-stimulating effects due to interactions between genetically diverse strains of Escherichia coli were monitored. Synergistic induction of biofilm formation resulting from...... the cocultivation of 403 undomesticated E. coli strains with a characterized E. coli K-12 strain was detected at a significant frequency. The survey suggests that different mechanisms underlie the observed stimulation, yet synergistic development of biofilm within the subset of E. coli isolates (n = 56) exhibiting...

Genetic diversity of K-antigen gene clusters of Escherichia coli and their molecular typing using a suspension array.

Science.gov (United States)

Yang, Shuang; Xi, Daoyi; Jing, Fuyi; Kong, Deju; Wu, Junli; Feng, Lu; Cao, Boyang; Wang, Lei

2018-04-01

Capsular polysaccharides (CPSs), or K-antigens, are the major surface antigens of Escherichia coli. More than 80 serologically unique K-antigens are classified into 4 groups (Groups 1-4) of capsules. Groups 1 and 4 contain the Wzy-dependent polymerization pathway and the gene clusters are in the order galF to gnd; Groups 2 and 3 contain the ABC-transporter-dependent pathway and the gene clusters consist of 3 regions, regions 1, 2 and 3. Little is known about the variations among the gene clusters. In this study, 9 serotypes of K-antigen gene clusters (K2ab, K11, K20, K24, K38, K84, K92, K96, and K102) were sequenced and correlated with their CPS chemical structures. On the basis of sequence data, a K-antigen-specific suspension array that detects 10 distinct CPSs, including the above 9 CPSs plus K30, was developed. This is the first report to catalog the genetic features of E. coli K-antigen variations and to develop a suspension array for their molecular typing. The method has a number of advantages over traditional bacteriophage and serum agglutination methods and lays the foundation for straightforward identification and detection of additional K-antigens in the future.

Genetic and physiological analysis of an envB spherelike mutant of Escherichia coli K-12 and characterization of its transductants

International Nuclear Information System (INIS)

Westling-Haggstrom, B.; Normark, S.

1975-01-01

The envB1 mutation mediating a distorted cell morphology of Escherichia coli K-12 was cotransducible with strA, aroE, aspB, and argG. The mapping data is consistent with a gene location for envB around 62.5 min. In partial diploids envB1 was recessive to its wild-type allele. The original envB mutant contained a second mutation in a locus denoted sloB close to strA. The following gene order is suggested: sloB-strA-aroE-envB-aspB-argG. The sloB1 mutation caused a marked reduction in the growth rate of both envB and envB + strains. Moreover, this mutation in the presence of envB1 appears to increase the ratio between deoxyribonucleic acid and protein in cells growing in rich medium. The phenotypic properties of envB1, sloB + , and envB + transductants were characterized. Cells with envB1, sloB + genotype were hypersensitive to several penicillins including the β-lactam compound, amidino penicillin. Penicillin hypersensitivity could not be explained by increased outer membrane penetrability. The original envB mutant (envB1, sloB1), as well as envB1, sloB1 or envB + , sloB1 transductants were resistant to amidino penicillin. Resistance was explained by the slow growth rate medicated by the sloB1 mutation. The similarity between envB cells and wild-type cells treated with sublethal concentrations of amidino penicillin was emphasized. (U.S.)

Low Temperature Mechanical Properties of Scandium-Modified Al-Zn-Mg-Cu Alloys

National Research Council Canada - National Science Library

Senkov, O

2002-01-01

Tensile properties of three wrought alloys, (1) Al-10Zn-3Mg-1.2Cu-0.15Zr, (2) Al-10Zn-3Mg-1.2Cu-0.15Zr-0.39Mn-0.49Sc, and (3) Al-12Zn-3Mg-1.2Cu-0.15Zr-0.39Mn-0.49Sc were studied in T6 and T7 conditions at 298K and 77K...

RESPONS ANTIBODI ANTI ETEC K99 PADA INDUK SAPI BUNTING SETELAH PEMBERIAN VAKSIN ESCHERICHIA COLI POLIVALEN

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Anita Esfandiari

2014-08-01

Full Text Available The objective of this experiment was to detect antibody (IgG against enterotoxigenic Escherichia coli (ETECK99 in the blood of cows vaccinated by Escherichia coli polyvalent vaccine. Eight dry cows were injectedsubcutaneously by polyvalent Escherichia coli twice prior to parturition. Before vaccinated, the cows were givenimmunomodulator orally for 3 days. Blood samples were drawn from coccigeal vein prior to the 1st vaccination,two week following the 1st vaccination and at 1, 2, and 4 weeks after the 2nd vaccination. Blood samples wereanalyzed for IgG and ETEC K99 using indirect Enzyme Linked Immunosorbent Assay (ELISA assay. Results of theexperiment indicated that absorbance values of all vaccinated cows before the first vaccination until third weekfollowing the 2nd vaccination were below cut off values. The absorbance values then increased and were above cutoff values at fourth week following the 2nd vaccination. In conclusion, antibody against ETEC K99, were detected inthe blood of cows, fourth week following the 2nd vaccination.

The effect of E coli virulence on bacterial translocation and systemic sepsis in the neonatal rabbit model.

Science.gov (United States)

Jackson, R J; Smith, S D; Wadowsky, R M; DePudyt, L; Rowe, M I

1991-04-01

In the surgical neonate, three factors that promote bacterial translocation and systemic infection are: (1) intestinal bacterial colonization and overgrowth; (2) compromised host defenses; and (3) disruption of the mucosal epithelial barrier. The newborn rabbit provides an excellent model to study these factors. Like the human, there is early closure of the gut mucosa to macromolecules, and nutrition can be maintained by breast or formula feeding. This study examines translocation and systemic sepsis after colonization with virulent K1 and avirulent K100 strains of Escherichia coli. New Zealand white rabbit pups (2 to 5 days old) were studied. The gastrointestinal tracts of 12 were colonized with K1 E coli; 14 were colonized with K100 E coli; 12 control animals were not inoculated. Mesenteric lymph node (MLN), liver, spleen, and colon homogenate were cultured 72 hours postinoculation. No bacteria were isolated from the colons of all but one control animal. Translocation or systemic sepsis did not occur. Translocation to the MLN was significantly increased (P less than .03) in K1 (50%) and K100 (36%) groups compared with controls (0%). Translocation to liver and spleen (systemic sepsis) was significantly increased (P less than .03) in K1 animals (67%) compared with K100 (0%) or controls (0%). Colonization by both strains of E coli led to translocation to the MLN, but only K1 E coli caused systemic sepsis. This suggests that although colonization by E coli in the newborn leads to translocation to the MLN, progression to systemic sepsis is the result of characteristics of the bacteria and/or neonatal host responses.

Mechanical properties and fracture mechanism of as-cast Mg{sub 77}TM{sub 12}Zn{sub 5}Y{sub 6} (TM = Cu, Ni) bulk amorphous matrix composites

Energy Technology Data Exchange (ETDEWEB)

Qiu, K.Q. [School of Materials Science and Engineering, Shenyang University of Technology, Shenyang 110178 (China)], E-mail: kqqiu@yahoo.com.cn; Hu, N.N.; Zhang, H.B. [School of Materials Science and Engineering, Shenyang University of Technology, Shenyang 110178 (China); Jiang, W.H. [Department of Materials Science and Engineering, The University of Tennessee, Knoxville, TN 37990 (United States); Ren, Y.L. [School of Materials Science and Engineering, Shenyang University of Technology, Shenyang 110178 (China); Liaw, P.K. [Department of Materials Science and Engineering, The University of Tennessee, Knoxville, TN 37990 (United States)

2009-06-10

Comparative investigations on the microstructures, thermal stability and mechanical properties of Mg{sub 77}Cu{sub 12}Zn{sub 5}Y{sub 6} and Mg{sub 77}Ni{sub 12}Zn{sub 5}Y{sub 6} bulk metallic glass matrix composites were carried out by using scanning electron microscopy (SEM), DSC and compressive tester. The results show that the microstructure of as-cast samples with 3 mm in diameter for Cu-containing alloy is consisted of Mg flakes and dotted Mg{sub 2}Cu phase in the amorphous matrix, while the as-cast Ni-containing alloy with the same diameter is mainly consisted of Mg flakes in the amorphous matrix. The glass transition temperature and supercooled liquid region are 413 K and 27 K for the Cu-containing, 443 K and 32 K for the Ni-containing amorphous matrix composites, respectively. The fracture strength, yield strength and plastic strain are 532 MPa, 390 MPa and 2.4% for the Cu-containing alloy, 667 MPa, 412 MPa and 7% for the Ni-containing alloy, respectively. Furthermore, the fracture mechanism for the amorphous matrix composites was discussed according to both the fracture surfaces and the stress-strain curves.

Cellular chain formation in Escherichia coli biofilms

DEFF Research Database (Denmark)

Vejborg, Rebecca Munk; Klemm, Per

2009-01-01

; type I fimbriae expression significantly reduced cellular chain formation, presumably by steric hindrance. Cellular chain formation did not appear to be specific to E coli K-12. Although many urinary tract infection (UTI) isolates were found to form rather homogeneous, flat biofilms, three isolates...

[Effect of Pseudomonas aeruginosa exometabolites on planktonic and biofilm cultures of Escherichia coli].

Science.gov (United States)

Kuznetsova, M V; Karpunina, T I; Maslennikova, I L; Nesterova, L Iu; Demakov, V A

2012-01-01

Study the effect of P. aeruginosa exometabolites on planktonic and biofilm cultures of bioluminescent E. coli strain. E. coli K12 TG1 (pF1 lux+ Ap(r)) recombinant bioluminescent strain, P. aeruginosa ATCC 27853 reference strain and 2 nosocomial isolates were used. Pyocyanin and pyoverdin content in supernatant of P. aeruginosa over-night cultures was evaluated according to E. Deziel et al. (2001). Planktonic and biofilm cultures of E. coli were obtained in 96-well plates (LB, statically, 37 degrees C), optical density of plankton, film biomass (OD600, OD580) and bioluminescence in plankton and biofilm were evaluated in microplate reader Infiniti M200 (Tecan, Austria). P. aeruginosa exometabolites increased the duration of lag-phase in E. coli, and short term exposition inhibited luminescence of planktonic cells. These effects are determined by bactericidal action ofpyocyanin and pyoverdin. Supernatants ofover-night cultures of P. aeruginosa inhibit formation of biofilm and disrupt the formed biofilm of E. coli. Effect of pyocyanin and pyoverdin on these processes is not established, other factors may have higher significance. Bioluminescence of E. coli K12 TGI that reflects the energetic status of the cell allows to evaluate and prognose the character of coexistence of P. aeruginosa in combined with E. coli planktonic and biofilm culture.

The Escherichia coli K-12 gntP gene allows E. coli F-18 to occupy a distinct nutritional niche in the streptomycin-treated mouse large intestine

DEFF Research Database (Denmark)

Sweeney, N.J.; Klemm, Per; McCormick, Beth A.

1996-01-01

Escherichia coli F-18 is a human fecal isolate that makes type 1 fimbriae, encoded by the fim gene cluster, and is an excellent colonizer of the streptomycin-treated mouse intestine. E. coli F-18 fimA::tet, lacking type 1 fimbriae, was constructed by bacteriophage P1 transduction of the fim regio...

Thermoluminescence of LiF: Mg, Ti between 77 and 315 K

International Nuclear Information System (INIS)

Rosa, L.A.R. da.

1989-01-01

A special thermoluminescent system was developed. It is able to operate right from liquid nitrogen temperature and also permits the determination of the sample thermoluminescent emission spectrum. Using this system, the thermoluminescence displayed by 77 K irradiated LiF:Mg,Ti (TLD-100), from the irradiation temperature to 315 K, was studied. In this temperature range seven glow peaks, at 139, 153, 194, 240, 260, 283 and 300 K, were determined. (author)

beta-Chloro-L-alanine inhibition of the Escherichia coli alanine-valine transaminase.

OpenAIRE

Whalen, W A; Wang, M D; Berg, C M

1985-01-01

beta-Chloro-L-alanine, an amino acid analog which inhibits a number of enzymes, reversibly inhibited the Escherichia coli K-12 alanine-valine transaminase, transaminase C. This inhibition, along with the inhibition of transaminase B, accounted for the isoleucine-plus-valine requirement of E. coli in the presence of beta-chloro-L-alanine.

Involvement of UV-inducible repair in pyrimidine dimer excision in Escherichia coli

International Nuclear Information System (INIS)

Masek, F.; Sedliakova, M.

1978-01-01

The influence of UV radiation on pyrimidine dimer excision in the cells of three excision-proficient E.coli strains was studied. For this purpose cells were irradiated with a first fluence of 300 ergs/mm 2 and at different time intervals with a second fluence of 500 ergs/mm 2 . After the second fluence dimer excision was found to be partly inhibited in E.coli B/r Hcr + and E.coli 15 555-7, but not in E.coli K12 SR20. (author)

Involvement of UV-inducible repair in pyrimidine dimer excision in Escherichia coli

Energy Technology Data Exchange (ETDEWEB)

Masek, F; Sedliakova, M [Slovenska Akademia Vied, Bratislava (Czechoslovakia)

1978-11-15

The influence of UV radiation on pyrimidine dimer excision in the cells of three excision-proficient E.coli strains was studied. For this purpose cells were irradiated with a first fluence of 300 ergs/mm/sup 2/ and at different time intervals with a second fluence of 500 ergs/mm/sup 2/. After the second fluence dimer excision was found to be partly inhibited in E.coli B/r Hcr/sup +/ and E.coli 15 555-7, but not in E.coli K12 SR20.

Lactobacillus rhamnosus GG Suppresses Meningitic E. coli K1 Penetration across Human Intestinal Epithelial Cells In Vitro and Protects Neonatal Rats against Experimental Hematogenous Meningitis

OpenAIRE

Sheng-He Huang; Lina He; Yanhong Zhou; Chun-Hua Wu; Ambrose Jong

2009-01-01

The purpose of this study was to examine prophylactic efficacy of probiotics in neonatal sepsis and meningitis caused by E. coli K1. The potential inhibitory effect of Lactobacillus rhamnosus GG (LGG) on meningitic E. coli K1 infection was examined by using (i) in vitro inhibition assays with E44 (a CSF isolate from a newborn baby with E. coli meningitis), and (ii) the neonatal rat model of E. coli sepsis and meningitis. The in vitro studies demonstrated that LGG blocked E44 adhesion, invasio...

Inhibition of Apoptosis by Escherichia coli K1 Is Accompanied by Increased Expression of BclXL and Blockade of Mitochondrial Cytochrome c Release in Macrophages

OpenAIRE

Sukumaran, Sunil K.; Selvaraj, Suresh K.; Prasadarao, Nemani V.

2004-01-01

Escherichia coli K1 survival in the blood is a critical step for the onset of meningitis in neonates. Therefore, the circulating bacteria are impelled to avoid host defense mechanisms by finding a niche to survive and multiply. Our recent studies have shown that E. coli K1 enters and survives in both monocytes and macrophages in the newborn rat model of meningitis as well as in macrophage cell lines. Here we demonstrate that E. coli K1 not only extends the survival of human and murine infecte...

NleB/SseK effectors from Citrobacter rodentium, Escherichia coli, and Salmonella enterica display distinct differences in host substrate specificity

DEFF Research Database (Denmark)

El Qaidi, Samir; Chen, Kangming; Halim, Adnan

2017-01-01

proteins with N-acetyl-D-glucosamine to inhibit antibacterial and inflammatory host responses. NleB is conserved among the attaching/effacing pathogens enterohemorrhagic E. coli (EHEC), enteropathogenic E. coli (EPEC), and Citrobacter rodentium. Moreover, Salmonella enterica strains encode up to three Nle......B orthologs named SseK1, SseK2, and SseK3. However, there are conflicting reports regarding the activities and host protein targets among the NleB/SseK orthologs. Therefore, here we performed in vitro glycosylation assays and cell culture experiments to compare the activities and substrate specificities...... of these effectors. SseK1, SseK3, EHEC NleB1, EPEC NleB1, and C. rodentium NleB blocked TNF-mediated NF-κB pathway activation, whereas SseK2 and NleB2 did not. C. rodentium NleB, EHEC NleB1, and SseK1 glycosylated host glyceraldehyde 3-phosphate dehydrogenase (GAPDH). C. rodentium NleB, EHEC NleB1, EPEC NleB1...

Evaluation of the Live Biotherapeutic Product, Asymptomatic Bacteriuria Escherichia coli 2-12, in Healthy Dogs and Dogs with Clinical Recurrent UTI.

Science.gov (United States)

Segev, G; Sykes, J E; Klumpp, D J; Schaeffer, A J; Antaki, E M; Byrne, B A; Yaggie, R E; Westropp, J L

2018-01-01

Antimicrobial resistance is an emerging problem. To investigate the safety and efficacy of a live biotherapeutic product, ASB E. coli 2-12 for UTI treatment. Six healthy research dogs; nine client-owned dogs with recurrent UTI. Prospective noncontrolled clinical trial. For safety data, research dogs were sedated, a urinary catheter was inserted into the bladder; 10 10 CFU/mL of ASB E. coli 2-12 was instilled. Urine was cultured on days 1, 3, and 8 post-instillation and dogs were observed for lower urinary tract signs (LUTS). For client-owned dogs, ASB E. coli 2-12 was instilled similarly and urine cultures analyzed on days 1, 7, and 14 days postinstillation. No LUTS were noted in any of the 6 research dogs after ASB E. coli 2-12 infusion. Pulse field gel electrophoresis (PFGE) studies confirmed the bacterial strains isolated matched that ASB E. coli 2-12 strain. Four of the nine client-owned dogs had complete or nearly complete clinical cures by day 14. Of these four dogs, 3 also had microbiologic cures at day 14; one of these dogs had subclinical bacteriuria (in addition to ASB E. coli 2-12). Three of these four dogs had ASB E. coli 2-12 isolated from their urine at day 14. With the exception of mild, temporary, self-limiting, hyporexia in two dogs on the day of biotherapeutic administration, there were no major adverse effects. These results suggest ASB E. coli 2-12 is safe and should be investigated in a larger controlled study evaluating clinical UTI in dogs. Copyright © 2017 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

Leaner and meaner genomes in Escherichia coli

DEFF Research Database (Denmark)

Ussery, David

2006-01-01

A 'better' Escherichia coli K-12 genome has recently been engineered in which about 15% of the genome has been removed by planned deletions. Comparison with related bacterial genomes that have undergone a natural reduction in size suggests that there is plenty of scope for yet more deletions....

Microstructures and Dehydrogenation Properties of Ball-milled MgH2-K2Ti6O13-Ni Composite Systems

Directory of Open Access Journals (Sweden)

ZHANG Jian

2016-11-01

Full Text Available The K2Ti6O13 whisker separate-doped and K2Ti6O13 whisker and Ni powder multi-doped MgH2 hydrogen storage composite systems were prepared by mechanical milling method. The microstructures and dehydrogenation properties of the prepared samples were characterized by some testing methods such as X-ray diffraction (XRD, scanning electron microscope (SEM and differential scanning calorimeter (DSC. The results show that the K2Ti6O13 whisker not only plays the roles in refining the MgH2 crystalline grain, but also inhibit the agglomeration of MgH2 particles in K2Ti6O13 whisker separate-doped system, which results in the decreased dehydrogenation temperature of MgH2 matrix. When the mass ratio of K2Ti6O13 to MgH2 is 3:7, the improvement effect on dehydrogenation properties of MgH2 is the most remarkable. As compared with pure ball-milled MgH2, the dehydrogenation temperature of MgH2 in K2Ti6O13 whisker separate-doped system is decreased by nearly 75℃. For K2Ti6O13 whisker and Ni powder multi-dopedsystem, the dehydrogenation temperature of MgH2 matrix is further decreased compared to K2Ti6O13 whisker separate-doped one due to the dual effects of refined MgH2 crystalline grain by K2Ti6O13 whisker and destabilized MgH2 lattice by Ni solution. As compared with pure ball-milled MgH2, the dehydrogenation temperature of MgH2 in K2Ti6O13 whisker and Ni powder multi-doped system is decreased by nearly 87℃.

Synthesis of avenanthramides using engineered Escherichia coli.

Science.gov (United States)

Lee, Su Jin; Sim, Geun Young; Kang, Hyunook; Yeo, Won Seok; Kim, Bong-Gyu; Ahn, Joong-Hoon

2018-03-22

Hydroxycinnamoyl anthranilates, also known as avenanthramides (avns), are a group of phenolic alkaloids with anti-inflammatory, antioxidant, anti-itch, anti-irritant, and antiatherogenic activities. Some avenanthramides (avn A-H and avn K) are conjugates of hydroxycinnamic acids (HC), including p-coumaric acid, caffeic acid, and ferulic acid, and anthranilate derivatives, including anthranilate, 4-hydroxyanthranilate, and 5-hydroxyanthranilate. Avns are primarily found in oat grain, in which they were originally designated as phytoalexins. Knowledge of the avns biosynthesis pathway has now made it possible to synthesize avns through a genetic engineering strategy, which would help to further elucidate their properties and exploit their beneficial biological activities. The aim of the present study was to synthesize natural avns in Escherichia coli to serve as a valuable resource. We synthesized nine avns in E. coli. We first synthesized avn D from glucose in E. coli harboring tyrosine ammonia lyase (TAL), 4-coumarate:coenzyme A ligase (4CL), anthranilate N-hydroxycinnamoyl/benzoyltransferase (HCBT), and anthranilate synthase (trpEG). A trpD deletion mutant was used to increase the amount of anthranilate in E. coli. After optimizing the incubation temperature and cell density, approximately 317.2 mg/L of avn D was synthesized. Avn E and avn F were then synthesized from avn D, using either E. coli harboring HpaBC and SOMT9 or E. coli harboring HapBC alone, respectively. Avn A and avn G were synthesized by feeding 5-hydroxyanthranilate or 4-hydroxyanthranilate to E. coli harboring TAL, 4CL, and HCBT. Avn B, avn C, avn H, and avn K were synthesized from avn A or avn G, using the same approach employed for the synthesis of avn E and avn F from avn D. Using different HCs, nine avns were synthesized, three of which (avn D, avn E, and avn F) were synthesized from glucose in E. coli. These diverse avns provide a strategy to synthesize both natural and unnatural avns

Structure of spinel at high temperature using in-situ XANES study at the Al and Mg K-edge

Energy Technology Data Exchange (ETDEWEB)

Ligny, D de [Universite Claude Bernard Lyon 1, LPCML, 69622 Villeurbanne (France); Neuville, D R [Physique des Mineraux et Magmas, Geochimie-Cosmochimie, CNRS-IPGP, 4 place Jussieu, 75005 Paris (France); Flank, A-M; Lagarde, P, E-mail: deligny@pcml.univ-lyon1.f [Synchrotron SOLEIL, L' Orme des Merisiers, Saint Aubin, 91192 France (France)

2009-11-15

We present structural information obtained on spinel at high temperature (298-2400 K) using in situ XANES at the Mg and Al K-edge. Spinel, {sup [4]}(Al{sub x},Mg{sub 1-x}){sup [6]}(Al{sub 2-x},Mg{sub x})O{sub 4}, with increasing temperature, show a substitution of Mg by Al and Al by Mg in their respective sites. This substitution corresponds to an inversion of the Mg and Al sites. Furthermore, both experiments at the Al and Mg K-edges are in good agreement with XANES calculation made using FDMNES code.

NarK is a nitrite-extrusion system involved in anaerobic nitrate respiration by Escherichia coli

NARCIS (Netherlands)

Rowe, John J.; Ubbink-Kok, Trees; Molenaar, Douwe; Konings, Wilhelmus; Driessen, Arnold J.M.

Escherichia coli can use nitrate as a terminal electron acceptor for anaerobic respiration. A polytopic membrane protein, termed NarK, has been implicated in nitrate uptake and nitrite excretion and is thought to function as a nitrate/nitrite antiporter. The longest-lived radioactive isotope of

K-12 Local Network (LAN) Design Guide

National Research Council Canada - National Science Library

Horton, Cody

1998-01-01

...) educators preparing to design and implement LANs in K-12 schools and libraries. Data was collected during the implementation of LANs in K-12 schools of the Monterey Peninsula Uniform School District (MPUSD...

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

Directory of Open Access Journals (Sweden)

Kateřina Nedbalcová

2015-01-01

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

Antibacterial Activities and Possible Modes of Action of Acacia nilotica (L. Del. against Multidrug-Resistant Escherichia coli and Salmonella

Directory of Open Access Journals (Sweden)

Muhammad Bilal Sadiq

2017-01-01

Full Text Available Medicinal plants are frequently used for the treatment of various infectious diseases. The objective of this study was to evaluate the antibacterial activity and mode of action of Acacia nilotica and the antibiogram patterns of foodborne and clinical strains of Escherichia coli and Salmonella. The mechanism of action of acacia extracts against E. coli and Salmonella was elucidated by observing morphological damages including cell integrity and cell membrane permeability, as well as changes in cell structures and growth patterns in kill-time experiments. The clinical isolates of E. coli and Salmonella were found resistant to more of the tested antibiotics, compared to food isolates. Minimum inhibitory concentration and minimum bactericidal concentration of acacia leaf extracts were in the ranges of 1.56–3.12 mg/mL and 3.12–6.25 mg/mL, respectively, whereas pods and bark extracts showed somewhat higher values of 3.12–6.25 mg/mL and 6.25–12.5 mg/mL, respectively, against all tested pathogens. The release of electrolytes and essential cellular constituents (proteins and nucleic acids indicated that acacia extracts damaged the cellular membrane of the pathogens. These changes corresponded to simultaneous reduction in the growth of viable bacteria. This study indicates that A. nilotica can be a potential source of new antimicrobials, effective against antibiotic-resistant strains of pathogens.

Creep behaviour of a casting titanium carbide reinforced AlSi12CuNiMg piston alloy at elevated temperatures; Hochtemperaturkriechverhalten der schmelzmetallurgisch hergestellten dispersionsverstaerkten Kolbenlegierung AlSi12CuNiMg

Energy Technology Data Exchange (ETDEWEB)

Michel, S.; Scholz, A. [Zentrum fuer Konstruktionswerkstoffe, TU Darmstadt (Germany); Tonn, B. [Institut fuer Metallurgie, TU Clausthal (Germany); Zak, H.

2012-03-15

This paper deals with the creep behaviour of the titanium carbide reinforced AlSi12CuNiMg piston alloy at 350 C and its comparison to the conventional AlSi12Cu4Ni2MgTiZr piston alloy. With only 0,02 vol-% TiC reinforcement the creep strength and creep rupture strength of the AlSi12CuNiMg piston alloy are significantly improved and reach the level of the expensive AlSi12Cu4Ni2MgTiZr alloy. (Copyright copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

CLINICAL ISOLATES OF MECA, METHICILLIN, VANCOMYCIN RESISTANCE S. AUREUS; ESBLs PRODUCING K.PNEUMONIA, E.COLI, P. AUREGENOSA FROM VARIOUS CLINICAL SOURCE AND ITS ANTIMICROBIAL RESISTANCE PATTERNS

Directory of Open Access Journals (Sweden)

Ismail Mahmud Ali, Amirthalingam R

2015-01-01

Full Text Available Background and Objective: Antimicrobial resistance has turned into a key medical and public health crisis globally since the injudicious use of magic bullets (drugs. Aim of this study is focused on the clinical isolate and their percentages of resistant to antibiotics in gram positive bacteria such as MRSA, VRSA, and MSSA are common causes of nosocomical, skin structure infections, bacteremia and infection of other systems; ESBLs producing Enterobacteriaceae (E. coli, Klebsiella spp. is common agent of urinary tract, bloodstream, pulmonary and intra-abdominal infections and carbapenem resistant P. aeruginosa with its complete antimicrobial patterns which are currently practiced in this population. Methods: There are one hundred and fourteen (114 various clinical isolates, isolated from various clinical samples like throat swab, urine, pus, sputum, and blood culture, identified as specific isolate with resistance patterns were analyzed by BD phoenix-100 the auto analyzer. Results: Off 114 clinical isolate, 6 mecA-mediated resistance (cefoxitin>8mgc/ml, 11 methicillin resistance, 18 β lactam/βlactamase inhibitor, 12 methicillin sensitive and 3 vancomycin (>16µg/ml resistance S. aureus have been isolated from overall 50 isolate of S.aureus. In addition, there are 27 P.aeruginosa, 15 ESBLs from overall of 25 K. pneumoniae and 7 ESBLs out of 12 Escherichia coli species have been isolated. The resistance and susceptibility pattern percentages have been graphically represented for each isolates. Conclusion: Current study revealed that the drug classes of β lactam/βlactamase inhibitor having high resistance rate with S.aureus, P.aureginosa, K. pneumoniae and E. coli isolate. Also, some of other drug classes such as cepham and tetracycline having higher resistance rate with P.aureginosa and K.pneumoniae. In addition, the vancomycin resistances S. aureus have been isolated and reported as first time in this population.

Homologous overexpression of RfaH in E. coli K4 improves the production of chondroitin-like capsular polysaccharide.

Science.gov (United States)

Cimini, Donatella; De Rosa, Mario; Carlino, Elisabetta; Ruggiero, Alessandro; Schiraldi, Chiara

2013-05-09

Glycosaminoglycans, such as hyaluronic acid, heparin, and chondroitin sulfate, are among the top ranked products in industrial biotechnology for biomedical applications, with a growing world market of billion dollars per year. Recently a remarkable progress has been made in the development of tailor-made strains as sources for the manufacturing of such products. The genetic modification of E. coli K4, a natural producer of chondroitin sulfate precursor, is challenging considering the lack of detailed information on its genome, as well as its mobilome. Chondroitin sulfate is currently used as nutraceutical for the treatment of osteoarthritis, and several new therapeutic applications, spanning from the development of skin substitutes to live attenuated vaccines, are under evaluation. E. coli K4 was used as host for the overexpression of RfaH, a positive regulator that controls expression of the polysaccharide biosynthesis genes and other genes necessary for the virulence of E. coli K4. Various engineering strategies were compared to investigate different types of expression systems (plasmid vs integrative cassettes) and integration sites (genome vs endogenous mobile element). All strains analysed in shake flasks on different media showed a capsular polysaccharide production improved by 40 to 140%, compared to the wild type, with respect to the final product titer. A DO-stat fed-batch process on the 2L scale was also developed for the best performing integrative strain, EcK4r3, yielding 5.3 g ∙ L(-1) of K4 polysaccharide. The effect of rfaH overexpression in EcK4r3 affected the production of lipopolysaccharide and the expression of genes involved in the polysaccharide biosynthesis pathway (kfoC and kfoA), as expected. An alteration of cellular metabolism was revealed by changes of intracellular pools of UDP-sugars which are used as precursors for polysaccharide biosynthesis. The present study describes the identification of a gene target and the application of a

Magnetic properties of Mg12O12 nanocage doped with transition metal atoms (Mn, Fe, Co and Ni): DFT study

Science.gov (United States)

Javan, Masoud Bezi

2015-07-01

Binding energy of the Mg12O12 nanocage doped with transition metals (TM=Mn, Fe, Co and Ni) in endohedrally, exohedrally and substitutionally forms were studied using density functional theory with the generalized gradient approximation exchange-correlation functional along 6 different paths inside and outside of the Mg12O12 nanocage. The most stable structures were determined with full geometry optimization near the minimum of the binding energy curves of all the examined paths inside and outside of the Mg12O12 nanocage. The results reveal that for all stable structures, the Ni atom has a larger binding energy than the other TM atoms. It is also found that for all complexes additional peaks contributed by TM-3d, 4s and 4p states appear in the highest occupied molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO) gap of the host MgO cluster. The mid-gap states are mainly due to the hybridization between TM-3d, 4s and 4p orbitals and the cage π orbitals. The magnetic moment of the endohedrally doped TM atoms in the Mg12O12 are preserved to some extent due to the interaction between the TM and Mg12O12 nanocage, in contrast to the completely quenched magnetic moment of the Fe and Ni atoms in the Mg11(TM)O12 complexes. Furthermore, charge population analysis shows that charge transfer occurs from TM atom to the cage for endohedrally and substitutionally doping.

YibK is the 2'-O-methyltransferase TrmL that modifies the wobble nucleotide in Escherichia coli tRNA(Leu) isoacceptors

DEFF Research Database (Denmark)

Benítez-Páez, Alfonso; Villarroya, Magda; Douthwaite, Stephen Roger

2010-01-01

to uncover candidate E. coli genes for the missing enzyme(s). Transfer RNAs from null mutants for candidate genes were analyzed by mass spectrometry and revealed that inactivation of yibK leads to loss of 2'-O-methylation at position 34 in both tRNA(Leu)(CmAA) and tRNA(Leu)(cmnm5UmAA). Loss of Yib...... of the wobble nucleotide; YibK recognition of this target requires a pyridine at position 34 and N⁶-(isopentenyl)-2-methylthioadenosine at position 37. YibK is one of the last remaining E. coli tRNA modification enzymes to be identified and is now renamed TrmL.......Transfer RNAs are the most densely modified nucleic acid molecules in living cells. In Escherichia coli, more than 30 nucleoside modifications have been characterized, ranging from methylations and pseudouridylations to more complex additions that require multiple enzymatic steps. Most...

Hydration Experiments and Physical Observations at 193 K and 243 K for Mg-Sulfates Relevant to Mars

Science.gov (United States)

Vaniman, D. T.; Chipera, S. J.; Carey, J. W.

2006-03-01

Hydration of kieserite and amorphous Mg-sulfate at 243 K progresses along simple pathways involving only hexahydrite and kieserite. Kieserite forms a duricrust-like cement, but the anhydrous precursor does not.

Radiosensitization and radioprotection of E. coli by alcohols

International Nuclear Information System (INIS)

Worm, K.-H.; Klimczak, U.; Schulte-Frohlinde, D.

1993-01-01

The survival of E. coli K12 strain AB1157 and the isogenic repair-deficient mutant E. coli AB2480 (recA13, uvrA6) was measured after γ-irradiation in the presence of various alcohols as well as after incubation and subsequent removal of the alcohols before irradiation. The authors conclude that alcohols protect predominantly by OH radical scavenging. The comparatively small protection of cell survival by the more hydrophobic alcohols can be attributed to the sensitizing effect of these alcohols. (author)

Spectroscopic information from (3He, 7Be) reaction on 12C and 24Mg

International Nuclear Information System (INIS)

Rahman, Md.A.; Sen Gupta, H.M.

1986-01-01

The reaction ( 3 He, 7 Be) on 12 C and 24 Mg has been analysed using four discrete potential families for 7 Be channel and one discrete potential family for 3 He channel to extract alpha spectroscopic factors. It is shown that the relative spectroscopic factors are reliable if they are calculated staying within one potential family (S( 24 Mg/ 12 C) approx. 0.12). But, changing the potential family between 12 C and 24 Mg, one obtains the extreme cases, such as S( 24 Mg/ 12 C) = 0.025 and 0.51, i.e. 1:20

Irradiation effect of escherichia coli O157 : H7 in meats

International Nuclear Information System (INIS)

Ito, Hitoshi; Harsojo

1998-01-01

Escherichia coli O157 : H7 is a rapidly emerging food-born pathogen which has been linked to outbreaks of hemorrhagic diarrhea in Japan, USA or many European countries. From this study, two strains of E. coli O157 : H7 were isolated from beef and chicken meats in each one sample of 4 replicates. Some of the biochemical characteristics of these isolates were different from type strain IID959. These isolates could grow quickly at 10degC on cultivation of nutrient agar. D 10 value of these isolates were obtained to be 0.06kGy in 0.067M phosphate buffer suspension which were highly sensitive than D 10 value of 0.12kGy on type strain IID959. On the irradiation effect of type strain IID959 in ground beef, D 10 value was obtained as 0.26kGy at fresh condition and 0.46kGy at frozen condition, respectively. From these results, necessary dose for elimination of E. coli O157 : H7 was decided as 1.5kGy for fresh meats and 3kGy for frozen meats. (author)

Thermal degradation products of saccharides: effect study over Escherichia coli K12S cells; Produtos de termodegradacao de sacarideos: estudo do efeito sobre celulas de Escherichia coli K12S

Energy Technology Data Exchange (ETDEWEB)

Oliveira, R.L.B.C. de

1981-12-31

The heat sterilization of reducing sugars, in the presence of phosphates, in alkaline pH, promotes caramelization reactions, yielding a serie of degradation products. Among them, aldehyde-like compounds seem to be responsible for the decrease in viability of DNA repair-proficient E.coli cells. A positive interaction between toxic solutions and UV-radiation effects is observed in these cells. The sinergism UV-toxic solutions varies in function of post-irradiation time and is dependent on UV dose, indicating the interference of repair processes in toxicity. The effect of non-reducing sugars on cellular viability is negligible, suggesting that toxic substances generation is linked to the presence of at least a free carbonyl group in sugar structure. All tested reducing sugars, when experimental conditions remained constant, have similarly shaped inactivation kinetics and their effects are equally inhibited by catalase activity, during incubation. (author).

Microwave Acid Extraction to Analyze K and Mg Reserves in the Clay Fraction of Soils

Directory of Open Access Journals (Sweden)

Araína Hulmann Batista

Full Text Available ABSTRACT: Extraction of K and Mg with boiling 1 mol L-1 HNO3 in an open system for predicting K and Mg uptake by plants is a method of low reproducibility. The aim of this study was to compare the extraction capacity of different acid methods relative to hydrofluoric acid extraction for K and Mg. A further objective was to develop a chemical extraction method using a closed system (microwave for nonexchangeable and structural forms of these nutrients in order to replace the traditional method of extraction with boiling HNO3 on a hot plate (open system. The EPA 3051A method can be used to estimate the total content of K in the clay fraction of soils developed from carbonate and phyllite/mica schist rocks. In the clay fraction of soils developed from basalt, recoveries of K by the EPA 3051A (pseudo-total method were higher than for the EPA 3052 (total hydrofluoric extraction method. The relative abundance of K and Mg for soils in carbonate rocks, phyllite/mica schist, granite/gneiss, and basalt determined by aqua regia digestion is unreliable. The method using 1 mol L-1 HNO3 in an closed system (microwave showed potential for replacing the classical method of extraction of nonexchangeable forms of K (boiling 1 mol L-1 HNO3 in an open system - hot plate and reduced the loss of Si by volatilization.

Phytochemical analysis and antimicrobial activity of baobab (Adansonia digitata leaves and stem bark extracts on Staphylococcus aureus and Escherichia coli

Directory of Open Access Journals (Sweden)

Mohammed Sani Sambo Datsugwai

2017-05-01

Full Text Available The phytochemical analysis and antibacterial activity of methanolic and ethanolic leaf and stem bark extracts of baobab tree on Escherichia coli and Staphylococcus aureus were carried out using agar well diffusion method. The clinical bacterial isolates of Escherichia coli and Staphylococcus aureus were obtained from Microbiology laboratory, Kaduna State University, Kaduna. The bacteria isolates were re-confirmed and identified based on their morphology, cultural characteristics and biochemical tests. The bacteria isolates were confirmed to be Escherichia coli and Staphylococcus aureus. The phytochemical analysis revealed the presence of Alkaloids, Saponins, Flavonoids, Tannins and Terpenoids. The methanolic leaf extract showed a wide range of activity on test isolates, with varying zones of inhibitions as 12 mm, 10 mm, 7 mm, and 4 mm against Staphylococcus aureus and 13 mm, 9 mm, 7 mm, and 3 mm against Escherichia coli at concentration of 1000 mg/ml, 500 mg/ml, 200 mg/ml and 100 mg/ml respectively. The ethanolic leaf extract also showed a wide range of activity on test isolates with varying zones of inhibitions, such as 11mm, 6mm, 5mm and 3mm against S. aureus and 8mm, 7mm, 5mm, and 4mm against E. coli at the concentration of 1000 mg/ml, 500mg/ml, 200 mg/ml and 100mg/ml for each respectively. The methanolic stem bark extract showed less antibacterial activity against the test isolates with the inhibition of 5mm and 4mm against S. aureus and 4mm and 3mm against E.coli at concentration of 1000 mg/ml and 500 mg/ml respectively with no zones of inhibition at concentration of 200 mg/ml and 100mg/ml. The ethanolic stem bark extract also showed no antibacterial activity with no zones of inhibition against the test isolates at concentration of 1000 mg/ml, 500 mg/ml, 200mg/ml and 100 mg/ml. The methanolic leaf extract inhibited the growth of S. aureus and E.coli at concentration of 100 mg/ml with minimum bactericidal concentration at 100 mg/ml. The

Effects of simulated Mars conditions on the survival and growth of Escherichia coli and Serratia liquefaciens.

Science.gov (United States)

Berry, Bonnie J; Jenkins, David G; Schuerger, Andrew C

2010-04-01

Escherichia coli and Serratia liquefaciens, two bacterial spacecraft contaminants known to replicate under low atmospheric pressures of 2.5 kPa, were tested for growth and survival under simulated Mars conditions. Environmental stresses of high salinity, low temperature, and low pressure were screened alone and in combination for effects on bacterial survival and replication, and then cells were tested in Mars analog soils under simulated Mars conditions. Survival and replication of E. coli and S. liquefaciens cells in liquid medium were evaluated for 7 days under low temperatures (5, 10, 20, or 30 degrees C) with increasing concentrations (0, 5, 10, or 20%) of three salts (MgCl(2), MgSO(4), NaCl) reported to be present on the surface of Mars. Moderate to high growth rates were observed for E. coli and S. liquefaciens at 30 or 20 degrees C and in solutions with 0 or 5% salts. In contrast, cell densities of both species generally did not increase above initial inoculum levels under the highest salt concentrations (10 and 20%) and the four temperatures tested, with the exception that moderately higher cell densities were observed for both species at 10% MgSO(4) maintained at 20 or 30 degrees C. Growth rates of E. coli and S. liquefaciens in low salt concentrations were robust under all pressures (2.5, 10, or 101.3 kPa), exhibiting a general increase of up to 2.5 orders of magnitude above the initial inoculum levels of the assays. Vegetative E. coli cells were maintained in a Mars analog soil for 7 days under simulated Mars conditions that included temperatures between 20 and -50 degrees C for a day/night diurnal period, UVC irradiation (200 to 280 nm) at 3.6 W m(-2) for daytime operations (8 h), pressures held at a constant 0.71 kPa, and a gas composition that included the top five gases found in the martian atmosphere. Cell densities of E. coli failed to increase under simulated Mars conditions, and survival was reduced 1 to 2 orders of magnitude by the interactive

Pathogenic Escherichia coli and food handlers in luxury hotels in Nairobi, Kenya.

Science.gov (United States)

Onyango, Abel O; Kenya, Eucharia U; Mbithi, John J N; Ng'ayo, Musa O

2009-11-01

The epidemiology and virulence properties of pathogenic Escherichia coli among food handlers in tourist destination hotels in Kenya are largely uncharacterized. This cross-sectional study among consenting 885 food handlers working in nine luxurious tourist hotels in Nairobi, Kenya determined the epidemiology, virulence properties, antibiotics susceptibility profiles and conjugation abilities of pathogenic Escherichia coli. Pathogenic Escherichia coli was detected among 39 (4.4%) subjects, including 1.8% enteroaggregative Escherichia coli (EAEC) harboring aggR genes, 1.2% enterotoxigenic Escherichia coli (ETEC) expressing both LT and STp toxins, 1.1% enteropathogenic Escherichia coli (EPEC) and 0.2% Shiga-like Escherichia coli (EHEC) both harboring eaeA and stx2 genes respectively. All the pathotypes had increased surface hydrophobicity. Using multivariate analyses, food handlers with loose stools were more likely to be infected with pathogenic Escherichia coli. Majority 53.8% of the pathotypes were resistant to tetracycline with 40.2% being multi-drug resistant. About 85.7% pathotypes trans-conjugated with Escherichia coli K12 F(-) NA(r) LA. The carriage of multi-drug resistant, toxin expressing pathogenic Escherichia coli by this population is of public health concern because exposure to low doses can result in infection. Screening food handlers and implementing public awareness programs is recommended as an intervention to control transmission of enteric pathogens.

Effect of durable γ-radiation on E.Coli

International Nuclear Information System (INIS)

Koudela, K.; Drashil, V.

1990-01-01

Effect of prolonged low intensity γ-radiation on changes of frequency of reversion mutations was studied in Escherichia Coli. Frequency of His + revertants was shown to depend on growth phase. Cellular DNA absorbed more energy in stationary than DNA in growth phase. K-12 AB2497 strain of Escherichia Coli D-37 comprised about 60 Gy. This dose wasn't absorbed under continuous irradiation at dose rate of 0.21 R/min in 5 hours. The dose rate was considered to be sufficient to induce SOS-system and thus to increase mutations number. 2 refs

DISTRIBUTION OF MAJOR ELEMENTS (NA, K, CA, MG) IN THE ...

African Journals Online (AJOL)

Levels of sodium, potassium, calcium and magnesium were determined in plant organs (bud, flowers, fruit, seed, leaves, stems, roots, cobs, styles, shaft, grains and efflorescences) of three Fadama farms located in Ifaki-Ekiti, Ado-Ekiti and Ikere-Ekiti of Ekiti State, Nigeria. The highest levels of Mg, K, Na and Ca were ...

K88 Fimbrial Adhesin Targeting of Microspheres Containing Gentamicin Made with Albumin Glycated with Lactose

Directory of Open Access Journals (Sweden)

Andre-i Sarabia-Sainz

2015-09-01

Full Text Available The formulation and characterization of gentamicin-loaded microspheres as a delivery system targeting enterotoxigenic Escherichia coli K88 (E. coli K88 was investigated. Glycated albumin with lactose (BSA-glucose-β (4-1 galactose was used as the microsphere matrix (MS-Lac and gentamicin included as the transported antibiotic. The proposed target strategy was that exposed galactoses of MS-Lac could be specifically recognized by E. coli K88 adhesins, and the delivery of gentamicin would inhibit bacterial growth. Lactosylated microspheres (MS-Lac1, MS-Lac2 and MS-Lac3 were obtained using a water-in-oil emulsion, containing gentamicin, followed by crosslinking with different concentrations of glutaraldehyde. Electron microscopy displayed spherical particles with a mean size of 10–17 µm. In vitro release of gentamicin from MS-Lac was best fitted to a first order model, and the antibacterial activity of encapsulated and free gentamicin was comparable. MS-Lac treatments were recognized by plant galactose-specific lectins from Ricinus communis and Sophora japonica and by E. coli K88 adhesins. Results indicate MS-Lac1, produced with 4.2 mg/mL of crosslinker, as the best treatment and that lactosylated microsphere are promising platforms to obtain an active, targeted system against E. coli K88 infections.

Nucleoli in human early erythroblasts (K2, K1, K1/2 cells).

Science.gov (United States)

Smetana, K; Jirásková, I; Klamová, H

2005-01-01

Human early erythroid precursors classified according to the nuclear size were studied to provide information on nucleoli in these cells using simple cytochemical procedures for demonstration of RNA and proteins of silver-stained nucleolar organizers. K2 cells with nuclear diameter larger than 13 microm and K1 cells with nuclear diameter larger than 9 microm corresponding to proerythroblasts and macroblasts (large basophilic erythroblasts) mostly possessed large irregularly shaped nucleoli with multiple fibrillar centres representing "active nucleoli". K1/2 cells with nuclear diameter smaller than 9 microm corresponding to small basophilic erythroblasts were usually characterized by the presence of micronucleoli representing "inactive nucleolar types". On the other hand, a few K1/2 cells contained large nucleoli with multiple fibrillar centres similar to those present in K2 cells and thus appeared as "microproerythroblasts". The nucleolar asynchrony expressed by the presence of large irregularly shaped nucleoli with multiple nucleoli (active nucleoli) and ring-shaped nucleoli (resting nucleoli) in one and the same nucleus of K2 or K1 cells was not exceptional and might reflect a larger resistance of these cells to negative factors influencing the erythropoiesis. The intranucleolar translocation of silver-stained nucleolus organized regions was noted in K2 cells and might indicate the premature aging of these cells without further differentiation. More studies, however, are required in this direction.

Affinity isolation and I-DIRT mass spectrometric analysis of the Escherichia coli O157:H7 Sakai RNA polymerase complex.

Science.gov (United States)

Lee, David J; Busby, Stephen J W; Westblade, Lars F; Chait, Brian T

2008-02-01

Bacteria contain a single multisubunit RNA polymerase that is responsible for the synthesis of all RNA. Previous studies of the Escherichia coli K-12 laboratory strain identified a group of effector proteins that interact directly with RNA polymerase to modulate the efficiency of transcription initiation, elongation, or termination. Here we used a rapid affinity isolation technique to isolate RNA polymerase from the pathogenic Escherichia coli strain O157:H7 Sakai. We analyzed the RNA polymerase enzyme complex using mass spectrometry and identified associated proteins. Although E. coli O157:H7 Sakai contains more than 1,600 genes not present in the K-12 strain, many of which are predicted to be involved in transcription regulation, all of the identified proteins in this study were encoded on the "core" E. coli genome.

Differential mechanism of Escherichia coli Inactivation by (+)-limonene as a function of cell physiological state and drug's concentration.

Science.gov (United States)

Chueca, Beatriz; Pagán, Rafael; García-Gonzalo, Diego

2014-01-01

(+)-limonene is a lipophilic antimicrobial compound, extracted from citrus fruits' essential oils, that is used as a flavouring agent and organic solvent by the food industry. A recent study has proposed a common and controversial mechanism of cell death for bactericidal antibiotics, in which hydroxyl radicals ultimately inactivated cells. Our objective was to determine whether the mechanism of Escherichia coli MG1655 inactivation by (+)-limonene follows that of bactericidal antibiotics. A treatment with 2,000 μL/L (+)-limonene inactivated 4 log10 cycles of exponentially growing E. coli cells in 3 hours. On one hand, an increase of cell survival in the ΔacnB mutant (deficient in a TCA cycle enzyme), or in the presence of 2,2'-dipyridyl (inhibitor of Fenton reaction by iron chelation), thiourea, or cysteamine (hydroxyl radical scavengers) was observed. Moreover, the ΔrecA mutant (deficient in an enzyme involved in SOS response to DNA damage) was more sensitive to (+)-limonene. Thus, this indirect evidence indicates that the mechanism of exponentially growing E. coli cells inactivation by 2,000 μL/L (+)-limonene is due to the TCA cycle and Fenton-mediated hydroxyl radical formation that caused oxidative DNA damage, as observed for bactericidal drugs. However, several differences have been observed between the proposed mechanism for bactericidal drugs and for (+)-limonene. In this regard, our results demonstrated that E. coli inactivation was influenced by its physiological state and the drug's concentration: experiments with stationary-phase cells or 4,000 μL/L (+)-limonene uncovered a different mechanism of cell death, likely unrelated to hydroxyl radicals. Our research has also shown that drug's concentration is an important factor influencing the mechanism of bacterial inactivation by antibiotics, such as kanamycin. These results might help in improving and spreading the use of (+)-limonene as an antimicrobial compound, and in clarifying the controversy about

NON-LTE INVERSIONS OF THE Mg ii h and k AND UV TRIPLET LINES

Energy Technology Data Exchange (ETDEWEB)

De la Cruz Rodríguez, Jaime; Leenaarts, Jorrit [Institute for Solar Physics, Dept. of Astronomy, Stockholm University, AlbaNova University Centre, SE-106 91 Stockholm Sweden (Sweden); Ramos, Andrés Asensio [Instituto de Astrofísica de Canarias, E-38205, La Laguna, Tenerife (Spain)

2016-10-20

The Mg ii h and k lines are powerful diagnostics for studying the solar chromosphere. They have become particularly popular with the launch of the Interface Region Imaging Spectrograph ( IRIS ) satellite, and a number of studies that include these lines have lead to great progress in understanding chromospheric heating, in many cases thanks to the support from 3D MHD simulations. In this study, we utilize another approach to analyze observations: non-LTE inversions of the Mg ii h and k and UV triplet lines including the effects of partial redistribution. Our inversion code attempts to construct a model atmosphere that is compatible with the observed spectra. We have assessed the capabilities and limitations of the inversions using the FALC atmosphere and a snapshot from a 3D radiation-MHD simulation. We find that Mg ii h and k allow reconstructing a model atmosphere from the middle photosphere to the transition region. We have also explored the capabilities of a multi-line/multi-atom setup, including the Mg ii h and k, the Ca ii 854.2 nm, and the Fe i 630.25 lines to recover the full stratification of physical parameters, including the magnetic field vector, from the photosphere to the chromosphere. Finally, we present the first inversions of observed IRIS spectra from quiet-Sun, plage, and sunspot, with very promising results.

Growth inhibitor of E. coli K-12 in a sample of 39KCl

International Nuclear Information System (INIS)

Luckey, T.D.

1984-01-01

Growth rates and total population of E. coli were reduced fourfold when natural KCl (/sup N/KCl) in the medium was replaced by 39 KCl from a particular source. A prolonged lag period was noted in cultures containing either /sup N/KCl or 39 KCl when inoculated with bacteria adapted to 39 KCl. These changes were not due to endogenous radiation because these differences were not observed when (a) the KCl concentrations were reduced from 50 to 5 mM and (b) the 39 KCl from the prime source was replaced with 39 KCl from a second source; also the addition of 40 KCl to 39 KCl did not improve growth. These results suggest that the 39 KCl from the primary source contained an unidentified inhibitor that is not readily detected by physical and chemical analyses. 8 references, 3 figures, 2 tables

Metabolic Engineering of Escherichia coli K12 for Homofermentative Production of L-Lactate from Xylose.

Science.gov (United States)

Jiang, Ting; Zhang, Chen; He, Qin; Zheng, Zhaojuan; Ouyang, Jia

2018-02-01

The efficient utilization of xylose is regarded as a technical barrier to the commercial production of bulk chemicals from biomass. Due to the desirable mechanical properties of polylactic acid (PLA) depending on the isomeric composition of lactate, biotechnological production of lactate with high optical pure has been increasingly focused in recent years. The main objective of this work was to construct an engineered Escherichia coli for the optically pure L-lactate production from xylose. Six chromosomal deletions (pflB, ldhA, ackA, pta, frdA, adhE) and a chromosomal integration of L-lactate dehydrogenase-encoding gene (ldhL) from Bacillus coagulans was involved in construction of E. coli KSJ316. The recombinant strain could produce L-lactate from xylose resulting in a yield of 0.91 g/g xylose. The chemical purity of L-lactate was 95.52%, and the optical purity was greater than 99%. Moreover, three strategies, including overexpression of L-lactate dehydrogenase, intensification of xylose catabolism, and addition of additives to medium, were designed to enhance the production. The results showed that they could increase the concentration of L-lactate by 32.90, 20.13, and 233.88% relative to the control, respectively. This was the first report that adding formate not only could increase the xylose utilization but also led to the fewer by-product levels.

UV irradiation alters deoxynucleoside triphosphate pools in Escherichia coli

International Nuclear Information System (INIS)

Das, S.K.; Loeb, L.A.

1984-01-01

UV irradiation of exponentially growing Escherichia coli increased intracellular concentration of dATP and dTTP without significantly changing the concentrations of dGTP and dCTP. These selective increases in dATP and dTTP pools are seen in wild-type E. coli K12 and AB1157, as well as in recA and umuC strains, and are proportional to UV dose. The possible significance of these findings with respect to induction of the SOS response and nontargeted mutagenesis are discussed. (orig.)

Magnetic properties of Mg{sub 12}O{sub 12} nanocage doped with transition metal atoms (Mn, Fe, Co and Ni): DFT study

Energy Technology Data Exchange (ETDEWEB)

Javan, Masoud Bezi, E-mail: javan.masood@gmail.com

2015-07-01

Binding energy of the Mg{sub 12}O{sub 12} nanocage doped with transition metals (TM=Mn, Fe, Co and Ni) in endohedrally, exohedrally and substitutionally forms were studied using density functional theory with the generalized gradient approximation exchange-correlation functional along 6 different paths inside and outside of the Mg{sub 12}O{sub 12} nanocage. The most stable structures were determined with full geometry optimization near the minimum of the binding energy curves of all the examined paths inside and outside of the Mg{sub 12}O{sub 12} nanocage. The results reveal that for all stable structures, the Ni atom has a larger binding energy than the other TM atoms. It is also found that for all complexes additional peaks contributed by TM-3d, 4s and 4p states appear in the highest occupied molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO) gap of the host MgO cluster. The mid-gap states are mainly due to the hybridization between TM-3d, 4s and 4p orbitals and the cage π orbitals. The magnetic moment of the endohedrally doped TM atoms in the Mg{sub 12}O{sub 12} are preserved to some extent due to the interaction between the TM and Mg{sub 12}O{sub 12} nanocage, in contrast to the completely quenched magnetic moment of the Fe and Ni atoms in the Mg{sub 11}(TM)O{sub 12} complexes. Furthermore, charge population analysis shows that charge transfer occurs from TM atom to the cage for endohedrally and substitutionally doping. - Highlights: • Binding energy of the Mg{sub 12}O{sub 12} nanocage doped with transition metals was studied. • The most stable structures were determined near the minimum of the binding energy. • The encapsulated Ni atom has a larger binding energy than the other TM atoms. • Magnetic moment of the endohedrally doped TM atoms in the Mg{sub 12}O{sub 12} are preserved.

Reaction 12C(16O,α)24Mg leading to nuclear molecular resonances

International Nuclear Information System (INIS)

Nagatani, K.; Shimoda, T.; Tanner, D.; Tribble, R.; Yamaya, T.

1979-01-01

The reactions 12 C( 16 O,α) 24 Mg and 13 C( 16 O,α) 25 Mg were investigated at an incident energy of 145 MeV. In the reaction with the 12 C target, broad peaks are observed at forward angles which correspond to the molecular resonance states of the 12 C+ 12 C system, while the spectra with 13 C target show only a smooth continuum

Sex-related effects of nutritional supplementation of Escherichia coli: relevance to eating disorders.

Science.gov (United States)

Tennoune, Naouel; Legrand, Romain; Ouelaa, Wassila; Breton, Jonathan; Lucas, Nicolas; Bole-Feysot, Christine; do Rego, Jean-Claude; Déchelotte, Pierre; Fetissov, Sergueï O

2015-03-01

The biological background of sex-related differences in the development of eating disorders (EDs) is unknown. Recent data showed that gut bacteria Escherichia coli induce autoantibodies against anorexigenic α-melanocyte-stimulating hormone (α-MSH) associated with psychopathology in ED. The aim of this study was to compare the effects of E. coli on feeding and autoantibodies against α-MSH and adrenocorticotropic hormone (ACTH), between female and male rats. Commensal E. coli K12 were given in a culture medium daily to adult Wistar rats by intragastric gavage over a 3-wk period; control rats received culture medium only. Before gavage, E. coli K12 DNA was detected in feces of female but not male rats. E. coli provision was accompanied by an increase in body weight gain in females, but a decrease in body weight gain and food intake in males. Independent of E. coli treatment, plasma levels of anti-α-MSH and ACTH immunoglobulin (Ig)G were higher in female than male rats. Females responded to E. coli by increasing α-MSH IgG levels and affinity, but males by increasing α-MSH IgM levels. Affinity of IgG for ACTH was increased in both E. coli-treated females and males, although with different kinetics. IgG from females stimulated more efficiently α-MSH-induced cyclic adenosine monophosphate production by melanocortin 4 receptor-expressing cells compared with IgG from males. Sex-related response to how E. coli affects feeding and anti-melanocortin hormone antibody production may depend on the presence of these bacteria in the gut before E. coli supplementation. These data suggest that sex-related presence of certain gut bacteria may represent a risk factor for ED development. Copyright © 2015 Elsevier Inc. All rights reserved.

Affinity Isolation and I-DIRT Mass Spectrometric Analysis of the Escherichia coli O157:H7 Sakai RNA Polymerase Complex▿

Science.gov (United States)

Lee, David J.; Busby, Stephen J. W.; Westblade, Lars F.; Chait, Brian T.

2008-01-01

Bacteria contain a single multisubunit RNA polymerase that is responsible for the synthesis of all RNA. Previous studies of the Escherichia coli K-12 laboratory strain identified a group of effector proteins that interact directly with RNA polymerase to modulate the efficiency of transcription initiation, elongation, or termination. Here we used a rapid affinity isolation technique to isolate RNA polymerase from the pathogenic Escherichia coli strain O157:H7 Sakai. We analyzed the RNA polymerase enzyme complex using mass spectrometry and identified associated proteins. Although E. coli O157:H7 Sakai contains more than 1,600 genes not present in the K-12 strain, many of which are predicted to be involved in transcription regulation, all of the identified proteins in this study were encoded on the “core” E. coli genome. PMID:18083804

Dead end metabolites--defining the known unknowns of the E. coli metabolic network.

Directory of Open Access Journals (Sweden)

Amanda Mackie

Full Text Available The EcoCyc database is an online scientific database which provides an integrated view of the metabolic and regulatory network of the bacterium Escherichia coli K-12 and facilitates computational exploration of this important model organism. We have analysed the occurrence of dead end metabolites within the database--these are metabolites which lack the requisite reactions (either metabolic or transport that would account for their production or consumption within the metabolic network. 127 dead end metabolites were identified from the 995 compounds that are contained within the EcoCyc metabolic network. Their presence reflects either a deficit in our representation of the network or in our knowledge of E. coli metabolism. Extensive literature searches resulted in the addition of 38 transport reactions and 3 metabolic reactions to the database and led to an improved representation of the pathway for Vitamin B12 salvage. 39 dead end metabolites were identified as components of reactions that are not physiologically relevant to E. coli K-12--these reactions are properties of purified enzymes in vitro that would not be expected to occur in vivo. Our analysis led to improvements in the software that underpins the database and to the program that finds dead end metabolites within EcoCyc. The remaining dead end metabolites in the EcoCyc database likely represent deficiencies in our knowledge of E. coli metabolism.

DNA microarray analysis of fim mutations in Escherichia coli

DEFF Research Database (Denmark)

Schembri, Mark; Ussery, David; Workman, Christopher

2002-01-01

Bacterial adhesion is often mediated by complex polymeric surface structures referred to as fimbriae. Type I fimbriae of Escherichia coli represent the archetypical and best characterised fimbrial system. These adhesive organelles mediate binding to D-mannose and are directly associated...... we have used DNA microarray analysis to examine the molecular events involved in response to fimbrial gene expression in E. coli K-12. Observed differential expression levels of the fim genes were in good agreement with our current knowledge of the stoichiometry of type I fimbriae. Changes in fim...

Expression of exo-inulinase gene from Aspergillus niger 12 in E. coli strain Rosetta-gami B (DE3) and its characterization.

Science.gov (United States)

Yedahalli, Shreyas S; Rehmann, Lars; Bassi, Amarjeet

2016-05-01

Inulin is a linear carbohydrate polymer of fructose subunits (2-60) with terminal glucose units, produced as carbon storage in selected plants. It cannot directly be taken up by most microorganisms due to its large size, unless prior hydrolysis through inulinase enzymes occurs. The hydrolyzed inulin can be taken up by microbes and/or recovered and used industrially for the production of high fructose syrup, inulo-oligosaccharides, biofuel, and nutraceuticals. Cell-free enzymatic hydrolysis would be desirable for industrial applications, hence the recombinant expression, purification and characterization of an Aspergillus niger derived exo-inulinase was investigated in this study. The eukaroyototic exo-inulinase of Aspergillus niger 12 has been expressed, for the first time, in an E. coli strain [Rosetta-gami B (DE3)]. The molecular weight of recombinant exo-inulinase was estimated to be ∼81 kDa. The values of Km and Vmax of the recombinant exo-inulinase toward inulin were 5.3 ± 1.1 mM and 402.1 ± 53.1 µmol min(-1)  mg(-1) protein, respectively. Towards sucrose the corresponding values were 12.20 ± 1.6 mM and 902.8 ± 40.2 µmol min(-1)  mg(-1) protein towards sucrose. The S/I ratio was 2.24 ± 0.7, which is in the range of native inulinase. The optimum temperature and pH of the recombinant exo-inulinase towards inulin was 55°C and 5.0, while they were 50°C and 5.5 towards sucrose. The recombinant exo-inulinase activity towards inulin was enhanced by Cu(2+) and reduced by Fe(2+) , while its activity towards sucrose was enhanced by Co(2+) and reduced by Zn(2+) . © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:629-637, 2016. © 2016 American Institute of Chemical Engineers.

The study of preparation for immobilized cells membranes of E. Coli. by radiation technique

International Nuclear Information System (INIS)

Cao Jin; Chen Pin; Yu Yi

1991-01-01

The paper described the preparation of immobilized cells membranes with E. Coli by radiation technique. The nylon 6 was grafted with HEMA, which as a matrix to prepare immobilized cells membranes with E. Coli. by radiation entrapment at low temperature. The results showed that the retentive activity possessed a maximum value for membranes with E. Coli. when the irradiation dose was at 10-12 kGy, the entrapped cells has 2.3 g/ml at 50% HEMA concentration, the optimum pH and optimum temperature for membranes with E. Coli. are as same the original cells

Analysis of L-glutamic acid fermentation by using a dynamic metabolic simulation model of Escherichia coli.

Science.gov (United States)

Nishio, Yousuke; Ogishima, Soichi; Ichikawa, Masao; Yamada, Yohei; Usuda, Yoshihiro; Masuda, Tadashi; Tanaka, Hiroshi

2013-09-22

Understanding the process of amino acid fermentation as a comprehensive system is a challenging task. Previously, we developed a literature-based dynamic simulation model, which included transcriptional regulation, transcription, translation, and enzymatic reactions related to glycolysis, the pentose phosphate pathway, the tricarboxylic acid (TCA) cycle, and the anaplerotic pathway of Escherichia coli. During simulation, cell growth was defined such as to reproduce the experimental cell growth profile of fed-batch cultivation in jar fermenters. However, to confirm the biological appropriateness of our model, sensitivity analysis and experimental validation were required. We constructed an L-glutamic acid fermentation simulation model by removing sucAB, a gene encoding α-ketoglutarate dehydrogenase. We then performed systematic sensitivity analysis for L-glutamic acid production; the results of this process corresponded with previous experimental data regarding L-glutamic acid fermentation. Furthermore, it allowed us to predicted the possibility that accumulation of 3-phosphoglycerate in the cell would regulate the carbon flux into the TCA cycle and lead to an increase in the yield of L-glutamic acid via fermentation. We validated this hypothesis through a fermentation experiment involving a model L-glutamic acid-production strain, E. coli MG1655 ΔsucA in which the phosphoglycerate kinase gene had been amplified to cause accumulation of 3-phosphoglycerate. The observed increase in L-glutamic acid production verified the biologically meaningful predictive power of our dynamic metabolic simulation model. In this study, dynamic simulation using a literature-based model was shown to be useful for elucidating the precise mechanisms involved in fermentation processes inside the cell. Further exhaustive sensitivity analysis will facilitate identification of novel factors involved in the metabolic regulation of amino acid fermentation.

Oral challenge with E.coli K88 as a tool to assess growth and health performance in feeding trials of-weaned pigs

Directory of Open Access Journals (Sweden)

P. Bonilauri

2011-03-01

Full Text Available The valuation of dietary solutions for weaning pigs is problematic. In field situations, an accurate control of replications is difficult and disturbing factors are hardly removed; in experimental farm, hygienic conditions are in general superior to practical farms. In the study of alternatives to in-feed antibiotics the challenge with K88 E.coli has been often proposed. The predisposition to this colibacillosis is, at least partially, genetically controlled and depends on the presence of intestinal receptors for the F4 fimbrial antigens of K88 E.coli...

Drought and Epidemic Typhus, Central Mexico, 1655–1918

Science.gov (United States)

Acuna-Soto, Rudofo; Stahle, David W.

2014-01-01

Epidemic typhus is an infectious disease caused by the bacterium Rickettsia prowazekii and transmitted by body lice (Pediculus humanus corporis). This disease occurs where conditions are crowded and unsanitary. This disease accompanied war, famine, and poverty for centuries. Historical and proxy climate data indicate that drought was a major factor in the development of typhus epidemics in Mexico during 1655–1918. Evidence was found for 22 large typhus epidemics in central Mexico, and tree-ring chronologies were used to reconstruct moisture levels over central Mexico for the past 500 years. Below-average tree growth, reconstructed drought, and low crop yields occurred during 19 of these 22 typhus epidemics. Historical documents describe how drought created large numbers of environmental refugees that fled the famine-stricken countryside for food relief in towns. These refugees often ended up in improvised shelters in which crowding encouraged conditions necessary for spread of typhus. PMID:24564928

Cloning and characterization of newly isolated lipase from Enterobacter sp. Bn12.

Science.gov (United States)

Farrokh, Parisa; Yakhchali, Bagher; Karkhane, Ali Asghar

2014-01-01

A mesophilic Enterobacter sp. Bn12 producing an alkaline thermostable lipase was isolated from soil in Tehran, Iran. The lipase gene (ELBn12) was identified from a genomic library. Sequence analysis of the DNA fragment revealed an open reading frame of 879 bp encoding a lipase with a molecular mass of 31.3 kDa. The deduced amino acid sequence showed 96% identity with a lipase of Enterobacter sp. Ag1 and the identity of their DNA sequences was 88.9%. ELBn12 belongs to the lipase subfamily I.1 and its catalytic triad consists of Ser82, Asp237 and His259. The lipase was expressed in Escherichia coli (BL21) pLysS and partially purified by anion exchange chromatography. The maximum activity of ELBn12 was obtained at temperature of 60 °C and pH 8.0 towards tricaprylin (C8) and its specific activity was around 2900 U/mg. ELBn12 was stable within a broad pH range from 6.0 to 11.0. The enzyme showed high stability in both polar and nonpolar organic solvents at 50% (v/v). The lipase activity was enhanced in the presence of 10 mM of Ca(2+), Mg(2+) and K(+), while heavy metals (Fe(3+) and Zn(2+)) had strong inhibitory effect. ELBn12 showed high activity in the presence of 1% (w/v) nonionic surfactants, however ionic surfactants inhibited the lipolytic activity. ELBn12 characteristics show that it has a potential to be used in various industrial processes.

Effect of bacterial components of mixed culture supernatants of planktonic and biofilm Pseudomonas aeruginosa with commensal Escherichia coli on the neutrophil response in vitro.

Science.gov (United States)

Maslennikova, Irina L; Kuznetsova, Marina V; Nekrasova, Irina V; Shirshev, Sergei V

2017-11-30

Pseudomonas aeruginosa (PA) responsible for acute and chronic infections often forms a well-organized bacterial population with different microbial species including commensal strains of Escherichia coli. Bacterial extracellular components of mixed culture can modulate the influence of bacteria on the neutrophil functions. The objective of this study was to compare the effect of pyocyanin, pyoverdine, LPS, exopolysaccharide of single species and mixed culture supernatants of PA strains and E. coli K12 on microbicidal, secretory activity of human neutrophils in vitro. Bacterial components of E. coli K12 in mixed supernatants with 'biofilm' PA strains (PA ATCC, PA BALG) enhanced short-term microbicidal mechanisms and inhibited neutrophil secretion delayed in time. The influence of 'planktonic' PA (PA 9-3) exometabolites in mixed culture is almost mimicked by E. coli K12 effect on functional neutrophil changes. This investigation may help to understand some of the mechanisms of neutrophil response to mixed infections of different PA with other bacteria species. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Dietary N-Carbamylglutamate Supplementation Boosts Intestinal Mucosal Immunity in Escherichia coli Challenged Piglets.

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Fengrui Zhang

Full Text Available N-carbamylglutamate (NCG has been shown to enhance performance in neonatal piglets. However, few studies have demonstrated the effect of NCG on the intestinal mucosal barrier. This study was conducted to determine the effects of dietary NCG supplementation on intestinal mucosal immunity in neonatal piglets after an Escherichia coli (E. coli challenge. New-born piglets (4 d old were assigned randomly to one of four treatments (n = 7, including (I sham challenge, (II sham challenge +50 mg/kg NCG, (III E. coli challenge, and (IV E. coli challenge +50 mg/kg NCG. On d 8, pigs in the E. coli challenge groups (III and IV were orally challenged with 5 mL of E. coli K88 (10(8 CFU/mL, whereas pigs in the sham challenge groups (I and II were orally dosed with an equal volume of water. On d 13, all piglets were sacrificed, and samples were collected and examined. The results show that average daily gain in the E. coli challenged piglets (III and IV was decreased (PE.coli<0.05. However, it tended to be higher in the NCG treated piglets (II and IV. Ileum secretory IgA, as well as IFN-γ, IL-2, IL-4 and IL-10 in ileal homogenates, were increased in E. coli challenged piglets (III and IV. Similarly, ileum SIgA and IL-10 levels, and CD4(+ percentage in NCG treated piglets (II and IV were higher than no-NCG treated piglets (PNCG<0.05. However, the IL-2 level was only decreased in the piglets of E. coli challenge + NCG group (IV compared with E. coli challenge group (III (P<0.05. No change in the IL-2 level of the sham challenged piglets (III was observed. In conclusion, dietary NCG supplementation has some beneficial effects on intestinal mucosal immunity in E. coli challenged piglets, which might be associated with stimulated lymphocyte proliferation and cytokine synthesis. Our findings have an important implication that NCG may be used to reduce diarrhea in neonatal piglets.

Identification of serotypes and virulence markers of Escherichia coli isolated from human stool and urine samples in Egypt

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K M Osman

2012-01-01

Full Text Available Purpose: Haemorrhagic colitis and haemolytic-uremic syndrome are associated with Shiga-toxin producing Escherichia coli (STEC. There are others DEC (Diarrhoeagenic E. coli pathotypes responsible for outbreaks and others toxins associated to these. Most clinical signs of disease arise as a consequence of the production of Shiga toxin 1 (Stx1, Stx2 or combinations of these toxins. Other major virulence factors include E. coli haemolysin (hlyA, and intimin, the product of the eaeA gene that is involved in the attaching and effacing adherence phenotype. Materials and Methods: In this study, the PCR assay was used to detect 12 E. coli genes associated with virulence (stx1, stx2, hylA, Flic h7 , stb, F41, K99, sta, F17, LT-I, LT-II and eaeA. Results: A total of 108 E. coli strains were serotyped into 64 typable strains. The investigated strains from the stool, 8/80 (10% strains were O 164:K, while the 56/110 strains isolated from the urine were O126:K71 (44/110, 40% and O 86:K 61 (12/110, 11%. The distribution pattern of the detected virulence genes was observed to be in the following order: F17 (10% from the stool and 44% from the urine, Sta (10% from the stool, hylA (10% from the stool and 44% from the urine, Stb (44% from the urine and stx1 (27% from the urine. The 8 faecal strains encoded a combination of the F17, Sta and hylA genes, while the 56 urine strains encoded a combination of the F17 0+ Stb + hylA (44/110, 40% and Stx1 only (12/60, 20%. Conclusion: This is the first report on the molecular characterization of E. coli diarrhoeagenic strains in Egypt and the first report on the potential role of E. coli in diarrhoea and urinary tract infections in a localized geographic area where the people engage in various occupational activities.

Electrochemical performance of Si-CeMg{sub 12} composites as anode materials for Li-ion batteries

Energy Technology Data Exchange (ETDEWEB)

Lu, Z.W. [Institute of New Energy Material Chemistry, Nankai University, Tianjin 300071 (China); Tianjin Institute of Power Sources, Tianjin 300381 (China); Wang, G.; Gao, X.P. [Institute of New Energy Material Chemistry, Nankai University, Tianjin 300071 (China); Liu, X.J.; Wang, J.Q. [Tianjin Institute of Power Sources, Tianjin 300381 (China)

2009-04-01

The Si-CeMg{sub 12} composites with 30 wt.%, 40 wt.% and 50 wt.% Si, were synthesized by directly ball milling Si and CeMg{sub 12} alloy. The microstructure of the Si-CeMg{sub 12} composites is confirmed by X-ray diffraction (XRD) and transmission electron microscopy (TEM), respectively. It is demonstrated from TEM images that active Si nanoparticles are distributed in the inactive CeMg{sub 12} matrix. The electrochemical performance of the Si-CeMg{sub 12} composites as a function of Si content is investigated. The maximum reversible (charge) capacities of the ball-milled Si-CeMg{sub 12} composites with 30 wt.%, 40 wt.% and 50 wt.% Si reach 470, 690 and 1080 mAh g{sup -1}, respectively, after full activations. It is found that the Si-CeMg{sub 12} composite with 40 wt.% Si delivers a larger reversible capacity and better cycle ability because the uniform distribution of active Si nanoparticles embedded in the CeMg{sub 12} matrix, which can accommodate the volume expansion of the composite during Li-alloying/dealloying processes. After subsequent cycles, the recrystallization of Si with lattice shrinkage is observed, which is unfavorable to the Li-alloying/dealloying reaction. The degeneration of CeMg{sub 12}-Si composites during repeated cycling is attributed not only to the Si pulverization led by the volume change, but partially also to the irreversible phase transformation of Si. (author)

Inhibition of Bifidobacterium Cell Wall 51.74 kDa Adhesin Isolated from Infants Feces Towards Adhesion of Enteric Phatogen E. coli on Enterocyte Balb/C Mice

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I Sukrama

2012-01-01

Full Text Available Objectives: To determine 51.74 kDa adhesin of Bifidobacterium sp cell wall isolated from infants feces as an anti adhesion of E. coli on enterocyte mice. Methods: Randomized Posttest-Only Control Group Design was employed to investigate adherence ability of this adhesin towards E.coli adhesion on mice entherocyte. Results: In this research, it was obtained, that the 51.74 kDa adhesin cell wall of Bifidobacterium sp has an ability to inhibit adhesion of E. coli on mice enterocyte. The ability was increased as an increase of adhsein concentration. Conclusions: that can be drawn from this research is the finding of 51.74 kDa adhesin cell wall of Bifidobacterium sp isolated from infants feces that can inhibit adhseion of E. coli on mice enterocyte. Future work that can be carried out are further researches concerning whether these protein can be applied to inhibit adherence of other pathogen bacteria

Metabolic engineering of Escherichia coli for the production of riboflavin

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2014-01-01

Background Riboflavin (vitamin B2), the precursor of the flavin cofactors flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), is used commercially as an animal feed supplement and food colorant. E. coli is a robust host for various genetic manipulations and has been employed for efficient production of biofuels, polymers, amino acids, and bulk chemicals. Thus, the aim of this study was to understand the metabolic capacity of E. coli for the riboflavin production by modification of central metabolism, riboflavin biosynthesis pathway and optimization of the fermentation conditions. Results The basic producer RF01S, in which the riboflavin biosynthesis genes ribABDEC from E. coli were overexpressed under the control of the inducible trc promoter, could accumulate 229.1 mg/L of riboflavin. Further engineering was performed by examining the impact of expression of zwf (encodes glucose 6-phosphate dehydrogenase) and gnd (encodes 6-phosphogluconate dehydrogenase) from Corynebacterium glutamicum and pgl (encodes 6-phosphogluconolactonase) from E. coli on riboflavin production. Deleting pgi (encodes glucose-6-phosphate isomerase) and genes of Entner-Doudoroff (ED) pathway successfully redirected the carbon flux into the oxidative pentose phosphate pathway, and overexpressing the acs (encodes acetyl-CoA synthetase) reduced the acetate accumulation. These modifications increased riboflavin production to 585.2 mg/L. By further modulating the expression of ribF (encodes riboflavin kinase) for reducing the conversion of riboflavin to FMN in RF05S, the final engineering strain RF05S-M40 could produce 1036.1 mg/L riboflavin in LB medium at 37°C. After optimizing the fermentation conditions, strain RF05S-M40 produced 2702.8 mg/L riboflavin in the optimized semi-defined medium, which was a value nearly 12-fold higher than that of RF01S, with a yield of 137.5 mg riboflavin/g glucose. Conclusions The engineered strain RF05S-M40 has the highest yield among all

Metabolic engineering of Escherichia coli for the production of riboflavin.

Science.gov (United States)

Lin, Zhenquan; Xu, Zhibo; Li, Yifan; Wang, Zhiwen; Chen, Tao; Zhao, Xueming

2014-07-16

Riboflavin (vitamin B2), the precursor of the flavin cofactors flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), is used commercially as an animal feed supplement and food colorant. E. coli is a robust host for various genetic manipulations and has been employed for efficient production of biofuels, polymers, amino acids, and bulk chemicals. Thus, the aim of this study was to understand the metabolic capacity of E. coli for the riboflavin production by modification of central metabolism, riboflavin biosynthesis pathway and optimization of the fermentation conditions. The basic producer RF01S, in which the riboflavin biosynthesis genes ribABDEC from E. coli were overexpressed under the control of the inducible trc promoter, could accumulate 229.1 mg/L of riboflavin. Further engineering was performed by examining the impact of expression of zwf (encodes glucose 6-phosphate dehydrogenase) and gnd (encodes 6-phosphogluconate dehydrogenase) from Corynebacterium glutamicum and pgl (encodes 6-phosphogluconolactonase) from E. coli on riboflavin production. Deleting pgi (encodes glucose-6-phosphate isomerase) and genes of Entner-Doudoroff (ED) pathway successfully redirected the carbon flux into the oxidative pentose phosphate pathway, and overexpressing the acs (encodes acetyl-CoA synthetase) reduced the acetate accumulation. These modifications increased riboflavin production to 585.2 mg/L. By further modulating the expression of ribF (encodes riboflavin kinase) for reducing the conversion of riboflavin to FMN in RF05S, the final engineering strain RF05S-M40 could produce 1036.1 mg/L riboflavin in LB medium at 37°C. After optimizing the fermentation conditions, strain RF05S-M40 produced 2702.8 mg/L riboflavin in the optimized semi-defined medium, which was a value nearly 12-fold higher than that of RF01S, with a yield of 137.5 mg riboflavin/g glucose. The engineered strain RF05S-M40 has the highest yield among all reported riboflavin production

Effect of K and Mg salts of aspartic acid on haemopoiesis and recovery from radiation damage in mice

International Nuclear Information System (INIS)

Pospisil, M.; Netikova, J.; Pipalova, I.; Mikeska, J.

1980-01-01

Male mice of non-inbred strain ''H'' were used to test the effect of a 10-day peroral administration of K and Mg aspartates on haemopoietic functions. The salts were proved to stimulate the proliferation and differentiation processes in the thymus, bone marrow and spleen tissues. Mice exposed to a single whole-body X-irradiation after pretreatment with K, Mg aspartate exhibited a more conspicuous postirradiation regeneration of haemopoietic organs and an increased postirradiation survival. The results suggest the possibility of using K, Mg aspartate for radioprotective purposes. (author)

Quasi-free K+ photo-production in 12C

International Nuclear Information System (INIS)

Maeda, K.; Yamazaki, H.; Asano, S.; Emura, T.; Endo, I.; Endo, S.; Ito, S.; Itoh, H.; Ifuku, K.; Konno, O.; Koike, M.; Maruyama, K.; Niki, K.; Niwa, K.; Okuno, H.; Sakaguchi, A.; Sasaki, T.; Suda, T.; Sumi, Y.; Takeya, M.; Terasawa, T.; Uchida, H.; Yamashita, H.; Yoshida, K.

1994-01-01

Quasi-free K + photo-production in the 12 C(γ,K + ) reaction has been investigated in a photon energy range of 0.7-1.1GeV. Differential cross sections for the quasi-free process of the 12 C(γ,K + ) reaction have been obtained and they are compared with a calculation of a quasi-free K + photo-production. The effective proton number Z eff =4.2±0.6 obtained from the experiment was in good agreement with a calculation of a semi-classical attenuation model. ((orig.))

A Numbers Game: Ribosome Densities, Bacterial Growth, and Antibiotic-Mediated Stasis and Death

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Bruce R. Levin

2017-02-01

Full Text Available We postulate that the inhibition of growth and low rates of mortality of bacteria exposed to ribosome-binding antibiotics deemed bacteriostatic can be attributed almost uniquely to these drugs reducing the number of ribosomes contributing to protein synthesis, i.e., the number of effective ribosomes. We tested this hypothesis with Escherichia coli K-12 MG1655 and constructs that had been deleted for 1 to 6 of the 7 rRNA (rrn operons. In the absence of antibiotics, constructs with fewer rrn operons have lower maximum growth rates and longer lag phases than those with more ribosomal operons. In the presence of the ribosome-binding “bacteriostatic” antibiotics tetracycline, chloramphenicol, and azithromycin, E. coli strains with 1 and 2 rrn operons are killed at a substantially higher rate than those with more rrn operons. This increase in the susceptibility of E. coli with fewer rrn operons to killing by ribosome-targeting bacteriostatic antibiotics is not reflected in their greater sensitivity to killing by the bactericidal antibiotic ciprofloxacin, which does not target ribosomes, but also to killing by gentamicin, which does. Finally, when such strains are exposed to these ribosome-targeting bacteriostatic antibiotics, the time before these bacteria start to grow again when the drugs are removed, referred to as the post-antibiotic effect (PAE, is markedly greater for constructs with fewer rrn operons than for those with more rrn operons. We interpret the results of these other experiments reported here as support for the hypothesis that the reduction in the effective number of ribosomes due to binding to these structures provides a sufficient explanation for the action of bacteriostatic antibiotics that target these structures.

[ACID-BASE MODULATION OF LYSOZYME ACTIVITY IN MEDIUM FOR CULTIVATION OF ENTEROBACTERIA].

Science.gov (United States)

Andryuschenko, S V; Perunova, N B

2015-01-01

Determination of modulating effect of acid-base state of medium for cultivation of enterobacteria on activity of C-type lysozyme. Escherichia coli BL21 (DE3) strain for protein expression, Escherichia coli K12 MG1655 model strain, Escherichia coli No. 242 strain, isolated from intestine biotope; 2 Klebsiella pneumoniae strains, one of those contained plasmid homologue of periplasmatic lysozyme inhibitor gene pliC; 1 typical Salmonella enterica ATCC 14028 strain and a Micrococcus luteus ATCC 15307 strain as a control--served as material for the study. The bacteria were cultivated for 24 hours in 2 ml of liquid medium LB at 37 degrees C, 250 rpm. Determination of antilysozyme activity (ALA) was carried out by a photonepehlometrical method according to O.V. Bukharin et al. (1999) with alterations. All the studied microorganisms, including Micrococcus luteus, at the specified conditions 24 hours after cultivation were established to change the pH of the liquid nutrient medium LB from the initial value of 6.6 ± 0.1 to 8.2 ± 0.2 units. ALA determination in the cultivation medium without buffer correction was accompanied by a decline of lysozyme activity at an order of magnitude. The effect was absent during ALA measurement by a standard technique. The local shift of acid-base state of biotope under the conditions of buffer system insufficiency results in a reversible alteration of antimicrobial activity of muramidase, that among other non-specific factors of the environment determines the background of interactions on the level of associative symbiosis. This aspect should be taken into consideration during development of models, that are close to real conditions of microsymbiocenotical interactions.

Influence of Na, K, Ca and Mg on lead atomization by tungsten coil atomic absorption spectrometry

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Oliveira Pedro V. de

2000-01-01

Full Text Available The atomization of lead in an electrothermal tungsten coil atomizer in the presence and absence of Na+, K+, Ca2+ and Mg2+ was investigated with the objective of understanding the interference processes. The lead atomization was less affected by Ca2+ and Mg2+ than by Na+ and K+. In the absence of concomitants, lead atomization efficiency was improved by the presence of H2 (10% v/v in the purge gas composition, during pyrolysis and atomization steps. The interference caused by Na+ and Ca2+ was negligible when the pyrolysis step was accomplished without H2 in the purge gas composition. The results showed that Na+, K+, Ca2+ and Mg2+ are directly involved in competition reactions for H2 in condensed phase.

Exigências de minerais para cabras durante a gestação: Na, K, Mg, S, Fe e Zn Minerals requirements of goats during the pregnancy: Na, K, Mg, S, F and Zn

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Roberto Germano Costa

2003-04-01

Full Text Available O trabalho foi realizado com o objetivo de estimar a retenção e a exigência líquida dos minerais Na, K, Mg, S, Fe e Zn durante a gestação de cabras com um ou dois fetos. A estimativa de retenção foi baseada na diferença entre o total de cada mineral depositado no feto, útero, membranas, fluídos fetais e glândula mamária dos animais nas diferentes etapas da gestação e o total de cada mineral armazenado nas cabras vazias, utilizando-se o modelo de predição ln=A+Bx+Cx2, em que x=tempo de gestação. Os conteúdos de Na, K, Mg, S, Fe e Zn, durante as gestações de um e dois fetos foram de: 13,2 e 21,4 mg; 13,3 e 21,3 g; 2,1 e 3,7 mg; 5,5 e 9,3 mg; 575,5 e 981,0 mg; 112,6 e 164,7 mg nas gestações, resultando em exigências líquidas diárias de 0,13 e 0,11 g; 0,21 e 0,31 g; 0,06 e 0,11g; 0,17 e 0,21 g; 22,94 e 40,51 mg; 2,63 e 2,78 mg, respectivamente.This work was carried out with the purpose of evaluating the retention and the requirement of Ca e P minerals during the pregnancy of goats with one or two foetus. The estimate of retention was based in the difference between the total of each mineral stored in the foetus, uterus, membranes, fetals fluids and mammary gland of animals in the differents phases of pregnancy and the total of each mineral stored in the empty goats, using the model of prediction ln=A+Bx+Cx2, where x=time of pregnancy. The comparison of the estimative with the real values obtained show that the suggested model explained with coherence and precision the biological behavior of minerals retention during all pregnancy. The contend of Na, K, Mg, S, Fe e Zn was: 13.2 and 21.4 mg; 13.3 and 21.3 g; 2.1 and 3.7 mg; 5.5 and 9.3 mg; 575.5 and 981.0 mg; 112.6 and 164.7 mg in the pregnancy of one and two foetus, respectively, that resulted in a diary liquid requirement of 0.13 and 0.11 g; 0.21 and 0.31 g 0.06 and 0.11g; 0.17 and 0.21 g; 22.94 and 40.51 mg; 2.63 and 2.78 mg, respectively.

A Parameter Study for Modeling Mg ii h and k Emission during Solar Flares

Energy Technology Data Exchange (ETDEWEB)

Rubio da Costa, Fatima [Department of Physics, Stanford University, Stanford, CA 94305 (United States); Kleint, Lucia, E-mail: frubio@stanford.edu [University of Applied Sciences and Arts Northwestern Switzerland, 5210, Windisch (Switzerland)

2017-06-20

Solar flares show highly unusual spectra in which the thermodynamic conditions of the solar atmosphere are encoded. Current models are unable to fully reproduce the spectroscopic flare observations, especially the single-peaked spectral profiles of the Mg ii h and k lines. We aim to understand the formation of the chromospheric and optically thick Mg ii h and k lines in flares through radiative transfer calculations. We take a flare atmosphere obtained from a simulation with the radiative hydrodynamic code RADYN as input for a radiative transfer modeling with the RH code. By iteratively changing this model atmosphere and varying thermodynamic parameters such as temperature, electron density, and velocity, we study their effects on the emergent intensity spectra. We reproduce the typical single-peaked Mg ii h and k flare spectral shape and approximate the intensity ratios to the subordinate Mg ii lines by increasing either densities, temperatures, or velocities at the line core formation height range. Additionally, by combining unresolved upflows and downflows up to ∼250 km s{sup −1} within one resolution element, we reproduce the widely broadened line wings. While we cannot unambiguously determine which mechanism dominates in flares, future modeling efforts should investigate unresolved components, additional heat dissipation, larger velocities, and higher densities and combine the analysis of multiple spectral lines.

Effect of different butyrate supplementations on growth and health of weaning pigs challenged or not with E. coli K88

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Paolo Trevisi

2010-01-01

Full Text Available In a full factorial design (4 diets X challenge, Yes/No, 72 weaning pigs were assigned to one of the diets: Control; experimental diets, obtained with the addition of 2 g/kg free sodium butyrate (fNaB, or 0.6 g/kg fat-protected sodium butyrate (pNaB, or 2 g/kg INVE-NutriAd commercial mixture (Mix, based on 75 g/kg protected butyrate. Oral challenge with Escherichia coli K88 was done on 2/3 of pigs on d 7. Pigs were slaughtered on d 13. The mortality in challenged pigs, tended to be higher in control group (50.0% than in the three supplemented groups (23.5%. Growth tended to be increased averagely by the supplements (p=0.100 after the challenge, that also significantly reduced growth. In general the diet did not affect the fecal shedding of Escherichia coli and Lactobacilli, the K88-specific IgA activity in blood, the morphology of oxyntic mucosa and the expression of H+/K+-ATPase gene. The supplementations tended to increase villous length of jejunum (p=0.101. On the whole, growth, villous height and surviving rate can be positively affected either when the supplementation is done by free butyrate, by protected butyrate or by the special Inve Nutri-Ad product and these effects are distributed both on pigs infected or not with Escherichia coli K88.

Laboratory Safety Guide for Arkansas K-12 Schools.

Science.gov (United States)

Arkansas State Dept. of Education, Little Rock.

This document presents laboratory safety rules for Arkansas K-12 schools which were developed by the Arkansas Science Teachers Association (ASTA) and the Arkansas Department of Education (ADE). Contents include: (1) "Laboratory Safety Guide for Arkansas K-12 Schools"; (2) "Safety Considerations"; (3) "Safety Standards for Science Laboratories";…

Development of a new fluorescent reporter:operator system: location of AraC regulated genes in Escherichia coli K-12.

Science.gov (United States)

Sellars, Laura E; Bryant, Jack A; Sánchez-Romero, María-Antonia; Sánchez-Morán, Eugenio; Busby, Stephen J W; Lee, David J

2017-08-03

In bacteria, many transcription activator and repressor proteins regulate multiple transcription units that are often distally distributed on the bacterial genome. To investigate the subcellular location of DNA bound proteins in the folded bacterial nucleoid, fluorescent reporters have been developed which can be targeted to specific DNA operator sites. Such Fluorescent Reporter-Operator System (FROS) probes consist of a fluorescent protein fused to a DNA binding protein, which binds to an array of DNA operator sites located within the genome. Here we have developed a new FROS probe using the Escherichia coli MalI transcription factor, fused to mCherry fluorescent protein. We have used this in combination with a LacI repressor::GFP protein based FROS probe to assess the cellular location of commonly regulated transcription units that are distal on the Escherichia coli genome. We developed a new DNA binding fluorescent reporter, consisting of the Escherichia coli MalI protein fused to the mCherry fluorescent protein. This was used in combination with a Lac repressor:green fluorescent protein fusion to examine the spatial positioning and possible co-localisation of target genes, regulated by the Escherichia coli AraC protein. We report that induction of gene expression with arabinose does not result in co-localisation of AraC-regulated transcription units. However, measurable repositioning was observed when gene expression was induced at the AraC-regulated promoter controlling expression of the araFGH genes, located close to the DNA replication terminus on the chromosome. Moreover, in dividing cells, arabinose-induced expression at the araFGH locus enhanced chromosome segregation after replication. Regions of the chromosome regulated by AraC do not colocalise, but transcription events can induce movement of chromosome loci in bacteria and our observations suggest a role for gene expression in chromosome segregation.

Phase relationships in the area of the beta aluminate of the system K{sub 2}O-MgO-AL{sub 2}O{sub 3}; Phasenbeziehungen im Bereich der Beta-Aluminate des Systems K{sub 2}O-MgO-Al{sub 2}O{sub 3}

Energy Technology Data Exchange (ETDEWEB)

Kroon, P de

1996-12-01

The aim of this work was to be able to make statements about the thermodynamic stability of K-{beta}``-Al{sub 2}O{sub 3} in the pseudo-binary system K{sub 2}O-Al{sub 2}O{sub 3} and in the pseudo-ternary system K{sub 2}O-MgO-Al{sub 2}O{sub 3} relative to the adjacent phases of KAlO{sub 2} {alpha}-Al{sub 2}O{sub 3}, MgAl{sub 2}O{sub 4} and K-{beta}-Al{sub 2}O{sub 3}. (orig./MM) [Deutsch] Ziel dieser Arbeit war es, Aussagen ueber die thermodynamische Stabilitaet von K-{beta}``-Al{sub 2}O{sub 3} im pseudobinaeren System K{sub 2}O-Al{sub 2}O{sub 3} und im pseudoternaeren System K{sub 2}O-MgO-Al{sub 2}O{sub 3} relativ zu den benachbarten Phasen KAlO{sub 2}, {alpha}-Al{sub 2}O{sub 3}, MgAl{sub 2}O{sub 4} und K-{beta}-Al{sub 2}O{sub 3} machen zu koennen. (orig./MM)

Hydriding and dehydriding rates of Mg, Mg-10TaF5, and Mg-10NbF5 prepared via reactive mechanical grinding

Science.gov (United States)

Song, Myoung Youp; Kwak, Young Jun; Lee, Seong Ho; Park, Hye Ryoung

2015-01-01

In this work, TaF5 and NbF5 were chosen as additives to enhance the hydriding and dehydriding rates of Mg. Mg, Mg-10TaF5, and Mg-10NbF5 samples were prepared by reactive mechanical grinding. The hydriding and dehydriding properties of the samples were then examined. Mg-10TaF5 had the largest amount of hydrogen absorbed for 30 min and the highest initial dehydriding rate after incubation period, followed in order by Mg-10NbF5, and Mg. At 593 K under 12 bar H2 at the first cycle, Mg-10TaF5 absorbed 3.63 wt% H for 5 min and 4.53 wt% H for 30 min. At 593 K under 1.0 bar H2 at the first cycle, Mg-10TaF5 desorbed 0 wt% H for 2.5 min, 0.59 wt% H for 5 min, 3.42 wt% H for 30 min, and 4.24 wt% H for 60 min. The reactive mechanical grinding of Mg with TaF5 or NbF5 is believed to have facilitated the nucleation and to have decreased the diffusion distances of hydrogen atoms. These two effects are believed to have increased the hydriding and dehydriding rates of Mg. The MgF2 and Ta2H formed in Mg-10TaF5, and the MgF2, NbH2, and NbF3 formed in Mg-10NbF5 are considered to have enhanced both of these effects.

Keeping Pace with K-12 Online Learning, 2016

Science.gov (United States)

Gemin, Butch; Pape, Larry

2017-01-01

"Keeping Pace with K-12 Online Learning 2016" marks the thirteenth consecutive year Evergreen has published its annual research of the K-12 education online learning market. The thirteen years of researching, writing and publishing this report represents a time of remarkable change. There has been a constant presence that has become the…

Interactions of Neuropathogenic Escherichia coli K1 (RS218) and Its Derivatives Lacking Genomic Islands with Phagocytic Acanthamoeba castellanii and Nonphagocytic Brain Endothelial Cells

Science.gov (United States)

Yousuf, Farzana Abubakar; Yousuf, Zuhair; Iqbal, Junaid; Siddiqui, Ruqaiyyah; Khan, Hafsa; Khan, Naveed Ahmed

2014-01-01

Here we determined the role of various genomic islands in E. coli K1 interactions with phagocytic A. castellanii and nonphagocytic brain microvascular endothelial cells. The findings revealed that the genomic islands deletion mutants of RS218 related to toxins (peptide toxin, α-hemolysin), adhesins (P fimbriae, F17-like fimbriae, nonfimbrial adhesins, Hek, and hemagglutinin), protein secretion system (T1SS for hemolysin), invasins (IbeA, CNF1), metabolism (D-serine catabolism, dihydroxyacetone, glycerol, and glyoxylate metabolism) showed reduced interactions with both A. castellanii and brain microvascular endothelial cells. Interestingly, the deletion of RS218-derived genomic island 21 containing adhesins (P fimbriae, F17-like fimbriae, nonfimbrial adhesins, Hek, and hemagglutinin), protein secretion system (T1SS for hemolysin), invasins (CNF1), metabolism (D-serine catabolism) abolished E. coli K1-mediated HBMEC cytotoxicity in a CNF1-independent manner. Therefore, the characterization of these genomic islands should reveal mechanisms of evolutionary gain for E. coli K1 pathogenicity. PMID:24818136

Interactions of Neuropathogenic Escherichia coli K1 (RS218 and Its Derivatives Lacking Genomic Islands with Phagocytic Acanthamoeba castellanii and Nonphagocytic Brain Endothelial Cells

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Farzana Abubakar Yousuf

2014-01-01

Full Text Available Here we determined the role of various genomic islands in E. coli K1 interactions with phagocytic A. castellanii and nonphagocytic brain microvascular endothelial cells. The findings revealed that the genomic islands deletion mutants of RS218 related to toxins (peptide toxin, α-hemolysin, adhesins (P fimbriae, F17-like fimbriae, nonfimbrial adhesins, Hek, and hemagglutinin, protein secretion system (T1SS for hemolysin, invasins (IbeA, CNF1, metabolism (D-serine catabolism, dihydroxyacetone, glycerol, and glyoxylate metabolism showed reduced interactions with both A. castellanii and brain microvascular endothelial cells. Interestingly, the deletion of RS218-derived genomic island 21 containing adhesins (P fimbriae, F17-like fimbriae, nonfimbrial adhesins, Hek, and hemagglutinin, protein secretion system (T1SS for hemolysin, invasins (CNF1, metabolism (D-serine catabolism abolished E. coli K1-mediated HBMEC cytotoxicity in a CNF1-independent manner. Therefore, the characterization of these genomic islands should reveal mechanisms of evolutionary gain for E. coli K1 pathogenicity.

Thermal degradation products of saccharides: effect study over Escherichia coli K12S cells

International Nuclear Information System (INIS)

Oliveira, R.L.B.C. de.

1980-01-01

The heat sterilization of reducing sugars, in the presence of phosphates, in alkaline pH, promotes caramelization reactions, yielding a serie of degradation products. Among them, aldehyde-like compounds seem to be responsible for the decrease in viability of DNA repair-proficient E.coli cells. A positive interaction between toxic solutions and UV-radiation effects is observed in these cells. The sinergism UV-toxic solutions varies in function of post-irradiation time and is dependent on UV dose, indicating the interference of repair processes in toxicity. The effect of non-reducing sugars on cellular viability is negligible, suggesting that toxic substances generation is linked to the presence of at least a free carbonyl group in sugar structure. All tested reducing sugars, when experimental conditions remained constant, have similarly shaped inactivation kinetics and their effects are equally inhibited by catalase activity, during incubation. (author)

Retaining K-12 Online Teachers: A Predictive Model for K-12 Online Teacher Turnover

Science.gov (United States)

Larkin, Ingle M.; Lokey-Vega, Anissa; Brantley-Dias, Laurie

2018-01-01

The purpose of this study was to measure and explore factors influencing K-12 online teachers' turnover intentions, with job satisfaction and organizational commitment serving as moderating variables. Using Fishbein and Ajzen's Theory of Reasoned Action and Planned Behavior (1975), this study was conducted in public, private, charter, for-profit,…

Energetic prediction on the stability of A2Mg12Si7, A2Mg4Si3, and AMgSi in the A2Si–Mg2Si system (A = Ca, Sr and Ba) and their calculated electronic structures

International Nuclear Information System (INIS)

Imai, Yoji; Mori, Yoshihisa; Nakamura, Shigeyuki; Takarabe, Ken-ichi

2014-01-01

Highlights: • Formation energies of A 2 Mg 4 Si 3 , A 2 Mg 12 Si 7 , and AMgSi (A = Ca,Sr,Ba) were calculated. • All AMgSi are quite stable compared to mixture of A 2 Si and Mg 2 Si. • Ba 2 Mg 4 Si 3 and Sr 2 Mg 4 Si 3 are predicted to be stable, but Ca 2 Mg 4 Si 3 is not. • Ca 2 Mg 12 Si 7 and Sr 2 Mg 12 Si 7 are energetically unstable. • Stability of Ba 2 Mg 12 Si 7 is a tender subject. -- Abstract: In order to evaluate the relative stability of A 2 Mg 4 Si 3 , A 2 Mg 12 Si 7 , and AMgSi (A = Ca, Sr, and Ba) in the A 2 Si–Mg 2 Si system, electronic energy changes in the formation of these compounds were calculated using a density-functional theory with the Perdew–Wang generalized gradient approximations. It was found that (1) AMgSi’s are quite stable compared to equi-molar mixture of A 2 Si and Mg 2 Si, (2) Ba 2 Mg 4 Si 3 and Sr 2 Mg 4 Si 3 are also stable, (3) Ca 2 Mg 4 Si 3 and Ca 2 Mg 12 Si 7 are less stable than the mixture of CaMgSi and Mg 2 Si, and (4) Stability of Ba 2 Mg 12 Si 7 is a tender subject and Sr 2 Mg 12 Si 7 is energetically unstable compared to the mixture of Sr 2 Mg 4 Si 3 (or, SrMgSi) and Mg 2 Si. The presence of Sr 2 Mg 12 Si 7 may be due to the vibrational and/or configurational entropy, which are not treated in the present study. From the calculated electronic densities of state, complex compounds of SrMgSi and Mg 2 Si have both p-type and n-type character, depending on the ratio of SrMgSi and Mg 2 Si in that compound

Ultrasound-assisted extraction of Ca, K and Mg from in vitro citrus culture

Directory of Open Access Journals (Sweden)

Arruda Sandra C. C.

2003-01-01

Full Text Available An ultrasound extraction procedure for Ca, K and Mg from in vitro plant cultures is proposed, comparing cultures of different embryogenic levels of Citrus sinensis and Citrus limonia, employing ultrasound energy. Parameters related to metals extraction, such as plant material sampling, acid concentration and sonication time were investigated. For accuracy check, the proposed ultrasound extraction procedure was compared with a microwave-assisted digestion procedure and no differences in the results were verified at 95% of the confidence level. With this simple and accurate extraction procedure, it was possible to determine differences in Ca, K and Mg concentrations during Citrus embryo formation/development and between cultures (embryogenic and non-embryogenic. Finally, the ultrasound extraction method demonstrated to be an excellent alternative for handless sampling and operational costs.

FimH adhesin of Escherichia coli K1 type 1 fimbriae activates BV-2 microglia

International Nuclear Information System (INIS)

Lee, Jongseok; Shin, Sooan; Teng, C.-H.; Hong, Suk Jin; Kim, Kwang Sik

2005-01-01

The generation of intense inflammation in the subarachnoid space in response to meningitis-causing bacteria contributes to brain dysfunction and neuronal injury in bacterial meningitis. Microglia, the major immune effector cells in the central nervous system (CNS), become activated by bacterial components to produce proinflammatory immune mediators. In this study, we showed that FimH adhesin, a tip component of type 1 fimbriae of meningitis-causing Escherichia coli K1, activated the murine microglial cell line, BV-2, which resulted in the production of nitric oxide and the release of tumor necrosis factor-α. Mitogen-activated protein kinases, ERK and p-38, and nuclear factor-κB were involved in FimH adhesin-mediated microglial activation. These findings suggest that FimH adhesin contributes to the CNS inflammatory response by virtue of activating microglia in E. coli meningitis

Detection and Classification of Live and Dead Escherichia coli by Laser-Induced Breakdown Spectroscopy

Science.gov (United States)

Sivakumar, P.; Fernández-Bravo, A.; Taleh, L.; Biddle, J.F.

2015-01-01

Abstract A common goal for astrobiology is to detect organic materials that may indicate the presence of life. However, organic materials alone may not be representative of currently living systems. Thus, it would be valuable to have a method with which to determine the health of living materials. Here, we present progress toward this goal by reporting on the application of laser-induced breakdown spectroscopy (LIBS) to study characteristics of live and dead cells using Escherichia coli (E. coli) strain K12 cells as a model organism since its growth and death in the laboratory are well understood. Our goal is to determine whether LIBS, in its femto- and/or nanosecond forms, could ascertain the state of a living organism. E. coli strain K12 cells were grown, collected, and exposed to one of two types of inactivation treatments: autoclaving and sonication. Cells were also kept alive as a control. We found that LIBS yields key information that allows for the discrimination of live and dead E. coli bacteria based on ionic shifts reflective of cell membrane integrity. Key Words: E. coli—Trace elements—Live and dead cells—Laser-induced breakdown spectroscopy—Atomic force microscopy. Astrobiology 15, 144–153. PMID:25683088

The gntP Gene of Escherichia coli Involved in Gluconate Uptake

DEFF Research Database (Denmark)

Klemm, Per; Tong, S.; Nielsen, Henrik

1996-01-01

The gntP gene, located between the fim and uxu loci in Escherichia coli K-12, has been cloned and characterized. Nucleotide sequencing of a region encompassing the gntP gene revealed an open reading frame of 447 codons with significant homology to the Bacillus subtilis gluconate permease. Northern...

High temperature in combination with UV irradiation enhances horizontal transfer of stx2 gene from E. coli O157:H7 to non-pathogenic E. coli.

Directory of Open Access Journals (Sweden)

Wan-Fu Yue

Full Text Available Shiga toxin (stx genes have been transferred to numerous bacteria, one of which is E. coli O157:H7. It is a common belief that stx gene is transferred by bacteriophages, because stx genes are located on lambdoid prophages in the E. coli O157:H7 genome. Both E. coli O157:H7 and non-pathogenic E. coli are highly enriched in cattle feedlots. We hypothesized that strong UV radiation in combination with high temperature accelerates stx gene transfer into non-pathogenic E. coli in feedlots.E. coli O157:H7 EDL933 strain were subjected to different UV irradiation (0 or 0.5 kJ/m(2 combination with different temperature (22, 28, 30, 32, and 37 °C treatments, and the activation of lambdoid prophages was analyzed by plaque forming unit while induction of Stx2 prophages was quantified by quantitative real-time PCR. Data showed that lambdoid prophages in E. coli O157:H7, including phages carrying stx2, were activated under UV radiation, a process enhanced by elevated temperature. Consistently, western blotting analysis indicated that the production of Shiga toxin 2 was also dramatically increased by UV irradiation and high temperature. In situ colony hybridization screening indicated that these activated Stx2 prophages were capable of converting laboratory strain of E. coli K12 into new Shiga toxigenic E. coli, which were further confirmed by PCR and ELISA analysis.These data implicate that high environmental temperature in combination with UV irradiation accelerates the spread of stx genes through enhancing Stx prophage induction and Stx phage mediated gene transfer. Cattle feedlot sludge are teemed with E. coli O157:H7 and non-pathogenic E. coli, and is frequently exposed to UV radiation via sunlight, which may contribute to the rapid spread of stx gene to non-pathogenic E. coli and diversity of shiga toxin producing E. coli.

Fast growth phenotype of E. coli K-12 from adaptive laboratory evolution does not require intracellular flux rewiring

DEFF Research Database (Denmark)

Long, Christopher P.; Gonzalez, Jacqueline E.; Feist, Adam M.

2017-01-01

and growth condition, to probe the limits of E. coli growth rate and gain insights into fast growth phenotypes. Previous studies have described up to 1.6-fold increases in growth rate following ALE, and have identified key causal genetic mutations and changes in transcriptional patterns. Here, we report...

Elucidating mechanisms of toxic action of dissolved organic chemicals in oil sands process-affected water (OSPW).

Science.gov (United States)

Morandi, Garrett D; Wiseman, Steve B; Guan, Miao; Zhang, Xiaowei W; Martin, Jonathan W; Giesy, John P

2017-11-01

Oil sands process-affected water (OSPW) is generated during extraction of bitumen in the surface-mining oil sands industry in Alberta, Canada, and is acutely and chronically toxic to aquatic organisms. It is known that dissolved organic compounds in OSPW are responsible for most toxic effects, but knowledge of the specific mechanism(s) of toxicity, is limited. Using bioassay-based effects-directed analysis, the dissolved organic fraction of OSPW has previously been fractionated, ultimately producing refined samples of dissolved organic chemicals in OSPW, each with distinct chemical profiles. Using the Escherichia coli K-12 strain MG1655 gene reporter live cell array, the present study investigated relationships between toxic potencies of each fraction, expression of genes and characterization of chemicals in each of five acutely toxic and one non-toxic extract of OSPW derived by use of effects-directed analysis. Effects on expressions of genes related to response to oxidative stress, protein stress and DNA damage were indicative of exposure to acutely toxic extracts of OSPW. Additionally, six genes were uniquely responsive to acutely toxic extracts of OSPW. Evidence presented supports a role for sulphur- and nitrogen-containing chemical classes in the toxicity of extracts of OSPW. Copyright © 2017 Elsevier Ltd. All rights reserved.

Clinically Relevant ESBL-Producing K. pneumoniae ST307 and E. coli ST38 in an Urban West African Rat Population

Directory of Open Access Journals (Sweden)

Katharina Schaufler

2018-02-01

Full Text Available High-risk ESBL-producing Enterobacteriaceae (ESBL-E have been described in wild birds and rodents worldwide. Rats are of special interest not only due to their indicator role for environmental pollution with multi-resistant bacteria but also as possible infection source. Data on the presence of high-risk ESBL-E in urban wildlife from Africa remain scarce, however. Twenty-nine animals from three different rat (Rattus species were captured in the city of Conakry (Guinea, West Africa in 2015. Rectal swabs were analyzed for ESBL-E using selective media. Species typing and phenotypic antimicrobial resistance analysis to broad-spectrum beta-lactams and other classes of antimicrobials was performed for Enterobacteriaceae-like isolates using the VITEK®2 system (BioMérieux, Germany. Confirmed ESBL-producing E. coli and K. pneumoniae were whole-genome sequenced and resistance genes, phylogenetic background and genes related to bacterial fitness and virulence were analyzed. In total, six of twenty-nine rats (20% carried ESBL-E (K. pneumoniae and E. coli. All ESBL-producers were multi-drug resistant with blaCTX−M−15 as the dominating ESBL-type. Interestingly, ESBL-associated clonal lineages E. coli ST38 and K. pneumoniae ST307 were found. The ESBL-plasmid in K. pneumoniae ST307 revealed high sequence similarities to pKPN3-307_TypeC, a >200 kbp IncFII plasmid originating from a human clinical ST307 isolate. This was in contrast to the core genome: the rat isolate was distantly related to the human clinical ST307 isolate (27 SNPs/Mbp. In addition, we identified π-fimbrial, capsule 2, and glycogen synthesis clusters in the rodent ST307 isolate, whose involvement in the adaptation to survival outside the host and in human urinary tracts has been suggested. Our results demonstrate the presence of clinically relevant, ESBL-producing K. pneumoniae ST307 and E. coli ST38 clonal lineages in an urban West African rat population. The human community is likely

Sensitivity of strains of Escherichia coli differing in repair capability to far UV, near UV and visible radiations

International Nuclear Information System (INIS)

Webb, R.B.; Brown, M.S.

1976-01-01

In stationary phase, strains of Escherichia coli deficient in excision (B/r Hcr) or recombination repair (K12 AB2463) were more sensitive than a repair proficient strain (B/r) to monochromatic near-ultraviolet (365nm) and visible (460 nm) radiations. The relative increase in sensitivity of mutants deficient in excision or recombination repair in comparison to the wildtype, was less at 365 nm than at 254 nm. However, a strain deficient in both excision and recombination repair (K12 AB2480) showed a large, almost equal, increase in sensitivity over mutants deficient in either excision or recombination repair at 365 nm and 254 nm. All strains tested were highly resistant to 650 nm radiation. Action spectra for lethality of strains B/r and B/r Hcr in stationary phase reveal small peaks or shoulders in the 330 to 340, 400 to 410 and 490 to 510 nm wavelength ranges. The presence of 5 micro g/ml acriflavine (an inhibitor of repair) in the plating medium greatly increased the sensitivity of strain B/r to radiation at 254, 365 and 460 nm, while strains E.coli B/r Hcr and K12 AB2463 were sensitized by small amounts. At each of the wavelengths tested, acriflavine in the plating medium had at most a small effect on E.coli K12 AB2480. Acriflavine failed to sensitize any strain tested at 650 nm. Evidence supports the interpretation that lesions induced in DNA by 365 nm and 460 nm radiations play the major role in the inactivation of E.coli by these wavelengths. Single-strand breaks (or alkali-labile bonds), but not pyrimidine dimers are candidates for the lethal DNA lesions in uvrA and repair proficient strains. At high fluences lethality may be enhanced by damage to the excision and recombination repair systems. (author)

Sensitivity of strains of Escherichia coli differing in repair capability to far uv, near uv and visible radiations

Energy Technology Data Exchange (ETDEWEB)

Webb, R B; Brown, M S [Argonne National Lab., Ill. (USA)

1976-11-01

In stationary phase, strains of Escherichia coli deficient in excision (B/r Hcr) or recombination repair (K12 AB2463) were more sensitive than a repair proficient strain (B/r) to monochromatic near-ultraviolet (365nm) and visible (460 nm) radiations. The relative increase in sensitivity of mutants deficient in excision or recombination repair in comparison to the wildtype, was less at 365 nm than at 254 nm. However, a strain deficient in both excision and recombination repair (K12 AB2480) showed a large, almost equal, increase in sensitivity over mutants deficient in either excision or recombination repair at 365 nm and 254 nm. All strains tested were highly resistant to 650 nm radiation. Action spectra for lethality of strains B/r and B/r Hcr in stationary phase reveal small peaks or shoulders in the 330 to 340, 400 to 410 and 490 to 510 nm wavelength ranges. The presence of 5 micro g/ml acriflavine (an inhibitor of repair) in the plating medium greatly increased the sensitivity of strain B/r to radiation at 254, 365 and 460 nm, while strains E.coli B/r Hcr and K12 AB2463 were sensitized by small amounts. At each of the wavelengths tested, acriflavine in the plating medium had at most a small effect on E.coli K12 AB2480. Acriflavine failed to sensitize any strain tested at 650 nm. Evidence supports the interpretation that lesions induced in DNA by 365 nm and 460 nm radiations play the major role in the inactivation of E.coli by these wavelengths. Single-strand breaks (or alkali-labile bonds), but not pyrimidine dimers are candidates for the lethal DNA lesions in uvrA and repair proficient strains. At high fluences lethality may be enhanced by damage to the excision and recombination repair systems.

Nucleotide sequence of the melA gene, coding for alpha-galactosidase in Escherichia coli K-12.

OpenAIRE

Liljeström, P L; Liljeström, P

1987-01-01

Melibiose uptake and hydrolysis in E.coli is performed by the MelB and MelA proteins, respectively. We report the cloning and sequencing of the melA gene. The nucleotide sequence data showed that melA codes for a 450 amino acid long protein with a molecular weight of 50.6 kd. The sequence data also supported the assumption that the mel locus forms an operon with melA in proximal position. A comparison of MelA with alpha-galactosidase proteins from yeast and human origin showed that these prot...

Membrane fluidity and the radiosensitivity of E. coli K1060

International Nuclear Information System (INIS)

Alper, T.; Cramp, W.A.; George, A.; Lunec, J.

1981-01-01

Escherichia coli K1060 is deficient in ability to synthesize unsaturated fatty acids, so that the composition of the membrane, and therefore its fluidity, can be changed. A discussion is presented of the results of George et al (1980) concerning the relation of radiosensitivity to membrane fluidity. The following speculations are made: 1) At ice temperatures the membrane of oleic grown bacteria is in the 'gel' state, whereas in elaidic grown bacteria the membrane is in an even more rigid configuration. As a result, lesions produced during irradiation in the presence of oxygen are more lethal than in the more fluid conditions prevailing at room temperature. 2) At room temperature it may be that the bacteria are conditioned by pre-irradiation anoxia so that they become more able to repair damage. When the temperature is decreased to ice levels in bacteria modified by growth in oleic and elaidic acid, reduced membrane fluidity may impair the metabolic activity required for this pre-irradiation conditioning. 3) The lack of temperature effects with the linoleic grown bacteria, that is, no sensitization under aerated conditions and no loss in shoulder under anoxic conditions, is consistent with the lower membrane transitions temperature (fluid to gel) associated with this fatty acid. (U.K.)

Analysis of l-glutamic acid fermentation by using a dynamic metabolic simulation model of Escherichia coli

Science.gov (United States)

2013-01-01

Background Understanding the process of amino acid fermentation as a comprehensive system is a challenging task. Previously, we developed a literature-based dynamic simulation model, which included transcriptional regulation, transcription, translation, and enzymatic reactions related to glycolysis, the pentose phosphate pathway, the tricarboxylic acid (TCA) cycle, and the anaplerotic pathway of Escherichia coli. During simulation, cell growth was defined such as to reproduce the experimental cell growth profile of fed-batch cultivation in jar fermenters. However, to confirm the biological appropriateness of our model, sensitivity analysis and experimental validation were required. Results We constructed an l-glutamic acid fermentation simulation model by removing sucAB, a gene encoding α-ketoglutarate dehydrogenase. We then performed systematic sensitivity analysis for l-glutamic acid production; the results of this process corresponded with previous experimental data regarding l-glutamic acid fermentation. Furthermore, it allowed us to predicted the possibility that accumulation of 3-phosphoglycerate in the cell would regulate the carbon flux into the TCA cycle and lead to an increase in the yield of l-glutamic acid via fermentation. We validated this hypothesis through a fermentation experiment involving a model l-glutamic acid-production strain, E. coli MG1655 ΔsucA in which the phosphoglycerate kinase gene had been amplified to cause accumulation of 3-phosphoglycerate. The observed increase in l-glutamic acid production verified the biologically meaningful predictive power of our dynamic metabolic simulation model. Conclusions In this study, dynamic simulation using a literature-based model was shown to be useful for elucidating the precise mechanisms involved in fermentation processes inside the cell. Further exhaustive sensitivity analysis will facilitate identification of novel factors involved in the metabolic regulation of amino acid fermentation. PMID

Catalytic properties of pure and K+-doped Cu O/Mg O system towards 2-propanol conversion

International Nuclear Information System (INIS)

El-Molla, S. A.; Amin, N. H.; Hammed, M. N.; Sultan, S. N.; El-Shobaky, G. A.

2013-01-01

Cu O/Mg O system having different compositions was prepared by impregnation method followed by calcination at 400-900 C. The effect of Cu O content, calcination temperature and doping with small amounts of K + species (1-3 mol %) on physicochemical, surface and catalytic properties of the system were investigated using X-ray diffraction, adsorption of N 2 at - 196 C, and conversion of isopropyl alcohol at 150-400 C using a flow technique. The results revealed that the solids having the formulae 0.2 and 0.3 Cu O/Mg O calcined at 400 C consisted of nano sized Mg O and Cu O as major phases together with Cu 2 O as minor phase. The Bet-surface areas of different absorbents are decreased by increasing Cu O content, calcination temperature and K + -doping. Mg O-support material showed very small catalytic activity in 2-propanol conversion. The investigated system behaved as selective catalyst for dehydrogenation of 2-propanol with selectivity >80%. The catalytic activity increased by increasing Cu O content and decreased by increasing the calcination temperature within 400-900 C. K + -doping increased the catalytic activity and catalytic durability. (Author)

MAGNETIC DIAGNOSTICS OF THE SOLAR CHROMOSPHERE WITH THE Mg II h–k LINES

Energy Technology Data Exchange (ETDEWEB)

Del Pino Alemán, T.; Casini, R. [High Altitude Observatory, National Center for Atmospheric Research, P.O. Box 3000, Boulder, CO 80307-3000 (United States); Manso Sainz, R. [Max-Planck-Institut für Sonnensystemforschung, Justus-von-Liebig-Weg 3, D-37077 Göttingen (Germany)

2016-10-20

We investigated the formation of the Mg ii h–k doublet in a weakly magnetized atmosphere (20–100 G) using a newly developed numerical code for polarized radiative transfer in a plane-parallel geometry, which implements a recent formulation of partially coherent scattering by polarized multi-term atoms in arbitrary magnetic-field regimes. Our results confirm the importance of partial redistribution effects in the formation of the Mg ii h and k lines, as pointed out by previous work in the non-magnetic case. We show that the presence of a magnetic field can produce measurable modifications of the broadband linear polarization even for relatively small field strengths (∼10 G), while the circular polarization remains well represented by the classical magnetograph formula. Both these results open an important new window for the weak-field diagnostics of the upper solar atmosphere.

Expression optimization and biochemical characterization of a recombinant gamma-glutamyltranspeptidase from Escherichia coli novablue.

Science.gov (United States)

Yao, Ya-Feng; Weng, Yih-Ming; Hu, Hui-Yu; Ku, Kuo-Lung; Lin, Long-Liu

2006-09-01

A truncated Escherichia coli Novablue gamma-glutamyltranspeptidase (EcGGT) gene lacking the first 48-bp coding sequence for part of the signal sequence was amplified by polymerase chain reaction and cloned into expression vector pQE-30 to generate pQE-EcGGT. The maximum production of His(6)-tagged enzyme by E. coli M15 (pQE-EcGGT) was achieved with 0.1 mM IPTG induction for 12 h at 20 degrees C. The overexpressed enzyme was purified to homogeneity by nickel-chelate chromatography to a specific transpeptidase activity of 4.25 U/mg protein and a final yield of 83%. The molecular masses of the subunits of the purified enzyme were estimated to be 41 and 21 kDa respectively by SDS-PAGE, indicating EcGGT still undergoes the post-translational cleavage even in the truncation of signal sequence. The optimum temperature and pH for the recombinant enzyme were 40 degrees C and 9, respectively. The apparent K (m) and V (max) values for gamma-glutamyl-p-nitroanilide as gamma-glutamyl donor in the transpeptidation reaction were 37.9 microM and 53.7 x 10(-3) mM min(-1), respectively. The synthesis of L -theanine was performed in a reaction mixture containing 10 mM L -Gln, 40 mM ethylamine, and 1.04 U His(6)-tagged EcGGT/ml, pH 10, and a conversion rate of 45% was obtained.

Effect of flux on the composition and luminescent properties of Ca{sub 0.68}Mg{sub 0.2}SiO{sub 3}:0.12Eu{sup 3+} red phosphor

Energy Technology Data Exchange (ETDEWEB)

Wang, Jin shan [College of Material Science and Engineering, Sichuan University, Chengdu 610065, Sichuan (China); Zhu, Da-chuan, E-mail: zdc89@163.com [College of Material Science and Engineering, Sichuan University, Chengdu 610065, Sichuan (China); Zheng, Qi [College of Material Science and Engineering, Sichuan University, Chengdu 610065, Sichuan (China); Han, Tao [Research Institute for New Materials Technology, Chongqing University of Arts and Sciences, Yongchuan 402160, Chongqing (China)

2016-11-15

Ca{sub 0.68}Mg{sub 0.2}SiO{sub 3}:0.12Eu{sup 3+} red-emitting phosphor was synthesized via one-step calcination process of the precursor prepared by chemical coprecipitation with different fluxes. Then X-ray diffraction and fluorescence spectrophotometry were adopted to investigate the structure and luminescent properties of Ca{sub 0.68}Mg{sub 0.2}SiO{sub 3}:0.12Eu{sup 3+}. The results indicated that adding fluxes increased the crystalline significantly while the phase composition of samples was not changed. Furthermore, the fluxes improved the intensity of emission peak and the quantum efficiency greatly. With the concentration of flux (Li{sub 2}CO{sub 3} or K{sub 2}CO{sub 3}) increasing, the emission intensity of Ca{sub 0.68}Mg{sub 0.2}SiO{sub 3}:0.12Eu{sup 3+} firstly increased and then decreased. Meanwhile, red-shift phenomenon was observed in the emission spectra. The optimal adding fraction of Li{sub 2}CO{sub 3} and K{sub 2}CO{sub 3} was 6% and 5% respectively, and the luminous intensity of samples calcined with the optimum adding amount of Li{sub 2}CO{sub 3}, K{sub 2}CO{sub 3} is 42 and 48 times that of the samples without flux. K{sub 2}CO{sub 3} showed a better effect on improving the emission intensity of the phosphors than Li{sub 2}CO{sub 3}.

Critical current density improvements in MgB2 superconducting bulk samples by K2CO3 additions  

DEFF Research Database (Denmark)

Grivel, J.-C.

2018-01-01

MgB2 bulk samples with potassium carbonate doping were made by means of reaction of elemental Mg and B powders mixed with various amounts of K2CO3. The Tc of the superconducting phase as well as its a-axis parameter were decreased as a result of carbon doping. Potassium escaped the samples during...... reaction. The critical current density of MgB2 was improved both in self field and under applied magnetic field for T ≤ 30 K, with optimum results for 1 mol% K2CO3 addition. The normalized flux pinning force (f(b)) shows that the flux pinning mechanism at low field is similar for all samples, following...

Role of UV-inducible proteins in repair of various wild-type Escherichia coli cells

International Nuclear Information System (INIS)

Sedliakova, M.; Slezarikova, V.; Brozmanova, J.; Masek, F.; Bayerova, V.

1980-01-01

3 wild-type strains of E. coli, namely K12 AB2497, B/r WP2 and 15 555-7, proficient in excision and post-replication repair, differ markedly in their UV resistance. To elucidate this difference, the influence was investigated of induction by application of inducing fluence (IF) before lethal fluence (LF) on repair processes after LF. In cells distinguished by low UV resistance (E. coli 15 555-7; E. coli B/r WP2), dimer excision was less complete in cultures irradiated with IF + LF than in cultures irradiated with LF only. The highly resistant E. coli K12 AB2497 performed complete excision both after IF + LF or after LF alone. All 3 types of cell survived better after IF + LF than after LF only. Because, in most strains so far investigated, the application of IF reduced dimer excision and increased survival, dimer excision per se does not appear important for survival. We conclude that the rate and completeness of dimer excision can serve as a measure of efficiency of the excision system whose action is necessary for repair of another lesion. Cells of all investigated strains could not resume DNA replication and died progressively when irradiated with LF and post-incubated with chloramphenicol (LF CAP + ). Thus, it appears that inducible proteins are necessary for repair in all wild-type E. coli cells given with potentially lethal doses of UV irradiation. (orig.)

50 K anomalies in superconducting MgB{sub 2} wires in copper and silver tubes

Energy Technology Data Exchange (ETDEWEB)

Majoros, M [Interdisciplinary Research Centre in Superconductivity, University of Cambridge, Cambridge (United Kingdom); Glowacki, B A [Interdisciplinary Research Centre in Superconductivity, University of Cambridge, Cambridge (United Kingdom); Department of Materials Science and Metallurgy, University of Cambridge, Cambridge (United Kingdom); Vickers, M E [Department of Materials Science and Metallurgy, University of Cambridge, Cambridge (United Kingdom)

2002-02-01

In situ and ex situ MgB{sub 2} wires were prepared by the powder-in-tube method. Copper and silver tubes were used as a cladding material. AC susceptibility measurements revealed a small anomalous decrease with onset around 50 K. This effect persisted also when the wires were ground into powders. Electron microscopy and x-ray studies were performed on copper clad samples. Spectroscopic measurements in a SEM showed that regions contained either Cu or Mg and B. X-ray diffraction gave the major crystalline phases as Cu, MgCu{sub 2} and MgB{sub 2}. Diffraction evidence for Cu substituting in the Mg position was inconclusive. (author)

Influence of type-I fimbriae and fluid shear stress on bacterial behavior and multicellular architecture of early Escherichia coli biofilms at single-cell resolution.

Science.gov (United States)

Wang, Liyun; Keatch, Robert; Zhao, Qi; Wright, John A; Bryant, Clare E; Redmann, Anna L; Terentjev, Eugene M

2018-01-12

Biofilm formation on abiotic surfaces in food and medical industry can cause severe contamination and infection, yet how biological and physical factors determine cellular architecture of early biofilms and bacterial behavior of the constituent cells remains largely unknown. In this study we examine the specific role of type-I fimbriae in nascent stages of biofilm formation and the response of micro-colonies to environmental flow shear at single-cell resolution. The results show that type-I fimbriae are not required for reversible adhesion from plankton, but critical for irreversible adhesion of Escherichia coli ( E.coli ) MG1655 forming biofilms on polyethylene terephthalate (PET) surfaces. Besides establishing a firm cell-surface contact, the irreversible adhesion seems necessary to initiate the proliferation of E.coli on the surface. After application of shear stress, bacterial retention is dominated by the 3D architecture of colonies independent of the population and the multi-layered structure could protect the embedded cells from being insulted by fluid shear, while cell membrane permeability mainly depends on the biofilm population and the duration time of the shear stress. Importance Bacterial biofilms could lead to severe contamination problems in medical devices and food processing equipment. However, biofilms are usually studied at a rough macroscopic level, thus little is known about how individual bacterial behavior within biofilms and multicellular architecture are influenced by bacterial appendages (e.g. pili/fimbriae) and environmental factors during early biofilm formation. We apply Confocal Laser Scanning Microscopy (CLSM) to visualize E.coli micro-colonies at single-cell resolution. Our findings suggest that type-I fimbriae are vital to the initiation of bacterial proliferation on surfaces and that the responses of biofilm architecture and cell membrane permeability of constituent bacteria to fluid shear stress are different, which are

How much territory can a single E. coli cell control?

Directory of Open Access Journals (Sweden)

Ziad W. El-Hajj

2015-04-01

Full Text Available Bacteria have been traditionally classified in terms of size and shape and are best known for their very small size. E. coli cells in particular are small rods, each 1-2 microns. However the size varies with the medium, and faster growing cells are larger because they must have more ribosomes to make more protoplasm per unit time, and ribosomes take up space. Indeed, Maaloe's experiments on how E. coli establishes its size began with shifts between rich and poor media.Recently much larger bacteria have been described, including Epulopiscium fishelsoni at 700 μm and Thiomargarita namibiensisis at 750 μm. These are not only much longer than E. coli cells but also much wider, necessitating considerable intracellular organization. Epulopiscium cells for instance, at 80 μm wide, enclose a large enough volume of cytoplasm to present it with major transport problems.This review surveys E. coli cells much longer than those which grow in nature and in usual lab cultures. These include cells mutated in a single gene (metK which are 2-4x longer than their nonmutated parent. This metK mutant stops dividing when slowly starved of S-adenosylmethionine but continues to elongate to 50 μm and more. FtsZ mutants have been routinely isolated as long cells which form during growth at 42°C. The SOS response is a well-characterized regulatory network that is activated in response to DNA damage and also results in cell elongation. Our champion elongated E. coli is a metK strain with a further, as yet unidentified mutation, which reaches 750 μm with no internal divisions and no increase in width.

Identification of the Serratia marcescens hemolysin determinant by cloning into Escherichia coli

International Nuclear Information System (INIS)

Braun, V.; Neuss, B.; Ruan, Y.; Schiebel, E.; Schoeffler, H.; Jander, G.

1987-01-01

A cosmid bank of Serratia marcescens was established from which DNA fragments were cloned into the plasmid pBR322, which conferred the chromosomally encoded hemolytic activity to Escherichia coli K-12. By transposon mutagenesis with Tn1000 and Tn5 IS50/sub L/::phoA (TnphoA), the coding region was assigned to a DNA fragment, designated hyl, comprising approximately 7 kilobases. Two proteins with molecular weights of 61,000 (61K protein) and 160,000 (160K protein) were expressed by the pBR322 derivatives and by a plasmid which contained the hly genes under the control of a phage T7 promoter and the T7 RNA polymerase. When strongly overexpressed the 160K protein was released by E. coli cells into the extracellular medium concomitant with hemolytic activity. The genes encoding the 61K and the 160K proteins were transcribed in the same direction. Mutants expressing a 160K protein truncated at the carboxyl-terminal end were partially hemolytic. Hemolysis was progressively inhibited by saccharides with increasing molecular weights from maltotriose (M/sub r/ 504) to maltoheptaose (M/sub r/ 1152) and as totally abolished by dextran 4 (M/sub r/ 4000). This result and the observed influx of [ 14 C]sucrose into erythrocytes in the presence of hemolytic E. coli transformants under osmotically protective conditions suggest the formation of defined transmembrane channels by the hemolysin

Genomic and Phenomic Study of Mammary Pathogenic Escherichia coli

Science.gov (United States)

Blum, Shlomo E.; Heller, Elimelech D.; Sela, Shlomo; Elad, Daniel; Edery, Nir; Leitner, Gabriel

2015-01-01

Escherichia coli is a major etiological agent of intra-mammary infections (IMI) in cows, leading to acute mastitis and causing great economic losses in dairy production worldwide. Particular strains cause persistent IMI, leading to recurrent mastitis. Virulence factors of mammary pathogenic E. coli (MPEC) involved pathogenesis of mastitis as well as those differentiating strains causing acute or persistent mastitis are largely unknown. This study aimed to identify virulence markers in MPEC through whole genome and phenome comparative analysis. MPEC strains causing acute (VL2874 and P4) or persistent (VL2732) mastitis were compared to an environmental strain (K71) and to the genomes of strains representing different E. coli pathotypes. Intra-mammary challenge in mice confirmed experimentally that the strains studied here have different pathogenic potential, and that the environmental strain K71 is non-pathogenic in the mammary gland. Analysis of whole genome sequences and predicted proteomes revealed high similarity among MPEC, whereas MPEC significantly differed from the non-mammary pathogenic strain K71, and from E. coli genomes from other pathotypes. Functional features identified in MPEC genomes and lacking in the non-mammary pathogenic strain were associated with synthesis of lipopolysaccharide and other membrane antigens, ferric-dicitrate iron acquisition and sugars metabolism. Features associated with cytotoxicity or intra-cellular survival were found specifically in the genomes of strains from severe and acute (VL2874) or persistent (VL2732) mastitis, respectively. MPEC genomes were relatively similar to strain K-12, which was subsequently shown here to be possibly pathogenic in the mammary gland. Phenome analysis showed that the persistent MPEC was the most versatile in terms of nutrients metabolized and acute MPEC the least. Among phenotypes unique to MPEC compared to the non-mammary pathogenic strain were uric acid and D-serine metabolism. This study

NSF GK-12 Fellows as Mentors for K-12 Teachers Participating in Field Research Experiences

Science.gov (United States)

Ellins, K.; Perry, E.

2005-12-01

The University of Texas Institute for Geophysics (UTIG) recognizes the value of providing educational opportunities to K-12 teachers who play a critical role in shaping the minds of young people who are the future of our science. To that end, UTIG established the "Texas Teachers in the Field" program in 2000 to formalize the participation of K-12 teachers in field programs that included UTIG scientists. In 2002, "Texas Teachers in the Field" evolved through UTIG's involvement in a University of Texas at Austin GK-12 project led by the Environmental Sciences Institute, which enabled UTIG to partner a subset of GK-12 Fellows with teachers participating in geophysical field programs. During the three years of the GK-12 project, UTIG successfully partnered four GK-12 Fellows with five K-12 teachers. The Fellows served as mentors to the teachers, as liaisons between UTIG scientists leading field programs and teachers and their students, and as resources in science, mathematics, and technology instruction. Specifically, Fellows prepared teachers and their students for the field investigations, supervised the design of individual Teacher Research Experience (TRE) projects, and helped teachers to develop standards-aligned curriculum resources related to the field program for use in their own classrooms, as well as broader distribution. Although all but one TRE occurred during the school year, Texas school districts and principals were willing to release teachers to participate because the experience and destinations were so extraordinary (i.e., a land-based program in Tierra del Fuego, Argentina; and research cruises to the Southeast Caribbean Sea and Hess Deep in the Pacific Ocean) and carried opportunities to work with scientists from around the world. This exceptional collaboration of GK-12 Fellows, K-12 teachers and research scientists enriches K-12 student learning and promotes greater enthusiasm for science. The level of mentoring, preparation and follow-up provided

Pertussis Toxin Exploits Host Cell Signaling Pathways Induced by Meningitis-Causing E. coli K1-RS218 and Enhances Adherence of Monocytic THP-1 Cells to Human Cerebral Endothelial Cells

Directory of Open Access Journals (Sweden)

Laura Julia Starost

2016-10-01

Full Text Available Pertussis toxin (PTx, the major virulence factor of the whooping cough-causing bacterial pathogen Bordetella pertussis, permeabilizes the blood–brain barrier (BBB in vitro and in vivo. Breaking barriers might promote translocation of meningitis-causing bacteria across the BBB, thereby facilitating infection. PTx activates several host cell signaling pathways exploited by the neonatal meningitis-causing Escherichia coli K1-RS218 for invasion and translocation across the BBB. Here, we investigated whether PTx and E. coli K1-RS218 exert similar effects on MAPK p38, NF-κB activation and transcription of downstream targets in human cerebral endothelial TY10 cells using qRT-PCR, Western blotting, and ELISA in combination with specific inhibitors. PTx and E. coli K1-RS218 activate MAPK p38, but only E. coli K1-RS218 activates the NF-κB pathway. mRNA and protein levels of p38 and NF-κB downstream targets including IL-6, IL-8, CxCL-1, CxCL-2 and ICAM-1 were increased. The p38 specific inhibitor SB203590 blocked PTx-enhanced activity, whereas E. coli K1-RS218’s effects were inhibited by the NF-κB inhibitor Bay 11-7082. Further, we found that PTx enhances the adherence of human monocytic THP-1 cells to human cerebral endothelial TY10 cells, thereby contributing to enhanced translocation. These modulations of host cell signaling pathways by PTx and meningitis-causing E. coli support their contributions to pathogen and monocytic THP-1 cells translocation across the BBB.

Low Z elements (Mg, Al, and Si) K-edge X-ray absorption spectroscopy in minerals and disordered systems

International Nuclear Information System (INIS)

Ildefonse, P.; Calas, G.; Flank, A.M.; Lagarde, P.

1995-01-01

Soft X-ray absorption near edge spectroscopy (XANES) and extended X-ray absorption fine structure (EXAFS) spectroscopy have been performed at the Mg-, Al- and Si-K edges in order to establish the ability of this spectroscopy to derive structural information in disordered solids such as glasses and gels. Mg- and Al-K XANES are good structural probes to determine the coordination state of these elements in important minerals, glasses and gels. In a CaO-MgO-2SiO 2 glass Mg XANES spectra differ from that found in the crystalline equivalent, with a significant shift of the edge maxima to lower energy, consistent with a CN lower than 6. Mg-EXAFS on the same sample are in agreement and indicate the presence of 5-coordinated Mg with Mg-O distances of 2.01 A. In aluminosilicate gels, Al-K XANES has been used to investigate the [4]Al/Al total ratios. These ratios increase as the Al/Si ratios decrease. Aluminosilicate and ferric-silicate gels were studied by using Si-K edge XANES. XANES spectra differ significantly among the samples studied. Aluminosilicate gels with Al/Si=1 present a different Al and Si local environment from that known in clay minerals with the same Al/Si ratio. The gel-to-mineral transformation thus implies a dissolution-recrystallization mechanism. On the contrary, ferric-silicate gel presents a Si local environment close to that found in nontronite which may be formed by a long range ordering of the initial gels. (orig.)

Repair of membrane damage in X-irradiated E. coli

International Nuclear Information System (INIS)

Gillies, N.E.; Ratnajothi, N.H.; Hewamanna, R.; Obioha, F.I.

1984-01-01

When E. coli B/r or E. coli K12 AB1157 were X-irradiated in the presence of oxygen and incubated immediately after irradiation in broth containing penicillin in concentration that on its own was not lethal to unirradiated bacteria, substantial additional killing was caused. When treatment with penicillin was delayed for increasing times after irradiation the additional killing became progressively less. These results were interpreted as demonstrating the repair or removal of oxygen-dependent radiation-induced lesions in the bacterial membranes. Removal of these lesions was inhibited by incubation of the irradiated bacteria at low temperature before treatment with penicillin or by exposing the cells to a non-lethal concentration of toluene before irradiation. These observations suggest that an enzymatic repair process may be involved in the removal of the membrane lesions. The fatty acid mutant E. coli K 1060 proved exceptional in that some additional killing by penicillin was detectable after anaerobic as well as aerobic irradiation. This points to the importance of membrane composition in the development of those radiation lesions that are brought to light by penicillin treatment. (author)

Heterologous expression of mannanase and developing a new Reporter gene system in Lactobacillus casei and Escherichia coli

DEFF Research Database (Denmark)

Lin, Jinzhong; Zou, Yexia; Ma, Chengjie

2015-01-01

Reporter gene systems are useful for studying bacterial molecular biology, including the regulation of gene expression and the histochemical analysis of protein products. Here, two genes, β-1,4-mannanase (manB) from Bacillus pumilus and β-glucuronidase (gusA) from Escherichia coli K12, were clone....... casei and E.coli....

Classifying K-12 Blended Learning

Science.gov (United States)

Staker, Heather; Horn, Michael B.

2012-01-01

The growth of online learning in the K-12 sector is occurring both remotely through virtual schools and on campuses through blended learning. In emerging fields, definitions are important because they create a shared language that enables people to talk about the new phenomena. The blended-learning taxonomy and definitions presented in this paper…

Interstellar Molecules in K-12 Education

Science.gov (United States)

Kuiper, T. B. H.; Hofstadter, M. D.; Levin, S. M.; MacLaren, D.

2006-12-01

The Lewis Center for Educational Research (LCER) and the Jet Propulsion Laboratory (JPL) collaborate in a K-12 educational project in which students conduct observations for several research programs led by radio astronomers. The Goldstone-Apple Valley Radio Telescope (GAVRT) program provides participating teachers with curriculum elements, based on the students' observing experiences, which support national and state academic standards. The current program is based on 2.2-GHz and 8.4-GHz radiometric observations of variable sources. The research programs monitor Jupiter, Uranus, and a selected set of quasars. The telescope is a decommissioned NASA Deep Space Network antenna at Goldstone, California. In the next three years, a second telescope will be added. This telescope will at least operate at the above frequencies as well as 6 GHz and 12 GHz. Possibly, it will operate in a continuous band from 1.2 GHz to 14 GHz. In either case, the telescope will be able to observe at least the 6.6-GHz and 12.2-GHz methanol maser lines. The success of the GAVRT program depends critically on the participation of scientists committed to the research who have the ability and enthusiasm for interacting with K-12 students, typically through teleconferences. The scientists will initially work with the LCER staff to create curriculum elements around their observing program.

K-12 Technology Accessibility: The Message from State Governments

Science.gov (United States)

Shaheen, Natalie L.; Lazar, Jonathan

2018-01-01

This study examined state education technology plans and technology accessibility statutes to attempt to answer the question--is K-12 instructional technology accessibility discussed in state-level technology accessibility statutes and education technology plans across the 50 United States? When a K-12 school district is planning the construction…

Magnetic anisotropy of thin sputtered MgB2 films on MgO substrates in high magnetic fields

Directory of Open Access Journals (Sweden)

Savio Fabretti

2014-03-01

Full Text Available We investigated the magnetic anisotropy ratio of thin sputtered polycrystalline MgB2 films on MgO substrates. Using high magnetic field measurements, we estimated an anisotropy ratio of 1.35 for T = 0 K with an upper critical field of 31.74 T in the parallel case and 23.5 T in the perpendicular case. Direct measurements of a magnetic-field sweep at 4.2 K show a linear behavior, confirmed by a linear fit for magnetic fields perpendicular to the film plane. Furthermore, we observed a change of up to 12% of the anisotropy ratio in dependence of the film thickness.

HPLC Analysis of Water-Soluble Vitamins (B2, B3, B6, B12, and C and Fat-Soluble Vitamins (E, K, D, A, and β-Carotene of Okra (Abelmoschus esculentus

Directory of Open Access Journals (Sweden)

Rokayya Sami

2014-01-01

Full Text Available Okra is consumed as a vegetable by populations in Africa and Asia and particularly in Egypt. In this study, we investigated some nutritional components of okra grown in four different geographical locations of Egypt. A comparative analysis of water-soluble vitamins (B2, B3, B6, B12, and C and fat-soluble vitamins (E, K, D, A, and β-carotene in okra pods was carried out. Results of principal component analysis (PCA showed three clusters of varieties. The first cluster included the Dakahlia (D and Kafr El-Sheikh (K varieties. The second and the third clusters separated out the Suez (S and Mansoura (M varieties independently. The S pod showed the highest contents of vitamins B6 (49.81 μg/100 g and E (1.47 mg/100 g but contained the lowest contents of vitamins B3 (1.42 μg/100 g and B12 (undetected. The K pod showed the lowest vitamin C content (11.60 mg/100 g. The M pod showed the highest contents of vitamins B3 (22.70 μg/100 g, B12 (91.20 μg/100 g, C (27.14 mg/100 g, and K (0.21 mg/100 g. The D pod showed the lowest contents of vitamins E (0.15 mg/100 g, K (0.05 mg/100 g, and B6 (11.50 μg/100 g. These findings could help develop meal planning at the community level by incorporating okra varieties with high vitamin content.

[Synthesis of vitamin K2 by isopentenyl transferase NovA in Pichia pastoris Gpn12].

Science.gov (United States)

Wu, Xihua; Li, Zhemin; Liu, Hui; Wang, Peng; Wang, Li; Fang, Xue; Sun, Xiaowen; Ni, Wenfeng; Yang, Qiang; Zheng, Zhiming; Zhao, Genhai

2018-01-25

The effect of methanol addition on the heterologous expression of isoprenyl transferase NovQ was studied in Pichia pastoris Gpn12, with menadione and isopentenol as precursors to catalyze vitamin K2 (MK-3) synthesis. The expression of NovQ increased by 36% when 2% methanol was added every 24 h. The influence of initial pH, temperature, methanol addition, precursors (menadione, isopentenol) addition, catalytic time and cetyltrimethyl-ammonium bromide (CTAB) addition were explored in the P. pastoris whole-cell catalytic synthesis process of MK-3 in shaking flask. Three significant factors were then studied by response surface method. The optimal catalytic conditions obtained were as follows: catalytic temperature 31.56 ℃, menadione 295.54 mg/L, catalytic time 15.87 h. Consistent with the response surface prediction results, the optimized yield of MK-3 reached 98.47 mg/L in shaking flask, 35% higher than that of the control group. On this basis, the production in a 30-L fermenter reached 189.67 mg/L when the cell catalyst of 220 g/L (dry weight) was used to catalyze the synthesis for 24 h. This method laid the foundation for the large-scale production of MK-3 by P. pastoris Gpn12.

GroEL and dnaK genes of Escherichia coli are induced by UV irradiation and nalidixic acid in an htpR+-dependent fashion

International Nuclear Information System (INIS)

Krueger, J.H.; Walker, G.C.

1984-01-01

Two proteins with molecular weights of 61,000 and 73,000 were found to be induced by UV light in Escherichia coli mutants in which the SOS responses are constitutively expressed. The induction of these proteins by UV light and nalidixic acid was shown to be independent of the recA + lexA + regulatory system. Analysis of these proteins by two-dimensional gel electrophoresis and comparison with the heat-shock proteins of E. coli revealed that the M/sub r/ 61,000 protein comigrated with the groEL gene product, that the M/sub r/ 73,000 protein comigrated with the dnaK gene product, and that other heat-shock proteins were also induced. The induction of groEL and dnaK by UV light and nalidixic acid is controlled by the htpR locus. The results suggest that the regulatory response of E. coli to agents such as UV light and nalidixic acid is more complex than previously thought. 35 references, 6 figures, 1 table

Valorization of Spent Escherichia coli Media Using Green Microalgae Chlamydomonas reinhardtii and Feedstock Production

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Jian-Guo Zhang

2017-06-01

Full Text Available The coupling of Chlamydomonas reinhardtii biomass production for nutrients removal of Escherichia coli anaerobic broth (EAB is thought to be an economically feasible option for the cultivation of microalgae. The feasibility of growing microalgae in using EAB high in nutrients for the production of more biomass was examined. EAB comprised of nutrient-abundant effluents, which can be used to produce microalgae biomass and remove environment pollutant simultaneously. In this study, C. reinhardtii 21gr (cc1690 was cultivated in different diluted E. coli anaerobic broth supplemented with trace elements under mixotrophic and heterotrophic conditions. The results showed that C. reinhardtii grown in 1×, 1/2×, 1/5× and 1/10×E. coli anaerobic broth under mixotrophic conditions exhibited specific growth rates of 2.71, 2.68, 1.45, and 1.13 day-1, and biomass production of 201.9, 184.2, 175.5, and 163.8 mg L-1, respectively. Under heterotrophic conditions, the specific growth rates were 1.80, 1.86, 1.75, and 1.02 day-1, and biomass production were 45.6, 29.4, 15.8, and 12.1 mg L-1, respectively. The removal efficiency of chemical oxygen demand, total-nitrogen and total-phosphorus from 1×E. coli anaerobic broth was 21.51, 22.41, and 15.53%. Moreover, the dry biomass had relatively high carbohydrate (44.3% and lipid content (18.7%. Therefore, this study provides an environmentally sustainable as well economical method for biomass production in promising model microalgae and subsequently paves the way for industrial use.

H-NS mediated repression of CRISPR-based immunity in Escherichia coli K12 can be relieved by the transcription activator LeuO

OpenAIRE

2010-01-01

Abstract The recently discovered prokaryotic CRISPR/Cas defense system provides immunity against viral infections and plasmid conjugation. It has been demonstrated that in Escherichia coli transcription of the Cascade genes (casABCDE) and to some extent the CRISPR array, is repressed by heat-stable nucleoid-structuring (H-NS) protein, a global transcriptional repressor. Here we elaborate on the control of the E. coli CRISPR/Cas system, and study the effect on CRISPR-based anti-vira...

D-arabinose metabolism in Escherichia coli B: induction and cotransductional mapping of the L-fucose-D-arabinose pathway enzymes.

Science.gov (United States)

Elsinghorst, E A; Mortlock, R P

1988-12-01

D-Arabinose is degraded by Escherichia coli B via some of the L-fucose pathway enzymes and a D-ribulokinase which is distinct from the L-fuculokinase of the L-fucose pathway. We found that L-fucose and D-arabinose acted as the apparent inducers of the enzymes needed for their degradation. These enzymes, including D-ribulokinase, appeared to be coordinately regulated, and mutants which constitutively synthesized the L-fucose enzymes also constitutively synthesized D-ribulokinase. In contrast to D-arabinose-positive mutants of E. coli K-12, in which L-fuculose-1-phosphate and D-ribulose-1-phosphate act as inducers of the L-fucose pathway, we found that these intermediates did not act as inducers in E. coli B. To further characterize the E. coli B system, some of the L-fucose-D-arabinose genes were mapped by using bacteriophage P1 transduction. A transposon Tn10 insertion near the E. coli B L-fucose regulon was used in two- and three-factor reciprocal crosses. The gene encoding D-ribulokinase, designated darK, was found to map within the L-fucose regulon, and the partial gene order was found to be Tn10-fucA-darK-fucI-fucK-thyA.

Physical and chemical properties of MgO ceramics treated in molten K2SO4 for a long period

International Nuclear Information System (INIS)

Iwasa, Mikio; Kose, Saburo; Korenaga, Sadayoshi; Furukawa, Mitsuhiko.

1978-01-01

The wall materials of MHD power generating channel are exposed to thermally, physically and chemically severe conditions, so that they have to withstand great damages, especially the attack of seed materials. Several kinds of ceramics proposed as the wall materials have been tested in the simulated MHD environment. In this paper, MgO ceramics were treated in molten K 2 SO 4 , a typical seed material, and the changes in their physical and chemical properties were investigated in comparison with those of Al 2 O 3 ceramics. four kinds of MgO ceramics, three sintered and one electric fused, were immersed in molten K 2 SO 4 at 1300 0 C for the periods up to 1000 h, and weight, volume, surface roughness, bending strength and hardness were measured. The changes in the microstructures and chemical compositions due to the K 2 SO 4 treatment were also investigated. MgO ceramics were attacked by molten K 2 SO 4 only at the grain boundaries on the surface, in contrast at Al 2 O 3 ceramics which were severely damaged to form β-Al 2 O 3 . It was found that SiO 2 and CaO in the grain boundaries had played important roles to the attack of K 2 SO 4 . Generally, the changes in the properties of MgO ceramics by the K 2 SO 4 treatment were very small compared with those of Al 2 O 3 ceramics. It was concluded that MgO ceramics are more stable than Al 2 O 3 ceramics in molten K 2 SO 4 and their properties do not show substantial drops for long periods. (author)

Crystal structure of the high-affinity Na+K+-ATPase-ouabain complex with Mg2+ bound in the cation binding site.

Science.gov (United States)

Laursen, Mette; Yatime, Laure; Nissen, Poul; Fedosova, Natalya U

2013-07-02

The Na(+),K(+)-ATPase maintains electrochemical gradients for Na(+) and K(+) that are critical for animal cells. Cardiotonic steroids (CTSs), widely used in the clinic and recently assigned a role as endogenous regulators of intracellular processes, are highly specific inhibitors of the Na(+),K(+)-ATPase. Here we describe a crystal structure of the phosphorylated pig kidney Na(+),K(+)-ATPase in complex with the CTS representative ouabain, extending to 3.4 Å resolution. The structure provides key details on CTS binding, revealing an extensive hydrogen bonding network formed by the β-surface of the steroid core of ouabain and the side chains of αM1, αM2, and αM6. Furthermore, the structure reveals that cation transport site II is occupied by Mg(2+), and crystallographic studies indicate that Rb(+) and Mn(2+), but not Na(+), bind to this site. Comparison with the low-affinity [K2]E2-MgF(x)-ouabain structure [Ogawa et al. (2009) Proc Natl Acad Sci USA 106(33):13742-13747) shows that the CTS binding pocket of [Mg]E2P allows deep ouabain binding with possible long-range interactions between its polarized five-membered lactone ring and the Mg(2+). K(+) binding at the same site unwinds a turn of αM4, dragging residues Ile318-Val325 toward the cation site and thereby hindering deep ouabain binding. Thus, the structural data establish a basis for the interpretation of the biochemical evidence pointing at direct K(+)-Mg(2+) competition and explain the well-known antagonistic effect of K(+) on CTS binding.

Transcriptome analysis of all two-component regulatory system mutants of Escherichia coli K-12.

Science.gov (United States)

Oshima, Taku; Aiba, Hirofumi; Masuda, Yasushi; Kanaya, Shigehiko; Sugiura, Masahito; Wanner, Barry L; Mori, Hirotada; Mizuno, Takeshi

2002-10-01

We have systematically examined the mRNA profiles of 36 two-component deletion mutants, which include all two-component regulatory systems of Escherichia coli, under a single growth condition. DNA microarray results revealed that the mutants belong to one of three groups based on their gene expression profiles in Luria-Bertani broth under aerobic conditions: (i) those with no or little change; (ii) those with significant changes; and (iii) those with drastic changes. Under these conditions, the anaeroresponsive ArcB/ArcA system, the osmoresponsive EnvZ/OmpR system and the response regulator UvrY showed the most drastic changes. Cellular functions such as flagellar synthesis and expression of the RpoS regulon were affected by multiple two-component systems. A high correlation coefficient of expression profile was found between several two-component mutants. Together, these results support the view that a network of functional interactions, such as cross-regulation, exists between different two-component systems. The compiled data are avail-able at our website (http://ecoli.aist-nara.ac.jp/xp_analysis/ 2_components).

Soil Science Society of America - K-12 Outreach

Science.gov (United States)

Lindbo, David L.; Loynachan, Tom; Mblia, Monday; Robinson, Clay; Chapman, Susan

2013-04-01

The Soil Science Society of America created its K12 Committee in 2006 in part to compliment the Dig It! The Secrets of Soil exhibit that opened in July 2008 at the Smithsonian's Institution's Nation Museum of Natural History (of which SSS was a founding sponsor). The committee's work began quickly with a website designed to provide resources for K12 teachers. The first accomplishments included reviewing and posting links to web based information already available to teachers. These links were sorted by subject and grade level to make it easier for teachers to navigate the web and find what they needed quickly. Several presentations and lessons designed for K12 teachers were also posted at this time. Concurrent with this effort a subcommittee review and organized the national teaching standards to show where soils could fit into the overall K12 curriculum. As the website was being developed another subcommittee developed a soils book (Soil! Get the Inside Scoop, 2008) to further compliment the Dig It! exhibit. This was a new endeavor for SSSA having never worked with the non-academic audience in developing a book. Peer-reviews of this book included not only scientist but also students in order to make sure the book was attractive to them. Once the book was published and the website developed it became clear more outreach was needed. SSSA K12 Committee has attended both the National Science Teachers Association (since 2008) the USA Science and Engineering Festival (since 2010) with exhibits and workshops. It has cooperated and contributed to the American Geologic Institutes' Earth Science Week materials with brochures and lesson plans and with National Association of Conservation Districts by providing peer-review and distribution of materials. The most recent developments from the committee include a web redesign that is more student and teacher friendly, the development of a peer-review system to publish K12 Lesson Plans, and finally the publication of a new soils

The Csr system regulates genome-wide mRNA stability and transcription and thus gene expression in Escherichia coli.

Science.gov (United States)

Esquerré, Thomas; Bouvier, Marie; Turlan, Catherine; Carpousis, Agamemnon J; Girbal, Laurence; Cocaign-Bousquet, Muriel

2016-04-26

Bacterial adaptation requires large-scale regulation of gene expression. We have performed a genome-wide analysis of the Csr system, which regulates many important cellular functions. The Csr system is involved in post-transcriptional regulation, but a role in transcriptional regulation has also been suggested. Two proteins, an RNA-binding protein CsrA and an atypical signaling protein CsrD, participate in the Csr system. Genome-wide transcript stabilities and levels were compared in wildtype E. coli (MG1655) and isogenic mutant strains deficient in CsrA or CsrD activity demonstrating for the first time that CsrA and CsrD are global negative and positive regulators of transcription, respectively. The role of CsrA in transcription regulation may be indirect due to the 4.6-fold increase in csrD mRNA concentration in the CsrA deficient strain. Transcriptional action of CsrA and CsrD on a few genes was validated by transcriptional fusions. In addition to an effect on transcription, CsrA stabilizes thousands of mRNAs. This is the first demonstration that CsrA is a global positive regulator of mRNA stability. For one hundred genes, we predict that direct control of mRNA stability by CsrA might contribute to metabolic adaptation by regulating expression of genes involved in carbon metabolism and transport independently of transcriptional regulation.

Spin measurement for symmetric fission states of sup 24 Mg. A new angle on the sup 12 C+ sup 12 C interaction

Energy Technology Data Exchange (ETDEWEB)

Fulton, B.R.; Bennett, S.J.; Freer, M.; Murgatroyd, J.T. (School of Physics and Space Research, Birmingham Univ. (United Kingdom)); Gyapong, G.J.; Jarvis, N.S.; Jones, C.D.; Watson, D.L. (Dept. of Physics, York Univ. (United Kingdom)); Brown, J.D.; Rae, W.D.M.; Smith, A.E. (Dept. of Nuclear Physics, Oxford Univ. (United Kingdom)); Lilley, J.S. (Daresbury Lab., Warrington (United Kingdom))

1991-09-19

A new high resolution measurement of the {sup 12}C({sup 24}Mg, {sup 12}C{sup 12}C){sup 12}C breakup reaction has been performed. Sequential breakup is observed from specific states in {sup 24}Mg at excitation energies ranging from 20 to 26 MeV. Angular correlation measurements indicate that states with spins ranging from J=4 to 8 are populated. These states are consistent with superdeformed shape isomeric bands predicted by cranked cluster model and Nilsson-Strutinsky calculations. (orig.).

High efficiency generalized transduction in Escherichia coli O157:H7 [v1; ref status: indexed, http://f1000r.es/8f

Directory of Open Access Journals (Sweden)

Martin G Marinus

2013-01-01

Full Text Available Genetic manipulation in enterohemorrhagic E. coli O157:H7 is currently restricted to recombineering, a method that utilizes the recombination system of bacteriophage lambda, to introduce gene replacements and base changes inter alia into the genome. Bacteriophage 933W is a prophage in E. coli O157:H7 strain EDL933, which encodes the genes (stx2AB for the production of Shiga toxin which is the basis for the potentially fatal Hemolytic Uremic Syndrome in infected humans. We replaced the stx2AB genes with a kanamycin cassette using recombineering. After induction of the prophage by ultra-violet light, we found that bacteriophage lysates were capable of transducing to wildtype, point mutations in the lactose, arabinose and maltose genes. The lysates could also transduce tetracycline resistant cassettes. Bacteriophage 933W is also efficient at transducing markers in E. coli K-12. Co-transduction experiments indicated that the maximal amount of transferred DNA was likely the size of the bacteriophage genome, 61 kB. All tested transductants, in both E. coli K-12 and O157:H7, were kanamycin-sensitive indicating that the transducing particles contained host DNA.

Low Z elements (Mg, Al, and Si) K-edge X-ray absorption spectroscopy in minerals and disordered systems

Science.gov (United States)

Ildefonse, Ph.; Calas, G.; Flank, A. M.; Lagarde, P.

1995-05-01

Soft X-ray absorption near edge spectroscopy (XANES) and extended X-ray absorption fine structure (EXAFS) spectroscopy have been performed at the Mg-, Al- and Si-K edges in order to establish the ability of this spectroscopy to derive structural information in disordered solids such as glasses and gels. Mg- and Al-K XANES are good structural probes to determine the coordination state of these elements in important minerals, glasses and gels. In a CaOsbnd MgOsbnd 2SiO2 glass Mg XANES spectra differ from that found in the crystalline equivalent, with a significant shift of the edge maxima to lower energy, consistent with a CN lower than 6. Mg-EXAFS on the same sample are in agreement and indicate the presence of 5-coordinated Mg with Mgsbnd O distances of 2.01Å. In aluminosilicate gels, Alsbnd K XANES has been used to investigate the [4]Al/Altotal ratios. These ratios increase as the Al/Si ratios decrease. Aluminosilicate and ferric-silicate gels were studied by using Sisbnd K edge XANES. XANES spectra differ significantly among the samples studied. Aluminosilicate gels with Al/Si= 1 present a different Al and Si local environment from that known in clay minerals with the same Al/Si ratio. The gel-to-mineral transformation thus implies a dissolution-recrystallization mechanism. On the contrary, ferric-silicate gel presents a Si local environment close to that found in nontronite which may be formed by a long range ordering of the initial gels.