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1

Induction of a tomato anionic peroxidase gene (tap1) by wounding in transgenic tobacco and activation of tap1/GUS and tap2/GUS chimeric gene fusions in transgenic tobacco by wounding and pathogen attack.  

PubMed

The anionic peroxidase genes of tomato, tap1 and tap2, are induced by wounding in tomato fruits and by elicitor treatment in cell suspension cultures. These homologous genes code for anionic peroxidases that are postulated to cause polymerization of the phenolic residues into wall polymers in wound-healing and pathogen-infected tissues. An expression construct containing the entire TAP1 gene with its 5' and 3' flanking sequences was introduced into tobacco by Agrobacterium tumefaciens-mediated gene transfer. Also, constructs containing the 5' upstream regions of tap1 and tap2 including sequences coding for their respective putative leader peptides fused translationally to the beta-glucuronidase (GUS) reporter gene were made and introduced into tobacco. Northern blot analysis of transcripts from wound-healing leaf tissues of transformants containing tap1 showed that the introduced gene was being transcribed in the heterologous host. The induction of tap1 transcripts in the wound-healing transgenic tobacco tissues was observed by 48 h and increased over time period of 84 h. Wounding also led to expression of GUS in tap1/GUS and tap2/GUS transformants and GUS activity was localized to the wound site. Activation of the tap1 and tap2 promoters in wound-healing transgenic tobacco tissues showed a GUS expression profile that correlated with the postulated role for anionic peroxidases in phenolic polymerization in suberizing tissues. Inoculation of tap1/GUS and tap2/GUS transformant leaves with fungal conidia from Fusarium solani f. sp. pisi caused expression of GUS in locally inoculated regions, and GUS expression increased over a period of four days. PMID:7678769

Mohan, R; Bajar, A M; Kolattukudy, P E

1993-01-01

2

Recombinant human AhR-mediated GUS reporter gene assays for PCB congeners in transgenic tobacco plants in comparison with recombinant mouse and guinea pig AhRs.  

PubMed

Four expression plasmids for recombinant human aryl hydrocarbon receptor (hAhR) consisting of a ligand binding domain of hAhR, a DNA-binding domain of LexA and a transactivation domain of VP16 as well as ?-glucuronidase (GUS) reporter genes were constructed. All the expression plasmids were transformed into tobacco plants. The selected transgenic tobacco plants were used to assay. PCB congeners showed GUS activity in a TEF-dependent manner. The selected transgenic tobacco plant XhD4V17 was compared with the transgenic tobacco plants XmD4V26 and XgD2V23 containing recombinant mouse (m) AhR-mediated GUS reporter gene expression cassette and recombinant guinea pig (g) AhR-mediated GUS reporter gene expression cassette for PCB congener-inducible GUS activity. The data revealed that the tobacco plant XgD2V23 was the most active in PCB congener-inducible GUS activity. In a 1:1 mixture of PCB126 and PCB80 a reduced PCB126-induced GUS activity was observed in plant XgD2V23, which could possibly be due to interaction between PCB126 and PCB80. PMID:20936562

Shimazu, Sayuri; Kawabata, Yukiko; Inayoshi, Akito; Inui, Hideyuki; Ashida, Hitoshi; Ohkawa, Hideo

2010-11-01

3

Tagging of a cryptic promoter that confers root-specific gus expression in Arabidopsis thaliana  

Microsoft Academic Search

A 2.1-kb sequence was isolated by promoter trapping from an Arabidopsis thaliana transformant (T80) obtained by Agrobacterium-mediated T-DNA insertion. This sequence directed strong ?-glucuronidase (GUS) expression specifically in roots. The promoter-gus fusion was used to transform other A. thaliana plants. Most of the transformants obtained exhibited stronger GUS activity in roots than the T80 line and a weak activity\\u000a in

P. Mollier; B. Hoffmann; M. Orsel; G. Pelletier

2000-01-01

4

Efficient gusA transient expression in Porphyra yezoensis protoplasts mediated by endogenous beta-tubulin flanking sequences  

NASA Astrophysics Data System (ADS)

Endogenous tubulin promoter has been widely used for expressing foreign genes in green algae, but the efficiency and feasibility of endogenous tubulin promoter in the economically important Porphyra yezoensis (Rhodophyta) are unknown. In this study, the flanking sequences of beta-tubulin gene from P. yezoensis were amplified and two transient expression vectors were constructed to determine their transcription promoting feasibility for foreign gene gusA. The testing vector pATubGUS was constructed by inserting 5'-and 3'-flanking regions ( Tub5' and Tub3') up-and down-stream of ?-glucuronidase (GUS) gene ( gusA), respectively, into pA, a derivative of pCAT®3-enhancer vector. The control construct, pAGUSTub3, contains only gusA and Tub3'. These constructs were electroporated into P. yezoensis protoplasts and the GUS activities were quantitatively analyzed by spectrometry. The results demonstrated that gusA gene was efficiently expressed in P. yezoensis protoplasts under the regulation of 5'-flanking sequence of the beta-tubulin gene. More interestingly, the pATubGUS produced stronger GUS activity in P. yezoensis protoplasts when compared to the result from pBI221, in which the gusA gene was directed by a constitutive CaMV 35S promoter. The data suggest that the integration of P. yezoensis protoplast and its endogenous beta-tubulin flanking sequences is a potential novel system for foreign gene expression.

Gong, Qianhong; Yu, Wengong; Dai, Jixun; Liu, Hongquan; Xu, Rifu; Guan, Huashi; Pan, Kehou

2007-01-01

5

ß-Glucuronidase (GUS) transposons for ecological and genetic studies of rhizobia and other Gram-negative bacteria  

Microsoft Academic Search

A series of transposons are described which contain the gusA gene, encoding ?-glucuronidase (GUS), expressed from a variety of promoters, both regulated and constitutive. The regulated promoters include the tac promoter which can be induced by IPTG, and nifH promoters which are symbiotically activated in legume nodules. One transposon contains gusA with a strong Shine-Dalgarno translation initiation context, but no

K. J. Wilson; A. Sessitsch; J. C. Corbo; K. E. Giller; A. D. L. Akkermans; R. A. Jefferson

1995-01-01

6

Gus Grissom & Milt Thompson With Paresev  

NASA Technical Reports Server (NTRS)

The Paresev 1-A standing Rogers Dry Lakebed at the NASA Flight Research Center, Edwards, California. Mercury Astronaut Gus Grissom is at left and NASA test pilot Milton Thompson is at right. The Paresev evaluated a potential replacement for parachutes used on spacecraft. The Paresev was steerable and was evaluated for use on the Gemini spacecraft. The idea was not workable, however.

1962-01-01

7

Peptoid analogues of anoplin show antibacterial activity.  

PubMed

We have synthesised nine analogues of the antibacterial peptide anoplin with a peptoid residue at position 5 (H-GLLKXIKTLL-NH(2)). The most active compounds showed MIC-values of 12.5 and 25 microM against E.coli and S.aureus. These MIC-values are comparable with anoplin which showed 23 microM and 11 microM against E. coli and S.aureus. However, the selectivity was reversed. Our results indicate that peptoid analogues of anoplin are promising lead structures for developing new antibacterial agents. PMID:19799550

Meinike, K; Hansen, P R

2009-01-01

8

Expression of three Arabidopsis cytokinin oxidase\\/dehydrogenase promoter::GUS chimeric constructs in tobacco: response to developmental and biotic factors  

Microsoft Academic Search

Expression patterns of three Arabidopsis thaliana cytokinin oxidase\\/dehydrogenase promoter::GUS reporter fusions were investigated in tobacco plants. While cytokinin oxidase\\/dehydrogenase promoter 2 showed no expression in tobacco, the cytokinin oxidase\\/dehydrogenase promoters 3 and 4 were active in various tissues throughout development of the tobacco. Recently, the 1452 bp promoter region of AtCKX3 was reported as almost inactive in Arabidopsis. In contrast, the

Ivan Gális; Kristin D. Bilyeu; Maria Joao G. Godinho; Paula E. Jameson

2005-01-01

9

GUS gene remains stable in transgenic citrus callus recovered from cryopreservation.  

PubMed

The conservation of transgenic materials is very important, paticularly for their potential future use in crop development. In this study, transgenic callus cultures of 'Newhall' navel orange (Citrus sinensis Osbeck) were cryopreserved by a vitrification method. Transgenic calluses survived cryopreservation and recovered under normal culture conditions. The results of PCR (Polymerase Chain Reaction) amplification, Southern blotting and SSCP (Single-Strand Conformation Polymorphism) assay showed that the GUS gene was still maintained in the genome of callus cultures recovered from cryopreservation. X-Gluc staining further indicated GUS gene expression in callus cultures recovered from cryopreservation. PMID:14671689

Hao, Yu-Jin; Cheng, Yun-Jiang; Deng, Xiu-Xin

2003-01-01

10

Review: Biosafety of E. coli ?-glucuronidase (GUS) in plants  

Microsoft Academic Search

The ß-glucuronidase (GUS) gene is to date the most frequently used reporter gene in plants. Marketing of crops containing this gene requires prior evaluation of their biosafety. To aid such evaluations of the GUS gene, irrespective of the plant into which the gene has been introduced, the ecological and toxicological aspects of the gene and gene product have been examined.

Luud J. W. Gilissen; Peter L. J. Metz; Willem J. Stiekema; Jan-Peter Nap

1998-01-01

11

Elucidation of Septoria tritici x Wheat Interactions Using GUS-Expressing Isolates.  

PubMed

ABSTRACT Isolate ISR398 of Septoria tritici (which produces none to few pycnidia on the wheat cv. Seri 82 and high coverage on cv. Shafir) and isolate ISR8036 (which is virulent on both cultivars) were genetically cotrans-formed using the selectable marker gene hph, which confers resistance to hygromycin B (hygB), and the reporter gene uidA, encoding beta-glucuronidase (GUS). Most of the genetically transformed isolates (98.8%) produced similar pycnidial coverage on seedlings of 'Seri 82' and 'Shafir' as the two wild-type isolates. Southern analysis of 25 randomly selected hygB(R)GUS(+) transformants probed with the uidA sequence revealed multiple insertion sites. GUS activity was determined fluorimetrically by measuring the conversion of 4-methylumbelliferyl beta-D-glucuronide (MUG) to 4-methylumbelliferone (MU). The high GUS-expressing transformants 398D97 and 8036E27 were used to elucidate fungal development within inoculated leaf tissue by using GUS activity to estimate the fungal proteins content in planta. Increase in fungal biomass was recorded in 'Shafir' inoculated with the GUS-expressing transformants 398D97 and 8036E27 following a 12-day latent period. A 15-day latent period was recorded in 'Seri 82' inoculated with 8036E27, whereas an 18-day latent period was recorded on 'Seri 82' inoculated with 398D97 and the two mixtures 398D97 + ISR8036 and ISR398 + 8036E27. The rate of fungal development and the estimated level of fungal proteins at the pycnidia maturation stage was high in leaves of 'Shafir' and moderate to low on 'Seri 82', even in cases in which no significant differences were recorded in pycnidial coverage. An endogenous capacity to hydrolyze beta-1,4-D-glucuronidase was recorded in leaves inoculated with wild-type isolates. The latent periods in MU production of the uidA-expressing transformants mimicked those recorded for the wild-type isolates. However, at all stages, the levels of MU produced in wheat inoculated with wild-type isolates were markedly lower than those produced by GUS-expressing transformants. The mode of interaction (compatible or incompatible) determined the onset of the induction, rate, and level of enzyme production. PMID:18944623

Pnini-Cohen, S; Zilberstein, A; Schuster, S; Sharon, A; Eyal, Z

2000-03-01

12

Directed Chromosomal Integration and Expression of the Reporter Gene gusA3 in Lactobacillus acidophilus NCFM ?  

PubMed Central

Lactobacillus acidophilus NCFM is a probiotic microbe that survives passage through the human gastrointestinal tract and interacts with the host epithelium and mucosal immune cells. The potential for L. acidophilus to express antigens at mucosal surfaces has been investigated with various antigens and plasmid expression vectors. Plasmid instability and antibiotic selection complicate the possibility of testing these constructs in human clinical trials. Integrating antigen encoding genes into the chromosome for expression is expected to eliminate selection requirements and provide genetic stability. In this work, a reporter gene encoding a ?-glucuronidase (GusA3) was integrated into four intergenic chromosomal locations. The integrants were tested for genetic stability and GusA3 activity. Two locations were selected for insertion downstream of constitutively highly expressed genes, one downstream of slpA (LBA0169), encoding a highly expressed surface-layer protein, and one downstream of phosphopyruvate hydratase (LBA0889), a highly expressed gene with homologs in other lactic acid bacteria. An inducible location was selected downstream of lacZ (LBA1462), encoding a ?-galactosidase. A fourth location was selected in a low-expression region. The expression of gusA3 was evaluated from each location by measuring GusA3 activity on 4-methyl-umbelliferyl-?-d-glucuronide (MUG). GusA3 activity from both highly expressed loci was more than three logs higher than the gusA3-negative parent, L. acidophilus NCK1909. GusA3 activity from the lacZ locus was one log higher in cells grown in lactose than in glucose. The differences in expression levels between integration locations highlights the importance of rational targeting with gene cassettes intended for chromosomal expression.

Douglas, Grace L.; Klaenhammer, Todd R.

2011-01-01

13

The IRIS-GUS Shuttle Borne Upper Stage System  

NASA Technical Reports Server (NTRS)

This paper describes the Italian Research Interim Stage - Gyroscopic Upper Stage (IRIS-GUS) upper stage system that will be used to launch NASA's Triana Observatory from the Space Shuttle. Triana is a pathfinder earth science mission being executed on rapid schedule and small budget, therefore the mission's upper stage solution had to be a system that could be fielded quickly at relatively low cost and risk. The building of the IRIS-GUS system wa necessary because NASA lost the capability to launch moderately sized upper stage missions fro the Space Shuttle when the PAM-D system was retired. The IRIS-GUS system restores this capability. The resulting system is a hybrid which mates the existing, flight proven IRIS (Italian Research Interim Stage) airborne support equipment to a new upper stage, the Gyroscopic Upper Stage (GUS) built by the GSFC for Triana. Although a new system, the GUS exploits flight proven hardware and design approaches in most subsystems, in some cases implementing proven design approaches with state-of-the-art electronics. This paper describes the IRIS-GUS upper stage system elements, performance capabilities, and payload interfaces.

Tooley, Craig; Houghton, Martin; Bussolino, Luigi; Connors, Paul; Broudeur, Steve (Technical Monitor)

2002-01-01

14

Transient and stable gene expression in the fungal maize pathogen Cochliobolus heterostrophus after transformation with the beta-glucuronidase (GUS) gene.  

PubMed

The bacterial GUS (beta-glucuronidase) gene has been used as a reporter gene in plants and bacteria and was recently expressed in filamentous fungi. Here, we report the application of GUS for the establishment of transient and stable gene expression systems in the phytopathogenic fungus Cochliobolus heterostrophus. The utility of the transient expression system is demonstrated in applications involving promoter analysis and in tests of various parameters of a transformation system, for comparing the rates of stable and transient transformation events using GUS as sole screening marker and for comparing different transformation systems using either GUS or a dominant selection marker. For these purposes two plasmids were constructed harbouring the GUS gene and the hph gene of Escherichia coli which confers resistance to the antibiotic hygromycin B (HygB), ligated either to the P1 or GPD1 (glyceraldehyde 3 phosphate dehydrogenase) promoter of C. heterostrophus. In transient expression studies the first appearance of GUS activity was observed within 2 h after transformation and maximal values were obtained after 7 or 10 h, depending on the promoter fused to the GUS gene. At peak activity, the GPD1 promoter was revealed to be five fold stronger than the P1 promoter. The same difference in promoter strength was observed when the vectors were stably integrated in the fungal genome. Using the GUS gene as a colour selection marker in plate assays, it was possible to detect transformants and monitor the process of transient gene expression visually. Blue transformants obtained by screening for the GUS phenotype were mitotically unstable. Transformants obtained by selecting for HygB resistance were mitotically stable and expressed the beta-glucuronidase gene constitutively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8232214

Mönke, E; Schäfer, W

1993-10-01

15

Isolation of GUS marker lines for genes expressed in Arabidopsis endosperm, embryo and maternal tissues  

Microsoft Academic Search

In order to identify marker lines expressing GUS in various endosperm compartments and at different developmental stages, a collection of Arabidopsis thaliana (L.) Heynh. promoter trap lines were screened. The screen identified 16 lines displaying GUS-reporter gene expression in the endosperm, embryo and other seed organs. The distinctive pat- terns of GUS expression in these lines provide molecular markers for

Biljana Stangeland; Zhian Salehian; Reidunn Aalen; Abul Mandal; Odd-Arne Olsen

2003-01-01

16

4. AERIAL OBLIQUE FROM EAST, SHOWING DREDGING, BULKHEAD, CONSTRUCTION ACTIVITY ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

4. AERIAL OBLIQUE FROM EAST, SHOWING DREDGING, BULKHEAD, CONSTRUCTION ACTIVITY AT BASE IN BACKGROUND. USN PHOTO, C. SEPTEMBER, 1940. - Quonset Point Naval Air Station, Roger Williams Way, North Kingstown, Washington County, RI

17

GUS expression in sweet oranges (Citrus sinensis L. Osbeck) driven by three different phloem-specific promoters.  

PubMed

Huanglongbing (HLB) is associated with Candidatus Liberibacter spp., endogenous, sieve tube-restricted bacteria that are transmitted by citrus psyllid insect vectors. Transgenic expression in the phloem of specific genes that might affect Ca. Liberibacter spp. growth and development may be an adequate strategy to improve citrus resistance to HLB. To study specific phloem gene expression in citrus, we developed three different binary vector constructs with expression cassettes bearing the ?-glucuronidase (GUS) reporter gene (uidA) under the control of one of the three different promoters: Citrus phloem protein 2 (CsPP2), Arabidopsis thaliana phloem protein 2 (AtPP2), and Arabidopsis thaliana sucrose transporter 2 (AtSUC2). Transgenic lines of 'Hamlin', 'Pera', and 'Valencia' sweet oranges [Citrus sinensis (L.) Osbeck] were produced via Agrobacterium tumefaciens transformation. The epicotyl segments collected from in vitro germinated seedlings were used as explants. The gene nptII, which confers resistance to the antibiotic kanamycin, was used for selection. The transformation efficiency was expressed as the number of GUS-positive shoots over the total number of explants and varied from 1.54 to 6.08 % among the three cultivars and three constructs studied. Several lines of the three sweet orange cultivars analyzed using PCR and Southern blot analysis were genetically transformed with the three constructs evaluated. The histological GUS activity in the leaves indicates that the uidA gene was preferentially expressed in the phloem, which suggests that the use of the three promoters might be adequate for producing HLB-resistant transgenic sweet oranges. The results reported here conclusively demonstrate the preferential expression of GUS in the phloem driven by two heterologous and one homologous gene promoters. Key message The results reported here conclusively demonstrate the preferential expression of GUS in the phloem driven by two heterologous and one homologous gene promoters. PMID:22801867

Miyata, Luzia Yuriko; Harakava, Ricardo; Stipp, Liliane Cristina Libório; Mendes, Beatriz Madalena Januzzi; Appezzato-da-Glória, Beatriz; de Assis Alves Mourão Filho, Francisco

2012-11-01

18

Apca Activities Continue to Show Growth and Expansion  

Microsoft Academic Search

The activities and accomplishments of the Air Pollution Control Association during 1974-1975 continued to show growth and expansion, although there was a decline in membership. We have a financial surplus to report in spite of the fact that our publications program was plagued with inflationary price increases in paper, printing and distribution.

Lewis H. Rogers

1975-01-01

19

Synthetic analogs of anoplin show improved antimicrobial activities.  

PubMed

We present the antimicrobial and hemolytic activities of the decapeptide anoplin and 19 analogs thereof tested against methicillin-resistant Staphylococcus aureus ATCC 33591 (MRSA), Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), vancomycin-resistant Enterococcus faecium (ATCC 700221) (VRE), and Candida albicans (ATCC 200955). The anoplin analogs contain substitutions in amino acid positions 2, 3, 5, 6, 8, 9, and 10. We use these peptides to study the effect of altering the charge and hydrophobicity of anoplin on activity against red blood cells and microorganisms. We find that increasing the charge and/or hydrophobicity improves antimicrobial activity and increases hemolytic activity. For each strain tested, we identify at least six anoplin analogs with an improved therapeutic index compared with anoplin, the only exception being Enterococcus faecium, against which only few compounds are more specific than anoplin. Both 2Nal(6) and Cha(6) show improved therapeutic index against all strains tested. PMID:24019229

Munk, Jens K; Uggerhøj, Lars Erik; Poulsen, Tanja J; Frimodt-Møller, Niels; Wimmer, Reinhard; Nyberg, Nils T; Hansen, Paul R

2013-11-01

20

Chitinase A from Stenotrophomonas maltophilia shows transglycosylation and antifungal activities.  

PubMed

Stenotrophomonas maltophilia chitinase (StmChiA and StmChiB) genes were cloned and expressed as soluble proteins of 70.5 and 41.6 kDa in Escherichia coli. Ni-NTA affinity purified StmChiA and StmChiB were optimally active at pH 5.0 and 7.0, respectively and exhibited broad range pH activity. StmChiA and StmChiB had an optimum temperature of 40°C and are stable up to 50 and 40°C, respectively. Hydrolytic activity on chitooligosaccharides indicated that StmChiA was an endo-acting enzyme releasing chitobiose and StmChiB was both exo/endo-acting enzyme with the release of GlcNAc as the final product. StmChiA showed higher preference to ?-chitin and exhibited transglycosylation on even chain length tetra- and hexameric substrates. StmChiA, and not StmChiB, was active on chitinous polymers and showed antifungal activity against Fusarium oxysporum. PMID:23428818

Suma, Katta; Podile, Appa Rao

2013-04-01

21

Human resistin, a proinflammatory cytokine, shows chaperone-like activity  

PubMed Central

Resistin, a cysteine-rich adipocytokine, proposed as a link between obesity and diabetes in mice, was shown as a proinflammatory molecule in humans. We earlier reported that human resistin (hRes), a trimer, was resistant to heat and urea denaturation, existed in an oligomeric polydispersed state, and showed a concentration-dependent conformational change. These properties and an intimate correlation of hRes expression with cellular stress prompted us to investigate hRes as a possible chaperone. Here, we show that recombinant human resistin was able to protect the heat-labile enzymes citrate synthase and Nde1 from thermal aggregation and inactivation and was able to refold and restore their enzymatic activities after heat/guanidinium chloride denaturation. Furthermore, recombinant human resistin could bind misfolded proteins only. Molecular dynamics-based association–dissociation kinetics of hRes subunits pointed to resistin being a molecular chaperone. Bis-ANS, which blocks surface hydrophobicity, abrogated the chaperone activity of hRes, establishing the importance of surface hydrophobicity for chaperone activity. Replacement of Phe49 with Tyr (F49YhRes), a critical residue within the hydrophobic patch of hRes, although it could prevent thermal aggregation of citrate synthase and Nde1, was unable to refold and restore their activities. Treatment of U937 cells with tunicamycin/thapsigargin resulted in reduced hRes secretion and concomitant localization in the endoplasmic reticulum. Escherichia coli transformants expressing hRes could be rescued from thermal stress, pointing to hRes’s chaperone-like function in vivo. HeLa cells transfected with hRes showed protection from thapsigargin-induced apoptosis. In conclusion, hRes, an inflammatory protein, additionally exhibited chaperone-like properties, suggesting a possible link between inflammation and cellular stress.

Suragani, Madhuri; Aadinarayana, Varma D.; Pinjari, Aleem Basha; Tanneeru, Karunakar; Guruprasad, Lalitha; Banerjee, Sharmistha; Pandey, Saurabh; Chaudhuri, Tapan K.; Ehtesham, Nasreen Zafar

2013-01-01

22

Sirt5 Deacylation Activities Show Differential Sensitivities to Nicotinamide Inhibition  

PubMed Central

Sirtuins are protein deacylases regulating metabolism and aging processes, and the seven human isoforms are considered attractive therapeutic targets. Sirtuins transfer acyl groups from lysine sidechains to ADP-ribose, formed from the cosubstrate NAD+ by release of nicotinamide, which in turn is assumed to be a general Sirtuin inhibitor. Studies on Sirtuin regulation have been hampered, however, by shortcomings of available assays. Here, we describe a mass spectrometry–based, quantitative deacylation assay not requiring any substrate labeling. Using this assay, we show that the deacetylation activity of human Sirt5 features an unusual insensitivity to nicotinamide inhibition. In contrast, we find similar values for Sirt5 and Sirt3 for the intrinsic NAD+ affinity as well as the apparent NAD+ affinity in presence of peptide. Structure comparison and mutagenesis identify an Arg neighboring to the Sirt5 nicotinamide binding pocket as a mediator of nicotinamide resistance, and statistical sequence analyses along with testing further Sirtuins reveal a network of coevolved residues likely defining a nicotinamide-insensitive Sirtuin deacetylase family. The same Arg was recently reported to render Sirt5 a preferential desuccinylase, and we find that this Sirt5 activity is highly sensitive to nicotinamide inhibition. Analysis of Sirt5 structures and activity data suggest that an Arg/succinate interaction is the molecular basis of the differential nicotinamide sensitivities of the two Sirt5 activities. Our results thus indicate a Sirtuin subfamily with nicotinamide-insensitive deacetylase activity and suggest that the molecular features determining nicotinamide sensitivity overlap with those dominating deacylation specificity, possibly suggesting that other subfamily members might also prefer other acylations than acetylations.

Suenkel, Benjamin; Lakshminarasimhan, Mahadevan; Schutkowski, Mike; Steegborn, Clemens

2012-01-01

23

Commercial production of ?-glucuronidase (GUS): a model system for the production of proteins in plants  

Microsoft Academic Search

We have generated transgenic maize seed containing ß-glucuronidase(GUS) for commercial production. While many other investigators have demonstrated the expression of GUS as a scoreable marker, this is one of the first cases where a detailed characterization of the transgenic plants and the protein were performed which are necessary to use this as a commercial source of GUS. The recombinant ß-glucuronidase

Derrick R. Witcher; Elizabeth E. Hood; Dave Peterson; Michele Bailey; Diane Bond; Ann Kusnadi; Roque Evangelista; Zivko Nikolov; Cynthia Wooge; Richard Mehigh; William Kappel; James Register; John A. Howard

1998-01-01

24

High-Resolution Observations of a Filament showing Activated Barb  

NASA Astrophysics Data System (ADS)

Analysis of a filament showing an activated barb using observations from the Dutch Open Telescope (DOT) on 2010 August 20 are presented. The DOT takes Doppler images in H?, among other wavelengths, in a region about 110 × 110 arcsec^{2} in area, at a cadence of 30~seconds. The offline image restoration technique of speckle reconstruction is applied to obtain diffraction limited images. The filament developed a new barb in 10~minutes, which disappeared within the next 35~minutes. Such a rapid formation and disappearance of a filament barb is unusual, and has not been reported earlier. Line-of-sight velocity maps were constructed from the Doppler images of the target filament. We observe flows in the filament spine towards the barb location prior to its formation, and flows in the barb towards the spine during its disappearance. Photospheric magnetograms from Heliospheric Magnetic Imager on board the Solar Dynamics Observatory, at a cadence of 45~seconds, were used to determine the changes in magnetic flux in the region surrounding the barb location. The variation of magnetic flux in this duration supports the view that barbs are rooted in minor magnetic polarity. Our analysis shows that barbs can be short-lived and formation and disappearance of the barb was associated with cancellation of magnetic flux.

Joshi, Anand; Martin, Sara F.; Mathew, Shibu; Srivastava, Nandita

2012-07-01

25

?-Glucuronidase activity during development of the male gametophyte from transgenic and non-transgenic plants  

Microsoft Academic Search

Beta-glucuronidase (GUS) was histochemically analyzed in anthers and pollen of potato, tobacco and tomato. GUS activity was determined in transgenic plants containing a chimaeric GUS gene and in untransformed plants. In anthers of transgenic plants at premeiotic and meiotic stages of sporogenous development, indigogenic precipitation indicative of GUS activity was consistently manifest in cells of the vascular cylinder, the connectivum

Leon Plegt; Raoul J. Bino

1989-01-01

26

?-Glucuronidase activity in seedlings of the parasitic angiosperm Cusctua pentagona : developmental impact of the ?-glucuronidase inhibitor saccharic acid 1,4-lactone  

Microsoft Academic Search

Endogenous plant ?-glucuronidase (?-GUS) activity was detected in germinating seeds, seedlings, stems, flowers and haustoria of the parasitic angiosperm Cuscuta pentagona Engelm. In vitro characterisation of this activity showed it to have an acidic pH optimum, similar to previously characterised plant activities, and a sensitivity to the ?-GUS inhibitor saccharic acid 1,4-lactone (SAL). Application of SAL to seeds immediately after

Mark A. SchoenbeckA; Gabriel A. SwansonA

2007-01-01

27

GH3::GUS reflects cell-specific developmental patterns and stress-induced changes in wood anatomy in the poplar stem.  

PubMed

GH3 genes related to the auxin-inducible Glycine max (L.) Merr. GmGH3 gene encode enzymes that conjugate amino acids to auxin. To investigate the role of GH3 enzymes in stress responses and normal wood development, Populus x canescens (Ait.) was transformed with the promoter-reporter construct GH3::GUS containing a GH3 promoter and the 5' UTR from soybean. beta-Glucuronidase (GUS) activity was present in the vascular tissues of leaves and in developing lateral roots and was inducible in silent tissues by external auxin application. A decrease in GUS activity from the stem apex to the bottom corresponded to decreases in auxin concentrations in these tissues. High auxin concentration and high GH3::GUS activity were present in the pith tissue, which may provide storage for auxin compounds. GH3 reporter was active in ray cells, paratracheal parenchyma cells, maturing vessels and in cells surrounding maturing phloem fibers but not in the cambium and immature phloem, despite high auxin concentrations in the latter tissues. However, the GH3 promoter in these tissues became active when the plants were exposed to abiotic stresses, like bending or salinity, causing changes in wood anatomy. We suggest that adjustment of the internal auxin balance in wood in response to environmental cues involves GH3 auxin conjugate synthases. PMID:18595842

Teichmann, Thomas; Bolu-Arianto, Waode Hamsinah; Olbrich, Andrea; Langenfeld-Heyser, Rosemarie; Göbel, Cornelia; Grzeganek, Peter; Feussner, Ivo; Hänsch, Robert; Polle, Andrea

2008-09-01

28

Thalidomide Shows Activity Against AIDS-Related Kaposi's Sarcoma  

Cancer.gov

The drug thalidomide, used in the 1950s and 1960s as a sedative which led to birth defects in newborns of mothers who took the drug, has now been shown to have clinical activity against Kaposi's sarcoma (KS).

29

Ultraviolet imaging spectroscopy shows an active saturnian system.  

PubMed

Neutral oxygen in the saturnian system shows variability, and the total number of oxygen atoms peaks at 4 x 10(34). Saturn's aurora brightens in response to solar-wind forcing, and the auroral spectrum resembles Jupiter's. Phoebe's surface shows variable water-ice content, and the data indicate it originated in the outer solar system. Saturn's rings also show variable water abundance, with the purest ice in the outermost A ring. This radial variation is consistent with initially pure water ice bombarded by meteors, but smaller radial structures may indicate collisional transport and recent renewal events in the past 10(7) to 10(8) years. PMID:15604361

Esposito, Larry W; Colwell, Joshua E; Larsen, Kristopher; McClintock, William E; Stewart, A Ian F; Hallett, Janet Tew; Shemansky, Donald E; Ajello, Joseph M; Hansen, Candice J; Hendrix, Amanda R; West, Robert A; Keller, H Uwe; Korth, Axel; Pryor, Wayne R; Reulke, Ralf; Yung, Yuk L

2005-02-25

30

Conjugation of ovotransferrin with catechin shows improved antioxidant activity.  

PubMed

Ovotransferrin (OTF), representing 12-13% of the total egg white, is a member of transferrin family with antimicrobial and antioxidant activities. Catechin is a polyphenolic antioxidant found in green tea. The objective of the study was to conjugate ovotransferrin with catechin to improve the antioxidant activity of OTF. Conjugates were prepared either by the free radical method using hydrogen peroxide-ascorbic acid as the initiator or by the alkaline method at pH of 9.0. The oxygen-radical-scavenging effect was increased from 3.95 mol trolox equivalent (TE)/mol of ovotransferrin to 22.80 and 17.14 mol TE/mol sample, respectively, in radical and alkaline prepared conjugates, which indicated that conjugation with catechin is an effective way to improve antioxidant activity of the protein. Conjugation between ovotransferrin and catechin was analyzed by fluorescence analyses, ultra performance liquid chromatography, matrix-assisted laser desorption ionization time-of-flight mass spectrometry and liquid chromatography coupled online to a tandem mass spectrometer. Catechin was covalently bound to lysine (residues 327) and glutamic acid (residues 186) in ovotransferrin. The ovotransferrin-catechin conjugate may have a potential application as a functional food and nutraceutical ingredient. PMID:24606536

You, Juan; Luo, Yongkang; Wu, Jianping

2014-03-26

31

Functional Analysis of Plant Promoter rpL34 Using the GUS Marker Gene in New Tr,tnsgene Expression Vector pZD428  

SciTech Connect

Optimization of the transgene expression system is one of the critical steps for the high level production of heterologous proteins in plants, where the promoter is a key component regulating transgene expression. In this study, the activity of the rpL34 promoter was analyzed in transgenic tobacco (Nicotiana tabacum) NTI calli. A DNA fragment containing the rpL34 promoter and the reporter gene B-D-glucuronidase (GUS) were cloned into binary vector pZD427 to generate the transgene expression vector pZD428. The insertion was verified by enzyme restriction digestion and agarose gel electrophoresis analyses. The DNA fragment containing the rpL34 promoter and GUS reporter gene was then integrated into the tobacco genomes via Agrobacterium funiefaciens-mediated NT suspension cell transformation. The transformed CaNi were induced on Murashige and Skoog (MS) plates containing proper amounts of 2,4-D, cefotoxime, and kanamycin. Two hundred and sixty transformed calli were harvested for GUS activity and protein concentration measurements. GUS activity analyses revealed the specific activity up to 278,358 units per milligram total soluble protein. The GUS activity under the control of the rpL34 promoter is much higher than that under the control of the cauliflower mosaic virus 35S promoter, a commonly used promoter in plant biology. These results suggest that the rpL34 promoter is one of the most active promoters that can be used for heterologous protein production in calli and suspension cells.

Mauzey-Amato, Jacqueline M. (ASSOC WESTERN UNIVERSITY); Dai, Ziyu (BATTELLE (PACIFIC NW LAB))

2000-11-01

32

Insurance Applications of Active Fault Maps Showing Epistemic Uncertainty  

NASA Astrophysics Data System (ADS)

Insurance loss modeling for earthquakes utilizes available maps of active faulting produced by geoscientists. All such maps are subject to uncertainty, arising from lack of knowledge of fault geometry and rupture history. Field work to undertake geological fault investigations drains human and monetary resources, and this inevitably limits the resolution of fault parameters. Some areas are more accessible than others; some may be of greater social or economic importance than others; some areas may be investigated more rapidly or diligently than others; or funding restrictions may have curtailed the extent of the fault mapping program. In contrast with the aleatory uncertainty associated with the inherent variability in the dynamics of earthquake fault rupture, uncertainty associated with lack of knowledge of fault geometry and rupture history is epistemic. The extent of this epistemic uncertainty may vary substantially from one regional or national fault map to another. However aware the local cartographer may be, this uncertainty is generally not conveyed in detail to the international map user. For example, an area may be left blank for a variety of reasons, ranging from lack of sufficient investigation of a fault to lack of convincing evidence of activity. Epistemic uncertainty in fault parameters is of concern in any probabilistic assessment of seismic hazard, not least in insurance earthquake risk applications. A logic-tree framework is appropriate for incorporating epistemic uncertainty. Some insurance contracts cover specific high-value properties or transport infrastructure, and therefore are extremely sensitive to the geometry of active faulting. Alternative Risk Transfer (ART) to the capital markets may also be considered. In order for such insurance or ART contracts to be properly priced, uncertainty should be taken into account. Accordingly, an estimate is needed for the likelihood of surface rupture capable of causing severe damage. Especially where a high deductible is in force, this requires estimation of the epistemic uncertainty on fault geometry and activity. Transport infrastructure insurance is of practical interest in seismic countries. On the North Anatolian Fault in Turkey, there is uncertainty over an unbroken segment between the eastern end of the Dazce Fault and Bolu. This may have ruptured during the 1944 earthquake. Existing hazard maps may simply use a question mark to flag uncertainty. However, a far more informative type of hazard map might express spatial variations in the confidence level associated with a fault map. Through such visual guidance, an insurance risk analyst would be better placed to price earthquake cover, allowing for epistemic uncertainty.

Woo, G.

2005-12-01

33

Construction of gusA transcriptional fusion vectors for Bacillus subtilis and their utilization for studies of spore formation  

Microsoft Academic Search

A series of gusA transcriptional fusion vectors is described for Bacillus subtilis (Bs). The series includes a vector for use with the amyE system of Shimotsu and Henner [Gene 43 (1986) 85-94-1, an integrative vector and vectors that provide gusA or gusA neo cassettes. The gusA fusions are compatible with lacZ fusion vectors that are widely used with Bs, and

Margaret L. Karow; Patrick J. Piggot

1995-01-01

34

A Rhizobium selenitireducens protein showing selenite reductase activity.  

PubMed

Biobarriers remove, via precipitation, the metalloid selenite (SeO??²) from groundwater; a process that involves the biological reduction of soluble SeO??² to insoluble elemental red selenium (Se?). The enzymes associated with this reduction process are poorly understood. In Rhizobium selenitireducens at least two enzymes are potentially involved; one, a nitrite reductase reduces SeO??² to Se? but another protein may also be involved which is investigated in this study. Proteins from R. selenitireducens cells were precipitated with ammonium sulfate and run on native electrophoresis gels. When these gels were incubated with NADH and SeO??² a band of precipitated Se? developed signifying the presence of a SeO??² reducing protein. Bands were cut from the gel and analyzed for peptides via LCMSMS. The amino acid sequences associated with the bands indicated the presence of an NADH:flavin oxidoreductase that resembles YP_001326930 from Sinorhizobium medicae. The protein is part of a protein family termed old-yellow-enzymes (OYE) that contain a flavin binding domain. OYE enzymes are often involved in protecting cells from oxidative stress and, due in part to an active site that has a highly accessible binding pocket, are generally active on a wide range of substrates. This report is the first of an OYE enzyme being involved in SeO??² reduction. PMID:24474405

Hunter, W J

2014-03-01

35

What Fraction of Active Galaxies Actually Show Outflows?  

NASA Astrophysics Data System (ADS)

Outflows from active galactic nuclei (AGNs) seem to be common and are thought to be important from a variety of perspectives: as an agent of chemical enhancement of the interstellar and intergalactic media, as an agent of angular momentum removal from the accreting central engine, and as an agent limiting star formation in starbursting systems by blowing out gas and dust from the host galaxy. To understand these processes, we must determine what fraction of AGNs feature outflows and understand what forms they take. We examine recent surveys of outflows detected in ultraviolet absorption over the entire range of velocities and velocity widths (i.e., broad absorption lines, associated absorption lines, and high-velocity narrow absorption lines). While the fraction of specific forms of outflows depends on AGN properties, the overall fraction displaying outflows is fairly constant, approximately 60%, over many orders of magnitude in luminosity. We discuss implications of this result and ways to refine our understanding of outflows. We acknowledge support from the US National Science Foundation through grant AST 05-07781.

Ganguly, Rajib; Brotherton, M. S.

2007-12-01

36

Replicase-based plasmid DNA shows anti-tumor activity  

PubMed Central

Background Double stranded RNA (dsRNA) has multiple anti-tumor mechanisms. Over the past several decades, there have been numerous attempts to utilize synthetic dsRNA to control tumor growth in animal models and clinical trials. Recently, it became clear that intracellular dsRNA is more effective than extracellular dsRNA on promoting apoptosis and orchestrating adaptive immune responses. To overcome the difficulty in delivering a large dose of synthetic dsRNA into tumors, we propose to deliver a RNA replicase-based plasmid DNA, hypothesizing that the dsRNA generated by the replicase-based plasmid in tumor cells will inhibit tumor growth. Methods The anti-tumor activity of a plasmid (pSIN-?) that encodes the sindbis RNA replicase genes (nsp1-4) was evaluated in mice with model tumors (TC-1 lung cancer cells or B16 melanoma cells) and compared to a traditional pCMV-? plasmid. Results In cell culture, transfection of tumor cells with pSIN-? generated dsRNA. In mice with model tumors, pSIN-? more effectively delayed tumor growth than pCMV-?, and in some cases, eradicated the tumors. Conclusion RNA replicase-based plasmid may be exploited to generate intracellular dsRNA to control tumor growth.

2011-01-01

37

Sulfur dioxide: episodic injection shows evidence for active venus volcanism.  

PubMed

Pioneer Venus ultraviolet spectra from the first 5 years of operation show a decline (by more than a factor of 10) in sulfur dioxide abundance at the cloud tops and in the amount of submicron haze above the clouds. At the time of the Pioneer Venus encounter, the values for both parameters greatly exceeded earlier upper limits. However, Venus had a similar appearance in the late 1950's, implying the episodic injection of sulfur dioxide possibly caused by episodic volcanism. The amount of haze in the Venus middle atmosphere is about ten times that found in Earth's stratosphere after the most recent major volcanic eruptions, and the thermal energy required for this injection on Venus is greater by about an order of magnitude than the largest of these recent Earth eruptions and about as large as the Krakatoa eruption of 1883. The episodic behavior of sulfur dioxide implies that steady-state models of the chemistry and dynamics of cloud-top regions may be of limited use. PMID:17830154

Esposito, L W

1984-03-01

38

Sulfur dioxide - Episodic injection shows evidence for active Venus volcanism  

NASA Astrophysics Data System (ADS)

Pioneer Venus ultraviolet spectra from the first 5 years of operation show a decline (by more than a factor of 10) in sulfur dioxide abundance at the cloud tops and in the amount of submicron haze above the clouds. At the time of the Pioneer Venus encounter, the values for both parameters greatly exceeded earlier upper limits. However, Venus had a similar appearance in the late 1950's, implying the episodic injection of sulfur dioxide possibly caused by episodic volcanism. The amount of haze in the Venus middle atmosphere is about ten times that found in earth's stratosphere after the most recent major volcanic eruptions, and the thermal energy required for this injection on Venus is greater by about an order of magnitude than the largest of these recent earth eruptions and about as large as the Krakatoa eruption of 1883. The episodic behavior of sulfur dioxide implies that steady-state models of the chemistry and dynamics of cloud-top regions may be of limited use.

Esposito, L. W.

1984-03-01

39

Sulfur dioxide - Episodic injection shows evidence for active Venus volcanism  

NASA Technical Reports Server (NTRS)

Pioneer Venus ultraviolet spectra from the first 5 years of operation show a decline (by more than a factor of 10) in sulfur dioxide abundance at the cloud tops and in the amount of submicron haze above the clouds. At the time of the Pioneer Venus encounter, the values for both parameters greatly exceeded earlier upper limits. However, Venus had a similar appearance in the late 1950's, implying the episodic injection of sulfur dioxide possibly caused by episodic volcanism. The amount of haze in the Venus middle atmosphere is about ten times that found in earth's stratosphere after the most recent major volcanic eruptions, and the thermal energy required for this injection on Venus is greater by about an order of magnitude than the largest of these recent earth eruptions and about as large as the Krakatoa eruption of 1883. The episodic behavior of sulfur dioxide implies that steady-state models of the chemistry and dynamics of cloud-top regions may be of limited use.

Esposito, L. W.

1984-01-01

40

Paresev on lakebed with Mercury astronaut Gus Grissom and Dryden test pilot Milt Thompson  

NASA Technical Reports Server (NTRS)

NASA Flight Research Center Paresev 1-A with Mercury Astronaut Gus Grissom (left) and NASA test pilot Milton Thompson. Do you suppose they are wondering if all those clouds will mean a canceled flight?

1962-01-01

41

Localized transient expression of GUS in leaf discs following cocultivation with Agrobacterium  

Microsoft Academic Search

A chimaeric gene has been constructed that expresses ß-D-glucuronidase (GUS) in transformed plant tissues, but not in bacterial cells. This gene has proved extremely useful for monitoring transformation during the period immediately following gene transfer from Agrobacterium tumefaciens. GUS expression was detectable 2 days after inoculation, peaked at 3–4 days and then declined; if selection was imposed expression increased again

Bart-Jan Janssen; Richard C. Gardner

1990-01-01

42

Quantitative Analysis of Bacterial Gene Expression by Using the gusA Reporter Gene System  

PubMed Central

An Azospirillum brasilense Sp7 strain containing a plasmid-borne translational cytN-gusA fusion was grown in a continuous culture to quantitatively evaluate the influence of extracellular signals (such as O2) on expression of the cytNOQP operon. The dissolved oxygen concentration was shifted at regular time intervals before the steady state was reached. The measured ?-glucuronidase activity was used to monitor cytN gene expression. However, as the ?-glucuronidase activity in the experimental setup not only depended on altered transcription of the hybrid gene when the signal was varied but was also influenced by cellular accumulation, degradation, and dilution of the hybrid fusion protein, a mathematical method was developed to describe the intrinsic properties of the dynamic bioprocess. After identification and validation of the mathematical model, the apparent specific rate of expression of the fusion, which was independent of the experimental setup, could be deduced from the model and used to quantify gene expression regulated by extracellular environmental signals. In principle, this approach can be generalized to assess the effects of external signals on bacterial gene expression.

Sun, Jun; Smets, Ilse; Bernaerts, Kristel; Van Impe, Jan; Vanderleyden, Jos; Marchal, Kathleen

2001-01-01

43

Transient and stable gene expression in the fungal maize pathogen Cochliobolus heterostrophus after transformation with the ?-glucuronidase (GUS) gene  

Microsoft Academic Search

The bacterial GUS (ß-glucuronidase) gene has been used as a reporter gene in plants and bacteria and was recently expressed in filamentous fungi. Here, we report the application of GUS for the establishment of transient and stable gene expression systems in the phytopathogenic fungus Cochliobolus heterostrophus. The utility of the transient expression system is demonstrated in applications involving promoter analysis

Enrico Miinke; Wilhelm Schäfer

1993-01-01

44

T-DNA insertional mutagenesis for activation tagging in rice.  

PubMed

We have developed a new T-DNA vector, pGA2715, which can be used for promoter trapping and activation tagging of rice (Oryza sativa) genes. The binary vector contains the promoterless beta-glucuronidase (GUS) reporter gene next to the right border. In addition, the multimerized transcriptional enhancers from the cauliflower mosaic virus 35S promoter are located next to the left border. A total of 13,450 T-DNA insertional lines have been generated using pGA2715. Histochemical GUS assays have revealed that the GUS-staining frequency from those lines is about twice as high as that from lines transformed with the binary vector pGA2707, which lacks the enhancer element. This result suggests that the enhancer sequence present in the T-DNA improves the GUS-tagging efficiency. Reverse transcriptase-PCR analysis of a subset of randomly selected pGA2715 lines shows that expression of the genes immediately adjacent to the inserted enhancer is increased significantly. Therefore, the large population of T-DNA-tagged lines transformed with pGA2715 could be used to screen for promoter activity using the gus reporter, as well as for creating gain-of-function mutants. PMID:12481047

Jeong, Dong-Hoon; An, Suyoung; Kang, Hong-Gyu; Moon, Sunok; Han, Jong-Jin; Park, Sunhee; Lee, Hyun Sook; An, Kyungsook; An, Gynheung

2002-12-01

45

26 CFR 1.513-3 - Qualified convention and trade show activity.  

Code of Federal Regulations, 2010 CFR

...including rental of exhibition space at a qualified convention or trade show where...organization's provision of exhibition space is not a qualified convention or trade show activity...derived from rentals of exhibition space to...

2010-04-01

46

26 CFR 1.513-3 - Qualified convention and trade show activity.  

Code of Federal Regulations, 2010 CFR

...including rental of exhibition space at a qualified convention or trade show where...organization's provision of exhibition space is not a qualified convention or trade show activity...derived from rentals of exhibition space to...

2009-04-01

47

T-DNA Insertional Mutagenesis for Activation Tagging in Rice1  

PubMed Central

We have developed a new T-DNA vector, pGA2715, which can be used for promoter trapping and activation tagging of rice (Oryza sativa) genes. The binary vector contains the promoterless ?-glucuronidase (GUS) reporter gene next to the right border. In addition, the multimerized transcriptional enhancers from the cauliflower mosaic virus 35S promoter are located next to the left border. A total of 13,450 T-DNA insertional lines have been generated using pGA2715. Histochemical GUS assays have revealed that the GUS-staining frequency from those lines is about twice as high as that from lines transformed with the binary vector pGA2707, which lacks the enhancer element. This result suggests that the enhancer sequence present in the T-DNA improves the GUS-tagging efficiency. Reverse transcriptase-PCR analysis of a subset of randomly selected pGA2715 lines shows that expression of the genes immediately adjacent to the inserted enhancer is increased significantly. Therefore, the large population of T-DNA-tagged lines transformed with pGA2715 could be used to screen for promoter activity using the gus reporter, as well as for creating gain-of-function mutants.

Jeong, Dong-Hoon; An, Suyoung; Kang, Hong-Gyu; Moon, Sunok; Han, Jong-Jin; Park, Sunhee; Lee, Hyun Sook; An, Kyungsook; An, Gynheung

2002-01-01

48

Specimen block counter-staining for localization of GUS expression in transgenic arabidopsis and tobacco  

NASA Technical Reports Server (NTRS)

A simple counter-staining procedure has been developed for comparative beta-glucuronidase (GUS) expression and anatomical localization in transgenic herbaceous arabidopsis and tobacco. This protocol provides good anatomical visualization for monitoring chimeric gene expression at both the organ and tissue levels. It can be used with different histochemical stains and can be extended to the study of woody species. The specimens are paraffin-embedded, the block is trimmed to reveal internal structure, safranin-O staining solution is briefly applied to the surface of the block, then washed off and, after drying, a drop of immersion oil is placed on the stained surface for subsequent photographic work. This gives tissue counter-staining with good structural preservation without loss of GUS staining product; moreover, sample observation is rapid and efficient compared to existing procedures.

Kim, M. K.; Choi, J-W; Jeon, J-H; Franceschi, V. R.; Davin, L. B.; Lewis, N. G.

2002-01-01

49

Elucidation of Septoria tritici ? Wheat Interactions Using GUS-Expressing Isolates  

Microsoft Academic Search

Pnini-Cohen, S., Zilberstein, A., Schuster, S., Sharon, A., and Eyal, Z. 2000. Elucidation of Septoria tritici ? wheat interactions using GUS-ex- pressing isolates. Phytopathology 90:297-304. Isolate ISR398 of Septoria tritici (which produces none to few pycnidia on the wheat cv. Seri 82 and high coverage on cv. Shafir) and isolate ISR8036 (which is virulent on both cultivars) were genetically cotrans-

S. Pnini-Cohen; A. Zilberstein; S. Schuster; A. Sharon; Z. Eyal

2000-01-01

50

A maize spermine synthase 1 PEST sequence fused to the GUS reporter protein facilitates proteolytic degradation.  

PubMed

Polyamines are low molecular weight aliphatic compounds involved in various biochemical, cellular and physiological processes in all organisms. In plants, genes involved in polyamine biosynthesis and catabolism are regulated at transcriptional, translational, and posttranslational level. In this research, we focused on the characterization of a PEST sequence (rich in proline, glutamic acid, serine, and threonine) of the maize spermine synthase 1 (ZmSPMS1). To this aim, 123 bp encoding 40 amino acids of the C-terminal region of the ZmSPMS1 enzyme containing the PEST sequence were fused to the GUS reporter gene. This fusion was evaluated in Arabidopsis thaliana transgenic lines and onion monolayers transient expression system. The ZmSPMS1 PEST sequence leads to specific degradation of the GUS reporter protein. It is suggested that the 26S proteasome may be involved in GUS::PEST fusion degradation in both onion and Arabidopsis. The PEST sequences appear to be present in plant spermine synthases, mainly in monocots. PMID:24642522

Maruri-López, Israel; Rodríguez-Kessler, Margarita; Rodríguez-Hernández, Aída Araceli; Becerra-Flora, Alicia; Olivares-Grajales, Juan Elías; Jiménez-Bremont, Juan Francisco

2014-05-01

51

Nodulation competitiveness of Rhizobium leguminosarum bv. phaseoli and Rhizobium tropici strains measured by glucuronidase ( gus ) gene fusion  

Microsoft Academic Search

The nodulation competitiveness of 17 Rhizobium leguminosarum bv. phaseoli and 3 R. tropici strains was analysed in growth pouches, at pH 5.2 and 6.4. All 20 strains were coinoculated with a gus+ strain of R. leguminosarum bv. phaseoli strain KIM5s. The gus+ phenotype, carrying the glucuronidase gene, was used to type nodules directly in the growth pouches. Nodule occupancy ranged

W. Streit; K. Kosch; D. Werner

1992-01-01

52

Cloning and characterization of Photobacterium damselae ssp. piscicida phospholipase: an enzyme that shows haemolytic activity.  

PubMed

A phospholipase gene of Photobacterium damselae ssp. piscicida (ppp) was cloned from a genomic library and its nucleotide sequence was determined. The open reading frame consisted of 1218 bp encoding a protein of 405 amino acids with a predicted molecular mass of 46 kDa. The PPP had identities (53-55%) with phospholipase and haemolysin of Vibrio spp., while it showed low identities (23-26%) with glycerophospholipid cholesterol acyltransferase of Aeromonas spp. A recombinant PPP (rPPP) with a His tag at the C-terminus expressed in Escherichia coli and purified showed phospholipase activity. The rPPP also showed lecithin-dependent haemolytic activity against mammalian erythrocytes and direct haemolytic activity against fish erythrocytes. The culture supernatant of wild-type P. damselae ssp. piscicida showed phospholipase activity, while that of a PPP gene knockout mutant did not. PMID:17958612

Naka, H; Hirono, I; Aoki, T

2007-11-01

53

A Lipase Isolated from the Silkworm Bombyx mori Shows Antiviral Activity against Nucleopolyhedrovirus  

Microsoft Academic Search

A protein showing strong antiviral activity against Bombyx mori nucleopolyhedrovirus (BmNPV) was purified from the digestive juice of B. mori larvae. A homology search of the deduced amino acid sequence of the protein cDNA revealed 56% homology with Drosophila melanogaster lipase and 21% homology with human lipase. As lipase activity of the protein was confirmed in vitro, this protein was

Kangayam M. Ponnuvel; Hiroshi Nakazawa; Seiichi Furukawa; Ai Asaoka; Jun Ishibashi; Hiromitsu Tanaka; Minoru Yamakawa

2003-01-01

54

The colchicine derivative CT20126 shows a novel microtubule-modulating activity with apoptosis  

PubMed Central

New colchicine analogs have been synthesized with the aim of developing stronger potential anticancer activities. Among the analogs, CT20126 has been previously reported to show immunosuppressive activities. Here, we report that CT20126 also shows potential anticancer effects via an unusual mechanism: the modulation of microtubule integrity and cell cycle arrest at the G2/M phase before apoptosis. When we treated COS-7 cells with CT20126 (5???), the normal thread-like microtubules were disrupted into tubulin dimers within 10?min and thereafter repolymerized into short, thick filaments. In contrast, cells treated with the same concentration of colchicine exhibited microtubule depolymerization after 20?min and never underwent repolymerization. Furthermore, optical density (OD) analysis (350?nm) with purified tubulin showed that CT20126 had a higher repolymerizing activity than that of Taxol, a potent microtubule-polymerizing agent. These results suggest that the effects of CT20126 on microtubule integrity differ from those of colchicine: the analog first destabilizes microtubules and then stabilizes the disrupted tubulins into short, thick polymers. Furthermore, CT20126 induced a greater level of apoptotic activity in Jurkat T cells than colchicine (assessed by G2/M arrest, caspase-3 activation and cell sorting). At 20?n?, CT20126 induced 47% apoptosis among Jurkat T cells, whereas colchicine induced only 33% apoptosis. Our results suggest that the colchicine analog CT20126 can potently induce apoptosis by disrupting microtubule integrity in a manner that differs from that of colchicine or Taxol.

Kim, Sung-Kuk; Cho, Sang-Min; Kim, Ho; Seok, Heon; Kim, Soon-Ok; Kyu Kwon, Taeg; Chang, Jong-Soo

2013-01-01

55

Vv-AMP1, a ripening induced peptide from Vitis vinifera shows strong antifungal activity  

Microsoft Academic Search

BACKGROUND: Latest research shows that small antimicrobial peptides play a role in the innate defense system of plants. These peptides typically contribute to preformed defense by developing protective barriers around germinating seeds or between different tissue layers within plant organs. The encoding genes could also be upregulated by abiotic and biotic stimuli during active defense processes. The peptides display a

Abré de Beer; Melané A Vivier

2008-01-01

56

Temporal and spatial regulation of the symbiotic genes of Rhizobium meliloti in planta revealed by transposon Tn5-gusA.  

PubMed

Tn5-gusA promoter/probe transposons have been constructed that fuse the Escherichia coli gusA reporter gene transcriptionally or translationally with a target promoter. These have been used to monitor expression of Rhizobium meliloti symbiotic genes within alfalfa nodules. Fusions in all 11 nod genes studied show the same pattern of expression: first on the root surface, then throughout the developing nodule, then mainly in the nodule meristem, falling off progressively through the central region, and then disappearing. In contrast, fusions in all five nif genes studied, all four fix genes, and syrM show a second, different pattern: expression beginning later, first throughout the nodule except for the meristem, strongest just behind the meristem, and falling off progressively through the central region. Novel features revealed by these studies include nod expression in the meristem, regulated in planta expression of control genes nodD1 and nodD3, disappearance of nod expression late in organogenesis, and properties of syrM. PMID:2159937

Sharma, S B; Signer, E R

1990-03-01

57

Genomic organization and sequence of the Gus-s/sup a/ allele of the murine. beta. -glucuronidase gene  

SciTech Connect

The Gus-s/sup ..cap alpha../ allele of the mouse ..beta..-glucuronidase gene exhibits a high degree of inducibility by androgens due to its linkage with the Gus-r/sup ..cap alpha../ regulatory locus. The authors isolated Gus-s/sup ..cap alpha../ on a 28-kilobase pair fragment of mouse chromosome 5 and found that it contains 12 exons and 11 intervening sequences spanning 14 kilobase pairs of this genomic segment. The mRNA cap site was identified by ribonuclease protection and primer extension analyses which revealed an unusually short 5' noncoding sequence of 12 nucleotides. Proximal regulatory sequences in the 5'-flanking DNA and the complete sequence of the Gus-s/sup ..cap alpha../ mRNA transcript were also determined. Comparison of the amino acid sequence determined from the Gus-s/sup ..cap alpha../ nucleotide sequence with that of human ..beta..-glucuronidase indicated that the two human mRNA species differ due to alternate splicing of an exon homologous to exon 6 of the mouse gene.

Funkenstein, B.; Leary, S.L.; Stein, J.C.; Catterall, J.F.

1988-03-01

58

Agrobacterium-mediated transient GUS gene expression in buffel grass (Cenchrus ciliaris L.).  

PubMed

The study was conducted to standardize a protocol for Agrobacterium-mediated genetic transformation of buffel grass (Cenchrus ciliaris L.). Embryogenic calli, produced from one-year-old mature seeds of buffel grass, were used as target cells for Agrobacterium-mediated transformation. A. tumefaciens strain LBA4404, harbouring pCAMBIA-1301 or pCAMBIA-2301, was used for co-cultivation with embryogenic calli from three genotypes (IG-3108, IG-9757 and IG-97101). Co-culturing of calli with Agrobacterium for 30 minutes, followed by co-cultivation with 0.1 mM acetosyringone for 3 days was found to be optimum for maximum transformation efficiency. Presence of acetosyringone during co-cultivation was found to be necessary for transformation. Transient GUS (beta-glucuronidase) gene expression was used to monitor T-DNA delivery into the target cells. Significant genotypic variations in response to transformation were observed among the tested genotypes. A very high frequency (63.3%) of GUS gene expression was obtained following Agrobacterium-mediated gene transfer into embryogenic calli. The standardized protocol would be useful for Agrobacterium-mediated genetic transformation of buffel grass with genes of agronomic importance. PMID:14617824

Batra, Shweta; Kumar, Suresh

2003-01-01

59

Large-scale characterization of promoters from grapevine (Vitis spp.) using quantitative anthocyanin and GUS assay systems.  

PubMed

Successful implementation of cisgenic/intragenic/ingenic technology for crop improvement necessitates a better understanding of the function of native promoters for driving desired gene expression in host plant. Although the genome of grapevine (Vitis vinifera) has been determined, efforts to explore promoter resources for the development of cisgenics are still lacking. Particularly, there is a shortage of constitutive promoters for marker and/or target gene expression in this species. In this work, we utilized an anthocyanin-based color histogram analysis method to evaluate quantitatively a large number of promoters for their ability to activate transgene expression. Promoter fragments corresponding to known genes were amplified from various genotypes and used to drive the VvMybA1 gene of 'Merlot' for anthocyanin production in non-pigmented somatic embryo (SE) explants to infer transcriptional activity. Results revealed that among 15 tested promoters belonging to seven ubiquitin genes, at least three promoters generated constitutive activities reaching up to 100% value of the d35S promoter. In particular, the high activity levels of VvUb6-1 and VvUb7-2 promoters were verified by transient GUS quantitative assay as well as stable anthocyanin expression in sepal and corolla of transgenic tobacco. Variations in promoter activity of different ubiquitin genes in grapevine did not correlate with the presence and sizes of 5' UTR intron, but seemed to be related positively and negatively to the number of positive cis-acting elements and root-specific elements respectively. In addition, several of the 13 promoters derived from a PR1 gene and a PAL gene produced a higher basal activity as compared to previously reported inducible promoters and might be useful for further identification of strong inducible promoters. Our study contributed invaluable information on transcriptional activity of many previously uncharacterized native promoters that could be used for genetic engineering of grapevine. PMID:23017908

Li, Zhijian T; Kim, Kyung-Hee; Jasinski, Jonathon R; Creech, Matthew R; Gray, Dennis J

2012-11-01

60

Plant crude extracts could be the solution: extracts showing in vivo antitumorigenic activity.  

PubMed

Screening active compounds from plants lead to discover new medicinal drugs which have efficient protection and treatment roles against various diseases including cancer. In our study, extracts from different plants represent seeds of: Gossypium barbadense, Ricinus communis, Sesamum indicum, Nigella sativa, Vinca rosea and Melia azedarah; fruits of: Xanthium occidental; flowers of: Atriplex nummularia; barks of: Cinnamomum zeylanicum; latex of: Ficus carica and rhizomes of: Curcuma longa and Zingiber officinale were tested in vivo using three subsequent bioassays: the BST (Brine Shrimp Toxicity bioassay), AWD (Agar well diffusion antimicrobial bioassay) and AtPDT (Agrobacterium tumefaciens Potato Disc Tumor bioassay). AWD technique omitted any extracts have antimicrobial activities while BST omitted any extract did not has physiological activity and determined the various LC(50) of each plant extract. For the first time, using a range of concentrations in the AtPDT modified protocol allowed the detection of tumor promotion caused by extract represented by A. nummularia. Using cluster analysis leads to classifying the different plant extracts activities to six groups regarding to their toxicity, antitumor activities and both of them. The extracts from edible plants represent 50% of the first and the second group which have the highest antitumor activities represented in F. caraica (group 1) and C. longa (group 2) as well as the non-edible plant extracts of Gossypium barbadense and Ricinus communis. A comparison study between the edible and herbaceous plants different extracts for their antitumor activities was performed. We recommended using the modified protocols used in this study for investigating more plants and using crude plant extracts which have antitumor activities in cancer treatment. Edible plants, which show in vivo antitumor activities, are recommended as save sources for antitumor compounds. PMID:18390447

Amara, A A; El-Masry, M H; Bogdady, H H

2008-04-01

61

Bovine chromaffin cells in culture show carboxylesterase activities sensitive to organophosphorus compounds.  

PubMed

Carboxylesterase activities are widely distributed in a great variety of tissues; however, the biological function of these enzymes remains unclear. Some organophosphorus compounds induce a neurodegenarative syndrome related to the covalent modification of a carboxylesterase known as neuropathy target esterase. We investigated the expression of neuropathy target esterase and related carboxylesterase in bovine chromaffin cells with the aim of developing a potential in vitro model for studying the cellular function of carboxylesterase enzymes and toxic effects of organophosphorus compounds. Total phenyl valerate esterase exhibited an activity of 1.27 +/- 0.19 mU/10(5) cells (SD, n = 15). From the phenyl valerate esterase paraoxon and mipafox inhibition curves the following activities have been determined: B-activity (resistant to 40 microM paraoxon), 1.05 +/- 0.08 mU/10(5) cells (n = 8); C-activity (resistant to 40 microM paraoxon plus 250 microM mipafox), 0.12 +/- 0.05 mU/10(5) cells (n = 8); and neuropathy target esterase, calculated by the difference between B- and C-activities, 0.93 +/- 0.08 mU/10(5) cells (n = 8). All of these activities increased linearly with the number of cells and time of incubation with the substrate. Most of the phenol product of the reaction was released and detected in the extracellular medium. None of the components of the reaction were shown to affect cell viability when assessed by trypan blue exclusion. The study shows that bovine chromaffin cells possess carboxylesterase activities and respond to inhibition by paraoxon and mipafox, thus facilitating the discrimination of neuropathy target esterase. In conclusion, bovine chromaffin cells are appropriate as an in vitro cell model for studying toxic effects of organophosphorus compounds. PMID:8930121

Sogorb, M A; Vilanova, E; Quintanar, J L; Viniegra, S

1996-09-01

62

Antiparkinson drug--Mucuna pruriens shows antioxidant and metal chelating activity.  

PubMed

Parkinson's disease is a neurodegenerative disorder for which no neurorestorative therapeutic treatment is currently available. Oxidative stress plays an important role in the pathophysiology of Parkinson's disease. The ancient Indian medical system, Ayurveda, traditionally uses Mucuna pruriens to treat Parkinson's disease. In our earlier studies, Mucuna pruriens has been shown to possess antiparkinson and neuroprotective effects in animal models of Parkinson's disease. The antioxidant activity of Mucuna pruriens was demonstrated by its ability to scavenge DPPH radicals, ABTS radicals and reactive oxygen species. Mucuna pruriens significantly inhibited the oxidation of lipids and deoxyribose sugar. Mucuna pruriens exhibited divalent iron chelating activity and did not show any genotoxic/mutagenic effect on the plasmid DNA. These results suggest that the neuroprotective and neurorestorative effect of Mucuna pruriens may be related to its antioxidant activity independent of the symptomatic effect. In addition, the drug appears to be therapeutically safe in the treatment of patients with Parkinson's disease. PMID:18064727

Dhanasekaran, Muralikrishnan; Tharakan, Binu; Manyam, Bala V

2008-01-01

63

Redox-active cysteines of a membrane electron transporter DsbD show dual compartment accessibility  

PubMed Central

The membrane-embedded domain of the unusual electron transporter DsbD (DsbD?) uses two redox-active cysteines to catalyze electron transfer between thioredoxin-fold polypeptides on opposite sides of the bacterial cytoplasmic membrane. How the electrons are transferred across the membrane is unknown. Here, we show that DsbD? displays an inherent functional and structural symmetry: first, the two cysteines of DsbD? can be alkylated from both the cytoplasm and the periplasm. Second, when the two cysteines are disulfide-bonded, cysteine scanning shows that the C-terminal halves of the cysteine-containing transmembrane segments 1 and 4 are exposed to the aqueous environment while the N-terminal halves are not. Third, proline residues located pseudo-symmetrically around the two cysteines are required for redox activity and accessibility of the cysteines. Fourth, mixed disulfide complexes, apparent intermediates in the electron transfer process, are detected between DsbD? and thioredoxin molecules on each side of the membrane. We propose a model where the two redox-active cysteines are located at the center of the membrane, accessible on both sides of the membrane to the thioredoxin proteins.

Cho, Seung-Hyun; Porat, Amir; Ye, Jiqing; Beckwith, Jon

2007-01-01

64

Two chitinase-like proteins abundantly accumulated in latex of mulberry show insecticidal activity  

PubMed Central

Background Plant latex is the cytoplasm of highly specialized cells known as laticifers, and is thought to have a critical role in defense against herbivorous insects. Proteins abundantly accumulated in latex might therefore be involved in the defense system. Results We purified latex abundant protein a and b (LA-a and LA-b) from mulberry (Morus sp.) and analyzed their properties. LA-a and LA-b have molecular masses of approximately 50 and 46 kDa, respectively, and are abundant in the soluble fraction of latex. Western blotting analysis suggested that they share sequence similarity with each other. The sequences of LA-a and LA-b, as determined by Edman degradation, showed chitin-binding domains of plant chitinases at the N termini. These proteins showed small but significant chitinase and chitosanase activities. Lectin RCA120 indicated that, unlike common plant chitinases, LA-a and LA-b are glycosylated. LA-a and LA-b showed insecticidal activities when fed to larvae of the model insect Drosophila melanogaster. Conclusions Our results suggest that the two LA proteins have a crucial role in defense against herbivorous insects, possibly by hydrolyzing their chitin.

2010-01-01

65

Two analogues of fenarimol show curative activity in an experimental model of Chagas disease.  

PubMed

Chagas disease, caused by the protozoan parasite Trypanosoma cruzi (T. cruzi), is an increasing threat to global health. Available medicines were introduced over 40 years ago, have undesirable side effects, and give equivocal results of cure in the chronic stage of the disease. We report the development of two compounds, 6 and (S)-7, with PCR-confirmed curative activity in a mouse model of established T. cruzi infection after once daily oral dosing for 20 days at 20 mg/kg 6 and 10 mg/kg (S)-7. Compounds 6 and (S)-7 have potent in vitro activity, are noncytotoxic, show no adverse effects in vivo following repeat dosing, are prepared by a short synthetic route, and have druglike properties suitable for preclinical development. PMID:24304150

Keenan, Martine; Chaplin, Jason H; Alexander, Paul W; Abbott, Michael J; Best, Wayne M; Khong, Andrea; Botero, Adriana; Perez, Catherine; Cornwall, Scott; Thompson, R Andrew; White, Karen L; Shackleford, David M; Koltun, Maria; Chiu, Francis C K; Morizzi, Julia; Ryan, Eileen; Campbell, Michael; von Geldern, Thomas W; Scandale, Ivan; Chatelain, Eric; Charman, Susan A

2013-12-27

66

An NGR-integrated and enediyne-energized apoprotein shows CD13-targeting antitumor activity.  

PubMed

Targeting and inhibiting angiogenesis is a promising strategy for treatment of cancer. NGR peptide motif is a tumor-homing peptide, which could bind with CD13 expressed on tumor blood vessels. Lidamycin is a highly potent antitumor antibiotic, which is composed of an apoprotein (LDP) and an active enediyne chromophore (AE). Here, an NGR-integrated and enediyne-energized apoprotein composed of cyclic NGR peptide and lidamycin was developed by a two-step procedure. Firstly, we prepared the fusion protein composed of NGR peptide and LDP by recombinant DNA technology. Then, AE was reloaded to the fusion protein to get NGR-LDP-AE. Our experiments showed that NGR-LDP could bind to CD13-expressing HT-1080 cells, whereas the recombinant LDP (rLDP) showed weak binding. NGR-LDP-AE exerted highly potent cytotoxicity to cultured tumor cells in vitro. In vivo antitumor activity was evaluated in murine hepatoma 22 (H22) model and human fibrosarcoma HT-1080 model. At the tolerable dose, NGR-LDP-AE and lidamycin inhibited H22 tumor growth by 94.8 and 66.9%, and the median survival time of the mice was 62 and 37 days, respectively. In the HT-1080 model, NGR-LDP-AE inhibited tumor growth by 88.6%, which was statistically different from that of lidamycin (74.5%). Immunohistochemical study showed that NGR-LDP could bind to tumor blood vessels. Conclusively, these results demonstrate that fusion of LDP with CNGRC peptide delivers AE to tumor blood vessels and improves its antitumor activity. PMID:23206754

Zheng, Yan-Bo; Shang, Bo-Yang; Li, Yi; Zhen, Yong-Su

2013-03-01

67

The Reverse Transcription Inhibitor Abacavir Shows Anticancer Activity in Prostate Cancer Cell Lines  

PubMed Central

Background Transposable Elements (TEs) comprise nearly 45% of the entire genome and are part of sophisticated regulatory network systems that control developmental processes in normal and pathological conditions. The retroviral/retrotransposon gene machinery consists mainly of Long Interspersed Nuclear Elements (LINEs-1) and Human Endogenous Retroviruses (HERVs) that code for their own endogenous reverse transcriptase (RT). Interestingly, RT is typically expressed at high levels in cancer cells. Recent studies report that RT inhibition by non-nucleoside reverse transcriptase inhibitors (NNRTIs) induces growth arrest and cell differentiation in vitro and antagonizes growth of human tumors in animal model. In the present study we analyze the anticancer activity of Abacavir (ABC), a nucleoside reverse transcription inhibitor (NRTI), on PC3 and LNCaP prostate cancer cell lines. Principal Findings ABC significantly reduces cell growth, migration and invasion processes, considerably slows S phase progression, induces senescence and cell death in prostate cancer cells. Consistent with these observations, microarray analysis on PC3 cells shows that ABC induces specific and dose-dependent changes in gene expression, involving multiple cellular pathways. Notably, by quantitative Real-Time PCR we found that LINE-1 ORF1 and ORF2 mRNA levels were significantly up-regulated by ABC treatment. Conclusions Our results demonstrate the potential of ABC as anticancer agent able to induce antiproliferative activity and trigger senescence in prostate cancer cells. Noteworthy, we show that ABC elicits up-regulation of LINE-1 expression, suggesting the involvement of these elements in the observed cellular modifications.

Molinari, Agnese; Parisi, Chiara; Bozzuto, Giuseppina; Toccacieli, Laura; Formisano, Giuseppe; De Orsi, Daniela; Paradisi, Silvia; Grober, OlI Maria Victoria; Ravo, Maria; Weisz, Alessandro; Arcieri, Romano; Vella, Stefano; Gaudi, Simona

2010-01-01

68

Children with High Functioning Autism show increased prefrontal and temporal cortex activity during error monitoring  

PubMed Central

Evidence exists for deficits in error monitoring in autism. These deficits may be particularly important because they may contribute to excessive perseveration and repetitive behavior in autism. We examined the neural correlates of error monitoring using fMRI in 8–12-year-old children with high-functioning autism (HFA, n=11) and typically developing children (TD, n=15) during performance of a Go/No-Go task by comparing the neural correlates of commission errors versus correct response inhibition trials. Compared to TD children, children with HFA showed increased BOLD fMRI signal in the anterior medial prefrontal cortex (amPFC) and the left superior temporal gyrus (STempG) during commission error (versus correct inhibition) trials. A follow-up region-of-interest analysis also showed increased BOLD signal in the right insula in HFA compared to TD controls. Our findings of increased amPFC and STempG activity in HFA, together with the increased activity in the insula, suggest a greater attention towards the internally-driven emotional state associated with making an error in children with HFA. Since error monitoring occurs across different cognitive tasks throughout daily life, an increased emotional reaction to errors may have important consequences for early learning processes.

Goldberg, Melissa C.; Spinelli, Simona; Joel, Suresh; Pekar, James J.; Denckla, Martha B.; Mostofsky, Stewart H.

2010-01-01

69

High nuclearity nickel compounds with three, four or five metal atoms showing antibacterial activity.  

PubMed

The effect on DNA and the antibacterial activity of a series of high nuclearity nickel compounds with three, four and five metal atoms were examined. The compounds have a mixed ligand composition with salicylhydroxamic acid and di-2-pyridyl-ketonoxime as chelate agents. In the trinuclear compound Ni(3)(shi)(2)(Hpko)(2)(py)(2)(1), two metal ions show a square planar geometry while the third one is in an octahedral environment. The compounds with four and five nickel atoms construct metallacrown cores with two distinct connectivities. The tetranuclear vacant metallacrown [12-MC(Ni(II)N(Hshi)2(pko)2)-4](2+) shows the connectivity pattern [-O-Ni-O-N-Ni-N-](2), while the pentanuclear ([Ni(II)][12-MC(Ni(II)N(shi)2(pko)2)-4])(2+) follows the pattern [-Ni-O-N-](4). Two distinct arrangements of the chelates around the ring metal ions were observed; a 6-5-6-5-6-5-6-5 arrangement for the [12-MC(Ni(II)N(Hshi)2(pko)2)-4] core and a 6-6-5-5-6-6-5-5 arrangement for the [12-MC(Ni(II)N(shi)2(pko)2)-4] core. Magnetic variable temperature susceptibility study of the trinuclear compound revealed the presence of one paramagnetic nickel(II) ion with strong crystal field dependence, with D=5.0(4) cm(-1), g(xy)=2.7(3) and g(z)=2.3(3). The effect of the synthesized Ni(II) complexes on the integrity and electrophoretic mobility of nucleic acids was examined. Only compounds 2, 3 and 4 altered the mobility of pDNA, forming high molecular weight concatamers at low concentrations or precipitates at higher concentrations. Antibacterial activity screening of the above compounds suggests that nickel compounds 2, 3 and 4 were the most active and can act as potent antibacterial agents. PMID:12576289

Alexiou, Maria; Tsivikas, Ioannis; Dendrinou-Samara, Catherine; Pantazaki, Anastasia A; Trikalitis, Pantelis; Lalioti, Nikolia; Kyriakidis, Dimitris A; Kessissoglou, Dimitris P

2003-01-15

70

Functional analysis of a recently originating, atypical presequence: mitochondrial import and processing of GUS fusion proteins in transgenic tobacco and yeast  

Microsoft Academic Search

A gene family of at least five members encodes the tobacco mitochondrial Rieske Fe-S protein (RISP). To determine whether all five RISPs are translocated to mitochondria, fusion proteins containing the putative presequences of tobacco RISPs and Escherichia coli ß-glucuronidase (GUS) were expressed in transgenic tobacco, and the resultant GUS proteins were localized by cell fractionation. The aminoterminal 75 and 71

Jintai Huang; Charles S. Levings

1995-01-01

71

Polyethylenimine-cyclodextrin-tegafur conjugate shows anti-cancer activity and a potential for gene delivery*  

PubMed Central

Polyethylenimine-cyclodextrin-tegafur (PEI-CyD-tegafur) conjugate was synthesized as a novel multifunctional prodrug of tegafur for co-delivery of chemotherapeutic agent tegafur and enhanced green fluorescent protein (EGFP) reporter plasmid DNA. Conjugation of tegafur to PEI-CyD via chemical linkage was characterized by 1H NMR spectrometry and ultraviolet (UV) spectrometry. PEI-CyD-tegafur was able to condense plasmid DNA into complexes of around 150 nm with positive charge at the N/P ratio of 25, in accordance with electron microscopy observation of compact and monodisperse nanoparticles. The results of in vitro experiments showed enhanced cytotoxicity and considerable transfection efficiency in B16F10 cell line. Therefore, PEI-CyD-tegafur may have great potential as a co-delivery system with anti-cancer activity and potential for gene delivery.

Hu, Qi-da; Fan, Hui; Lou, Wei-jian; Wang, Qing-qing; Tang, Gu-ping

2011-01-01

72

A modified HSP70 inhibitor shows broad activity as an anticancer agent  

PubMed Central

The stress-induced heat shock protein 70 (HSP70) is an ATP-dependent molecular chaperone that plays a key role in refolding misfolded proteins and promoting cell survival following stress. HSP70 is marginally expressed in non-transformed cells, but is greatly overexpressed in tumor cells. Silencing HSP70 is uniformly cytotoxic to tumor but not normal cells; therefore, there has been great interest in the development of HSP70 inhibitors for cancer therapy. Here we report that the HSP70 inhibitor 2-phenylethynesulfonamide (PES) binds to the substrate-binding domain of HSP70, and requires the C-terminal helical ‘lid’ of this protein (amino acids 573-616) in order to bind. Using molecular modeling and in silico docking, we have identified a candidate binding site for PES in this region of HSP70, and we identify point mutants that fail to interact with PES. A preliminary structure-activity relationship analysis has revealed a derivative of PES, 2-(3-chlorophenyl) ethynesulfonamide (PES-Cl), which shows increased cytotoxicity and ability to inhibit autophagy, along with significantly improved ability to extend the life of mice with pre-B cell lymphoma, compared to the parent compound (p=0.015). Interestingly, we also show that these HSP70 inhibitors impair the activity of the Anaphase Promoting Complex/Cyclosome (APC/C) in cell-free extracts, and induce G2/M arrest and genomic instability in cancer cells. PES-Cl is thus a promising new anti-cancer compound with several notable mechanisms of action.

Balaburski, Gregor M.; Leu, Julia I-Ju; Beeharry, Neil; Hayik, Seth; Andrake, Mark D.; Zhang, Gao; Herlyn, Meenhard; Villanueva, Jessie; Dunbrack, Roland L.; Yen, Tim; George, Donna L.; Murphy, Maureen E.

2013-01-01

73

Synthesis and conformational analysis of a series of galactosyl enkephalin analogues showing high analgesic activity.  

PubMed Central

Two galactosyl derivatives of [DMet2,Pro5] enkephalin-amide (compound 1), namely [DMet2,Pro5] enkephalin [N1.5-beta-D-galactopyranosyl] amide (compound 2) and O1.5-(beta-D-galactopyranosyl) [DMet2,Hyp5] enkephalin-amide (compound 3) have been synthesized. Such glycosylpeptides have been shown to be extremely potent analgesic agonists. The conformational analysis of these three compounds in DMSO-d6 solution has been carried out using two-dimensional NMR methods. Both the parent compound (1) and the beta N-galactosyl derivative (2) show similar NMR parameters which are consistent with fairly rigid beta-strands at both the N-terminus and C-terminus, connected by a glycine residue that displays a mixture between multiple conformational states. Thus, although the beta N-galactosyl derivative (2) has been shown to be significantly more potent than the parent compound (1) in the tail immersion and paw pressure tests of analgesia, no correlation can be established between the conformation of (1) and (2) in DMSO and the difference in analgesic activity. In contrast, important conformational differences with respect to (1) and (2) have been detected in the beta O-galactosyl derivative (3). In this case, only one of the likely conformations for (1) and (2) are consistent with the experimental data. These data show that the position of the galactose residue in compound (3) causes Gly3 to loose flexibility leading to a more rigid folded conformation. Such a change in conformation could be related to the difference in analgesic activity between (2) and (3). Images

Torres, J L; Pepermans, H; Valencia, G; Reig, F; Garcia-Anton, J M; Van Binst, G

1989-01-01

74

The postcentral gyrus shows sustained fMRI activation during the tactile motion aftereffect.  

PubMed

The tactile motion aftereffect (tMAE) is a perceptual illusion in which a stationary stimulus feels as though it is moving when presented following adaptation to a unidirectionally moving tactile stimulus. Using functional magnetic resonance imaging (fMRI), we localized the brain areas responsive to tactile motion and then investigated whether these areas underlie the tMAE. Tactile stimulation was delivered to the glabrous surface of the right hand by means of a plastic cylinder with a square-wave patterned surface. In the tactile motion localizer, we contrasted periods in which the cylinder rotated at 15 rpm with periods of rest (stationary contact). Activation was observed in the contralateral (left) thalamus, postcentral gyrus, and parietal operculum. In the tMAE experiment, the cylinder rotated at 15 or 60 rpm for 2 min. The 60-rpm speed induced reliable tMAEs, whereas the 15-rpm speed did not. Of the areas activated by the tactile motion localizer, only the postcentral gyrus showed a sustained fMRI response following the offset of 60-rpm (but not 15-rpm) stimulation, presumably reflecting the illusory perception of motion. PMID:22120108

Planetta, Peggy J; Servos, Philip

2012-02-01

75

Regulation of the activity of Korean radish cationic peroxidase promoter during dedifferentiation and differentiation.  

PubMed

Studies of the regulation of the activity of the Korean radish cationic peroxidase (KRCP) promoter during dedifferentiation and redifferentiation are reported here. Histochemical staining with 5-bromo-4-chloro-indolyl glucuronide (X-gluc) showed that only dedifferentiated marginal cells of leaf discs of the transgenic plants, but not of the interior region, were stained blue, as leaf discs were incubated on dedifferentiation-inducing medium from 5 days after callus induction (DACI). The levels of cationic peroxidase activity and of KRCP transcripts in Korean radish seedlings (Raphanus sativus L. F1 Handsome Fall) were also upregulated by a low ratio of cytokinin to auxin, but not by high concentrations of cytokinin. To identify important cis-regulatory regions controlling callus-specific expression, a series of 5' promoter deletions was carried out with KRCP::GUS gene fusion systems. The data suggest that at least two positively regulatory regions are involved in the KRCP::GUS expression during dedifferentiation induced by a low ratio of cytokinin to auxin: one from -471 to -242 and another from -241 to +196. GUS expression, however, was quickly decreased to a basal level during regeneration of root and shoot. Thus, the downstream region between +197 and +698 seems to be enough to suppress GUS expression of all constructs during regeneration. We further show that the 142-bp fragment (-471 to -328) has at least one cis-element to bind to the nuclear proteins from Korean radish seedlings induced by dedifferentiation. PMID:15596095

Kim, Soung Soo; Choi, Suh-Yeon; Park, Jin-Hyoun; Lee, Dong Ju

2004-12-01

76

Crystal structure of plant light-harvesting complex shows the active, energy-transmitting state  

PubMed Central

Plants dissipate excess excitation energy as heat by non-photochemical quenching (NPQ). NPQ has been thought to resemble in vitro aggregation quenching of the major antenna complex, light harvesting complex of photosystem II (LHC-II). Both processes are widely believed to involve a conformational change that creates a quenching centre of two neighbouring pigments within the complex. Using recombinant LHC-II lacking the pigments implicated in quenching, we show that they have no particular role. Single crystals of LHC-II emit strong, orientation-dependent fluorescence with an emission maximum at 680 nm. The average lifetime of the main 680 nm crystal emission at 100 K is 1.31 ns, but only 0.39 ns for LHC-II aggregates under identical conditions. The strong emission and comparatively long fluorescence lifetimes of single LHC-II crystals indicate that the complex is unquenched, and that therefore the crystal structure shows the active, energy-transmitting state of LHC-II. We conclude that quenching of excitation energy in the light-harvesting antenna is due to the molecular interaction with external pigments in vitro or other pigment–protein complexes such as PsbS in vivo, and does not require a conformational change within the complex.

Barros, Tiago; Royant, Antoine; Standfuss, Jorg; Dreuw, Andreas; Kuhlbrandt, Werner

2009-01-01

77

Piper betle shows antioxidant activities, inhibits MCF-7 cell proliferation and increases activities of catalase and superoxide dismutase  

PubMed Central

Background Breast cancer is the most common form of cancer and the focus on finding chemotherapeutic agents have recently shifted to natural products. Piper betle is a medicinal plant with various biological activities. However, not much data is available on the anti-cancer effects of P. betle on breast cancer. Due to the current interest in the potential effects of antioxidants from natural products in breast cancer treatment, we investigated the antioxidant activities of the leaves of P. betle and its inhibitory effect on the proliferation of the breast cancer cell line, MCF-7. Methods The leaves of P. betle were extracted with solvents of varying polarities (water, methanol, ethyl acetate and hexane) and their phenolic and flavonoid content were determined using colorimetric assays. Phenolic composition was characterized using HPLC. Antioxidant activities were measured using FRAP, DPPH, superoxide anion, nitric oxide and hyroxyl radical scavenging assays. Biological activities of the extracts were analysed using MTT assay and antioxidant enzyme (catalase, superoxide dismutase, glutathione peroxidase) assays in MCF-7 cells. Results Overall, the ethyl acetate extract showed the highest ferric reducing activity and radical scavenging activities against DPPH, superoxide anion and nitric oxide radicals. This extract also contained the highest phenolic content implying the potential contribution of phenolics towards the antioxidant activities. HPLC analyses revealed the presence of catechin, morin and quercetin in the leaves. The ethyl acetate extract also showed the highest inhibitory effect against the proliferation of MCF-7 cells (IC50=65 ?g/ml). Treatment of MCF-7 cells with the plant extract increased activities of catalase and superoxide dismutase. Conclusions Ethyl acetate is the optimal solvent for the extraction of compounds with antioxidant and anti-proliferative activities. The increased activities of catalase and superoxide dismutase in the treated cells could alter the antioxidant defense system, potentially contributing towards the anti-proliferative effect. There is great potential for the ethyl acetate extract of P. betle leaf as a source of natural antioxidants and to be developed as therapeutics in cancer treatment.

2012-01-01

78

HATS-2b: A transiting extrasolar planet orbiting a K-type star showing starspot activity  

NASA Astrophysics Data System (ADS)

We report the discovery of HATS-2b, the second transiting extrasolar planet detected by the HATSouth survey. HATS-2b is moving on a circular orbit around a V = 13.6 mag, K-type dwarf star (GSC 6665-00236), at a separation of 0.0230 ± 0.0003 AU and with a period of 1.3541 days. The planetary parameters have been robustly determined using a simultaneous fit of the HATSouth, MPG/ESO 2.2 m/GROND, Faulkes Telescope South/Spectral transit photometry, and MPG/ESO 2.2 m/FEROS, Euler 1.2 m/CORALIE, AAT 3.9 m/CYCLOPS radial-velocity measurements. HATS-2b has a mass of 1.37 ± 0.16 MJ, a radius of 1.14 ± 0.03 RJ, and an equilibrium temperature of 1567 ± 30 K. The host star has a mass of 0.88 ± 0.04 M? and a radius of 0.89 ± 0.02 R?, and it shows starspot activity. We characterized the stellar activity by analyzing two photometric follow-up transit light curves taken with the GROND instrument, both obtained simultaneously in four optical bands (covering the wavelength range of 3860-9520 Å). The two light curves contain anomalies compatible with starspots on the photosphere of the host star along the same transit chord. Tables of the individual photometric measurements are only available at the CDS via anonymous ftp to http://cdsarc.u-strasbg.fr (ftp://130.79.128.5) or via http://cdsarc.u-strasbg.fr/viz-bin/qcat?J/A+A/558/A55

Mohler-Fischer, M.; Mancini, L.; Hartman, J. D.; Bakos, G. Á.; Penev, K.; Bayliss, D.; Jordán, A.; Csubry, Z.; Zhou, G.; Rabus, M.; Nikolov, N.; Brahm, R.; Espinoza, N.; Buchhave, L. A.; Béky, B.; Suc, V.; Csák, B.; Henning, T.; Wright, D. J.; Tinney, C. G.; Addison, B. C.; Schmidt, B.; Noyes, R. W.; Papp, I.; Lázár, J.; Sári, P.; Conroy, P.

2013-10-01

79

Using analogy role-play activity in an undergraduate biology classroom to show central dogma revision.  

PubMed

For the study of biology in an undergraduate classroom, a classroom exercise was developed: an analogy role-play to learn mechanisms of gene transcription and protein translation (central dogma). To develop the central dogma role-play exercise, we made DNA and mRNA using paper sheets, tRNA using a wire dress hanger, and amino acids using Lego® blocks (Lego System A/S, Denmark). Students were studying in the course of mathematics, physics, or chemistry, so biology was not among their usual studies. In this exercise, students perform the central dogma role-play and respectively act out nuclear matrix proteins, a transcription factor, an RNA polymerase II, an mRNA transport protein, nuclear pore proteins, a large ribosomal subunit, a small ribosomal subunit, and several amino-acyl tRNA synthetases. Questionnaire results obtained after the activity show that this central dogma role-play analogy holds student interest in the practical molecular biological processes of transcription and translation. © 2014 by The International Union of Biochemistry and Molecular Biology, 42(4):351-356, 2014. PMID:24898113

Takemura, Masaharu; Kurabayashi, Mario

2014-07-01

80

A nanoparticle depot formulation of 4-(N)-stearoyl gemcitabine shows a strong antitumor activity  

PubMed Central

Objectives Depot formulation as a carrier for cytotoxic chemotherapeutic drugs is not well studied. The objective of the present study is to test the feasibility of using a subcutaneous depot formulation to administer a cytotoxic anticancer drug for systemic therapy. Methods A fatty acid amide prodrug of the nucleoside analog gemcitabine (4-(N)-stearoyl gemcitabine (GemC18)) was incorporated into poly(lactic-co-glycolic acid) (PLGA) nanoparticles or microspheres. A GemC18 solution was used as a control. The antitumor activities were evaluated after subcutaneous injection of the different formulations in C57BL/6 mice with pre-established model tumors. The clearance of GemC18 from the injection site was determined by measuring the percentage of GemC18 remaining at the injection site at different time after the injection. Key findings The depot formulation based on the GemC18-loaded PLGA nanoparticles showed the strongest antitumor, likely due to the proper ‘release’ of GemC18 from the injection site. Conclusions It is feasible to dose cytotoxic anticancer drugs as a nanoparticle-based depot formulation, especially when combined with an advanced prodrug strategy.

Zhu, Saijie; Li, Xinran; Lansakara-P, Dharmika S.P.; Kumar, Amit; Cui, Zhengrong

2012-01-01

81

Hydroxyeicosapentaenoic acids from the Pacific krill show high ligand activities for PPARs.  

PubMed

PPARs regulate the expression of genes for energy metabolism in a ligand-dependent manner. PPARs can influence fatty acid oxidation, the level of circulating triglycerides, glucose uptake and insulin sensitivity. Here, we demonstrate that 5-hydroxyeicosapentaenoic acid (HEPE), 8-HEPE, 9-HEPE, 12-HEPE and 18-HEPE (hydroxylation products of EPA) obtained from methanol extracts of Pacific krill (Euphausia pacifica) can act as PPAR ligands. Two of these products, 8-HEPE and 9-HEPE, enhanced the transcription levels of GAL4-PPARs to a significantly greater extent than 5-HEPE, 12-HEPE, 18-HEPE, EPA, and EPA ethyl-ester. 8-HEPE also activated significantly higher transcription of GAL4-PPAR?, GAL4-PPAR?, and GAL4-PPAR? than EPA at concentrations greater than 4, 64, and 64 ?M, respectively. We also demonstrated that 8-HEPE increased the expression levels of genes regulated by PPARs in FaO, 3T3-F442A, and C2C12 cells. Furthermore, 8-HEPE enhanced adipogenesis and glucose uptake. By contrast, at the same concentrations, EPA showed weak or little effect, indicating that 8-HEPE was the more potent inducer of physiological effects. PMID:24668940

Yamada, Hidetoshi; Oshiro, Eriko; Kikuchi, Sayaka; Hakozaki, Mayuka; Takahashi, Hideyuki; Kimura, Ken-Ichi

2014-05-01

82

26 CFR 1.513-3 - Qualified convention and trade show activity.  

Code of Federal Regulations, 2013 CFR

...certain activities, including rental of exhibition space at a qualified convention or...Certain activities â(1) Rental of exhibition space. The rental of display space...situation, however, the rental of exhibition space to suppliers is not...

2013-04-01

83

Mice lacking brain-type creatine kinase activity show defective thermoregulation.  

PubMed

The cytosolic brain-type creatine kinase and mitochondrial ubiquitous creatine kinase (CK-B and UbCKmit) are expressed during the prepubescent and adult period of mammalian life. These creatine kinase (CK) isoforms are present in neural cell types throughout the central and peripheral nervous system and in smooth muscle containing tissues, where they have an important role in cellular energy homeostasis. Here, we report on the coupling of CK activity to body temperature rhythm and adaptive thermoregulation in mice. With both brain-type CK isoforms being absent, the body temperature reproducibly drops ~1.0 degrees C below normal during every morning (inactive) period in the daily cycle. Facultative non-shivering thermogenesis is also impaired, since CK--/-- mice develop severe hypothermia during 24 h cold exposure. A relationship with fat metabolism was suggested because comparison of CK--/-- mice with wildtype controls revealed decreased weight gain associated with less white and brown fat accumulation and smaller brown adipocytes. Also, circulating levels of glucose, triglycerides and leptin are reduced. Extensive physiological testing and uncoupling protein1 analysis showed, however, that the thermogenic problems are not due to abnormal responsiveness of brown adipocytes, since noradrenaline infusion produced a normal increase of body temperature. Moreover, we demonstrate that the cyclic drop in morning temperature is also not related to altered rhythmicity with reduced locomotion, diminished food intake or increased torpor sensitivity. Although several integral functions appear altered when CK is absent in the brain, combined findings point into the direction of inefficient neuronal transmission as the dominant factor in the thermoregulatory defect. PMID:19419668

Streijger, Femke; Pluk, Helma; Oerlemans, Frank; Beckers, Gaby; Bianco, Antonio C; Ribeiro, Miriam O; Wieringa, Bé; Van der Zee, Catharina E E M

2009-04-20

84

Water Works: A Great Show on Earth. Classroom Activities for Third and Fourth Grades.  

ERIC Educational Resources Information Center

This curriculum guide is divided into five lessons, each containing several activities that reflect the natural path of inquiry that third or fourth grade students might take in considering the water that arrives in their bathroom sinks each morning. Starting from the familiar faucet, the students are encouraged to reflect on their own habits and…

McClure, Judy; Clark, Neil

85

Constituents of stem bark of Callistemon rigidus showing inhibitory effects on mouse alpha-amylase activity.  

PubMed

From stem bark of Callistemon rigidus (Myrtaceae), piceatannol and scirpusin B were isolated as components that exhibit inhibitory effects on alpha-amylase activity in isolated mouse plasma. In particular, scirpusin B also inhibited alpha-amylase in mouse gastrointestinal tract. Thus, we expect the depressive effect on the elevation of postprandial blood glucose may be a new medicinal use of this compound as well as the plant itself. PMID:16755033

Kobayashi, Kyoko; Ishihara, Tamaki; Khono, Eriko; Miyase, Toshio; Yoshizaki, Fumihiko

2006-06-01

86

New targeted therapy for advanced prostate cancer shows anti-tumor activity in clinical trials  

Cancer.gov

At the 24th EORTC-NCI-AACR Symposium on Molecular Targets and Cancer Therapeutics in Dublin, Ireland, researchers from a multi-center study reported that a new drug that specifically targets a protein found on the surface of prostate cancer cells has performed well in a phase I clinical trial, and a phase II trial has started. The drug reduced levels of circulating tumor cells and levels of prostate specific antigen, a marker for tumor activity, in patients who had already failed previous chemotherapy and hormone treatments.

87

Heterocyclic Aromatic Hydrocarbons Show Estrogenic Activity upon Metabolization in a Recombinant Transactivation Assay.  

PubMed

Heterocyclic aromatic hydrocarbons (hetero-PAHs) are increasingly studied at contaminated sites; especially at former industrial facilities where coal tar-oil was handled, e.g., wood treatment plants, high concentrations of hetero-PAHs are frequently detected in groundwater plumes. In previous studies, fractions of groundwater with high estrogenic activity contained hetero-PAHs and their hydroxylated metabolites. To evaluate this preliminary evidence, selected hetero-PAHs were screened for their estrogenic activity in lyticase yeast estrogen screen (LYES) and ER CALUX. All tested substances were inactive in the LYES. Hetero-PAHs such as acridine, xanthene, indole, 2-methylbenzofuran, 2,3-dimethylbenzofuran, dibenzofuran, dibenzothiophene, quinoline, and 6-methylquinoline were positive in the ER CALUX, with estradiol equivalence factors (EEFs) from 2.85 × 10(-7) to 3.18 × 10(-5). The EEF values of these substances were comparable to those of other xenoestrogens (e.g., alkylphenols or bisphenol A) that are sometimes found in surface water. Chemical analyses revealed that T47Dluc cells could metabolize most of the substances. Among the metabolites (tentatively) identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) were hydroxides and their keto tautomers, sulfates, sulfoxides, and N-oxides. Because of their high concentrations measured in groundwater, we conclude that hetero-PAHs and metabolites may be a potential risk and should be the subject of further research. PMID:24724806

Brinkmann, Markus; Maletz, Sibylle; Krauss, Martin; Bluhm, Kerstin; Schiwy, Sabrina; Kuckelkorn, Jochen; Tiehm, Andreas; Brack, Werner; Hollert, Henner

2014-05-20

88

Updated clinical results show experimental agent ibrutinib as highly active in CLL patients  

Cancer.gov

Updated results from a Phase Ib/II clinical trial led by the Ohio State University Comprehensive Cancer Center – Arthur G. James Cancer Hospital and Richard J. Solove Research Institute indicates that a novel therapeutic agent for chronic lymphocytic leukemia (CLL) is highly active and well tolerated in patients who have relapsed and are resistant to other therapy. The agent, ibrutinib (PCI-32765), is the first drug designed to target Bruton's tyrosine kinase (BTK), a protein essential for CLL-cell survival and proliferation. CLL is the most common form of leukemia, with about 15,000 new cases annually in the U.S. About 4,400 Americans die of the disease each year.

89

Delayed recovery of ?-glucuronidase activity driven by an Arabidopsis heat shock promoter in heat-stressed transgenic Nicotiana plumbaginifolia  

Microsoft Academic Search

The expression of the Arabidopsis heat shock protein (HSP) 18.2 promoter-?-d-glucuronidase (GUS) chimera gene was investigated in transgenic Nicotiana plumbaginifolia plants during the recovery phase at normal temperatures (20–22??°C) after a heat shock (HS) treatment. GUS activity increased\\u000a during the recovery phase after HS at 42??°C for 2?h, and maximal GUS activity was observed after 12?h at normal temperatures,\\u000a at

M. Moriwaki; T. Yamakawa; T. Washino; T. Kodama; Y. Igarashi

1999-01-01

90

Beta-adrenoceptor activation shows high-frequency fatigue in skeletal muscle fibers of the rat.  

PubMed

The effect of terbutaline (a beta 2-adrenergic agonist) on high-frequency fatigue (HFF) was studied in small bundles of rat soleus muscle fibers. HFF, the decline in force during continuous stimulation (50 Hz for 20 s), was reduced by 10-20% with 10 microM terbutaline. A similar reduction in HFF with 2 mM dibutyryl-adenosine 3',5'-cyclic monophosphate (DBcAMP) implicated adenosine 3',5'-cyclic monophosphate (cAMP) as the second messenger in the terbutaline effect. Sodium (Na-K)-pump inhibition with 1 mM ouabain depressed peak tetanic force but did not significantly alter either the subsequent fatigue or the effect of terbutaline on fatigue. This suggested that the pump was neither rate limiting in HFF nor involved in the terbutaline effect. Nevertheless, a significant hyperpolarization recorded with terbutaline implied that beta 2-adrenoceptor activation stimulated the Na-K pump at rest. Caffeine (1 mM) slowed HFF and prevented additional effects with terbutaline. Caffeine is known to potentiate Ca2+ release from the sarcoplasmic reticulum (SR), and we suggest that terbutaline, acting via cAMP, facilitates Ca2+ release from the SR to better maintain myoplasmic Ca2+ concentration during continuous tetanic stimulation. PMID:8203484

Cairns, S P; Dulhunty, A F

1994-05-01

91

KefF, the regulatory subunit of the potassium efflux system KefC, shows quinone oxidoreductase activity.  

PubMed

Escherichia coli and many other Gram-negative pathogenic bacteria protect themselves from the toxic effects of electrophilic compounds by using a potassium efflux system (Kef). Potassium efflux is coupled to the influx of protons, which lowers the internal pH and results in immediate protection. The activity of the Kef system is subject to complex regulation by glutathione and its S conjugates. Full activation of KefC requires a soluble ancillary protein, KefF. This protein has structural similarities to oxidoreductases, including human quinone reductases 1 and 2. Here, we show that KefF has enzymatic activity as an oxidoreductase, in addition to its role as the KefC activator. It accepts NADH and NADPH as electron donors and quinones and ferricyanide (in addition to other compounds) as acceptors. However, typical electrophilic activators of the Kef system, e.g., N-ethyl maleimide, are not substrates. If the enzymatic activity is disrupted by site-directed mutagenesis while retaining structural integrity, KefF is still able to activate the Kef system, showing that the role as an activator is independent of the enzyme activity. Potassium efflux assays show that electrophilic quinones are able to activate the Kef system by forming S conjugates with glutathione. Therefore, it appears that the enzymatic activity of KefF diminishes the redox toxicity of quinones, in parallel with the protection afforded by activation of the Kef system. PMID:21742892

Lyngberg, Lisbeth; Healy, Jessica; Bartlett, Wendy; Miller, Samantha; Conway, Stuart J; Booth, Ian R; Rasmussen, Tim

2011-09-01

92

KefF, the Regulatory Subunit of the Potassium Efflux System KefC, Shows Quinone Oxidoreductase Activity ? †  

PubMed Central

Escherichia coli and many other Gram-negative pathogenic bacteria protect themselves from the toxic effects of electrophilic compounds by using a potassium efflux system (Kef). Potassium efflux is coupled to the influx of protons, which lowers the internal pH and results in immediate protection. The activity of the Kef system is subject to complex regulation by glutathione and its S conjugates. Full activation of KefC requires a soluble ancillary protein, KefF. This protein has structural similarities to oxidoreductases, including human quinone reductases 1 and 2. Here, we show that KefF has enzymatic activity as an oxidoreductase, in addition to its role as the KefC activator. It accepts NADH and NADPH as electron donors and quinones and ferricyanide (in addition to other compounds) as acceptors. However, typical electrophilic activators of the Kef system, e.g., N-ethyl maleimide, are not substrates. If the enzymatic activity is disrupted by site-directed mutagenesis while retaining structural integrity, KefF is still able to activate the Kef system, showing that the role as an activator is independent of the enzyme activity. Potassium efflux assays show that electrophilic quinones are able to activate the Kef system by forming S conjugates with glutathione. Therefore, it appears that the enzymatic activity of KefF diminishes the redox toxicity of quinones, in parallel with the protection afforded by activation of the Kef system.

Lyngberg, Lisbeth; Healy, Jessica; Bartlett, Wendy; Miller, Samantha; Conway, Stuart J.; Booth, Ian R.; Rasmussen, Tim

2011-01-01

93

Phylogenetic distribution of genes encoding ?-glucuronidase activity in human colonic bacteria and the impact of diet on faecal glycosidase activities.  

PubMed

Bacterial ?-glucuronidase in the human colon plays an important role in cleaving liver conjugates of dietary compounds and xenobiotics, while other glycosidase activities are involved in the conversion of dietary plant glycosides. Here we detected an increase in ?-glucuronidase activity in faecal samples from obese volunteers following a high-protein moderate carbohydrate weight-loss diet, compared with a weight maintenance diet, but little or no changes were observed when the type of fermentable carbohydrate was varied. Other faecal glycosidase activities showed little or no change over a fivefold range of dietary NSP intake, although ?-glucosidase increased on a resistant starch-enriched diet. Two distinct groups of gene, gus and BG, have been reported to encode ?-glucuronidase activity among human colonic bacteria. Degenerate primers were designed against these genes. Overall, Firmicutes were found to account for 96% of amplified gus sequences, with three operational taxonomic units particularly abundant, whereas 59% of amplified BG sequences belonged to Bacteroidetes and 41% to Firmicutes. A similar distribution of operational taxonomic units was found in a published metagenome dataset involving a larger number of volunteers. Seven cultured isolates of human colonic bacteria that carried only the BG gene gave relatively low ?-glucuronidase activity that was not induced by 4-nitrophenyl-?-D-glucuronide. By comparison, in three of five isolates that possessed only the gus gene, ?-glucuronidase activity was induced. PMID:22364273

McIntosh, Freda M; Maison, Nathalie; Holtrop, Grietje; Young, Pauline; Stevens, Valerie J; Ince, Jennifer; Johnstone, Alexandra M; Lobley, Gerald E; Flint, Harry J; Louis, Petra

2012-08-01

94

Demonstration of vaginal colonization with GusA-expressing Lactobacillus jensenii following oral delivery in rhesus macaques  

PubMed Central

The vaginal microbiome, which harbors beneficial Lactobacillus strains, is believed to be a major host defense mechanism for preventing infections of the urogenital tract. It has been suggested that the gastrointestinal tract serves as a reservoir for lactobacilli that colonize the vagina. Using rhesus macaques, we examined whether oral delivery of human vaginal Lactobacillus jensenii-1153-1646, a GusA-producing strain, would result in colonization of the rectum and the vagina. Lactobacilli were identified from the vagina tracts of three macaques on the basis of ?-glucuronidase enzyme production, 16S rRNA gene sequence and DNA homology using a repetitive sequence-based polymerase chain reaction.

Lagenaur, Laurel A; Lee, Peter P; Hamer, Dean H; Sanders-Beer, Brigitte E

2012-01-01

95

Recombinant PNPLA3 protein shows triglyceride hydrolase activity and its I148M mutation results in loss of function.  

PubMed

The patatin-like phospholipase domain containing 3 (PNPLA3, also called adiponutrin, ADPN) is a membrane-bound protein highly expressed in the liver. The genetic variant I148M (rs738409) was found to be associated with progression of chronic liver disease. We aimed to establish a protein purification protocol in a yeast system (Pichia pastoris) and to examine the human PNPLA3 enzymatic activity, substrate specificity and the I148M mutation effect. hPNPLA3 148I wild type and 148M mutant cDNA were cloned into P. pastoris expression vectors. Yeast cells were grown in 3L fermentors. PNPLA3 protein was purified from membrane fractions by Ni-affinity chromatography. Enzymatic activity was assessed using radiolabeled substrates. Both 148I wild type and 148M mutant proteins are localized to the membrane. The wild type protein shows a predominant lipase activity with mild lysophosphatidic acid acyl transferase activity (LPAAT) and the I148M mutation results in a loss of function of both these activities. Our data show that PNPLA3 has a predominant lipase activity and I148M mutation results in a loss of function. PMID:24369119

Pingitore, Piero; Pirazzi, Carlo; Mancina, Rosellina M; Motta, Benedetta M; Indiveri, Cesare; Pujia, Arturo; Montalcini, Tiziana; Hedfalk, Kristina; Romeo, Stefano

2014-04-01

96

Pomalidomide shows significant therapeutic activity against CNS lymphoma with a major impact on the tumor microenvironment in murine models.  

PubMed

Primary CNS lymphoma carries a poor prognosis. Novel therapeutic agents are urgently needed. Pomalidomide (POM) is a novel immunomodulatory drug with anti-lymphoma activity. CNS pharmacokinetic analysis was performed in rats to assess the CNS penetration of POM. Preclinical evaluation of POM was performed in two murine models to assess its therapeutic activity against CNS lymphoma. The impact of POM on the CNS lymphoma immune microenvironment was evaluated by immunohistochemistry and immunofluorescence. In vitro cell culture experiments were carried out to further investigate the impact of POM on the biology of macrophages. POM crosses the blood brain barrier with CNS penetration of ~ 39%. Preclinical evaluations showed that it had significant therapeutic activity against CNS lymphoma with significant reduction in tumor growth rate and prolongation of survival, that it had a major impact on the tumor microenvironment with an increase in macrophages and natural killer cells, and that it decreased M2-polarized tumor-associated macrophages and increased M1-polarized macrophages when macrophages were evaluated based on polarization status. In vitro studies using various macrophage models showed that POM converted the polarization status of IL4-stimulated macrophages from M2 to M1, that M2 to M1 conversion by POM in the polarization status of lymphoma-associated macrophages is dependent on the presence of NK cells, that POM induced M2 to M1 conversion in the polarization of macrophages by inactivating STAT6 signaling and activating STAT1 signaling, and that POM functionally increased the phagocytic activity of macrophages. Based on our findings, POM is a promising therapeutic agent for CNS lymphoma with excellent CNS penetration, significant preclinical therapeutic activity, and a major impact on the tumor microenvironment. It can induce significant biological changes in tumor-associated macrophages, which likely play a major role in its therapeutic activity against CNS lymphoma. POM should be further evaluated in clinical trials. PMID:23940785

Li, Zhimin; Qiu, Yushi; Personett, David; Huang, Peng; Edenfield, Brandy; Katz, Jason; Babusis, Darius; Tang, Yang; Shirely, Michael A; Moghaddam, Mehran F; Copland, John A; Tun, Han W

2013-01-01

97

Dillapiole, isolated from Peperomia pellucida, shows gastroprotector activity against ethanol-induced gastric lesions in Wistar rats.  

PubMed

Peperomia pellucida is a plant used in traditional medicine to treat gastric ulcers. Although this gastroprotective activity was reported, the active compounds have not been identified. Therefore, the aim herein was to identify the most active compound in the gastroprotective activity of P. pellucida using an ethanol-induced gastric ulcer experimental rat model. A gastroprotective effect was observed when the hexane and dichloromethane extracts were tested, with the higher effect being obtained with the dichloromethane extract (82.3 ± 5.6%) at 100 mg/kg. Dillapiole was identified as the most active compound in this extract. Although there have been previous reports on dillapiole, this is the first on its gastroprotective activity. Rats treated with this compound at 3, 10, 30 and 100 mg/kg showed 23.1, 56.1, 73.2 and 85.5% gastroprotection, respectively. The effect elicited by dillapiole at 100 mg/kg was not attenuated by pretreatment with indomethacin (10 mg/kg, s.c.), a prostaglandin synthesis blocker, NG-nitro-l-arginine methyl ester (70 mg/kg, i.p.), a nitric oxide (NO) synthase inhibitor, or N-ethylmaleimide (10 mg/kg, s.c.), a blocker of sulfhydryl groups. This suggests that the gastroprotective mechanism of action of dillapiole does not involve prostaglandins, NO or sulfhydryl groups. PMID:24064453

Rojas-Martínez, Raúl; Arrieta, Jesús; Cruz-Antonio, Leticia; Arrieta-Baez, Daniel; Velázquez-Méndez, Antonio Magdiel; Sánchez-Mendoza, María Elena

2013-01-01

98

Peptide Nucleic Acids Conjugated to Short Basic Peptides Show Improved Pharmacokinetics and Antisense Activity in Adipose Tissue  

PubMed Central

A peptide nucleic acid (PNA) targeting a splice junction of the murine PTEN primary transcript was covalently conjugated to various basic peptides. When systemically administered to healthy mice, the conjugates displayed sequence-specific alteration of PTEN mRNA splicing as well as inhibition of full length PTEN protein expression. Correlating activity with drug concentration in various tissues indicated strong tissue-dependence with highest levels of activity observed in adipose tissue. While the presence of a peptide carrier was found to be crucial for efficient delivery to tissue, little difference was observed between the various peptides evaluated. A second PNA-conjugate targeting the murine insulin receptor primary transcript showed a similar activity profile suggesting that short basic peptides can generally be used to effectively deliver peptide nucleic acids to adipose tissue.

Wancewicz, Edward V.; Maier, Martin A.; Siwkowski, Andrew M.; Albertshofer, Klaus; Winger, Theodore M.; Berdeja, Andres; Gaus, Hans; Vickers, Timothy A.; Bennett, C. Frank; Monia, Brett P.; Griffey, Richard H.; Nulf, Christopher J.; Hu, Jiaxin; Corey, David R.; Swayze, Eric E.; Kinberger, Garth A.

2010-01-01

99

Backbone cyclised peptides from plants show molluscicidal activity against the rice pest Pomacea canaliculata (golden apple snail).  

PubMed

Golden apple snails ( Pomacea canaliculata) are serious pests of rice in South East Asia. Cyclotides are backbone cyclized peptides produced by plants from Rubiaceae and Violaceae. In this study, we investigated the molluscicidal activity of cyclotides against golden apple snails. Crude cyclotide extracts from both Oldenlandia affinis and Viola odorata plants showed molluscicidal activity comparable to the synthetic molluscicide metaldehyde. Individual cyclotides from each extract demonstrated a range of molluscicidal activities. The cyclotides cycloviolacin O1, kalata B1, and kalata B2 were more toxic to golden apple snails than metaldehyde, while kalata B7 and kalata B8 did not cause significant mortality. The toxicity of the cyclotide kalata B2 on a nontarget species, the Nile tilapia ( Oreochromis niloticus), was three times lower than the common piscicide rotenone. Our findings suggest that the existing diversity of cyclotides in plants could be used to develop natural molluscicides. PMID:18557620

Plan, Manuel Rey R; Saska, Ivana; Cagauan, Arsenia G; Craik, David J

2008-07-01

100

Prion Protein-Deficient Cells Show Altered Response to Oxidative Stress Due to Decreased SOD1 Activity  

Microsoft Academic Search

The cellular function of the prion protein (PrPC), a cell surface glycoprotein expressed in neurones and astrocytes, has not been elucidated. Cell culture experiments reveal that cerebellar cells lacking PrPCare more sensitive to oxidative stress and undergo cell death more readily than wild-type cells. This effect is reversible by treatment with vitamin E.In vivostudies show that the activity of Cu\\/Zn

David R. Brown; Walter J. Schulz-Schaeffer; Bernhard Schmidt; Hans A. Kretzschmar

1997-01-01

101

Thiophene-degrading Escherichia coli mutants possess sulfone oxidase activity and show altered resistance to sulfur-containing antibiotics.  

PubMed

We have previously isolated mutants of Escherichia coli which show increased oxidation of heterocyclic furan and thiophene substrates. We have now found that strains carrying the thdA mutation express a novel enzyme activity which oxidizes a variety of substrates containing a sulfone (SO2) moiety. Both heterocyclic sulfones (e.g., tetramethylene sulfone) and simple aliphatic sulfones (e.g., ethyl sulfone) were oxidized. The thdA mutants were more resistant than wild-type strains to aromatic sulfone antibiotics such as dapsone. In contrast they showed increased susceptibility to thiolutin, a cyclic antibiotic containing sulfur at the sulfide level of oxidation. Several new thdA mutant alleles were isolated by selecting for increased oxidation of various aliphatic sulfur compounds. These new thdA mutants showed similar sulfone oxidase activity and the same map location (at 10.7 min) as the original thdA1 mutation. The constitutive fadR mutation was required for the phenotypic expression of thdA-mediated oxidation of sulfur compounds. However, the thdA-directed expression of sulfone oxidase activity was not fadR dependent. The thdC and thdD mutations probably protect against the toxicity of thiophene derivatives rather than conferring improved metabolic capability. PMID:2285321

Juhl, M J; Clark, D P

1990-10-01

102

Rosmarinic Acid from Eelgrass Shows Nematicidal and Antibacterial Activities against Pine Wood Nematode and Its Carrying Bacteria  

PubMed Central

Pine wilt disease (PWD), a destructive disease for pine trees, is caused by the pine wood nematode (PWN), Bursaphelenchus xylophilus and additional bacteria. In this study, extracts of Zostera marina showed a high nematicidal activity against PWN and some of the bacteria that it carries. Light yellow crystals were obtained from extracts of Z. marina through solvent extraction, followed by chromatography on AB-8 resin and crystallization. The NMR and HPLC analysis showed that the isolated compound was rosmarinic acid (RosA). RosA showed effective nematicidal activity, of which the LC50 (50% lethal concentration) to PWN at 24 h, 48 h and 72 h was 1.18 mg/g, 1.05 mg/g and 0.95 mg/g, respectively. To get a high yield rate of RosA from Z. marina, single factor experiments and an L9 (34) orthogonal experiment were performed. This extraction process involved 70% ethanol for 3 h at 40 °C. The extraction dosage was 1:50 (w/v). The highest yield of RosA from Zostera was 3.13 mg/g DW (dried weight). The crude extracts of Zostera marina (10 mg/mL) and RosA (1 mg/mL) also showed inhibitory effects to some bacterial strains carried by PWN: Klebsiella sp., Stenotrophomonas maltophilia, Streptomyces sp. and Pantoea agglomerans. The results of these studies provide clues for preparing pesticide to control PWD from Z. marina.

Wang, Jingyu; Pan, Xueru; Han, Yi; Guo, Daosen; Guo, Qunqun; Li, Ronggui

2012-01-01

103

Rosmarinic acid from eelgrass shows nematicidal and antibacterial activities against pine wood nematode and its carrying bacteria.  

PubMed

Pine wilt disease (PWD), a destructive disease for pine trees, is caused by the pine wood nematode (PWN), Bursaphelenchus xylophilus and additional bacteria. In this study, extracts of Zostera marina showed a high nematicidal activity against PWN and some of the bacteria that it carries. Light yellow crystals were obtained from extracts of Z. marina through solvent extraction, followed by chromatography on AB-8 resin and crystallization. The NMR and HPLC analysis showed that the isolated compound was rosmarinic acid (RosA). RosA showed effective nematicidal activity, of which the LC?? (50% lethal concentration) to PWN at 24 h, 48 h and 72 h was 1.18 mg/g, 1.05 mg/g and 0.95 mg/g, respectively. To get a high yield rate of RosA from Z. marina, single factor experiments and an L? (3?) orthogonal experiment were performed. This extraction process involved 70% ethanol for 3 h at 40 °C. The extraction dosage was 1:50 (w/v). The highest yield of RosA from Zostera was 3.13 mg/g DW (dried weight). The crude extracts of Zostera marina (10 mg/mL) and RosA (1 mg/mL) also showed inhibitory effects to some bacterial strains carried by PWN: Klebsiella sp., Stenotrophomonas maltophilia, Streptomyces sp. and Pantoea agglomerans. The results of these studies provide clues for preparing pesticide to control PWD from Z. marina. PMID:23201594

Wang, Jingyu; Pan, Xueru; Han, Yi; Guo, Daosen; Guo, Qunqun; Li, Ronggui

2012-12-01

104

Isopimaric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureus.  

PubMed

The diterpene isopimaric acid was extracted from the immature cones of Pinus nigra (Arnold) using bioassay-guided fractionation of a crude hexane extract. Isopimaric acid was assayed against multidrug-resistant (MDR) and methicillin-resistant Staphylococcus aureus (MRSA). The minimum inhibitory concentrations (MIC) were 32-64 microg/mL and compared with a commercially obtained resin acid, abietic acid, with MICs of 64 microg/mL. Resin acids are known to have antibacterial activity and are valued in traditional medicine for their antiseptic properties. These results show that isopimaric acid is active against MDR and MRSA strains of S. aureus which are becoming increasingly resistant to antibiotics. Both compounds were evaluated for modulation activity in combination with antibiotics, but did not potentiate the activity of the antibiotics tested. However, the compounds were also assayed in combination with the efflux pump inhibitor reserpine, to see if inhibition of the TetK or NorA efflux pump increased their activity. Interestingly, rather than a potentiation of activity by a reduction in MIC, a two to four-fold increase in MIC was seen. It may be that isopimaric acid and abietic acid are not substrates for these efflux pumps, but it is also possible that an antagonistic interaction with reserpine may render the antibiotics inactive. 1H-NMR of abietic acid and reserpine taken individually and in combination, revealed a shift in resonance of some peaks for both compounds when mixed together compared with the spectra of the compounds on their own. It is proposed that this may be due to complex formation between abietic acid and reserpine and that this complex formation is responsible for a reduction in activity and elevation of MIC. PMID:16114093

Smith, Eileen; Williamson, Elizabeth; Zloh, Mire; Gibbons, Simon

2005-06-01

105

Cloning and characterization of a novel amidase from Paracoccus sp. M-1, showing aryl acylamidase and acyl transferase activities.  

PubMed

A novel amidase gene, designated pamh, was cloned from Paracoccus sp. M-1. Site-directed mutagenesis and bioinformatic analysis showed that the PamH protein belonged to the amidase signature enzyme family. PamH was expressed in Escherichia coli, purified, and characterized. The molecular mass of PamH was determined to be 52 kDa with an isoelectric point of 5.13. PamH displayed its highest enzymatic activity at 45°C and at pH 8.0 and was stable within a pH range of 5.0-10.0. The PamH enzyme exhibited amidase activity, aryl acylamidase activity, and acyl transferase activity, allowing it to function across a very broad substrate spectrum. PamH was highly active on aromatic and short-chain aliphatic amides (benzamide and propionamide), moderately active on amino acid amides, and possessed weak urease activity. Of the anilides examined, only propanil was a good substrate for PamH. For propanil, the k (cat) and K (m) were 2.8 s(-1) and 158 ?M, respectively, and the catalytic efficiency value (k (cat)/K (m)) was 0.018 ?M(-1) s(-1). In addition, PamH was able to catalyze the acyl transfer reaction to hydroxylamine for both amide and anilide substrates, including acetamide, propanil, and 4-nitroacetanilide; the highest reaction rate was shown with isobutyramide. These characteristics make PamH an excellent candidate for environmental remediation and an important enzyme for the biosynthesis of novel amides. PMID:22101784

Shen, Weiliang; Chen, Honghong; Jia, Kaizhi; Ni, Jun; Yan, Xin; Li, Shunpeng

2012-05-01

106

Quercetin and quercetin 3-O-glycosides from Bauhinia longifolia (Bong.) Steud. show anti-Mayaro virus activity  

PubMed Central

Background The arthropod-borne Mayaro virus (MAYV) causes ‘Mayaro fever’, a disease of medical significance, primarily affecting individuals in permanent contact with forested areas in tropical South America. Recently, MAYV has attracted attention due to its likely urbanization. Currently, there are no licensed drugs against most mosquito-transmitted viruses. Here, we investigated the in vitro anti-MAYV activity of the flavonoids quercetin and its derivatives from the Brazilian shrub Bauhinia longifolia (Bong.) Steud. Methods Flavonoids were purified by chromatographic fractionation from leaf extracts of B. longifolia and chemically identified as quercetin and quercetin glycosides using spectroscopic techniques. Cytotoxicity of purified flavonoids and of EtOAc- and n-BuOH-containing flavonoid mixtures was measured by the dye-uptake assay while their antiviral activity was evaluated by a virus yield inhibition assay. Results The following flavonoids were purified from B. longifolia leaves: non-glycosylated quercetin and its glycosides guaijaverin, quercitrin, isoquercitrin, and hyperin. EtOAc and n-BuOH fractions containing these flavonoids demonstrated the highest antiviral activity of all tested substances, while quercetin had the highest antiviral activity amongst purified flavonoids. Quercetin, EtOAc, or n-BuOH fractions inhibited MAYV production by more than 90% at 25 ?g/mL, displaying a stronger antiviral effect than the licensed antiviral ribavirin. A mixture of the isomers isoquercitrin and hyperin had a modest antiviral effect (IC90?=?104.9), while guaijaverin and quercitrin did not show significant antiviral activity. Conclusions B. longifolia is a good source of flavonoids with anti-Mayaro virus activity. This is the first report of the activity of quercetin and its derivatives against an alphavirus.

2014-01-01

107

Repressible extracellular phosphodiesterases showing cyclic 2',3'- and cyclic 3',5'-nucleotide phosphodiesterase activities in Neurospora crassa.  

PubMed Central

Two molecular species of repressible extracellular phosphodiesterases showing cyclic 2',3'- and cyclic 3',5'-nucleotide phosphodiesterase activities were detected in mycelial culture media of wild-type Neurospora crassa and purified. The two molecular species were found to be monomeric and polymeric forms of an enzyme constituted of identical subunits having molecular weights of 50,000. This enzyme had the same electrophoretic mobility as repressible acid phosphatase. The enzyme designated repressible cyclic phosphodiesterase showed pH optima of 3.2 to 4.0 with a cyclic 3',5'-AMP substrate and 5.0 to 5.6 with a cyclic 2',3'-AMP substrate. Repressible cyclic phosphodiesterase was activated by MnCl2 and CoCl2 with cyclic 2',3'-AMP as substrate and was slightly activated by MnCl2 with cyclic 3',5'-AMP. The enzyme hydrolyzed cyclic 3',5'- and cyclic 2',3'-nucleotides, in addition to bis-rho-nitrophenyl phosphate, but not certain 5' -and 3'-nucleotides. 3'-GMP and 3'-CMP were hydrolyzed less efficiently. Mutant strains A1 (nuc-1) and B1 (nuc-2), which cannot utilize RNA or DNA as a sole source of phosphorus, were unable to produce repressible cyclic phosphodiesterase. The wild type (74A) and a heterocaryon between strains A1 and B1 produced the enzyme and showed growth on orthophosphate-free media containing cyclic 2',3'-AMP or cyclic 3',5'-AMP, whereas both mutants showed little or no growth on these media. Images

Hasunuma, K

1983-01-01

108

"The Show"  

ERIC Educational Resources Information Center

For the past 16 years, the blue-collar city of Huntington, West Virginia, has rolled out the red carpet to welcome young wrestlers and their families as old friends. They have come to town chasing the same dream for a spot in what many of them call "The Show". For three days, under the lights of an arena packed with 5,000 fans, the state's best…

Gehring, John

2004-01-01

109

The pronociceptive dorsal reticular nucleus contains mostly tonic neurons and shows a high prevalence of spontaneous activity in block preparation.  

PubMed

Despite the importance and significant clinical impact of understanding information processing in the nociceptive system, the functional properties of neurons in many parts of this system are still unknown. In this work we performed whole cell patch-clamp recording in rat brain stem blocks to characterize the electrophysiological properties of neurons in the dorsal reticular nucleus (DRt), a region known to be involved in pronociceptive modulation. We also compared properties of DRt neurons with those in the adjacent parvicellular reticular nucleus and in neighboring regions outside the reticular formation. We found that neurons in the DRt and parvicellular reticular nucleus had similar electrophysiological properties and exhibited mostly toniclike firing patterns, whereas neurons outside the reticular formation showed a larger diversity of firing patterns. Interestingly, more than one-half of the neurons also showed spontaneous activity. While the general view of the reticular formation, being a loosely associated mesh of groups of neurons with diverse function, and earlier reports suggests more electrophysiological heterogeneity, we showed that this is indeed not the case. Our results indicate that functional difference of neurons in the reticular formation may mostly be determined by their connectivity profiles and not by their intrinsic electrophysiological properties. The dominance of tonic neurons in the DRt supports previous conclusions that these neurons encode stimulus intensity through their firing frequency, while the high prevalence of spontaneous activity most likely shapes nociceptive modulation by this brain stem region. PMID:24431401

Sousa, Mafalda; Szucs, Peter; Lima, Deolinda; Aguiar, Paulo

2014-04-01

110

Discovery of LPMO activity on hemicelluloses shows the importance of oxidative processes in plant cell wall degradation.  

PubMed

The recently discovered lytic polysaccharide monooxygenases (LPMOs) are known to carry out oxidative cleavage of glycoside bonds in chitin and cellulose, thus boosting the activity of well-known hydrolytic depolymerizing enzymes. Because biomass-degrading microorganisms tend to produce a plethora of LPMOs, and considering the complexity and copolymeric nature of the plant cell wall, it has been speculated that some LPMOs may act on other substrates, in particular the hemicelluloses that tether to cellulose microfibrils. We demonstrate that an LPMO from Neurospora crassa, NcLPMO9C, indeed degrades various hemicelluloses, in particular xyloglucan. This activity was discovered using a glycan microarray-based screening method for detection of substrate specificities of carbohydrate-active enzymes, and further explored using defined oligomeric hemicelluloses, isolated polymeric hemicelluloses and cell walls. Products generated by NcLPMO9C were analyzed using high performance anion exchange chromatography and multidimensional mass spectrometry. We show that NcLPMO9C generates oxidized products from a variety of substrates and that its product profile differs from those of hydrolytic enzymes acting on the same substrates. The enzyme particularly acts on the glucose backbone of xyloglucan, accepting various substitutions (xylose, galactose) in almost all positions. Because the attachment of xyloglucan to cellulose hampers depolymerization of the latter, it is possible that the beneficial effect of the LPMOs that are present in current commercial cellulase mixtures in part is due to hitherto undetected LPMO activities on recalcitrant hemicellulose structures. PMID:24733907

Agger, Jane W; Isaksen, Trine; Várnai, Anikó; Vidal-Melgosa, Silvia; Willats, William G T; Ludwig, Roland; Horn, Svein J; Eijsink, Vincent G H; Westereng, Bjørge

2014-04-29

111

Cytochrome c oxidase subunit 4 isoform 2-knockout mice show reduced enzyme activity, airway hyporeactivity, and lung pathology  

PubMed Central

Cytochrome c oxidase (COX) is the terminal enzyme of the mitochondrial electron transport chain. The purpose of this study was to analyze the function of lung-specific cytochrome c oxidase subunit 4 isoform 2 (COX4i2) in vitro and in COX4i2-knockout mice in vivo. COX was isolated from cow lung and liver as control and functionally analyzed. COX4i2-knockout mice were generated and the effect of the gene knockout was determined, including COX activity, tissue energy levels, noninvasive and invasive lung function, and lung pathology. These studies were complemented by a comprehensive functional screen performed at the German Mouse Clinic (Neuherberg, Germany). We show that isolated cow lung COX containing COX4i2 is about twice as active (88 and 102% increased activity in the presence of allosteric activator ADP and inhibitor ATP, respectively) as liver COX, which lacks COX4i2. In COX4i2-knockout mice, lung COX activity and cellular ATP levels were significantly reduced (?50 and ?29%, respectively). Knockout mice showed decreased airway responsiveness (60% reduced Penh and 58% reduced airway resistance upon challenge with 25 and 100 mg methacholine, respectively), and they developed a lung pathology deteriorating with age that included the appearance of Charcot-Leyden crystals. In addition, there was an interesting sex-specific phenotype, in which the knockout females showed reduced lean mass (?12%), reduced total oxygen consumption rate (?8%), improved glucose tolerance, and reduced grip force (?14%) compared to wild-type females. Our data suggest that high activity lung COX is a central determinant of airway function and is required for maximal airway responsiveness and healthy lung function. Since airway constriction requires energy, we propose a model in which reduced tissue ATP levels explain protection from airway hyperresponsiveness, i.e., absence of COX4i2 leads to reduced lung COX activity and ATP levels, which results in impaired airway constriction and thus reduced airway responsiveness; long-term lung pathology develops in the knockout mice due to impairment of energy-costly lung maintenance processes; and therefore, we propose mitochondrial oxidative phosphorylation as a novel target for the treatment of respiratory diseases, such as asthma.—Hüttemann, M., Lee, I., Gao, X., Pecina, P., Pecinova, A., Liu, J., Aras, S., Sommer, N., Sanderson, T. H., Tost, M., Neff, F., Aguilar-Pimentel, J. A., Becker, L., Naton, B., Rathkolb, B., Rozman, J., Favor, J., Hans, W., Prehn, C., Puk, O., Schrewe, A., Sun, M., Höfler, H., Adamski, J., Bekeredjian, R., Graw, J., Adler, T., Busch, D. H., Klingenspor, M., Klopstock, T., Ollert, M., Wolf, E., Fuchs, H., Gailus-Durner, V., Hrab? de Angelis, M., Weissmann, N., Doan, J. W., Bassett, D. J. P., Grossman, L. I. Cytochrome c oxidase subunit 4 isoform 2-knockout mice show reduced enzyme activity, airway hyporeactivity, and lung pathology.

Huttemann, Maik; Lee, Icksoo; Gao, Xiufeng; Pecina, Petr; Pecinova, Alena; Liu, Jenney; Aras, Siddhesh; Sommer, Natascha; Sanderson, Thomas H.; Tost, Monica; Neff, Frauke; Aguilar-Pimentel, Juan Antonio; Becker, Lore; Naton, Beatrix; Rathkolb, Birgit; Rozman, Jan; Favor, Jack; Hans, Wolfgang; Prehn, Cornelia; Puk, Oliver; Schrewe, Anja; Sun, Minxuan; Hofler, Heinz; Adamski, Jerzy; Bekeredjian, Raffi; Graw, Jochen; Adler, Thure; Busch, Dirk H.; Klingenspor, Martin; Klopstock, Thomas; Ollert, Markus; Wolf, Eckhard; Fuchs, Helmut; Gailus-Durner, Valerie; Hrabe de Angelis, Martin; Weissmann, Norbert; Doan, Jeffrey W.; Bassett, David J. P.; Grossman, Lawrence I.

2012-01-01

112

Map showing recently active breaks along the San Andreas Fault between Pt. Delgada and Bolinas Bay, California  

USGS Publications Warehouse

This strip map is one of a series of maps showing recently active fault breaks along the San Andreas and other active faults in California. It is designed to inform persons who are concerned with land use near the fault of the location of those fault breaks that have moved recently. The lines on the map are lines of rupture and creep that can be identified by field evidence and that clearly affect the present surface of the land. Map users should keep in mind that these lines are intended primarily as guides to help locate the fault; the mapped lines are not necessarily shown with the precision demanded by some engineering or land utilization needs.

Brown, Robert D., Jr.; Wolfe, Edward W.

1970-01-01

113

Schiff base oxovanadium(IV) complexes of phenanthroline bases showing DNA photocleavage activity at near-IR light and photocytotoxicity  

Microsoft Academic Search

Oxovanadium(IV) complexes [VO(L)(B)](ClO4) (1–3) of N-2-pyridylmethylidine-2-hydroxyphenylamine (HL) Schiff base and phenanthroline bases (B), viz. 1,10-phenanthroline (phen in 1), dipyrido[3,2-d:2?,3?-f]quinoxaline (dpq in 2) or dipyrido[3,2-a:2?,3?-c]phenazine (dppz in 3), were prepared, characterized and their DNA binding property, photo-induced DNA cleavage activity and photocytotoxicity in HeLa cells studied. The crystal structure of 1 shows the presence of a VO2+ moiety in VO2N4 coordination

Puja Prasad; Pijus K. Sasmal; Imran Khan; Paturu Kondaiah; Akhil R. Chakravarty

2011-01-01

114

Origanum vulgare subsp. hirtum essential oil prevented biofilm formation and showed antibacterial activity against planktonic and sessile bacterial cells.  

PubMed

Essential oils from six different populations of Origanum vulgare subsp. hirtum were compared for their antibiofilm properties. The six essential oils (A to F) were characterized by a combination of gas chromatography with flame ionization detector and gas chromatography with mass spectrometer detector analyses. All oils showed weak activity against the planktonic form of a group of Staphylococcus aureus strains and against a Pseudomonas aeruginosa ATCC 15442 reference strain. The ability to inhibit biofilm formation was investigated at sub-MIC levels of 200, 100, and 50 m g/ml by staining sessile cells with safranin. Sample E showed the highest average effectiveness against all tested strains at 50 m g/ml and had inhibition percentages ranging from 30 to 52%. In the screening that used preformed biofilm from the reference strain P. aeruginosa, essential oils A through E were inactive at 200 m g/ml; F was active with a percentage of inhibition equal to 53.2%. Oregano essential oil can inhibit the formation of biofilms of various food pathogens and food spoilage organisms. PMID:24112575

Schillaci, Domenico; Napoli, Edoardo Marco; Cusimano, Maria Grazia; Vitale, Maria; Ruberto, Andgiuseppe

2013-10-01

115

Do different implant surfaces exposed in the oral cavity of humans show different biofilm compositions and activities?  

PubMed

Osseointegrated dental implants play an important role in restorative dentistry. However, plaque accumulation may cause inflammatory reactions around the implants, sometimes leading to implant failure. In this in vivo study the influence of two physical hard coatings on bacterial adhesion was examined in comparison with a pure titanium surface. Thin glass sheets coated with titanium nitride (TiN), zirconium nitride (ZrN) or pure titanium were mounted on removable intraoral splints in two adults. After 60 h of intraoral exposure, the biofilms were analyzed to determine the number of bacteria, the types of bacteria [by applying single-strand conformation polymorphism (SSCP analysis) of 16S rRNA genes], and whether or not the bacteria were active (by SSCP analysis of 16S rRNA). The results showed that bacterial cell counts were higher on the pure titanium-coated glass sheets than on the glass sheets coated with TiN or ZrN. The lowest number of bacterial cells was present on theZrN-coated glass. However, the metabolic activity (RNA fingerprints) of bacteria on TiN- and ZrN-coated glass sheets seemed to be lower than the activity of bacteria on the titanium-coated surfaces, whereas SSCP fingerprints based on 16S rDNA revealed that the major 16S bands are common to all of the fingerprints, independently of the surface coating. PMID:15560835

Groessner-Schreiber, Birte; Hannig, Matthias; Dück, Alexander; Griepentrog, Michael; Wenderoth, Dirk F

2004-12-01

116

C(5) modified uracil derivatives showing antiproliferative and erythroid differentiation inducing activities on human chronic myelogenous leukemia K562 cells  

PubMed Central

The K562 cell line has been proposed as a useful experimental system to identify anti-tumor compounds acting by inducing terminal erythroid differentiation. K562 cells exhibit a low proportion of hemoglobin-synthesizing cells under standard cell growth conditions, but are able to undergo terminal erythroid differentiation when treated with a variety of anti-tumor compounds. In this paper we report a screening study on a set of different modified C(5) uracil derivatives for the evaluation of their antiproliferative effect in connection with erythroid differentiation pathways, and for defining a new class of drug candidates for the treatment of chronic myelogenous leukemia. Activity of the derivatives tested can be classified in two effect: an antiproliferative effect linked to a high level of erythroid differentiation activity and an antiproliferative effect without activation of gamma globin genes The highest antiproliferative effect and erythroid induction was shown by compound 9, a thymine derivative bearing a n-octyl chain on nitrogen N(1), whereas thymine did not show any effect, suggesting the importance of the linear alkyl chain in position N(1). To our knowledge this compound should be considered among the most efficient inducers of erythroid differentiation of K562 cells. This work is the starting point for the quest of more effective and specific drugs for the induction of terminal erythroid differentiation, for leading new insights in the treatment of neoplastic diseases with molecules acting by inducing differentiation rather than by simply exerting cytotoxic effects.

Brognara, Eleonora; Lampronti, Ilaria; Breveglieri, Giulia; Accetta, Alessandro; Corradini, Roberto; Manicardi, Alex; Borgatti, Monica; Canella, Alessandro; Multineddu, Chiara; Marchelli, Rosangela; Gambari, Roberto

2011-01-01

117

Fusion of protegrin-1 and plectasin to MAP30 shows significant inhibition activity against dengue virus replication.  

PubMed

Dengue virus (DENV) broadly disseminates in tropical and sub-tropical countries and there are no vaccine or anti-dengue drugs available. DENV outbreaks cause serious economic burden due to infection complications that requires special medical care and hospitalization. This study presents a new strategy for inexpensive production of anti-DENV peptide-fusion protein to prevent and/or treat DENV infection. Antiviral cationic peptides protegrin-1 (PG1) and plectasin (PLSN) were fused with MAP30 protein to produce recombinant antiviral peptide-fusion protein (PG1-MAP30-PLSN) as inclusion bodies in E. coli. High yield production of PG1-MAP30-PLSN protein was achieved by solubilization of inclusion bodies in alkaline buffer followed by the application of appropriate refolding techniques. Antiviral PG1-MAP30-PLSN protein considerably inhibited DENV protease (NS2B-NS3pro) with half-maximal inhibitory concentration (IC50) 0.5±0.1 ?M. The real-time proliferation assay (RTCA) and the end-point proliferation assay (MTT assay) showed that the maximal-nontoxic dose of the peptide-fusion protein against Vero cells is approximately 0.67±0.2 ?M. The cell-based assays showed considerable inhibition of the peptide-fusion protein against binding and proliferating stages of DENV2 into the target cells. The peptide-fusion protein protected DENV2-challeged mice with 100% of survival at the dose of 50 mg/kg. In conclusion, producing recombinant antiviral peptide-fusion protein by combining short antiviral peptide with a central protein owning similar activity could be useful to minimize the overall cost of short peptide production and take advantage of its synergistic antiviral activities. PMID:24722532

Rothan, Hussin A; Bahrani, Hirbod; Mohamed, Zulqarnain; Abd Rahman, Noorsaadah; Yusof, Rohana

2014-01-01

118

Fusion of Protegrin-1 and Plectasin to MAP30 Shows Significant Inhibition Activity against Dengue Virus Replication  

PubMed Central

Dengue virus (DENV) broadly disseminates in tropical and sub-tropical countries and there are no vaccine or anti-dengue drugs available. DENV outbreaks cause serious economic burden due to infection complications that requires special medical care and hospitalization. This study presents a new strategy for inexpensive production of anti-DENV peptide-fusion protein to prevent and/or treat DENV infection. Antiviral cationic peptides protegrin-1 (PG1) and plectasin (PLSN) were fused with MAP30 protein to produce recombinant antiviral peptide-fusion protein (PG1-MAP30-PLSN) as inclusion bodies in E. coli. High yield production of PG1-MAP30-PLSN protein was achieved by solubilization of inclusion bodies in alkaline buffer followed by the application of appropriate refolding techniques. Antiviral PG1-MAP30-PLSN protein considerably inhibited DENV protease (NS2B-NS3pro) with half-maximal inhibitory concentration (IC50) 0.5±0.1 ?M. The real-time proliferation assay (RTCA) and the end-point proliferation assay (MTT assay) showed that the maximal-nontoxic dose of the peptide-fusion protein against Vero cells is approximately 0.67±0.2 ?M. The cell-based assays showed considerable inhibition of the peptide-fusion protein against binding and proliferating stages of DENV2 into the target cells. The peptide-fusion protein protected DENV2-challeged mice with 100% of survival at the dose of 50 mg/kg. In conclusion, producing recombinant antiviral peptide-fusion protein by combining short antiviral peptide with a central protein owning similar activity could be useful to minimize the overall cost of short peptide production and take advantage of its synergistic antiviral activities.

Rothan, Hussin A.; Bahrani, Hirbod; Mohamed, Zulqarnain; Abd Rahman, Noorsaadah; Yusof, Rohana

2014-01-01

119

Nanoencapsulation of tacrolimus in lipid-core nanocapsules showed similar immunosuppressive activity after oral and intraperitoneal administrations.  

PubMed

Tacrolimus is widely used in the prophylaxis of solid-organ transplant rejection. Several studies have reported that tacrolimus has variable and poor bioavailability after oral administration, apart from adverse effects such as gastrointestinal disorders, hyperglycemia, nephro- and neurotoxicity. The aim of this work was to encapsulate tacrolimus (TAC) in lipid-core nanocapsules (LNC) as an oral strategy to deliver the drug. To validate our hypothesis, the pharmacodynamic effect of TAC-LNC was determined after oral and intraperitoneal (i.p.) administrations to mice. TAC-LNC had z-average diameter of 210 nm (unimodal), and 99.5% of encapsulation efficiency. In vitro sustained release was determined for TAC-LNC fitting an anomalous transport mechanism (n = 0.8). TAC-LNC demonstrated higher immunosuppressive activity after oral and i.p. administrations, when compared to the drug solution. TAC-LNC administered at 6.0 mg kg(-1) day(-1) showed equivalent percent reduction in lymphocyte when both routes of administration were used. After oral administration, drug nanoencapsulation allows reducing the dose by at least 40%. Furthermore, the nanoencapsulation of TAC in lipid-core nanocapsules showed pharmacodynamic effect similar for the oral and the i.p. routes. In conclusion, the lipid-core nanocapsules were able to improve the TAC deliver across the oral absorption barrier. PMID:25016659

Friedrich, R B; Dimer, F A; Guterres, S S; Beck, R C R; Pohlmann, A R

2014-08-01

120

Molecular and biochemical characterization of AtPAP15, a purple acid phosphatase with phytase activity, in Arabidopsis.  

PubMed

Purple acid phosphatase (PAP) catalyzes the hydrolysis of phosphate monoesters and anhydrides to release phosphate within an acidic pH range. Among the 29 PAP-like proteins in Arabidopsis (Arabidopsis thaliana), AtPAP15 (At3g07130) displays a greater degree of amino acid identity with soybean (Glycine max; GmPHY) and tobacco (Nicotiana tabacum) PAP (NtPAP) with phytase activity than the other AtPAPs. In this study, transgenic Arabidopsis that expressed an AtPAP15 promoterbeta-glucuronidase (GUS) fusion protein showed that AtPAP15 expression was developmentally and temporally regulated, with strong GUS staining at the early stages of seedling growth and pollen germination. The expression was also organ/tissue specific, with strongest GUS staining in the vasculature, pollen grains, and roots. The recombinant AtPAP purified from transgenic tobacco exhibited broad substrate specificity with moderate phytase activity. AtPAP15 T-DNA insertion lines exhibited a lower phytase and phosphatase activity in seedling and germinating pollen and lower pollen germination rate compared with the wild type and their complementation lines. Therefore, AtPAP15 likely mobilizes phosphorus reserves in plants, particularly during seed and pollen germination. Since AtPAP15 is not expressed in the root hair or in the epidermal cells, it is unlikely to play any role in external phosphorus assimilation. PMID:19633233

Kuang, Ruibin; Chan, Kam-Ho; Yeung, Edward; Lim, Boon Leong

2009-09-01

121

Evaluation of About Being Active, an online lesson about physical activity shows that perception of being physically active is higher in eating competent low-income women  

PubMed Central

Background Eating competence (EC) has been associated with positive health outcomes such as reduced cardiovascular risk and higher diet quality. This study compared reported physical activity and EC in 512 low-income women participating in an online program that included a physical activity lesson and assessed response to this lesson. Methods Educational intervention and surveys were completed online. EC was assessed with the Satter Eating Competence Inventory for Low-Income (ecSI/LI). Results Participants were mostly white, <31 years, overweight/obese (60%), and food insecure (58%). EC was higher for those who self-reported being physically active (30.1 ± 8.3 vs. 24.9 ± 8.1; P<0.001) and were active for ? 30 minutes/day (29.9 ± 8.3 vs. 26.3 ± 8.6), even with age, weight satisfaction, and BMI controlled. EC of obese physically active persons was higher than normal weight, but physically inactive women. The physical activity module was well received with responses unrelated to time involved or physical activity level. Conclusions Low-income women were interested in learning about physical activity and responded positively to online delivery. Overall EC levels were low, but higher for physically active women, supporting efforts to enhance EC. Additional research is needed to determine if EC is associated with responses to physical activity education.

2013-01-01

122

An EGFR/CD13 bispecific fusion protein and its enediyne-energized analog show potent antitumor activity.  

PubMed

Targeting to two or more objectives simultaneously has been pursued as a strategy to potentially increase the efficiency and selectivity of targeted drugs to certain cancers. In this study, an epidermal growth factor receptor (EGFR)/CD13-targeting, bispecific fusion protein ER(Fv)-LDP-NGR consisting of an anti-EGFR single-chain variable fragment (scFv), an apoprotein (LDP) of lidamycin (LDM), and a tri-CNGRC (Cys-Asn-Gly-Arg-Cys) peptide against CD13 was constructed, and then an enediyne-energized analog ER(Fv)-LDP-NGR-AE was generated by integration with an enediyne chomophore (AE) derived from LDM. The apoprotein LDP was used as a 'scaffold' to connect the scFv fragment and the tri-CNGRC peptide and also served as a specific 'carrier' for the extremely potent cytotoxic enediyne chromophore of LDM. Compared with its monospecific counterparts, ER(Fv)-LDP and LDP-NGR, the bispecific fusion protein ER(Fv)-LDP-NGR showed higher affinity to EGFR/CD13-overexpressed tumor cells. Determined by the MTT assay, the bispecific, enediyne-energized ER(Fv)-LDP-NGR-AE showed highly potent cytotoxicity to EGFR/CD13-overexpressed MCF-7 cells, with an IC50 value of 3.4×10 mol/l, whereas for the EGFR-overexpressed A431 cells, the IC50 value was 2.2×10 mol/l. For MCF-7 cells, the bispecific ER(Fv)-LDP-NGR-AE was more potent in cytotoxicity than the corresponding monospecific energized fusion proteins. In athymic mice models, the bispecific fusion protein ER(Fv)-LDP-NGR presented stronger inhibitory activity than the monospecific ER(Fv)-LDP and LDP-NGR. For the enediyne-energized fusion proteins, ER(Fv)-LDP-NGR-AE significantly inhibited the growth of EGFR/CD13-overexpressed MCF-7 xenograft and EGFR-overexpressed A431 xenograft by 86.3 and 81.4%, respectively. In addition, the bispecific ER(Fv)-LDP-NGR-AE showed much higher efficacy than its monospecific analogs ER(Fv)-LDP-AE and LDP-NGR-AE in both MCF-7 and A431 xenograft models. The results show that EGFR/CD13 bitargeting effectively improved the antitumor efficacy. Both the bispecific fusion protein and its enediyne-energized analog are highly effective in athymic mice bearing xenografts, and the latter exerts more marked efficacy. Generation of a pair of bispecific antibody-based therapeutics and its corresponding antibody-drug conjugate simultaneously may be a feasible strategy for the development of new targeted drugs for cancer therapy. PMID:24100279

Sheng, Weijin; Shang, Yue; Li, Liang; Zhen, Yongsu

2014-01-01

123

Telavancin shows superior activity to vancomycin with multidrug-resistant Staphylococcus aureus in a range of in vitro biofilm models.  

PubMed

The activity of telavancin was compared with vancomycin against methicillin-resistant Staphylococcus aureus (MRSA) in planktonic culture and biofilms grown using a range of in vitro models. Antibiotic efficacy was determined using 24 clinical isolates, including healthcare-associated (HA)-MRSA, community-associated (CA)-MRSA and isolates with reduced (intermediate) susceptibility to vancomycin (VISA). Activity against biofilms was compared using three models: 96-peg plates, 96-well flat-bottom plates and a flow-cell system. Cell death was evaluated using a metabolic dye and Live/Dead staining. The planktonic minimum inhibitory concentration (MIC) range for telavancin was lower than that for vancomycin (0.06-0.25 mg/l and 0.5-8 mg/l, respectively). Vancomycin (100?×?MIC) killed, on average, 59% of cells in HA-MRSA biofilms grown on 96-peg plates, 44% of cells in CA-MRSA biofilms and 26% of cells in VISA biofilms. Telavancin (100?×?MIC) killed, on average, 63%, 49% and 41% of cells, respectively. The antibiotics showed similar efficacy against MRSA biofilms but telavancin was more effective against those formed by VISA isolates. In the flow-cell system, antibiotic cell killing was enhanced with both antibiotics, killing up to 80% of biofilm-associated cells. The variance in cell killing displayed when biofilms were grown using different systems highlights the importance of selecting an appropriate model for antimicrobial efficacy tests. The flow-cell system more closely reflects conditions encountered during infection and is possibly more clinically relevant than a 96-well plate system. Despite differences between the models evaluated, telavancin typically demonstrated improved efficacy over vancomycin, indicating the potential value of the agent in the treatment of biofilm-mediated infections caused by S. aureus, especially multidrug-resistant isolates. PMID:23624635

Smith, K; Gemmell, C G; Lang, S

2013-10-01

124

Microfluidic Single Cell Analysis Show Porcine Induced Pluripotent Stem Cell-Derived Endothelial Cells Improve Myocardial Function by Paracrine Activation  

PubMed Central

Rationale Induced pluripotent stem cells (iPSCs) hold great promise for the development of patient-specific therapies for cardiovascular disease. However, clinical translation will require preclinical optimization and validation of large animal iPSC models. Objective To successfully derive endothelial cells from porcine iPSCs and demonstrate their potential utility for the treatment of myocardial ischemia. Methods and Results Porcine adipose stromal cells were reprogrammed to generate porcine iPSCs (piPSCs). Immunohistochemistry, quantitative PCR, microarray hybridization, and angiogenic assays confirmed that piPSC-derived endothelial cells (piPSC-ECs) shared similar morphological and functional properties as endothelial cells isolated from the autologous pig aorta. To demonstrate their therapeutic potential, piPSC-ECs were transplanted into mice with myocardial infarction (MI). Compared to control, animals transplanted with piPSC-ECs showed significant functional improvement measured by echocardiography (fractional shortening at week 4: 27.2±1.3% vs. 22.3±1.1%; P<0.001) and magnetic resonance imaging (ejection fraction at week 4: 45.8±1.3% vs. 42.3±0.9%; P<0.05). Quantitative protein assays and microfluidic single cell PCR profiling showed that piPSC-ECs released pro-angiogenic and anti-apoptotic factors in the ischemic microenvironment, which promoted neovascularization and cardiomyocyte survival, respectively. Release of paracrine factors varied significantly among subpopulations of transplanted cells, suggesting that transplantation of specific cell populations may result in greater functional recovery. Conclusion In summary, this is the first study to successfully differentiate piPSCs-ECs from piPSCs and demonstrate that transplantation of piPSC-ECs improved cardiac function following MI via paracrine activation. Further development of these large animal iPSC models will yield significant insights into their therapeutic potential and accelerate the clinical translation of autologous iPSC-based therapy.

Gu, Mingxia; Nguyen, Patricia K.; Lee, Andrew S.; Xu, Dan; Hu, Shijun; Plews, Jordan R; Han, Leng; Huber, Bruno C.; Lee, Won Hee; Gong, Yongquan; de Almeida, Patricia E.; Lyons, Jennifer; Ikeno, Fumi; Pacharinsak, Cholawat; Connolly, Andrew J.; Gambhir, Sanjiv S.; Robbins, Robert C.; Longaker, Michael T.; Wu, Joseph C.

2012-01-01

125

Topically Delivered Adipose Derived Stem Cells Show an Activated-Fibroblast Phenotype and Enhance Granulation Tissue Formation in Skin Wounds  

PubMed Central

Multipotent mesenchymal stem cells (MSCs) are found in various tissues and can proliferate extensively in vitro. MSCs have been used in preclinical animal studies and clinical trials in many fields. Adipose derived stem cells (ASCs) have several advantages compared to other MSCs for use in cell-based treatments because they are easy to isolate with relative abundance. However, quantitative approaches for wound repair using ASCs have been limited because of lack of animal models which allow for quantification. Here, we addressed the effect of topically delivered ASCs in wound repair by quantitative analysis using the rabbit ear model. We characterized rabbit ASCs, and analyzed their multipotency in comparison to bone marrow derived-MSCs (BM-MSCs) and dermal fibroblasts (DFs) in vitro. Topically delivered ASCs increased granulation tissue formation in wounds when compared to saline controls, whereas BM-MSCs or DFs did not. These studies suggest that ASCs and BM-MSCs are not identical, though they have similar surface markers. We found that topically delivered ASCs are engrafted and proliferate in the wounds. We showed that transplanted ASCs exhibited activated fibroblast phenotype, increased endothelial cell recruitment, and enhanced macrophage recruitment in vivo.

Hong, Seok Jong; Xu, Wei; Leung, Kai P.; Mustoe, Thomas A.; Galiano, Robert D.

2013-01-01

126

Sertaconazole Nitrate Shows Fungicidal and Fungistatic Activities against Trichophyton rubrum, Trichophyton mentagrophytes, and Epidermophyton floccosum, Causative Agents of Tinea Pedis?  

PubMed Central

The fungistatic and fungicidal activities of sertaconazole against dermatophytes were evaluated by testing 150 clinical isolates of causative agents of tinea pedis, Trichophyton rubrum, Trichophyton mentagrophytes, and Epidermophyton floccosum. The overall geometric means for fungistatic and fungicidal activities of sertaconazole against these isolates were 0.26 and 2.26 ?g/ml, respectively, although values were higher for T. mentagrophytes than for the others. This is the first comprehensive demonstration of the fungicidal activity of sertaconazole against dermatophytes.

Carrillo-Munoz, Alfonso J.; Tur-Tur, Cristina; Cardenes, Delia C.; Estivill, Dolors; Giusiano, Gustavo

2011-01-01

127

Sertaconazole nitrate shows fungicidal and fungistatic activities against Trichophyton rubrum, Trichophyton mentagrophytes, and Epidermophyton floccosum, causative agents of tinea pedis.  

PubMed

The fungistatic and fungicidal activities of sertaconazole against dermatophytes were evaluated by testing 150 clinical isolates of causative agents of tinea pedis, Trichophyton rubrum, Trichophyton mentagrophytes, and Epidermophyton floccosum. The overall geometric means for fungistatic and fungicidal activities of sertaconazole against these isolates were 0.26 and 2.26 ?g/ml, respectively, although values were higher for T. mentagrophytes than for the others. This is the first comprehensive demonstration of the fungicidal activity of sertaconazole against dermatophytes. PMID:21746955

Carrillo-Muñoz, Alfonso J; Tur-Tur, Cristina; Cárdenes, Delia C; Estivill, Dolors; Giusiano, Gustavo

2011-09-01

128

The expression pattern of alfalfa flavanone 3-hydroxylase promoter-gus fusion in Nicotiana benthamiana correlates with the presence of flavonoids detected in situ  

Microsoft Academic Search

Flavanone 3-hydroxylase is an enzyme acting in the central part of the flavonoid biosynthesis pathway. It is generally encoded by a single gene and seems to have a key position for the regulation in this pathway. These two features make a single f3h promoter-gus fusion a suitable tool to study both the f3h expression and the regulation of this pathway.

Bénédicte Charrier; Christine Leroux; Adam Kondorosi; Pascal Ratet

1996-01-01

129

Archive of SID Isolation and characterization of thermophilic alkaline proteases resistant to sodium dodecyl sulfate and ethyl- ene diamine tetraacetic acid from Bacillus sp. GUS1  

Microsoft Academic Search

Thermophilic Bacillus sp. GUS1, isolated from a soil sample obtained from citrus garden, produced at least three proteases as detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and zymogram analysis. The enzymes were stable in the alkaline pH range (8.0-12.0), with the optimum temper- ature and pH range of the proteases being 70ºC and 6.0-12.0, respectively. All three proteases

Sara Seifzadeh; Reza Hassan Sajedi; Reyhaneh Sariri

2008-01-01

130

Bifunctional gfp-and gusA-containing mini-Tn 5 transposon derivatives for combined gene expression and bacterial localization studies  

Microsoft Academic Search

The gfp gene, encoding the green fluorescent protein, was combined with the gusA gene, coding for the ?-glucuronidase enzyme, in mini-Tn5 transposon derivatives for use in Gram-negative bacteria. These mini-Tn5 elements allow simultaneously monitoring of gene expression and localization of the marked bacteria. Introduction of the resultant mini-Tn5 transposons into Rhizobium etli, Azospirillum brasilense and Pseudomonas stutzeri allowed us to

Chuanwu Xi; Mark Lambrecht; Jos Vanderleyden; Jan Michiels

1999-01-01

131

Development of gusA reporter gene constructs for cereal transformation: Availability of plant transformation vectors from the CAMBIA Molecular Genetic Resource Service  

Microsoft Academic Search

The use of reporter genes to characterise sequence elements that act to regulate gene expression in transgenic plants has been vital to the development of foreign gene expression strategies for use in cereal transformation. ThegusA locus ofEscherichia coli, which encodes the enzymeß-glucuronidase (GUS), is by far the most popular reporter gene used in plant transformation. In this paper we extend

David McElroy; Douglas A. Chamberlain; Eunpyo Moon; Kate J. Wilson

1995-01-01

132

Spatial and temporal patterns of GUS expression directed by 5? regions of the Arabidopsis thaliana farnesyl diphosphate synthase genes FPS1 and FPS2  

Microsoft Academic Search

Farnesyl diphosphate synthase (FPS), the enzyme that catalyses the synthesis of farnesyl diphosphate (FPP) from isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), is considered a regulatory enzyme of plant isoprenoid biosynthesis. The promoter regions of the FPS1 and FPS2 genes controlling the expression of isoforms FPS1S and FPS2, respectively, were fused to the ß-glucuronidase (GUS) reporter gene and introduced into

Núria Cunillera; Albert Boronat; Albert Ferrer

2000-01-01

133

Mice Deficient for the Ets Transcription Factor Elk1 Show Normal Immune Responses and Mildly Impaired Neuronal Gene Activation  

Microsoft Academic Search

The transcription factor Elk-1 belongs to the ternary complex factor (TCF) subfamily of Ets proteins. TCFs interact with serum response factor to bind jointly to serum response elements in the promoters of immediate- early genes (IEGs). TCFs mediate the rapid transcriptional response of IEGs to various extracellular stimuli which activate mitogen-activated protein kinase signaling. To investigate physiological functions of Elk-1

Francesca Cesari; Stephan Brecht; Kristina Vintersten; Lam Giang Vuong; Matthias Hofmann; Karin Klingel; Jens-Jorg Schnorr; Sergei Arsenian; Hansjorg Schild; Thomas Herdegen; Franziska F. Wiebel; Alfred Nordheim

2004-01-01

134

Imidazopyridine-Based Fatty Acid Synthase Inhibitors That Show Anti-HCV Activity and in Vivo Target Modulation  

PubMed Central

Potent imidazopyridine-based inhibitors of fatty acid synthase (FASN) are described. The compounds are shown to have antiviral (HCV replicon) activities that track with their biochemical activities. The most potent analogue (compound 19) also inhibits rat FASN and inhibits de novo palmitate synthesis in vitro (cell-based) as well as in vivo.

2012-01-01

135

Immunomodulatory Activity of Dietary Fiber: Arabinoxylan and Mixed-Linked Beta-Glucan Isolated from Barley Show Modest Activities in Vitro  

PubMed Central

High intake of dietary fiber is claimed to protect against development of colorectal cancer. Barley is a rich source of dietary fiber, and possible immunomodulatory effects of barley polysaccharides might explain a potential protective effect. Dietary fiber was isolated by extraction and enzyme treatment. A mixed-linked ?-glucan (WSM-TPX, 96.5% ?-glucan, Mw 886 kDa), an arabinoxylan (WUM-BS-LA, 96.4% arabinoxylan, Mw 156 kDa), a mixed-linked ?-glucan rich fraction containing 10% arabinoxylan (WSM-TP) and an arabinoxylan rich fraction containing 30% mixed-linked ?-glucan (WUM-BS) showed no significant effect on IL-8 secretion and proliferation of two intestinal epithelial cell lines, Caco-2 and HT-29, and had no significant effect on the NF-?B activity in the monocytic cell line U937-3?B-LUC. Further enriched arabinoxylan fractions (WUM-BS-LA) from different barley varieties (Tyra, NK96300, SB94897 and CDCGainer) were less active than the mixed-linked ?-glucan rich fractions (WSM-TP and WSM-TPX) in the complement-fixing test. The mixed-linked ?-glucan rich fraction from NK96300 and CDCGainer showed similar activities as the positive control while mixed-linked ?-glucan rich fractions from Tyra and SB94897 were less active. From these results it is concluded that the isolated high molecular weight mixed-linked ?-glucans and arabinoxylans from barley show low immunological responses in selected in vitro test systems and thus possible anti-colon cancer effects of barley dietary fiber cannot be explained by our observations.

Samuelsen, Anne Berit; Rieder, Anne; Grimmer, Stine; Michaelsen, Terje E.; Knutsen, Svein H.

2011-01-01

136

Gender-linked Differences in the Toys, Television Shows, Computer Games, and Outdoor Activities of 5- to 13-year-old Children  

Microsoft Academic Search

This study was designed to compare how 5- to 13-year-old children's leisure activity preferences differ with age and gender. Responses from 60 boys and 60 girls about their favorite toys, television shows, computer games, and outdoor activities were compared across leisure categories. The results showed that gender was a significant factor. Overall, boys spent more time in these leisure activities

Isabelle D. Cherney; Kamala London

2006-01-01

137

Ultrastructural characterization of cationic liposome-DNA complexes showing enhanced stability in serum and high transfection activity in vivo  

Microsoft Academic Search

We have investigated the morphology and transfection activity of cationic liposome-DNA complexes (CLDC) under conditions relevant to both in vivo and in vitro studies. Moreover we have attempted to establish structure-function relationships relevant for high transfection activities under both conditions. CLDC were composed of dimethyldioctadecylammonium bromide with either 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) or cholesterol (Chol) interacting either with pre-condensed DNA or with

Brigitte Sternberg; Keelung Hong; Weiwen Zheng; Demetrios Papahadjopoulos

1998-01-01

138

Outer membrane protein A deficient Escherichia coli activates neutrophils to produce superoxide and shows increased susceptibility to antibacterial peptides  

Microsoft Academic Search

The outer membrane protein A (OmpA) of Gram-negative bacteria has been ascribed multiple functions including maintenance of structural membrane integrity and porin activity. OmpA has also been implicated in various host defense processes in that it contributes to bacterial serum resistance and activates certain immune cells. Recently, OmpA was shown to be the molecular target for neutrophil elastase (NE), and

Huamei Fu; Abderr Azzaq Belaaouaj; Claes Dahlgren; Johan Bylund

2003-01-01

139

In Vitro Analysis of Albendazole Sulfoxide Enantiomers Shows that (+)-(R)-Albendazole Sulfoxide Is the Active Enantiomer against Taenia solium  

PubMed Central

Albendazole is an anthelmintic drug widely used in the treatment of neurocysticercosis (NCC), an infection of the brain with Taenia solium cysts. However, drug levels of its active metabolite, albendazole sulfoxide (ABZSO), are erratic, likely resulting in decreased efficacy and suboptimal cure rates in NCC. Racemic albendazole sulfoxide is composed of ABZSO (+)-(R)- and (?)-(S) enantiomers that have been shown to differ in pharmacokinetics and activity against other helminths. The antiparasitic activities of racemic ABZSO and its (+)-(R)- and (?)-(S) enantiomers against T. solium cysts were evaluated in vitro. Parasites were collected from naturally infected pigs, cultured, and exposed to the racemic mixture or to each enantiomer (range, 10 to 500 ng/ml) or to praziquantel as a reference drug. The activity of each compound against cysts was assayed by measuring the ability to evaginate and inhibition of alkaline phosphatase (AP) and parasite antigen release. (+)-(R)-ABZSO was significantly more active than (?)-(S)-ABZSO in suppressing the release of AP and antigen into the supernatant in a dose- and time-dependent manner, indicating that most of the activity of ABZSO resides in the (+)-(R) enantiomer. Use of this enantiomer alone may lead to increased efficacy and/or less toxicity compared to albendazole.

Paredes, Adriana; de Campos Lourenco, Tiago; Marzal, Miguel; Rivera, Andrea; Dorny, Pierre; Mahanty, Siddhartha; Guerra-Giraldez, Cristina; Garcia, Hector H.; Cass, Quezia B.

2013-01-01

140

Structure-activity relationship study of antimicrobial dermaseptin S4 showing the consequences of peptide oligomerization on selective cytotoxicity.  

PubMed

To understand how peptide organization in aqueous solution might affect the activity of antimicrobial peptides, the potency of various dermaseptin S4 analogs was assessed against human red blood cells (RBC), protozoa, and several Gram-negative bacteria. Dermaseptin S4 had weak antibacterial activity but potent hemolytic or antiprotozoan effects. K(4)K(20)-S4 was 2-3-fold more potent against protozoa and RBC, yet K(4)K(20)-S4 was more potent by 2 orders of magnitude against bacteria. K(4)-S4 had similar behavior as K(4)K(20)-S4, but K(20)-S4 and analogous negative charge substitutions were as active as dermaseptin S4 or had reduced activity. Binding experiments suggested that potency enhancement was not the result of increased affinity to target cells. In contrast, potency correlated well with aggregation properties. Fluorescence studies indicated that K(20)-S4 and all negative charge substitutions were as aggregated as dermaseptin S4, whereas K(4)-S4 and K(4)K(20)-S4 were clearly less aggregated. Overall, the data indicated that N-terminal domain interaction between dermaseptin S4 monomers is responsible for the peptide's oligomerization in solution and, hence, for its limited spectrum of action. Moreover, bell-shaped dose-response profiles obtained with bacteria but not with protozoa or RBC implied that aggregation can have dramatic consequences on antibacterial activity. Based on these results, we tested the feasibility of selectivity reversal in the activity of dermaseptin S4. Tampering with the composition of the hydrophobic domains by reducing hydrophobicity or by increasing the net positive charge affected dramatically the peptide's activity and resulted in various analogs that displayed potent antibacterial activity but reduced hemolytic activity. Among these, maximal antibacterial activity was displayed by a 15-mer version that was more potent by 2 orders of magnitude compared with native dermaseptin S4. These results emphasize the notion that peptide-based antibiotics represent a highly modular synthetic antimicrobial system and provide indications of how the peptide's physico-chemical properties affect potency and selectivity. PMID:10660589

Feder, R; Dagan, A; Mor, A

2000-02-11

141

HIV-1 matrix protein p17 binds to the IL-8 receptor CXCR1 and shows IL-8-like chemokine activity on monocytes through Rho/ROCK activation.  

PubMed

Exogenous HIV-1 matrix protein p17 was found to deregulate biologic activities of many different immune cells that are directly or indirectly involved in AIDS pathogenesis after binding to unknown cellular receptor(s). In particular, p17 was found to induce a functional program in monocytes related to activation and inflammation. In the present study, we demonstrate that CXCR1 is the receptor molecule responsible for p17 chemokine-like activity on monocytes. After CXCR1 binding, p17 was capable of triggering rapid adhesion and chemotaxis of monocytes through a pathway that involved Rho/ROCK. Moreover, CXCR1-silenced primary monocytes lost responsiveness to p17 chemoattraction, whereas CXCR1-transfected Jurkat cells acquired responsiveness. Surface plasmon resonance studies confirmed the capacity of p17 to bind CXCR1 and showed that the p17/CXCR1 interaction occurred with a low affinity compared with that measured for IL-8, the physiologic CXCR1 ligand. In all of its activities, p17 mimicked IL-8, the natural high-affinity ligand of CXCR1. Recent studies have highlighted the role of IL-8 and CXCR1 in HIV-1 replication and AIDS pathogenesis. Our findings herein call for an exploration of the therapeutic potential of blocking the p17/IL-8/CXCR1 axis in HIV-1 infection. PMID:22262769

Giagulli, Cinzia; Magiera, Anna K; Bugatti, Antonella; Caccuri, Francesca; Marsico, Stefania; Rusnati, Marco; Vermi, William; Fiorentini, Simona; Caruso, Arnaldo

2012-03-01

142

Healthy children show gender differences in correlations between nonverbal cognitive ability and brain activation during visual perception.  

PubMed

Humans perceive textual and nontextual information in visual perception, and both depend on language. In childhood education, students exhibit diverse perceptual abilities, such that some students process textual information better and some process nontextual information better. These predispositions involve many factors, including cognitive ability and learning preference. However, the relationship between verbal and nonverbal cognitive abilities and brain activation during visual perception has not yet been examined in children. We used functional magnetic resonance imaging to examine the relationship between nonverbal and verbal cognitive abilities and brain activation during nontextual visual perception in large numbers of children. A significant positive correlation was found between nonverbal cognitive abilities and brain activation in the right temporoparietal junction, which is thought to be related to attention reorienting. This significant positive correlation existed only in boys. These findings suggested that male brain activation differed from female brain activation, and that this depended on individual cognitive processes, even if there was no gender difference in behavioral performance. PMID:24937269

Asano, Kohei; Taki, Yasuyuki; Hashizume, Hiroshi; Sassa, Yuko; Thyreau, Benjamin; Asano, Michiko; Takeuchi, Hikaru; Kawashima, Ryuta

2014-08-01

143

UCLA study shows prostate cancer finds a way to circumvent hormone therapy by activating a survival cell signaling pathway:  

Cancer.gov

In a study at UCLA’s Jonsson Comprehensive Cancer Center, researchers found that when a common type of prostate cancer was treated with conventional hormone ablation therapy blocking androgen production or androgen receptor function, the cancer was able to adapt and compensate by activating a survival cell signaling pathway, effectively circumventing the roadblock put up by this treatment.

144

Schizophrenic subjects show aberrant fMRI activation of dorsolateral prefrontal cortex and basal ganglia during working memory performance  

Microsoft Academic Search

Background: Working memory (WM) deficits in schizophrenia have been associated with dorsolateral prefrontal cortex (DLPFC) dysfunction in neuroimaging studies. We previously found increased DLPFC activation in schizophrenic versus normal subjects during WM performance (Manoach et al 1999b). We now have investigated whether schizophrenic subjects recruit different brain regions, particularly the basal ganglia and thalamus, components of frontostriatal circuitry thought to

Dara S. Manoach; Randy L. Gollub; Etienne S. Benson; Meghan M. Searl; Donald C. Goff; Elkan Halpern; Clifford B. Saper; Scott L. Rauch

2000-01-01

145

Parthenolide, a sesquiterpene lactone from the medical herb feverfew, shows anticancer activity against human melanoma cells in vitro.  

PubMed

Metastatic melanoma is a highly life-threatening disease. The lack of response to radiotherapy and chemotherapy highlights the critical need for novel treatments. Parthenolide, an active component of feverfew (Tanacetum parthenium), inhibits proliferation and kills various cancer cells mainly by inducing apoptosis. The aim of the study was to examine anticancer effects of parthenolide in melanoma cells in vitro. The cytotoxicity of parthenolide was tested in melanoma cell lines and melanocytes, as well as melanoma cells directly derived from a surgical excision. Adherent cell proliferation was measured by tetrazolium derivative reduction assay. Loss of the plasma membrane integrity, hypodiploid events, reactive oxygen species generation, mitochondrial membrane potential dissipation, and caspase-3 activity were assessed by flow cytometric analysis. Microscopy was used to observe morphological changes and cell detachment. Parthenolide reduced the number of viable adherent cells in melanoma cultures. Half maximal inhibitory concentration values around 4 mumol/l were determined. Cell death accompanied by mitochondrial membrane depolarization and caspase-3 activation was observed as the result of parthenolide application. Interestingly, the melanoma cells from vertical growth phase and melanocytes were less susceptible to parthenolide-induced cell death than metastatic cells when drug concentration was at least 6 mumol/l. Reactive oxygen species level was not significantly increased in melanoma cells. However, preincubation of parthenolide with the thiol nucleophile N-acetyl-cysteine protected melanoma cells from parthenolide-induced cell death suggesting the reaction with intracellular thiols as the mechanism responsible for parthenolide activity. In conclusion, the observed anticancer activity makes parthenolide an attractive drug candidate for further testing in melanoma therapy. PMID:19949351

Lesiak, Karolina; Koprowska, Kamila; Zalesna, Izabela; Nejc, Dariusz; Düchler, Markus; Czyz, Malgorzata

2010-02-01

146

The human anti-CD30 antibody 5F11 shows in vitro and in vivo activity against malignant lymphoma  

Microsoft Academic Search

CD30 is a promising target for antibody- based immunotherapy of Hodgkin lym- phoma (HL) and anaplastic large cell lym- phoma. To overcome the limitations from currently available murine anti-CD30 monoclonal antibodies (mAbs), a new fully human anti-CD30 antibody was gener- ated. Binding properties were evaluated by recombinant CD30 capture enzyme- linked immunosorbent assay (ELISA) and fluorescence-activated cell-sorter (FACS) flow cytometry.

Peter Borchmann; John F. Treml; Hinrich Hansen; Claudia Gottstein; Roland Schnell; Oliver Staak; Hui-fen Zhang; Thomas Davis; Tibor Keler; Volker Diehl; Robert F. Graziano; Andreas Engert

2003-01-01

147

Rare Germ Line CHEK2 Variants Identified in Breast Cancer Families Encode Proteins That Show Impaired Activation  

Microsoft Academic Search

Germ line mutations in CHEK2, the gene that encodes the Chk2 serine\\/threonine kinase activated in response to DNA damage, have been found to confer an increased risk of some cancers. We have previously reported the presence of the common deleterious 1100delC and four rare CHEK2 mutations in inherited breast cancer. Here, we report that predictions made by bioinformatic analysis on

Nayanta Sodha; Tine S. Mantoni; Sean V. Tavtigian; Rosalind Eeles; Michelle D. Garrett

148

In Vitro and In Vivo Antimalarial Activity Assays of Seeds from Balanites aegyptiaca: Compounds of the Extract Show Growth Inhibition and Activity against Plasmodial Aminopeptidase  

PubMed Central

Balanites aegyptiaca (Balanitaceae) is a widely grown desert plant with multiuse potential. In the present paper, a crude extract from B. aegyptiaca seeds equivalent to a ratio of 1?:?2000 seeds to the extract was screened for antiplasmodial activity. The determined IC50 value for the chloroquine-susceptible Plasmodium falciparum NF54 strain was 68.26??g/?L ± 3.5. Analysis of the extract by gas chromatography-mass spectrometry detected 6-phenyl-2(H)-1,2,4-triazin-5-one oxime, an inhibitor of the parasitic M18 Aspartyl Aminopeptidase as one of the compounds which is responsible for the in vitro antiplasmodial activity. The crude plant extract had a Ki of 2.35??g/?L and showed a dose-dependent response. After depletion of the compound, a significantly lower inhibition was determined with a Ki of 4.8??g/?L. Moreover, two phenolic compounds, that is, 2,6-di-tert-butyl-phenol and 2,4-di-tert-butyl-phenol, with determined IC50 values of 50.29??M ± 3 and 47.82??M ± 2.5, respectively, were detected. These compounds may contribute to the in vitro antimalarial activity due to their antioxidative properties. In an in vivo experiment, treatment of BALB/c mice with the aqueous Balanite extract did not lead to eradication of the parasites, although a reduced parasitemia at day 12 p.i. was observed.

Kusch, Peter; Deininger, Susanne; Specht, Sabine; Maniako, Rudeka; Haubrich, Stefanie; Pommerening, Tanja; Lin, Paul Kong Thoo; Hoerauf, Achim; Kaiser, Annette

2011-01-01

149

In Vitro and In Vivo Antimalarial Activity Assays of Seeds from Balanites aegyptiaca: Compounds of the Extract Show Growth Inhibition and Activity against Plasmodial Aminopeptidase.  

PubMed

Balanites aegyptiaca (Balanitaceae) is a widely grown desert plant with multiuse potential. In the present paper, a crude extract from B. aegyptiaca seeds equivalent to a ratio of 1?:?2000 seeds to the extract was screened for antiplasmodial activity. The determined IC(50) value for the chloroquine-susceptible Plasmodium falciparum NF54 strain was 68.26??g/?L ± 3.5. Analysis of the extract by gas chromatography-mass spectrometry detected 6-phenyl-2(H)-1,2,4-triazin-5-one oxime, an inhibitor of the parasitic M18 Aspartyl Aminopeptidase as one of the compounds which is responsible for the in vitro antiplasmodial activity. The crude plant extract had a K(i) of 2.35??g/?L and showed a dose-dependent response. After depletion of the compound, a significantly lower inhibition was determined with a K(i) of 4.8??g/?L. Moreover, two phenolic compounds, that is, 2,6-di-tert-butyl-phenol and 2,4-di-tert-butyl-phenol, with determined IC(50) values of 50.29??M ± 3 and 47.82??M ± 2.5, respectively, were detected. These compounds may contribute to the in vitro antimalarial activity due to their antioxidative properties. In an in vivo experiment, treatment of BALB/c mice with the aqueous Balanite extract did not lead to eradication of the parasites, although a reduced parasitemia at day 12 p.i. was observed. PMID:21687598

Kusch, Peter; Deininger, Susanne; Specht, Sabine; Maniako, Rudeka; Haubrich, Stefanie; Pommerening, Tanja; Lin, Paul Kong Thoo; Hoerauf, Achim; Kaiser, Annette

2011-01-01

150

Benzochalcones bearing pyrazoline moieties show anti-colorectal cancer activities and selective inhibitory effects on aurora kinases.  

PubMed

Colorectal cancer is the third and fourth leading cause of cancer in males and females, respectively. Flavonoids, including chalcones, are secondary metabolites in plants that exhibit diverse biological activities, including antibacterial, antimalarial, and antitumor activities. In order to find potent and novel chemotherapy drugs for colorectal cancer, a series of benzochalcone derivatives, in which an ?,?-unsaturated carbonyl group was replaced with a pyrazoline, was designed and synthesized. A clonogenic survival assay was performed with each derivative to evaluate antitumor activity. 1-(5-(2,4-Dimethoxyphenyl)-4,5-dihydro-1H-pyrazol-3-yl)naphthalen-2-ol (derivative 7) had the most potent inhibitory effect on the long-term clonogenicity of HCT116 human colorectal cancer cells (IC50=2.4 ?M). The results of Western blot and flow cytometric analyses suggested that derivative 7 could inhibit the proliferation of colorectal cancer cells through inhibition of cell cycle progression and induction of apoptosis. To elucidate its molecular mechanism, in vitro kinase binding assays were carried out, which demonstrated that derivative 7 inhibited aurora kinases A and B selectively. The binding modes between the compound and aurora kinases were interpreted using in silico docking experiments to explain the selective inhibitory effects on aurora kinases A and B. These findings will facilitate the design of potent novel benzochalcones as anticancer agents. PMID:24095020

Shin, Soon Young; Yoon, Hyuk; Hwang, Doseok; Ahn, Seunghyun; Kim, Dong-Wook; Koh, Dongsoo; Lee, Young Han; Lim, Yoongho

2013-11-15

151

Patients with ARDS show improvement but not normalisation of alveolar surface activity with surfactant treatment: putative role of neutral lipids  

PubMed Central

Background Extensive biochemical and biophysical changes of the pulmonary surfactant system occur in the acute respiratory distress syndrome (ARDS). Methods The effect of intrabronchial administration of a recombinant surfactant protein C?based surfactant preparation (Venticute) on gas exchange, surfactant composition and function was investigated in 31 patients with ARDS in a randomised controlled phase I/II clinical pilot trial. Bronchoalveolar lavage fluids for surfactant analysis were obtained 3?h before and 48 and 120?h after the first surfactant application. Potentially deleterious effects of surfactant neutral lipids in patients with ARDS were also identified. Results Before treatment all patients had marked abnormalities in the surfactant phospholipid and protein composition. In response to surfactant treatment, gas exchange improved and surfactant phospholipid and protein content were almost normalised. Alveolar surface activity was dramatically impaired before treatment and only partially improved after surfactant administration. Further analysis of the bronchoalveolar lavage fluids revealed a twofold increase in neutral lipid content and altered neutral lipid profile in patients with ARDS compared with healthy controls. These differences persisted even after administration of large amounts of Venticute. Supplementation of Venticute or natural surfactant with a synthetic neutral lipid preparation, mimicking the profile in ARDS, caused a dose?dependent deterioration of surface activity in vitro. Conclusion Intrabronchial surfactant treatment improves gas exchange in ARDS, but the efficacy may be limited by increased concentration and altered neutral lipid profile in surfactant under these conditions.

Markart, Philipp; Ruppert, Clemens; Wygrecka, Malgorzata; Colaris, Thorsten; Dahal, Bhola; Walmrath, Dieter; Harbach, Heinz; Wilhelm, Jochen; Seeger, Werner; Schmidt, Reinhold; Guenther, Andreas

2007-01-01

152

Monounsaturated fatty acid ether oligomers formed during heating of virgin olive oil show agglutination activity against human red blood cells.  

PubMed

The present work focuses on the characterization of molecules formed when virgin olive oil is heated at 130 °C for 24 h open in air, which are found to be strong agglutinins. The hemagglutinating activity of the newly formed molecule isolated from the heated virgin olive oil sample was estimated against human red blood cells (RBCs). Dimers and polymers (high molecular weight molecules) were identified through thin layer chromatography (TLC) of the oil mixture. (1)H and (13)C nuclear magnetic resonance (NMR) and gas chromatography-mass spectroscopy (GC-MS) were the methods used for structural characterization. Among others, oligomerization of at least two monounsaturated fatty acids (FA) by an ether linkage between the hydrocarbon chains is involved. Light microscopy was used to characterize and visualize the agglutination process. Agglutination without fusion or lysis was observed. It was concluded that the heating of virgin olive oil open in air, among other effects, produces oligomerization as well as polymerization of unsaturated FA, possibly of monohydroxy, monounsaturated FA that is associated with strong hemagglutinating activity against human RBCs. The nutritional value and the effects on human health of such oligomers are not discussed in the literature and remain to be investigated. PMID:24410166

Patrikios, Ioannis S; Mavromoustakos, Thomas M

2014-01-29

153

Purification and properties of a novel latent proteinase showing myosin heavy chain-degrading activity from threadfin-bream muscle.  

PubMed

A novel latent proteinase of which activity was induced by heating in the presence of NaCl was purified to homogeneity from threadfin-bream muscle by a combination of DEAE-cellulose, Con A-Sepharose, Arg-Sepharose, and Shim-pack HAC chromatographies. This proteinase was a glycoprotein having a monomeric subunit structure; Mr was estimated to be 77,000 on SDS-PAGE analysis. The proteinase hydrolyzed Boc-Leu-Thr-Arg-MCA as well as myosin heavy chain in the presence of 2-4% NaCl at pH 7.0 and at 60 degrees C, optimally. The proteinase was classified as serine proteinase based on the effects of soybean trypsin inhibitor, leupeptin, and antipain. PMID:2358432

Kinoshita, M; Toyohara, H; Shimizu, Y

1990-04-01

154

Ferrocenyl-l-amino acid copper(ii) complexes showing remarkable photo-induced anticancer activity in visible light.  

PubMed

Ferrocene-conjugated copper(ii) complexes [Cu(Fc-aa)(aip)](ClO4) () and [Cu(Fc-aa)(pyip)](ClO4) () of l-amino acid reduced Schiff bases (Fc-aa), 2-(9-anthryl)-1H-imidazo[4,5-f][1,10]phenanthroline (aip) and 2-(1-pyrenyl)-1H-imidazo[4,5-f][1,10]phenanthroline (pyip), where Fc-aa is ferrocenylmethyl-l-tyrosine (Fc-Tyr in , ), ferrocenylmethyl-l-tryptophan (Fc-Trp in , ) and ferrocenylmethyl-l-methionine (Fc-Met in , ), were prepared and characterized, and their photocytotoxicity was studied (Fc = ferrocenyl moiety). Phenyl analogues, viz. [Cu(Ph-Met)(aip)](ClO4) () and [Cu(Ph-Met)(pyip)](ClO4) (), were prepared and used as control compounds. The bis-imidazophenanthroline copper(ii) complexes, viz. [Cu(aip)2(NO3)](NO3) () and [Cu(pyip)2(NO3)](NO3) (), were also prepared and used as controls. Complexes having a redox inactive cooper(ii) center showed the Fc(+)-Fc redox couple at ?0.5 V vs. SCE in DMF-0.1 mol [Bu(n)4N](ClO4). The copper(ii)-based d-d band was observed near 600 nm in DMF-Tris-HCl buffer (1?:?1 v/v). The ferrocenyl complexes showed low dark toxicity, but remarkably high photocytotoxicity in human cervical HeLa and human breast adenocarcinoma MCF-7 cancer cells giving an excellent photo-dynamic effect while their phenyl analogues were inactive. The photo-exposure caused significant morphological changes in the cancer cells when compared to the non-irradiated ones. The photophysical processes were rationalized from the theoretical studies. Fluorescence microscopic images showed and localizing predominantly in the endoplasmic reticulum (ER) of the cancer cells, thus minimizing any undesirable effects involving nuclear DNA. PMID:24971754

Goswami, Tridib K; Gadadhar, Sudarshan; Balaji, Babu; Gole, Bappaditya; Karande, Anjali A; Chakravarty, Akhil R

2014-07-15

155

Homemade Laser Show  

NSDL National Science Digital Library

With a laser pointer and some household items, learners can create their own laser light show. They can explore diffuse reflection, refraction and diffraction. The webpage includes a video which shows how to set up the activity and also includes scientific explanation. Because this activity involves lasers, it requires adult supervision.

Houston, Children'S M.

2011-01-01

156

Glutaminase 2 negatively regulates the PI3K/AKT signaling and shows tumor suppression activity in human hepatocellular carcinoma  

PubMed Central

The tumor suppressor p53 and its signaling pathway play a critical role in tumor prevention. As a direct p53 target gene, the role of glutaminase 2 (GLS2) in tumorigenesis is unclear. In this study, we found that GLS2 expression is significantly decreased in majority of human hepatocellular carcinoma (HCC). Restoration of GLS2 expression in HCC cells inhibits the anchorage-independent growth of cells and reduces the growth of HCC xenograft tumors. Interestingly, we found that GLS2 negatively regulates the PI3K/AKT signaling, which is frequently activated in HCC. Blocking the PI3K/AKT signaling in HCC cells largely abolishes the inhibitory effect of GLS2 on the anchorage-independent cell growth and xenograft tumor growth. The GLS2 promoter is hypermethylated in majority of HCC samples. CpG methylation of GLS2 promoter inhibits GLS2 transcription, whereas reducing the methylation of GLS2 promoter induces GLS2 expression. Taken together, our results demonstrate that GLS2 plays an important role in tumor suppression in HCC, and the negative regulation of PI3K/AKT signaling contributes greatly to this function of GLS2. Furthermore, hypermethylation of GLS2 promoter is an important mechanism contributing to the decreased GLS2 expression in HCC.

Liu, Juan; Zhang, Cen; Lin, Meihua; Zhu, Wei; Liang, Yingjian; Hong, Xuehui; Zhao, Yuhan; Young, Ken H.; Hu, Wenwei; Feng, Zhaohui

2014-01-01

157

IgVH-genes analysis from psoriatic arthritis shows involvement of antigen-activated synovial B-lymphocytes.  

PubMed

The pathogenesis of psoriatic arthritis (PA), which occurs in 5-7% of patients with psoriasis vulgaris, is enigmatic. There are no molecular data about synovial B-cells of chronic synovitis of PA. In order to understand the B-cell response in PA, we analysed IgVH genes and specified replacement to silent mutation (R/S) ratios of the complementarity determining regions (CDR) and framework regions (FR) as well as VH families. To prove the existence of a common pattern we additionally analysed the IgVH genes at the amino-acid level. From 5 PA patients we took cryo-tissue sections with somatically mutated IgVH genes (sum R/sum S in the CDRs: 2.5-6.8), 62 of which were analysed. In one patient two cases of clonally related IgVH genes were observed. Identical amino-acid replacements in IgVH1 and IgVH4 were found at the same mutational "cold spot". These data indicate that antigen-activated B-cells participate in the formation of chronic synovitis of PA. Since, neither histopathologically nor immunohistochemically, no germinal centres could be detected, the clonally related IgVH genes may be interpreted as residues of a germinal centre reaction that occurred before synovectomy. The existence of identical amino-acid replacement mutations in IgVH1 and IgVH4 genes suggests that a limited number of antigens are common to all PA patients analysed. The recombinant expression of the IgVH could help to define B-cell specificities underlying the pathogenesis of chronic synovitis and dermatitis of PA. PMID:12491138

Gerhard, N; Krenn, V; Magalhães, R; Morawietz, L; Brändlein, S; König, A

2002-12-01

158

Testing the Durability of Polymyxin B Immobilization on a Polymer Showing Antimicrobial Activity: A Novel Approach with the Ion-Step Method  

Microsoft Academic Search

A polymyxin B (PMB) modified copolymer of ethyl acrylate and 2-hydroxyethyl methacrylate is reported that shows antibacterial activity towards E. coli. The polymer is designed for biosensor interfaces to provide protection against microbial contamination. However, leaching of a biomolecule from its solid support is a well-known problem in immobilization chemistry. It has been proposed that antimicrobial activity results from release

Achilles Tzoris; Elizabeth A. H. Hall; Geert A. J. Besselink; Piet Bergveld

2003-01-01

159

Showing What They Know  

ERIC Educational Resources Information Center

Having students show their skills in three dimensions, known as performance-based assessment, dates back at least to Socrates. Individual schools such as Barrington High School--located just outside of Providence--have been requiring students to actively demonstrate their knowledge for years. The Rhode Island's high school graduating class became…

Cech, Scott J.

2008-01-01

160

The Ozone Show.  

ERIC Educational Resources Information Center

Uses a talk show activity for a final assessment tool for students to debate about the ozone hole. Students are assessed on five areas: (1) cooperative learning; (2) the written component; (3) content; (4) self-evaluation; and (5) peer evaluation. (SAH)

Mathieu, Aaron

2000-01-01

161

Bi-functional gfp- and gusA-containing mini-Tn5 transposon derivatives for combined gene expression and bacterial localization studies.  

PubMed

The gfp gene, encoding the green fluorescent protein, was combined with the gusA gene, coding for the beta-glucuronidase enzyme, in mini-Tn5 transposon derivatives for use in Gram-negative bacteria. These mini-Tn5 elements allow simultaneously monitoring of gene expression and localization of the marked bacteria. Introduction of the resultant mini-Tn5 transposons into Rhizobium etli, Azospirillum brasilense and Pseudomonas stutzeri allowed us to visualise the interaction of these bacteria with their host plant. The dual-marker mini-Tn5 transposons constitute a powerful new tool for studying gene expression and ecology of bacteria in the environment and during the interaction with plants. PMID:10076635

Xi, C; Lambrecht, M; Vanderleyden, J; Michiels, J

1999-02-01

162

Gold(I) analogues of a platinum-acridine antitumor agent are only moderately cytotoxic but show potent activity against Mycobacterium tuberculosis.  

PubMed

Cationic gold(I) complexes containing 1-[2-(acridin-9-ylamino)ethyl]-1,3-dimethylthiourea (1), [AuL(1)](n+) (where L is Cl(-), Br(-), SCN(-), PEt(3), PPh(3), or 1), derived from a class of analogous platinum(II) antitumor agents, have been synthesized. Unlike platinum, gold does not form permanent adducts with DNA, and its complexes are 2 orders of magnitude less cytotoxic in non-small-cell lung cancer cells than the most active platinum-based agent. Instead, several gold analogues show submicromolar and selective antimicrobial activity against Mycobacterium tuberculosis. PMID:19803526

Eiter, Lauren C; Hall, Nathan W; Day, Cynthia S; Saluta, Gilda; Kucera, Gregory L; Bierbach, Ulrich

2009-11-12

163

Gold(I) Analogues of a Platinum-Acridine Antitumor Agent are only Moderately Cytotoxic but Show Potent Activity Against Mycobacterium Tuberculosis  

PubMed Central

A series of cationic gold(I) complexes containing 1-[2-(acridin-9-ylamino)ethyl]-1,3-dimethylthiourea (1), [AuL(1)]n+ (where L is Cl?, Br-, SCN?, PEt3, PPh3, or 1), derived from a class of analogous platinum(II) antitumor agents, has been synthesized. Unlike platinum, gold does not form permanent adducts with DNA, and its complexes are two orders of magnitude less cytotoxic in non-small-cell lung cancer cells than the most active platinum-based agent. Instead, several gold analogues show submicromolar and selective antimicrobial activity against Mycobacterium tuberculosis.

Eiter, Lauren C.; Hall, Nathan W.; Day, Cynthia S.; Saluta, Gilda; Kucera, Gregory L.; Bierbach, Ulrich

2011-01-01

164

The major indole alkaloid N,?-D-glucopyranosyl vincosamide from leaves of Psychotria leiocarpa Cham. & Schltdl. is not an antifeedant but shows broad antioxidant activity.  

PubMed

N,?-D-glucopyranosyl vincosamide (GPV), a major alkaloid of Psychotria leiocarpa, constitutes up to 2.5% of the dry weight in leaves. Alkaloid content was not elicited by mechanical wounding or jasmonate. At concentrations found in natural conditions or 2.5 fold higher, GPV did not inhibit herbivory in two unrelated generalist models (Helix aspersa and Spodoptera frugiperda) or in a specific interaction model (Heliconius erato fed with Passiflora suberosa). In situ staining assay showed quenching activity of hydrogen peroxide by GPV. Exposure of P. leiocarpa to acute UV-B stress did not change GPV or chlorophyll content, indicating high tolerance to this stress by the species. In vitro antioxidant tests against singlet oxygen, superoxide anions and hydroxyl radicals showed efficient quenching activity of the alkaloid. GPV was not effective as antifeedant, but it may act indirectly in P. leiocarpa protection against oxidative stress generated upon wounding, UV exposure and perhaps other environmental stresses. PMID:22891663

Matsuura, Hélio Nitta; Fett-Neto, Arthur Germano

2013-03-01

165

Cloning and characterization of a novel Athspr promoter specifically active in vascular tissue.  

PubMed

The vascular system--xylem, phloem and the cambium--is essential for water supply, nutrient transport, and physical support in higher plants. Although it is known that vascular-specific gene expression is regulated by cis-acting regulatory sequences in promoters, it is largely unknown how many regulatory elements exist and what their roles are in promoters. To understand the regulatory elements of vascular-specific promoters and their roles in vascular development, a T-DNA insertion mutant showing delayed growth and diminished resistance to environmental stress was isolated using promoter trap strategy. The novel gene, Arabidopsis thaliana heat shock protein-related (Athspr), was cloned from Arabidopsis ecotype C24. Strong GUS (?-glucuronidase) staining in the original promoter trap line was found in the vascular tissues of all organs in the mutant. The Athspr promoter was cloned and fused with GUS and eGFP (enhanced green fluorescent protein) reporter genes to verify its vascular-specific expression in Arabidopsis. Further histochemical analysis in transgenic plants demonstrated a similar GUS expression pattern in the vascular tissues. In addition, ATHSPR-eGFP driven by Athspr promoter was observed in vascular bundles of the transgenic seedling roots. Finally, comparative analysis with promoter motifs from 37 genes involved in vascular development revealed that Athspr and all other promoters active in vascular tissues contained regulatory elements responding to phytohormones, light, biotic and abiotic stresses, as well as those regulating tissue-specific expression. These results demonstrated that the Athspr promoter has a vascular tissue-specific activity and Athspr may have multiple functions in vascular development and resistance against various stresses. PMID:24675528

Zhang, Liang; Yang, Tao; Li, Xiaoying; Hao, Hongyan; Xu, Shengtao; Cheng, Wei; Sun, Yingli; Wang, Chongying

2014-05-01

166

Calcium-sensitive cls mutants of Saccharomyces cerevisiae showing a Pet- phenotype are ascribable to defects of vacuolar membrane H(+)-ATPase activity.  

PubMed

Ca(2+)-sensitive mutants of the yeast Saccharomyces cerevisiae showing a Pet- phenotype (cls7-cls11) have lesions in a system for maintaining intracellular Ca2+ homeostasis (Ohya, Y., Ohsumi, Y., and Anraku, Y. (1986) J. Gen. Microbiol. 132, 979-988). Genetic and biochemical studies have demonstrated that these Pet- cls mutants are related to defects in vacuolar membrane H(+)-ATPase. CLS7 and CLS8 were found to be identical with the structural genes encoding subunit c (VMA3) and subunit a (VMA1), respectively, of the enzyme. In addition, these five mutants all had vma defects; no vacuolar membrane ATPase activity was detected in the cls cells, and the cls mutants showed a loss of ability to acidify the vacuole in vivo. Measurements of the cytosolic free Ca2+ concentration [( Ca2+]i) in individual cells showed that the average [Ca2+]i in wild-type cells was 150 +/- 80 nM, whereas that in five Pet- cls cells was 900 +/- 100 nM. These data are consistent with the observation that vacuolar membrane vesicles prepared from the Pet- cls cells have lost ATP-dependent Ca2+ uptake activities. The cls defects of vacuolar membrane H(+)-ATPase resulted in pleiotropic effects on several cellular activities, including Ca2+ homeostasis, glycerol metabolism, and phospholipid metabolism. The mutants showed an inositol-dependent phenotype, possibly due to alteration in regulation of phospholipid biosynthesis; the phosphatidylserine decarboxylase activities of the mutants were 15-50% of that of the wild-type cells and were not repressed by the addition of inositol. In contrast to the majority of previously isolated pet mutants (Tzagoloff, A., and Dieckmann, C. L. (1990) Microbiol. Rev. 54, 211-225), the Pet- cls mutants showed no detectable mitochondrial defects. Taking all these findings into account, we suggest that at least six genes, VMA1 (CLS8, subunit a), VMA2 (subunit b), VMA3 (CLS7, subunit c), VMA11 (CLS9), VMA12 (CLS10), and VMA13 (CLS11), are required for expression of the vacuolar membrane H(+)-ATPase activity. PMID:1830311

Ohya, Y; Umemoto, N; Tanida, I; Ohta, A; Iida, H; Anraku, Y

1991-07-25

167

B-Ring-Aryl Substituted Luotonin A Analogues with a New Binding Mode to the Topoisomerase 1-DNA Complex Show Enhanced Cytotoxic Activity.  

PubMed

Topoisomerase 1 inhibition is an important strategy in targeted cancer chemotherapy. The drugs currently in use acting on this enzyme belong to the family of the camptothecins, and suffer severe limitations because of their low stability, which is associated with the hydrolysis of the ?-lactone moiety in their E ring. Luotonin A is a natural camptothecin analogue that lacks this functional group and therefore shows a much-improved stability, but at the cost of a lower activity. Therefore, the development of luotonin A analogues with an increased potency is important for progress in this area. In the present paper, a small library of luotonin A analogues modified at their A and B rings was generated by cerium(IV) ammonium nitrate-catalyzed Friedländer reactions. All analogues showed an activity similar or higher than the natural luotonin A in terms of topoisomerase 1 inhibition and some compounds had an activity comparable to that of camptothecin. Furthermore, most compounds showed a better activity than luotonin A in cell cytotoxicity assays. In order to rationalize these results, the first docking studies of luotonin-topoisomerase 1-DNA ternary complexes were undertaken. Most compounds bound in a manner similar to luotonin A and to standard topoisomerase poisons such as topotecan but, interestingly, the two most promising analogues, bearing a 3,5-dimethylphenyl substituent at ring B, docked in a different orientation. This binding mode allows the hydrophobic moiety to be shielded from the aqueous environment by being buried between the deoxyribose belonging to the G(+1) guanine and Arg364 in the scissile strand and the surface of the protein and a hydrogen bond between the D-ring carbonyl and the basic amino acid. The discovery of this new binding mode and its associated higher inhibitory potency is a significant advance in the design of new topoisomerase 1 inhibitors. PMID:24830682

González-Ruiz, Víctor; Pascua, Irene; Fernández-Marcelo, Tamara; Ribelles, Pascual; Bianchini, Giulia; Sridharan, Vellaisamy; Iniesta, Pilar; Ramos, M Teresa; Olives, Ana I; Martín, M Antonia; Menéndez, J Carlos

2014-01-01

168

Eupafolin and Ethyl Acetate Fraction of Kalanchoe gracilis Stem Extract Show Potent Antiviral Activities against Enterovirus 71 and Coxsackievirus A16  

PubMed Central

Enterovirus 71 (EV71) and coxsackievirus A16 (CoxA16) are main pathogens of hand-foot-and-mouth disease, occasionally causing aseptic meningitis and encephalitis in tropical and subtropical regions. Kalanchoe gracilis, Da-Huan-Hun, is a Chinese folk medicine for treating pain and inflammation, exhibiting antioxidant and anti-inflammatory activities. Our prior report (2012) cited K. gracilis leaf extract as moderately active against EV71 and CoxA16. This study further rates antienteroviral potential of K. gracilis stem (KGS) extract to identify potent antiviral fractions and components. The extract moderately inhibits viral cytopathicity and virus yield, as well as in vitro replication of EV71 (IC50 = 75.18??g/mL) and CoxA16 (IC50 = 81.41??g/mL). Ethyl acetate (EA) fraction of KGS extract showed greater antiviral activity than that of n-butanol or aqueous fraction: IC50 values of 4.21??g/mL against EV71 and 9.08??g/mL against CoxA16. HPLC analysis, UV-Vis absorption spectroscopy, and plaque reduction assay indicate that eupafolin is a vital component of EA fraction showing potent activity against EV71 (IC50 = 1.39??M) and CoxA16 (IC50 = 5.24??M). Eupafolin specifically lessened virus-induced upregulation of IL-6 and RANTES by inhibiting virus-induced ERK1/2, AP-1, and STAT3 signals. Anti-enteroviral potency of KGS EA fraction and eupafolin shows the clinical potential against EV71 and CoxA16 infection.

Wang, Ching-Ying; Huang, Shun-Chueh; Lai, Zhen-Rung; Ho, Yu-Ling; Jou, Yu-Jen; Kung, Szu-Hao; Zhang, Yongjun; Chang, Yuan-Shiun; Lin, Cheng-Wen

2013-01-01

169

Eupafolin and Ethyl Acetate Fraction of Kalanchoe gracilis Stem Extract Show Potent Antiviral Activities against Enterovirus 71 and Coxsackievirus A16.  

PubMed

Enterovirus 71 (EV71) and coxsackievirus A16 (CoxA16) are main pathogens of hand-foot-and-mouth disease, occasionally causing aseptic meningitis and encephalitis in tropical and subtropical regions. Kalanchoe gracilis, Da-Huan-Hun, is a Chinese folk medicine for treating pain and inflammation, exhibiting antioxidant and anti-inflammatory activities. Our prior report (2012) cited K. gracilis leaf extract as moderately active against EV71 and CoxA16. This study further rates antienteroviral potential of K. gracilis stem (KGS) extract to identify potent antiviral fractions and components. The extract moderately inhibits viral cytopathicity and virus yield, as well as in vitro replication of EV71 (IC50 = 75.18? ? g/mL) and CoxA16 (IC50 = 81.41? ? g/mL). Ethyl acetate (EA) fraction of KGS extract showed greater antiviral activity than that of n-butanol or aqueous fraction: IC50 values of 4.21? ? g/mL against EV71 and 9.08? ? g/mL against CoxA16. HPLC analysis, UV-Vis absorption spectroscopy, and plaque reduction assay indicate that eupafolin is a vital component of EA fraction showing potent activity against EV71 (IC50 = 1.39? ? M) and CoxA16 (IC50 = 5.24? ? M). Eupafolin specifically lessened virus-induced upregulation of IL-6 and RANTES by inhibiting virus-induced ERK1/2, AP-1, and STAT3 signals. Anti-enteroviral potency of KGS EA fraction and eupafolin shows the clinical potential against EV71 and CoxA16 infection. PMID:24078828

Wang, Ching-Ying; Huang, Shun-Chueh; Lai, Zhen-Rung; Ho, Yu-Ling; Jou, Yu-Jen; Kung, Szu-Hao; Zhang, Yongjun; Chang, Yuan-Shiun; Lin, Cheng-Wen

2013-01-01

170

B-Ring-Aryl Substituted Luotonin A Analogues with a New Binding Mode to the Topoisomerase 1-DNA Complex Show Enhanced Cytotoxic Activity  

PubMed Central

Topoisomerase 1 inhibition is an important strategy in targeted cancer chemotherapy. The drugs currently in use acting on this enzyme belong to the family of the camptothecins, and suffer severe limitations because of their low stability, which is associated with the hydrolysis of the ?-lactone moiety in their E ring. Luotonin A is a natural camptothecin analogue that lacks this functional group and therefore shows a much-improved stability, but at the cost of a lower activity. Therefore, the development of luotonin A analogues with an increased potency is important for progress in this area. In the present paper, a small library of luotonin A analogues modified at their A and B rings was generated by cerium(IV) ammonium nitrate-catalyzed Friedländer reactions. All analogues showed an activity similar or higher than the natural luotonin A in terms of topoisomerase 1 inhibition and some compounds had an activity comparable to that of camptothecin. Furthermore, most compounds showed a better activity than luotonin A in cell cytotoxicity assays. In order to rationalize these results, the first docking studies of luotonin-topoisomerase 1-DNA ternary complexes were undertaken. Most compounds bound in a manner similar to luotonin A and to standard topoisomerase poisons such as topotecan but, interestingly, the two most promising analogues, bearing a 3,5-dimethylphenyl substituent at ring B, docked in a different orientation. This binding mode allows the hydrophobic moiety to be shielded from the aqueous environment by being buried between the deoxyribose belonging to the G(+1) guanine and Arg364 in the scissile strand and the surface of the protein and a hydrogen bond between the D-ring carbonyl and the basic amino acid. The discovery of this new binding mode and its associated higher inhibitory potency is a significant advance in the design of new topoisomerase 1 inhibitors.

Gonzalez-Ruiz, Victor; Pascua, Irene; Fernandez-Marcelo, Tamara; Ribelles, Pascual; Bianchini, Giulia; Sridharan, Vellaisamy; Iniesta, Pilar; Ramos, M. Teresa; Olives, Ana I.; Martin, M. Antonia; Menendez, J. Carlos

2014-01-01

171

Two Genetic Elements Regulate Murine ?-Glucuronidase Synthesis following Transcript Accumulation  

PubMed Central

Mutant alleles of two genetic regulatory elements, which underlie a three- to sixfold reduction in ?-glucuronidase (GUS) activity levels, distinguish mice of the H haplotype from those of the other two common GUS haplotypes, A and B. Both elements are tightly linked to the GUS structural gene over which they exert control. One (Gus-u) exerts a cis-active effect upon GUS activity levels in all tissues at all times while the other (Gus-t) regulates GUS activity in trans after the 12th postnatal day in certain tissues. While previous studies show that differences in the rate of GUS synthesis account for the combined effects of these two elements in liver of adult mice, we demonstrate the separate effects of each on GUS synthesis at times during early postnatal development when their individual expressions can be distinguished. Assessments of the relative levels of S1 nuclease protection of a radiolabeled GUS antisense RNA probe after hybridization with total liver RNA preparations from adult mice of A and H haplotypes reveal no differences. These results argue that Gus-u and Gus-t exert their control of GUS expression subsequent to the accumulation of processed GUS transcripts.

Wawrzyniak, C. J.; Meredith, S. A.; Ganschow, R. E.

1989-01-01

172

Novel Antiviral Characteristics of Nanosized Copper(I) Iodide Particles Showing Inactivation Activity against 2009 Pandemic H1N1 Influenza Virus  

PubMed Central

We investigated the antiviral activity of nanosized copper(I) iodide (CuI) particles having an average size of 160 nm. CuI particles showed aqueous stability and generated hydroxyl radicals, which were probably derived from monovalent copper (Cu+). We confirmed that CuI particles showed antiviral activity against an influenza A virus of swine origin (pandemic [H1N1] 2009) by plaque titration assay. The virus titer decreased in a dose-dependent manner upon incubation with CuI particles, with the 50% effective concentration being approximately 17 ?g/ml after exposure for 60 min. SDS-PAGE analysis confirmed the inactivation of the virus due to the degradation of viral proteins such as hemagglutinin and neuraminidase by CuI. Electron spin resonance (ESR) spectroscopy revealed that CuI generates hydroxyl radicals in aqueous solution, and radical production was found to be blocked by the radical scavenger N-acetylcysteine. Taken together, these findings indicate that CuI particles exert antiviral activity by generating hydroxyl radicals. Thus, CuI may be a useful material for protecting against viral attacks and may be suitable for applications such as filters, face masks, protective clothing, and kitchen cloths.

Sato, Tetsuya; Hayata, Taishi; Nagao, Tomokazu; Nakayama, Mikio; Nakayama, Tsuruo; Sugamata, Ryuichi; Suzuki, Kazuo

2012-01-01

173

Structural and functional characterization of pronyl-lysine, a novel protein modification in bread crust melanoidins showing in vitro antioxidative and phase I/II enzyme modulating activity.  

PubMed

Application of an in vitro antioxidant assay to solvent fractions isolated from bread crust, bread crumb, and flour, respectively, revealed the highest antioxidative potential for the dark brown, ethanol solubles of the crust, whereas corresponding crumb and flour fractions showed only minor activities. To investigate whether these browning products may also act as antioxidants in biological systems, their modulating activity on detoxification enzymes was investigated as a functional parameter in intestinal Caco-2 cells. The bread crust and, in particular, the intensely brown, ethanolic crust fraction induced a significantly elevated glutathione S-transferase (GST) activity and a decreased phase I NADPH-cytochrome c reductase (CCR) activity compared to crumb-exposed cells. Antioxidant screening of Maillard-type model mixtures, followed by structure determination, revealed the pyrrolinone reductones 1 and 2 as the key antioxidants formed from the hexose-derived acetylformoin and N(alpha)-acetyl-L-lysine methyl ester or glycine methyl ester, chosen as model substances to mimic nonenzymatic browning reactions with the lysine side chain or the N terminus of proteins, respectively. Quantitation of protein-bound pyrrolinone reductonyl-lysine, abbreviated pronyl-lysine, revealed high amounts in the bread crust (62.2 mg/kg), low amounts in the crumb (8.0 mg/kg), and the absence of this compound in untreated flour. Exposing Caco-2 cells for 48 h to either synthetically pronylated albumin or purified pronyl-glycine (3) significantly increased phase II GST activity by 12 or 34%, respectively, thus demonstrating for the first time that "pronylated" proteins as part of bread crust melanoidins act as monofunctional inducers of GST, serving as a functional parameter of an antioxidant, chemopreventive activity in vitro. PMID:12428950

Lindenmeier, Michael; Faist, Veronika; Hofmann, Thomas

2002-11-20

174

Organ-specific expression of ?-glucuronidase activity driven by the Arabidopsis heat-shock promoter in heat-stressed transgenic Nicotiana plumbaginifolia  

Microsoft Academic Search

The expression of the Arabidopsis heat-shock protein (HSP) 18.2 promoter ?-d-glucuronidase (GUS) chimera gene was studied in various organs of the transgenic Nicotiana plumbaginifolia during the recovery phase at normal temperatures (20–22??C) following heat-shock (HS) treatment. The optimum HS temperature\\u000a for GUS activity in the anthers, petals and capsules was 42??C, but in immature seeds and the placentas of capsules

M. Moriwaki; T. Yamakawa; T. Washino; T. Kodama; Y. Igarashi

1999-01-01

175

The environmental chemical tributyltin chloride (TBT) shows both estrogenic and adipogenic activities in mice which might depend on the exposure dose  

SciTech Connect

Exposure during early development to chemicals with hormonal action may be associated with weight gain during adulthood because of altered body homeostasis. It is known that organotins affect adipose mass when exposure occurs during fetal development, although no knowledge of effects are available for exposures after birth. Here we show that the environmental organotin tributyltin chloride (TBT) exerts adipogenic action when peripubertal and sexually mature mice are exposed to the chemical. The duration and extent of these effects depend on the sex and on the dose of the compound, and the effects are relevant at doses close to the estimated human intake (0.5 {mu}g/kg). At higher doses (50-500 {mu}g/kg), TBT also activated estrogen receptors (ERs) in adipose cells in vitro and in vivo, based on results from acute and longitudinal studies in ERE/luciferase reporter mice. In 3T3-L1 cells (which have no ERs), transiently transfected with the ERE-dependent reporter plus or minus ER{alpha} or ER{beta}, TBT (in a dose range of 1-100 nM) directly targets each ER subtype in a receptor-specific manner through a direct mechanism mediated by ER{alpha} in undifferentiated preadipocytic cells and by ER{beta} in differentiating adipocytes. The ER antagonist ICI-182,780 inhibits this effect. In summary, the results of this work suggest that TBT is adipogenic at all ages and in both sexes and that it might be an ER activator in fat cells. These findings might help to resolve the apparent paradox of an adipogenic chemical being also an estrogen receptor activator by showing that the two apparently opposite actions are separated by the different doses to which the organism is exposed. - Research Highlights: > The environmental organotin tributyltin chloride shows dose-dependent estrogenic and adipogenic activities in mice. > The duration and extent of these effects depend on the sex and the dose of the compound. > The estrogenic and adipogenic effects of TBT occur at doses closed to the estimated human intake. > TBT activates the estrogen receptors (ER{alpha} and ER{beta}) in 3T3-L1 cells at nM concentrations.

Penza, M.; Jeremic, M.; Marrazzo, E. [Laboratory of Biotechnology and Department of Laboratory Medicine, Civic Hospital of Brescia (Italy); Maggi, A.; Ciana, P. [Centre of Excellence on Neurodegenerative Diseases, University of Milan (Italy); Rando, G. [Center for Integrative Genomics, University of Lausanne (Switzerland); Grigolato, P.G. [Human Pathology, School of Medicine, University of Brescia (Italy); Di Lorenzo, D., E-mail: dilorenzodiego@yahoo.it [Laboratory of Biotechnology and Department of Laboratory Medicine, Civic Hospital of Brescia (Italy)

2011-08-15

176

A Novel Small Molecule FL118 That Selectively Inhibits Survivin, Mcl-1, XIAP and cIAP2 in a p53-Independent Manner, Shows Superior Antitumor Activity  

PubMed Central

Drug/radiation resistance to treatment and tumor relapse are major obstacles in identifying a cure for cancer. Development of novel agents that address these challenges would therefore be of the upmost importance in the fight against cancer. In this regard, studies show that the antiapoptotic protein survivin is a central molecule involved in both hurdles. Using cancer cell-based survivin-reporter systems (US 7,569,221 B2) via high throughput screening (HTS) of compound libraries, followed by in vitro and in vivo analyses of HTS-derived hit-lead compounds, we identified a novel anticancer compound (designated FL118). FL118 shows structural similarity to irinotecan. However, while the inhibition of DNA topoisomerase 1 activity by FL118 was no better than the active form of irinotecan, SN-38 at 1 µM, FL118 effectively inhibited cancer cell growth at less than nM levels in a p53 status-independent manner. Moreover, FL118 selectively inhibited survivin promoter activity and gene expression also in a p53 status-independent manner. Although the survivin promoter-reporter system was used for the identification of FL118, our studies revealed that FL118 not only inhibits survivin expression but also selectively and independently inhibits three additional cancer-associated survival genes (Mcl-1, XIAP and cIAP2) in a p53 status-independent manner, while showing no inhibitory effects on control genes. Genetic silencing or overexpression of FL118 targets demonstrated a role for these targets in FL118’s effects. Follow-up in vivo studies revealed that FL118 exhibits superior antitumor efficacy in human tumor xenograft models in comparison with irinotecan, topotecan, doxorubicin, 5-FU, gemcitabine, docetaxel, oxaliplatin, cytoxan and cisplatin, and a majority of mice treated with FL118 showed tumor regression with a weekly × 4 schedule. FL118 induced favorable body-weight-loss profiles (temporary and reversible) and was able to eliminate large tumors. Together, the molecular targeting features of FL118 plus its superior antitumor activity warrant its further development toward clinical trials.

Cheng, Qiuying; Keefe, James T.; Rustum, Youcef M.; Li, Fengzhi

2012-01-01

177

Antiviral lectins from red and blue-green algae show potent in vitro and in vivo activity against hepatitis C virus.  

PubMed

Hepatitis C virus (HCV) infection is a significant public health problem with over 170,000,000 chronic carriers and infection rates increasing worldwide. Chronic HCV infection is one of the leading causes of hepatocellular carcinoma which was estimated to result in ?10,000 deaths in the United States in the year 2011. Current treatment options for HCV infection are limited to PEG-ylated interferon alpha (IFN-?), the nucleoside ribavirin and the recently approved HCV protease inhibitors telaprevir and boceprevir. Although showing significantly improved efficacy over the previous therapies, treatment with protease inhibitors has been shown to result in the rapid emergence of drug-resistant virus. Here we report the activity of two proteins, originally isolated from natural product extracts, which demonstrate low or sub-nanomolar in vitro activity against both genotype I and genotype II HCV. These proteins inhibit viral infectivity, binding to the HCV envelope glycoproteins E1 and E2 and block viral entry into human hepatocytes. In addition, we demonstrate that the most potent of these agents, the protein griffithsin, is readily bioavailable after subcutaneous injection and shows significant in vivo efficacy in reducing HCV viral titers in a mouse model system with engrafted human hepatocytes. These results indicate that HCV viral entry inhibitors can be an effective component of anti-HCV therapy and that these proteins should be studied further for their therapeutic potential. PMID:23700478

Takebe, Yutaka; Saucedo, Carrie J; Lund, Garry; Uenishi, Rie; Hase, Saiki; Tsuchiura, Takayo; Kneteman, Norman; Ramessar, Koreen; Tyrrell, D Lorne J; Shirakura, Masayuki; Wakita, Takaji; McMahon, James B; O'Keefe, Barry R

2013-01-01

178

SCH 2047069, a novel oral kinesin spindle protein inhibitor, shows single-agent antitumor activity and enhances the efficacy of chemotherapeutics.  

PubMed

Kinesin spindle protein (KSP) is a mitotic kinesin required for the formation of the bipolar mitotic spindle, and inhibition of this motor protein results in mitotic arrest and cell death. KSP inhibitors show preclinical antitumor activity and are currently undergoing testing in clinical trials. These agents have been dosed intravenously using various dosing schedules. We sought to identify a KSP inhibitor that could be delivered orally and thus provide convenience of dosing as well as the ability to achieve more continuous exposure via the use of dose-dense administration. We discovered SCH 2047069, a potent KSP inhibitor with oral bioavailability across species and the ability to cross the blood-brain barrier. The compound induces mitotic arrest characterized by a monaster spindle and is associated with an increase in histone H3 and mitotic protein monoclonal 2 phosphorylation both in vitro and in vivo. SCH 2047069 showed antitumor activity in a variety of preclinical models as a single agent and in combination with paclitaxel, gemcitabine, or vincristine. PMID:20978164

Basso, Andrea D; Liu, Ming; Dai, Chaoyang; Gray, Kimberly; Nale, Lissette; Tevar, Seema; Lee, Suining; Liang, Lianzhu; Ponery, Abdul; Yaremko, Bohdan; Smith, Elizabeth; Tang, Huadong; Sheth, Payal R; Siddiqui, M Arshad; Hicklin, Daniel J; Kirschmeier, Paul

2010-11-01

179

Pyrethroid and their metabolite, 3-phenoxybenzoic acid showed similar (anti)estrogenic activity in human and rat estrogen receptor ?-mediated reporter gene assays.  

PubMed

Pyrethroids are commonly used as pesticides which are suspected as endocrine disruptors in many studies; however, still we do not know their effects on different species. To compare their effects on human estrogen receptor (hER?) and rat estrogen receptor (rER?), we developed a hER? and rER? mediated luciferase reporter assay to investigate the (anti)estrogenic activities of three frequently used pyrethroids (fenvalerate, cypermethrin, permethrin) and their metabolite 3-phenoxybenzoic acid (3-PBA). All three pyrethroids significantly induced expression of the luciferase, while none of them were antagonistic to 1 nM E? mediated induction. Interestingly, 3-PBA, showed antagonist activity by decreasing the effect of 1 nM E? to 55.12% in hER? assay and to 45.12% in rER? assay. Our results firstly demonstrated that pyrethroids and 3-PBA showed similar response in the hER? and rER? mediated reporter gene assay, which indicated that data derived from reporter gene assay or other receptor mediated assay systems with rat ER system might be used to predict to estrogenic or anti-estrogenic effects to human systems. PMID:24388911

Sun, Hong; Chen, Wen; Xu, Xiaolin; Ding, Zhen; Chen, Xiaodong; Wang, Xinru

2014-01-01

180

Antiviral Lectins from Red and Blue-Green Algae Show Potent In Vitro and In Vivo Activity against Hepatitis C Virus  

PubMed Central

Hepatitis C virus (HCV) infection is a significant public health problem with over 170,000,000 chronic carriers and infection rates increasing worldwide. Chronic HCV infection is one of the leading causes of hepatocellular carcinoma which was estimated to result in ?10,000 deaths in the United States in the year 2011. Current treatment options for HCV infection are limited to PEG-ylated interferon alpha (IFN-?), the nucleoside ribavirin and the recently approved HCV protease inhibitors telaprevir and boceprevir. Although showing significantly improved efficacy over the previous therapies, treatment with protease inhibitors has been shown to result in the rapid emergence of drug-resistant virus. Here we report the activity of two proteins, originally isolated from natural product extracts, which demonstrate low or sub-nanomolar in vitro activity against both genotype I and genotype II HCV. These proteins inhibit viral infectivity, binding to the HCV envelope glycoproteins E1 and E2 and block viral entry into human hepatocytes. In addition, we demonstrate that the most potent of these agents, the protein griffithsin, is readily bioavailable after subcutaneous injection and shows significant in vivo efficacy in reducing HCV viral titers in a mouse model system with engrafted human hepatocytes. These results indicate that HCV viral entry inhibitors can be an effective component of anti-HCV therapy and that these proteins should be studied further for their therapeutic potential.

Takebe, Yutaka; Saucedo, Carrie J.; Lund, Garry; Uenishi, Rie; Hase, Saiki; Tsuchiura, Takayo; Kneteman, Norman; Ramessar, Koreen; Tyrrell, D. Lorne J.; Shirakura, Masayuki; Wakita, Takaji; McMahon, James B.; O'Keefe, Barry R.

2013-01-01

181

Catechins and procyanidins of Ginkgo biloba show potent activities towards the inhibition of ?-amyloid peptide aggregation and destabilization of preformed fibrils.  

PubMed

Catechins and procyanidins, together with flavonoid glycosides and terpene trilactones, are three important categories of components in the standard extract of Ginkgo biloba leaves (EGb761). In this research, catechins and proanthocyanidins were found to exist in both the extract of Ginkgo leaves and Ginkgo products. By comparing with reference compounds, six of them were identified as (+)-catechin, (-)-epicatechin, (-)-gallocatechin, (-)-epigallocatechin and procyanidins B1 and B3. The activities of these polyphenols in the inhibition of A?42 aggregation and the destabilization of preformed fibrils were evaluated using biochemical assays, which showed that all six of the polyphenols, as well as a fraction of the extract of Ginkgo biloba leaves (EGb) containing catechins and procyanidins, exerted potent inhibitory activities towards A?42 aggregation and could also destabilize the performed fibrils. Catechins and procyanidins can therefore be regarded as the potent active constituents of EGb761 in terms of their inhibition of A?42 aggregation and destabilization of the fibrils. Although quantitative mass spectroscopic analysis revealed that the catechins and procyanidins are only present in low concentrations in EGb761, these components should be studied in greater detail because of their potent inhibitory effects towards A?42 aggregation and their ability to destabilize preformed fibrils, especially during the quality control of Ginkgo leaves and the manufacture of Ginkgo products. PMID:24759072

Xie, Haiyan; Wang, Jing-Rong; Yau, Lee-Fong; Liu, Yong; Liu, Liang; Han, Quan-Bin; Zhao, Zhongzhen; Jiang, Zhi-Hong

2014-01-01

182

Part I-mechanism of adaptation: high nitric oxide adapted A549 cells show enhanced DNA damage response and activation of antiapoptotic pathways.  

PubMed

Our previous studies demonstrate that A549, a human lung adenocarcinoma line, could be adapted to the free radical nitric oxide (NO([Symbol: see text])). NO([Symbol: see text]) has been shown to be overexpressed in human tumors. The original cell line, A549 (parent), and the newly adapted A549-HNO (which has a more aggressive phenotype) serves as a useful model system to study the role of NO([Symbol: see text]) in tumor biology. It is well known that DNA damage response (DDR) is altered in cancer cells and NO([Symbol: see text]) is known to cause DNA damage. Modulations in molecular mechanisms involved in DNA damage response in A549-HNO cells can provide better insights into the enhanced growth behavior of these cells. Thus, here, we carried out a series of time course experiments by treating A549 and A549-HNO cells with NO([Symbol: see text]) donor and examining levels of proteins involved in the DDR pathway. We observed induced expression of key components of DDR pathway in A549-HNO cells. The HNO cells showed sustained expression of key proteins involved in both nonhomologous end joining (NHEJ) and homologous recombination pathways, whereas parent cells only expressed low levels of NHEJ pathway proteins. Further with prolonged NO([Symbol: see text]) exposure, ATR, Chk1, and p53 were activated and upregulated in HNO cells. Activation of p53 results in inhibition of apoptosis through induced Mcl1 expression. It also leads to cell cycle modulation. Interestingly, several reports show that cancer stem cells have enhanced expression of proteins involved in DNA damage response and also activated an antiapoptotic response. Our results here suggest that our HNO adapted A549 cells have increased activation of DNA damage response pathway proteins which can lead to better DNA repair function. Enhanced DDR leads to activation of antiapoptosis response and modulation in the cell cycle which may lead to better survival of these cells under harsh conditions. Thus, our present investigation further supports the hypothesis that HNO exposure leads to survival of these cells. PMID:24241898

Aqil, Madeeha; Elseth, Kim M; Vesper, Benjamin J; Deliu, Zane; Aydogan, Bulent; Xue, Jiaping; Radosevich, James A

2014-03-01

183

Alpha-momorcharin, a RIP produced by bitter melon, enhances defense response in tobacco plants against diverse plant viruses and shows antifungal activity in vitro.  

PubMed

Alpha-momorcharin (?-MMC) is type-1 ribosome inactivating proteins (RIPs) with molecular weight of 29 kDa and has lots of biological activity. Our recent study indicated that the ?-MMC purified from seeds of Momordica charantia exhibited distinct antiviral and antifungal activity. Tobacco plants pre-treated with 0.5 mg/mL ?-MMC 3 days before inoculation with various viruses showed less-severe symptom and less reactive oxygen species (ROS) accumulation compared to that inoculated with viruses only. Quantitative real-time PCR analysis revealed that the replication levels of viruses were lower in the plants treated with the ?-MMC than control plants at 15 days post inoculation. Moreover, the coat protein expression of viruses was almost completely inhibited in plants which were treated with the ?-MMC compared with control plants. Furthermore, the SA-responsive defense-related genes including non-expressor of pathogenesis-related genes 1 (NPR1), PR1, PR2 were up-regulated and activities of some antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) were increased after the ?-MMC treatment. In addition, the ?-MMC (500 ?g/mL) revealed remarkable antifungal effect against phytopathogenic fungi, in the growth inhibition range 50.35-67.21 %, along with their MIC values ranging from 100 to 500 ?g/mL. The ?-MMC had also a strong detrimental effect on spore germination of all the tested plant pathogens along with concentration as well as time-dependent kinetic inhibition of Sclerotinia sclerotiorum. The ?-MMC showed a remarkable antiviral and antifungal effect and hence could possibly be exploited in crop protection for controlling certain important plant diseases. PMID:22983699

Zhu, Feng; Zhang, Ping; Meng, Yan-Fa; Xu, Fei; Zhang, Da-Wei; Cheng, Jian; Lin, Hong-Hui; Xi, De-Hui

2013-01-01

184

Tissue-specific microvascular endothelial cells show distinct capacity to activate NK cells: implications for the pathophysiology of granulomatosis with polyangiitis.  

PubMed

The relevance of tissue specificity of microvascular endothelial cells (MECs) in the response to inflammatory stimuli and sensitivity to immune cell-mediated injury is not well defined. We hypothesized that such MEC characteristics might shape their interaction with NK cells through the use of different adhesion molecules and NK cell receptor ligands or the release of different soluble factors and render them more or less vulnerable to NK cell injury during autoimmune vasculitis, such as granulomatosis with polyangiitis (GPA). To generate a comprehensive expression profile of human MECs of renal, lung, and dermal tissue origin, we characterized, in detail, their response to inflammatory cytokines and to proteinase 3, a major autoantigen in GPA, and analyzed the effects on NK cell activation. In this study, we show that renal MECs were more susceptible than lung and dermal MECs to the effect of inflammatory signals, showing upregulation of ICAM-1 and VCAM-1 on their surface, as well as release of CCL2, soluble fractalkine, and soluble VCAM-1. Proteinase 3-stimulated renal and lung MECs triggered CD107a degranulation in control NK cell. Notably, NK cells from GPA patients expressed markers of recent in vivo activation (CD69, CD107a), degranulated more efficiently than did control NK cells in the presence of renal MECs, and induced direct killing of renal MECs in vitro. These results suggest that, upon inflammatory conditions in GPA, renal MECs may contribute to the recruitment and activation of NK cells in the target vessel wall, which may participate in the necrotizing vasculitis of the kidney during this disease. PMID:24600034

Tognarelli, Sara; Gayet, Julie; Lambert, Marion; Dupuy, Stéphanie; Karras, Alexandre; Cohen, Pascal; Guillevin, Loïc; de Menthon, Mathilde; Caillat-Zucman, Sophie

2014-04-01

185

A novel esterase from Burkholderia gladioli which shows high deacetylation activity on cephalosporins is related to beta-lactamases and DD-peptidases.  

PubMed

The gene (estB) encoding for a novel esterase (EstB) from Burkholderia gladioli (formerly Pseudomonas marginata) NCPPB 1891 was cloned in Escherichia coli. Sequence analysis showed an open reading frame encoding a polypeptide of 392 amino acid residues, with a molecular mass of about 42 kDa. Comparison of the amino acid sequence with those of other homologous enzymes indicated homologies to beta-lactamases, penicillin binding proteins and DD-peptidases. The serine residue (Ser(75)) which is located within a present class A beta-lactamase motif ([F,Y]-X-[L,I,V,M,F,Y]-X-S-[T,V]-X-K-X-X-X-X-[A,G,L]-X-X-[L,C]) was identified by site-directed mutagenesis to represent the active nucleophile. A second serine residue (Ser(149)) which is located within a G-x-S-x-G motif which is typically found in esterases and lipases was demonstrated not to play a significant role in enzyme function. The estB gene was overexpressed in E. coli using a tac promoter-based expression system. Investigation of EstB protein with respect to the ability to hydrolyse beta-lactam substrates clearly demonstrated that this protein has no beta-lactamase activity. The recombinant enzyme is active on triglycerides and on nitrophenyl esters with acyl chain lengths up to C6. The preference for short chain length substrates indicated that EstB is a typical carboxylesterase. As a special feature EstB esterase was found to have high deacetylation activity on cephalosporin derivatives. PMID:11472796

Petersen, E I; Valinger, G; Sölkner, B; Stubenrauch, G; Schwab, H

2001-07-26

186

Novel phosphatidylinositol 3-kinase inhibitor NVP-BKM120 induces apoptosis in myeloma cells and shows synergistic anti-myeloma activity with dexamethasone.  

PubMed

NVP-BKM120 is a novel phosphatidylinositol 3-kinase (PI3K) inhibitor and is currently being investigated in phase I clinical trials in solid tumors. This study aimed to evaluate the therapeutic efficacy of BKM120 in multiple myeloma (MM). BKM120 induces cell growth inhibition and apoptosis in both MM cell lines and freshly isolated primary MM cells. However, BKM120 only shows limited cytotoxicity toward normal lymphocytes. The presence of MM bone marrow stromal cells, insulin-like growth factor, or interleukin-6 does not affect BKM120-induced tumor cell apoptosis. More importantly, BKM120 treatment significantly inhibits tumor growth in vivo and prolongs the survival of myeloma-bearing mice. In addition, BKM120 shows synergistic cytotoxicity with dexamethasone in dexamethasone-sensitive MM cells. Low doses of BKM120 and dexamethasone, each of which alone has limited cytotoxicity, induce significant cell apoptosis in MM.1S and ARP-1. Mechanistic study shows that BKM120 exposure causes cell cycle arrest by upregulating p27 (Kip1) and downregulating cyclin D1 and induces caspase-dependent apoptosis by downregulating antiapoptotic XIAP and upregulating expression of cytotoxic small isoform of Bim, BimS. In summary, our findings demonstrate the in vitro and in vivo anti-MM activity of BKM120 and suggest that BKM120 alone or together with other MM chemotherapeutics, particularly dexamethasone, may be a promising treatment for MM. PMID:22207485

Zheng, Yuhuan; Yang, Jing; Qian, Jianfei; Zhang, Liang; Lu, Yong; Li, Haiyan; Lin, Heather; Lan, Yongsheng; Liu, Zhiqiang; He, Jin; Hong, Sungyoul; Thomas, Sheeba; Shah, Jatin; Baladandayuthapani, Veera; Kwak, Larry W; Yi, Qing

2012-06-01

187

Localization of DIR1 at the tissue, cellular and subcellular levels during Systemic Acquired Resistance in Arabidopsis using DIR1:GUS and DIR1:EGFP reporters  

PubMed Central

Background Systemic Acquired Resistance (SAR) is an induced resistance response to pathogens, characterized by the translocation of a long-distance signal from induced leaves to distant tissues to prime them for increased resistance to future infection. DEFECTIVE in INDUCED RESISTANCE 1 (DIR1) has been hypothesized to chaperone a small signaling molecule to distant tissues during SAR in Arabidopsis. Results DIR1 promoter:DIR1-GUS/dir1-1 lines were constructed to examine DIR1 expression. DIR1 is expressed in seedlings, flowers and ubiquitously in untreated or mock-inoculated mature leaf cells, including phloem sieve elements and companion cells. Inoculation of leaves with SAR-inducing avirulent or virulent Pseudomonas syringae pv tomato (Pst) resulted in Type III Secretion System-dependent suppression of DIR1 expression in leaf cells. Transient expression of fluorescent fusion proteins in tobacco and intercellular washing fluid experiments indicated that DIR1's ER signal sequence targets it for secretion to the cell wall. However, DIR1 expressed without a signal sequence rescued the dir1-1 SAR defect, suggesting that a cytosolic pool of DIR1 is important for the SAR response. Conclusions Although expression of DIR1 decreases during SAR induction, the protein localizes to all living cell types of the vasculature, including companion cells and sieve elements, and therefore DIR1 is well situated to participate in long-distance signaling during SAR.

2011-01-01

188

Introduction of a NIa proteinase cleavage site between the reporter gene and HC-Pro only partially restores the biological properties of GUS- or GFP-tagged LMV.  

PubMed

Lettuce mosaic virus (LMV) isolates LMV-E and LMV-0 differ in their virulence on lettuce varieties carrying the mo1(2) resistance gene, which reduces viral accumulation and blocks the expression of symptoms after infection with avirulent isolates such as LMV-0. Previous work had indicated that reporter genes such as GUS or GFP affect the biological properties of recombinant LMV isolates in both susceptible and resistant lettuce varieties when fused to the N-terminus of the viral protein HC-Pro. The impact of the addition of a cleavage site for the NIa proteinase between the reporter gene and HC-Pro was evaluated, in an effort to recover the full spectrum of the biological properties of parental isolates. Symptoms, accumulation, cell-to-cell and long distance movement of the recombinant viruses containing the NIa cleavage site were studied in susceptible and mo1(2) lettuce varieties. Both LMV-0 and LMV-E recombinant viruses recovered the behaviour of their wild-type parent in susceptible plants upon addition of the NIa cleavage site. While the recombinant LMV-E modified in this way recovered the breaking properties of its wild-type counterpart in mo1(2) plants, similar modification of the LMV-0 derived recombinants failed to rescue a severe inhibition in systemic accumulation in mo1(2) plants, despite the fact that neither cell-to-cell movement nor phloem loading or unloading seemed to be severely affected. PMID:14659562

German-Retana, Sylvie; Redondo, Elise; Tavert-Roudet, Geneviève; Le Gall, Olivier; Candresse, Thierry

2003-12-01

189

Tentacles of in vitro-grown round-leaf sundew (Drosera rotundifolia L.) show induction of chitinase activity upon mimicking the presence of prey.  

PubMed

Induction of plant-derived chitinases in the leaves of a carnivorous plant was demonstrated using aseptically grown round-leaf sundew (Drosera rotundifolia L.). The presence of insect prey was mimicked by placing the chemical inducers gelatine, salicylic acid and crustacean chitin on leaves. In addition, mechanical stirring of tentacles was performed. Chitinase activity was markedly increased in leaf exudates upon application of notably chitin. Application of gelatine increased the proteolytic activity of leaf exudates, indicating that the reaction of sundew leaves depends on the molecular nature of the inducer applied. In situ hybridization of sundew leaves with a Drosera chitinase probe showed chitinase gene expression in different cell types of non-treated leaves, but not in the secretory cells of the glandular heads. Upon induction, chitinase mRNA was also present in the secretory cells of the sundew leaf. The combined results indicate that chitinase is likely to be involved in the decomposition of insect prey by carnivorous plants. This adds a novel role to the already broad function of chitinases in the plant kingdom and may contribute to our understanding of the molecular mechanisms behind the ecological success of carnivorous plants in nutritionally poor environments. PMID:16049675

Matusíková, Ildikó; Salaj, Ján; Moravcíková, Jana; Mlynárová, Ludmila; Nap, Jan-Peter; Libantová, Jana

2005-12-01

190

Reduced Expression of the Retinoblastoma Protein Shows That the Related Signaling Pathway Is Essential for Mediating the Antineoplastic Activity of Erufosine  

PubMed Central

Erufosine is a new antineoplastic agent of the group of alkylphosphocholines, which interferes with signal transduction and induces apoptosis in various leukemic and tumor cell lines. The present study was designed to examine for the first time the mechanism of resistance to erufosine in malignant cells with permanently reduced expression of the retinoblastoma (Rb) protein. Bearing in mind the high number of malignancies with reduced level of this tumor-suppressor, this investigation was deemed important for using erufosine, alone or in combination, in patients with compromised RB1 gene expression. For this purpose, clones of the leukemic T-cell line SKW-3 were used, which had been engineered to constantly express differently low Rb levels. The alkylphosphocholine induced apoptosis, stimulated the expression of the cyclin dependent kinase inhibitor p27Kip1 and inhibited the synthesis of cyclin D3, thereby causing a G2 phase cell cycle arrest and death of cells with wild type Rb expression. In contrast, Rb-deficiency impeded the changes induced by eru-fosine in the expression of these proteins and abrogated the induction of G2 arrest, which was correlated with reduced antiproliferative and anticlonogenic activities of the compound. In conclusion, analysis of our results showed for the first time that the Rb signaling pathway is essential for mediating the antineoplastic activity of erufosine and its efficacy in patients with malignant diseases may be predicted by determining the Rb status.

Zaharieva, Maya M.; Kirilov, Milen; Chai, Minquang; Berger, Stefan M.; Konstantinov, Spiro; Berger, Martin R.

2014-01-01

191

Comparing routes of delivery for nanoliposomal irinotecan shows superior anti-tumor activity of local administration in treating intracranial glioblastoma xenografts  

PubMed Central

Background Liposomal drug packaging is well established as an effective means for increasing drug half-life, sustaining drug activity, and increasing drug efficacy, whether administered locally or distally to the site of disease. However, information regarding the relative effectiveness of peripheral (distal) versus local administration of liposomal therapeutics is limited. This issue is of importance with respect to the treatment of central nervous system cancer, for which the blood-brain barrier presents a significant challenge in achieving sufficient drug concentration in tumors to provide treatment benefit for patients. Methods We compared the anti-tumor activity and efficacy of a nanoliposomal formulation of irinotecan when delivered peripherally by vascular route with intratumoral administration by convection-enhanced delivery (CED) for treating intracranial glioblastoma xenografts in athymic mice. Results Our results show significantly greater anti-tumor activity and survival benefit from CED of nanoliposomal irinotecan. In 2 of 3 efficacy experiments, there were animal subjects that experienced apparent cure of tumor from local administration of therapy, as indicated by a lack of detectable intracranial tumor through bioluminescence imaging and histopathologic analysis. Results from investigating the effectiveness of combination therapy with nanoliposomal irinotecan plus radiation revealed that CED administration of irinotecan plus radiation conferred greater survival benefit than did irinotecan or radiation monotherapy and also when compared with radiation plus vascularly administered irinotecan. Conclusions Our results indicate that liposomal formulation plus direct intratumoral administration of therapeutic are important for maximizing the anti-tumor effects of irinotecan and support clinical trial evaluation of this therapeutic plus route of administration combination.

Chen, Pin-Yuan; Ozawa, Tomoko; Drummond, Daryl C.; Kalra, Ashish; Fitzgerald, Jonathan B.; Kirpotin, Dmitri B.; Wei, Kuo-Chen; Butowski, Nicholas; Prados, Michael D.; Berger, Mitchel S.; Forsayeth, John R.; Bankiewicz, Krystof; James, C. David

2013-01-01

192

Background Synaptic Activity in Rat Entorhinal Cortex Shows a Progressively Greater Dominance of Inhibition over Excitation from Deep to Superficial Layers  

PubMed Central

The entorhinal cortex (EC) controls hippocampal input and output, playing major roles in memory and spatial navigation. Different layers of the EC subserve different functions and a number of studies have compared properties of neurones across layers. We have studied synaptic inhibition and excitation in EC neurones, and we have previously compared spontaneous synaptic release of glutamate and GABA using patch clamp recordings of synaptic currents in principal neurones of layers II (L2) and V (L5). Here, we add comparative studies in layer III (L3). Such studies essentially look at neuronal activity from a presynaptic viewpoint. To correlate this with the postsynaptic consequences of spontaneous transmitter release, we have determined global postsynaptic conductances mediated by the two transmitters, using a method to estimate conductances from membrane potential fluctuations. We have previously presented some of this data for L3 and now extend to L2 and L5. Inhibition dominates excitation in all layers but the ratio follows a clear rank order (highest to lowest) of L2>L3>L5. The variance of the background conductances was markedly higher for excitation and inhibition in L2 compared to L3 or L5. We also show that induction of synchronized network epileptiform activity by blockade of GABA inhibition reveals a relative reluctance of L2 to participate in such activity. This was associated with maintenance of a dominant background inhibition in L2, whereas in L3 and L5 the absolute level of inhibition fell below that of excitation, coincident with the appearance of synchronized discharges. Further experiments identified potential roles for competition for bicuculline by ambient GABA at the GABAA receptor, and strychnine-sensitive glycine receptors in residual inhibition in L2. We discuss our results in terms of control of excitability in neuronal subpopulations of EC neurones and what these may suggest for their functional roles.

Greenhill, Stuart David; Chamberlain, Sophie Elizabeth Lyn; Lench, Alex; Massey, Peter Vernon; Yuill, Kathryn Heather; Woodhall, Gavin Lawrence; Jones, Roland Spencer Gwynne

2014-01-01

193

Structural analysis of the alcohol acyltransferase protein family from Cucumis melo shows that enzyme activity depends on an essential solvent channel.  

PubMed

Alcohol acyltransferases (AAT) play a key role in ester biosynthesis. In Cucumis melo var. cantalupensis, AATs are encoded by a gene family of four members (CmAAT1-4). CmAAT1, CmAAT3 and CmAAT4 are capable of synthesizing esters, with CmAAT1 the most active. CmAAT2 is inactive and has an Ala268 residue instead of a threonine which is present in all other active AATs, although the role of this residue is still unclear. The present work aims to understand the molecular mechanism involved in ester biosynthesis in melon fruit and to clarify the importance of the Ala268 residue. First, structural models for each protein were built by comparative modelling methodology. Afterwards, conformational interaction between the protein and several ligands, alcohols and acyl-CoAs was explored by molecular docking and molecular dynamics simulation. Structural analysis showed that CmAATs share a similar structure. Also, well-defined solvent channels were described in the CmAATs except for CmAAT2 which does not have a proper channel and instead has a small pocket around Ala268. Residues of the catalytic HxxxD motif interact with substrates within the solvent channel, with Ser363 also important. Strong binding interaction energies were described for the best substrate couple of each CmAAT (hexyl-, benzyl- and cinnamyl-acetate for CmAAT1, 3 and 4 respectively). CmAAT1 and CmAAT2 protein surfaces share similar electrostatic potentials; nevertheless the entrance channels for the substrates differ in location and electrostatic character, suggesting that Ala268 might be responsible for that. This could partly explain the major differences in activity reported for these two enzymes. PMID:23311868

Galaz, Sebastián; Morales-Quintana, Luis; Moya-León, María Alejandra; Herrera, Raúl

2013-03-01

194

The fruiting body and its caterpillar host of Cordyceps sinensis show close resemblance in main constituents and anti-oxidation activity.  

PubMed

Cordyceps (summer-grass, winter-worm), one of the most valued traditional Chinese medicines, is used commonly for the replenishment of body health. It consists of the dried fungus Cordyceps sinensis growing on caterpillar larvae. For medication, the fruiting body (fungus) and the worm (caterpillar) are used together. However, the pharmacological efficiency and the main constituents of the individual parts have not been determined. In the present study the water extracts from the fruiting body and worm of natural Cordyceps were analyzed for their content of nucleosides and polysaccharides; the results showed that the worm had chemical composition similar to the fruiting body. In addition, both the fruiting body and worm of Cordyceps showed similar potency in their anti-oxidation activities in the xanthine oxidase assay, the induction of hemolysis assay and the lipid-peroxidation assay. These results suggest that the function of the worm in Cordyceps is to provide a growth medium for the fruiting body, and that eventually, the worm is totally invaded by C. sinensis mycelia. PMID:12120813

Li, S P; Su, Z R; Dong, T T X; Tsim, K W K

2002-05-01

195

Macrophages in T cell/histiocyte rich large B cell lymphoma strongly express metal-binding proteins and show a bi-activated phenotype.  

PubMed

Abundant macrophage infiltration in tumors often correlates with a poor prognosis. T cell/histiocyte rich large B cell lymphoma (THRLBCL) is a distinct aggressive B cell lymphoma entity showing a high macrophage content. To further elucidate the role of tumor-associated macrophages in THRLBCL, we performed gene expression profiling of microdissected histiocyte subsets of THRLBCL, nodular lymphocyte predominant Hodgkin lymphoma (NLPHL), Piringer lymphadenitis, sarcoidosis, nonspecific lymphadenitis and monocytes from peripheral blood. In a supervised principal component analysis, histiocytes from THRLBCL were most closely related to epithelioid cells from NLPHL, with both types of cells expressing genes related to proinflammatory and regulatory macrophage activity. Moreover, histiocytes from THRLBCL strongly expressed metal-binding proteins like MT2A, by which histiocytes of THRLBCL can be distinguished from the other histiocyte subsets investigated. Interestingly, the validation at the protein level showed a strong expression of TXN, CXCL9, MT2A and SOD2 not only in macrophages of THRLBCL but also in the tumor cells of NLPHL and classical Hodgkin lymphoma (cHL). Overall, the present findings indicate that macrophages in the microenvironment of THRLBCL have acquired a distinct gene expression pattern that is characterized by a mixed M1/M2 phenotype and a strong expression of several metal binding proteins. The microenvironments in NLPHL and THRLBCL appear to have a similar influence on the macrophage phenotype. The high expression of metal binding proteins in histiocytes of THRLBCL may be diagnostically useful, but a potential pathophysiological role remains to be identified. PMID:23686423

Hartmann, Sylvia; Tousseyn, Thomas; Döring, Claudia; Flüchter, Patricia; Hackstein, Holger; Herreman, An; Ponzoni, Maurilio; de Wolf-Peeters, Chris; Facchetti, Fabio; Gascoyne, Randy D; Küppers, Ralf; Steidl, Christian; Hansmann, Martin-Leo

2013-12-01

196

A socio-ecological approach promoting physical activity and limiting sedentary behavior in adolescence showed weight benefits maintained 2.5 years after intervention cessation.  

PubMed

Background:Obesity in youth remains a major public health issue. Yet no effective long-term preventive strategy exists. We previously showed that a school-based socio-ecological approach targeting behavior and social/environmental influences on physical activity (PA) prevented 4-year excessive weight gain in 12-year olds. In this study, we investigated if this efficacy persists 30 months after intervention cessation.Methods and Findings:The program targeted students, family, school and the living environment to promote/support PA and prevent sedentary behavior (SB). A total of 732 students from eight randomized middle schools completed the 4-year trial. At the 30-month post-trial follow-up, body mass index (BMI), fat mass index (FMI), leisure PA (LPA), home/school/workplace active commuting, TV/video time (TVT), and attitudes toward PA were measured in 531 adolescents. The beneficial effects of the intervention on the excess BMI increase (+0.01 vs +0.34?kg?m(-2) in the intervention and control groups, respectively) and on the overweight incidence in initially non-overweight students (4.3% vs 8.6%; odds ratio=0.48 (95% confidence interval: 0.23-1.01)) were maintained at the post-trial follow-up. LPA was not maintained at the level achieved during the trial. However, we still observed a prevention of the age-related decrease of the adolescents' percentage reporting regular LPA (-14.4% vs -26.5%) and a higher intention to exercise in the intervention group. The intervention promoted lower TVT (-14.0 vs +13.6?min per day) and higher active commuting changes (+11.7% vs -4.8%). Trends in higher BMI reduction in students with high initial TVT and in the least wealthy group were noted. TVT changes throughout the follow-up predicted excess BMI and FMI changes.Conclusions:Long-term multilevel approach targeting PA and SB prevents excessive weight gain up to 30 months after intervention cessation. The efficacy may be higher in the most sedentary and least wealthy adolescents. Healthy PA-related behavior inducing long-lasting weight effects can be promoted in youth providing that an ecological approach is introduced in the prevention strategy. PMID:24509504

Simon, C; Kellou, N; Dugas, J; Platat, C; Copin, N; Schweitzer, B; Hausser, F; Bergouignan, A; Lefai, E; Blanc, S

2014-07-01

197

Phenobarbital and propiconazole toxicogenomic profiles in mice show major similarities consistent with the key role that constitutive androstane receptor (CAR) activation plays in their mode of action  

PubMed Central

Toxicogenomics (TGx) is employed frequently to investigate underlying molecular mechanisms of the compound of interest and, thus, has become an aid to mode of action determination. However, the results and interpretation of a TGx dataset are influenced by the experimental design and methods of analysis employed. This article describes an evaluation and reanalysis, by two independent laboratories, of previously published TGx mouse liver microarray data for a triazole fungicide, propiconazole (PPZ), and the anticonvulsant drug phenobarbital (PB). Propiconazole produced an increase incidence of liver tumors in male CD-1 mice only at a dose that exceeded the maximum tolerated dose (2500 ppm). Firstly, we illustrate how experimental design differences between two in vivo studies with PPZ and PB may impact the comparisons of TGx results. Secondly, we demonstrate that different researchers using different pathway analysis tools can come to different conclusions on specific mechanistic pathways, even when using the same datasets. Finally, despite these differences the results across three different analyses also show a striking degree of similarity observed for PPZ and PB treated livers when the expression data are viewed as major signaling pathways and cell processes affected. Additional studies described here show that the postulated key event of hepatocellular proliferation was observed in CD-1 mice for both PPZ and PB, and that PPZ is also a potent activator of the mouse CAR nuclear receptor. Thus, with regard to the events which are hallmarks of CAR-induced effects that are key events in the mode of action (MOA) of mouse liver carcinogenesis with PB, PPZ-induced tumors can be viewed as being promoted by a similar PB-like CAR-dependent MOA.

Currie, Richard A.; Peffer, Richard C.; Goetz, Amber K.; Omiecinski, Curtis J.; Goodman, Jay I.

2014-01-01

198

Phenobarbital and propiconazole toxicogenomic profiles in mice show major similarities consistent with the key role that constitutive androstane receptor (CAR) activation plays in their mode of action.  

PubMed

Toxicogenomics (TGx) is employed frequently to investigate underlying molecular mechanisms of the compound of interest and, thus, has become an aid to mode of action determination. However, the results and interpretation of a TGx dataset are influenced by the experimental design and methods of analysis employed. This article describes an evaluation and reanalysis, by two independent laboratories, of previously published TGx mouse liver microarray data for a triazole fungicide, propiconazole (PPZ), and the anticonvulsant drug phenobarbital (PB). Propiconazole produced an increase incidence of liver tumors in male CD-1 mice only at a dose that exceeded the maximum tolerated dose (2500 ppm). Firstly, we illustrate how experimental design differences between two in vivo studies with PPZ and PB may impact the comparisons of TGx results. Secondly, we demonstrate that different researchers using different pathway analysis tools can come to different conclusions on specific mechanistic pathways, even when using the same datasets. Finally, despite these differences the results across three different analyses also show a striking degree of similarity observed for PPZ and PB treated livers when the expression data are viewed as major signaling pathways and cell processes affected. Additional studies described here show that the postulated key event of hepatocellular proliferation was observed in CD-1 mice for both PPZ and PB, and that PPZ is also a potent activator of the mouse CAR nuclear receptor. Thus, with regard to the events which are hallmarks of CAR-induced effects that are key events in the mode of action (MOA) of mouse liver carcinogenesis with PB, PPZ-induced tumors can be viewed as being promoted by a similar PB-like CAR-dependent MOA. PMID:24675475

Currie, Richard A; Peffer, Richard C; Goetz, Amber K; Omiecinski, Curtis J; Goodman, Jay I

2014-07-01

199

The novel toluidine sulphonamide EL102 shows pre-clinical in vitro and in vivo activity against prostate cancer and circumvents MDR1 resistance  

PubMed Central

Background: Taxanes are routinely used for the treatment of prostate cancer, however the majority of patients eventually develop resistance. We investigated the potential efficacy of EL102, a novel toluidine sulphonamide, in pre-clinical models of prostate cancer. Methods: The effect of EL102 and/or docetaxel on PC-3, DU145, 22Rv1 and CWR22 prostate cancer cells was assessed using cell viability, cell cycle analysis and PARP cleavage assays. Tubulin polymerisation and immunofluorescence assays were used to assess tubulin dynamics. CWR22 xenograft murine model was used to assess effects on tumour proliferation. Multidrug-resistant lung cancer DLKPA was used to assess EL102 in a MDR1-mediated drug resistance background. Results: EL102 has in vitro activity against prostate cancer, characterised by accumulation in G2/M, induction of apoptosis, inhibition of Hif1?, and inhibition of tubulin polymerisation and decreased microtubule stability. In vivo, a combination of EL102 and docetaxel exhibits superior tumour inhibition. The DLKP cell line and multidrug-resistant DLKPA variant (which exhibits 205 to 691-fold greater resistance to docetaxel, paclitaxel, vincristine and doxorubicin) are equally sensitive to EL102. Conclusion: EL102 shows potential as both a single agent and within combination regimens for the treatment of prostate cancer, particularly in the chemoresistance setting.

Toner, A P; McLaughlin, F; Giles, F J; Sullivan, F J; O'Connell, E; Carleton, L A; Breen, L; Dunne, G; Gorman, A M; Lewis, J D; Glynn, S A

2013-01-01

200

CKR-L3, a deletion version CCR6-isoform shows coreceptor-activity for limited human and simian immunodeficiency viruses  

PubMed Central

Background The chemokine receptors (CKRs), mainly CCR5 and CXCR4 function as major coreceptors in infections caused by human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV). Approximately 20 G protein-coupled receptors (GPCRs) have been identified as minor coreceptors, alike CCR6 that we reported recently. Since CKR-L3 is indentified as a natural isoform of CCR6, we attempted in this study to explore the coreceptor function of CKR-L3. Methods NP-2 cells transduced with CD4-receptor (NP-2/CD4) normally remain resistant to HIV or SIV infection. However, the introduction of functional coreceptors can make these cells susceptible to these viruses. NP-2/CD4/CKR-L3 cells were produced to examine the coreceptor activity of CKR-L3. Likely, CCR6-isoform and the major coreceptors, CCR5 and CXCR4 were also examined in parallel. Presence of viral antigen in infected NP-2/CD4/coreceptor cells was detected by indirect immunofluorescence assay (IFA). The results were validated by detection of syncytia, proviral DNA and by measuring reverse transcriptase (RT) activities. Results HIV-2MIR and SIVsmE660 were found to infect NP-2/CD4/CKR-L3 cells, indicative of the coreceptor function of CKR-L3. Viral antigens appeared faster in NP-2/CD4/CKR-L3 cells than in NP-2/CD4/CCR6, indicating that CKR-L3 is a more efficient coreceptor. Moreover, syncytia formation was more rapid and RT release evidenced earlier and at higher levels with CKR-L3 than with CCR6. Sequence analysis in the C2-V3 envelope region of HIV-2MIR replicated through CKR-L3 and CCR6 coreceptor showed two and three amino acid substitutions respectively, in the C2 region compared to the CCR5-variant. The SIVsmE660-CKRL3 variant showed three amino acid substitutions in the V1 region, one change in the V2 and two changes in the C2 region. The SIVsmE660-CCR6 variant produced two changes in the V1 region, and three in the C2 region. Conclusions Isoform CKR-L3 exhibited coreceptor activity for limited primary HIV and SIV isolates with better efficiency than the parent CCR6-isoform. Amino acid substitutions in the envelope region of these viruses may confer selective pressure towards CKR-L3-use. CKR-L3 with other minor coreceptors may contribute to HIV and SIV pathogenesis including dissemination, trafficking and latency especially when major coreceptors become compromised. However, further works will be required to determine its clinical significance in HIV and SIV infection.

2014-01-01

201

Show Me Your Menu.  

ERIC Educational Resources Information Center

Describes the importance of eating properly. Presents two middle school level activities: the first activity has students record what they eat for nine days; the second activity includes listing the foods according to the food pyramid and discussing their decisions. Recommends three Web sites. (YDS)

Allen, Barbara

2000-01-01

202

Peripheral blood mononuclear cells isolated from patients with diabetic nephropathy show increased activation of the oxidative-stress sensitive transcription factor NF-kB  

Microsoft Academic Search

Summary   Increased oxidative stress and subsequent activation of the transcription factor NF-kB has been linked to the development\\u000a of late diabetic complications. To determine whether oxidative stress dependent NF-kB activation is evident in patients with\\u000a diabetic nephropathy we used an Electrophoretic Mobility Shift Assay based semiquantitative detection system which enabled\\u000a us to determine NF-kB activation in ex vivo isolated peripheral

M. A. Hofmann; S. Schiekofer; B. Isermann; M. Kanitz; M. Henkels; M. Joswig; A. Treusch; M. Morcos; T. Weiss; V. Borcea; A. K. M. Abdel Khalek; J. Amiral; H. Tritschler; E. Ritz; P. Wahl; R. Ziegler; A. Bierhaus; P. P. Nawroth

1999-01-01

203

Phenacetin O-deethylase: an activity of a cytochrome P-450 showing genetic linkage with that catalysing the 4-hydroxylation of debrisoquine?  

PubMed Central

Phenacetin O-deethylase activity was impaired, both in vivo and in vitro, in poor metabolisers of debrisoquine, consistent with the work of others. No impairment was observed in the oxidation of acetanilide, amylobarbitone or antipyrine in the PM phenotype. There was a good correlation (r = 0.804) between the high affinity component of phenacetin O-deethylase and debrisoquine 4-hydroxylase activities. No such correlation was observed with the low affinity component of phenacetin O-deethylase activity. Although debrisoquine was a competitive inhibitor of phenacetin O-deethylase activity, phenacetin was without effect on debrisoquine 4-hydroxylation. There was also marked differences in the effects of sparteine, guanoxan and alpha-naphthoflavone on the two activities. Cigarette smoking was associated with a significant, two-fold, increase in phenacetin O-deethylase activity whilst debrisoquine 4-hydroxylase activity was not affected. It is concluded that the high affinity component of phenacetin O-deethylase and debrisoquine 4-hydroxylase activities are catalysed by different isozymes of cytochrome P-450 but that these are most probably regulated by closely linked genes.

Kahn, G C; Boobis, A R; Brodie, M J; Toverud, E L; Murray, S; Davies, D S

1985-01-01

204

The maize autonomous element Activator ( Ac ) shows a minimal germinal excision frequency of 0.2%–0.5% in transgenic Arabidopsis thaliana plants  

Microsoft Academic Search

The autonomous mobile element Activator from Zea mays was introduced into Arabidopsis thaliana via Agrobacterium-mediated gene transfer. The use of a chimaeric construct, where the Ac element is located in the leader of the neomycin phosphotransferase (NPT II) gene, enabled the excision of Ac to be monitored by assaying for the reconstitution of NPT II gene activity. Using this approach,

Renate Schmidt; Lothar Willmitzer

1989-01-01

205

Stress Sensitive Healthy Females Show Less Left Amygdala Activation in Response to Withdrawal-Related Visual Stimuli under Passive Viewing Conditions  

ERIC Educational Resources Information Center

The amygdalae are key players in the processing of a variety of emotional stimuli. Especially aversive visual stimuli have been reported to attract attention and activate the amygdalae. However, as it has been argued that passively viewing withdrawal-related images could attenuate instead of activate amygdalae neuronal responses, its role under…

Baeken, Chris; Van Schuerbeek, Peter; De Raedt, Rudi; Vanderhasselt, Marie-Anne; De Mey, Johan; Bossuyt, Axel; Luypaert, Robert

2012-01-01

206

A 'Propagating' Active Across-Arc Normal Fault Shows Rupture Process of the Basement: the Case of the Southwestern Ryukyu Arc  

NASA Astrophysics Data System (ADS)

Ryukyu Arc is located on the southwestern extension of Japanese Island-arc towards the east of Taiwan Island along the margin of the Asian continent off China. The island-arc forms an arcuate trench-arc-backarc system. A NW-ward subduction of the Philippine Sea Plate (PSP)at a rate of 6-8 cm/y relative to the Eurasian Plate (EP) causes frequent earthquakes. The PSP is subducting almost normally in the north-central area and more obliquely around the southwestern area. Behind the arc-trench system, the Okinawa Trough (OT) was formed by back-arc rifting, where active hydrothermal vent systems have been discovered. Several across-arc submarine faults are located in the central and southern Ryukyu Arc. The East Ishigaki Fault (EIF) is one of the across-arc normal faults located in the southwestern Ryukyu Arc, ranging by 44km and extending from SE to NW. This fault was surveyed by SEABAT8160 multibeam echo sounder and by ROV Hyper-Dolphin in 2005 and 2008. The result shows that the main fault consists of five fault segments. A branched segment from the main fault was also observed. The southernmost segment is most mature (oldest but still active) and the northernmost one is most nascent. This suggests the north-westward propagation of the fault rupture corresponding to the rifting of the southwestern OT and the southward retreat of the arc-trench system. Considering that the fault is segmented and in some part branched, propagation might take place episodically rather than continuously from SE to NW. The ROV survey also revealed the rupture process of the limestone basement along this fault from the nascent stage to the mature stage. Most of the rock samples collected from the basement outcrop were limestone blocks (or calcareous sedimentary rocks). Limestone basement was observed to the west on the hanging wall far away from the main fault scarp. Then fine-grained sand with ripple marks was observed towards the main scarp. Limestone basement was observed on the main scarp and on the footwall. These suggest that basically the both sides are composed of the same material, that the whole study area is characterised by Ryukyu limestone exposure and that the basement was split by the across-arc normal fault. Coarse-grained sand and gravels/rubbles were observed towards and on the trough of the fault. On the main scarp an outcrop of limestone basement was exposed and in some part it was broken into rubbles. These facts suggest that crash of the basement due to rupturing is taking place repeatedly on the scarp and the trough. The observed fine-grained sand on the hanging wall might be the final product by the process of the crash of the limestone basement.

Matsumoto, T.; Shinjo, R.; Nakamura, M.; Kubo, A.; Doi, A.; Tamanaha, S.

2011-12-01

207

Television Quiz Show Simulation  

ERIC Educational Resources Information Center

This article explores the simulation of four television quiz shows for students in China studying English as a foreign language (EFL). It discusses the adaptation and implementation of television quiz shows and how the students reacted to them.

Hill, Jonnie Lynn

2007-01-01

208

213Bi-PAI2 conjugate selectively induces apoptosis in PC3 metastatic prostate cancer cell line and shows anti-cancer activity in a xenograft animal model  

PubMed Central

A novel ?-particle emitting (213Bi) plasminogen activator inhibitor type 2 construct, which targets the membrane-bound urokinase plasminogen activator on prostate cancer cells, was prepared and evaluated in vitro and in a xenograft animal model. The PC3 prostate cancer cell line expresses urokinase plasminogen activator which binds to its receptor on the cell membrane; plasminogen activator inhibitor type 2 is bound to urokinase plasminogen activator/urokinase plasminogen activator receptor to form stable complexes. In vitro, the cytotoxicity of 213Bi-plasminogen activator inhibitor type 2 against prostate cancer cells was tested using the MTS assay and apoptosis was documented using terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphosphate nick end-labelling (TUNEL) assay. In vivo, antiproliferative effects for tumours and prostate cancer lymph node metastasis were carried out in an athymic nude mouse model with a subcutaneous xenograft of PC3 cells. 213Bi-plasminogen activator inhibitor type 2 was specifically cytotoxic to PC3 cells in a concentration-dependent fashion, causing the cells to undergo apoptosis. A single local or i.p. injection of 213Bi-plasminogen activator inhibitor type 2 was able to completely regress the growth of tumours and lymph node metastases 2 days post subcutaneous inoculation, and obvious tumour regression was achieved in the therapy groups compared with control groups with 213Bi-plasminogen activator inhibitor type 2 when the tumours measured 30–40?mm3 and 85–100?mm3. All control animals and one of five (20%) mice treated with 3?mCi?kg?1 213Bi-plasminogen activator inhibitor type 2 developed metastases in the lymph nodes while no lymphatic spread of cancer was found in the 6?mCi?kg?1 treated groups at 2 days and 2 weeks post-cell inoculation. These results demonstrate that this novel 213Bi-plasminogen activator inhibitor type 2 conjugate selectively targets prostate cancer in vitro and in vivo, and could be considered for further development for the therapy of prostate cancer, especially for the control of micro-metastases or in minimal residual disease. British Journal of Cancer (2002) 86, 1197–1203. DOI: 10.1038/sj/bjc/6600179 www.bjcancer.com © 2002 Cancer Research UK

Li, Y; Rizvi, S M A; Ranson, M; Allen, B J

2002-01-01

209

WRKY Transcription Factors Involved in Activation of SA Biosynthesis Genes  

PubMed Central

Background Increased defense against a variety of pathogens in plants is achieved through activation of a mechanism known as systemic acquired resistance (SAR). The broad-spectrum resistance brought about by SAR is mediated through salicylic acid (SA). An important step in SA biosynthesis in Arabidopsis is the conversion of chorismate to isochorismate through the action of isochorismate synthase, encoded by the ICS1 gene. Also AVRPPHB SUSCEPTIBLE 3 (PBS3) plays an important role in SA metabolism, as pbs3 mutants accumulate drastically reduced levels of SA-glucoside, a putative storage form of SA. Bioinformatics analysis previously performed by us identified WRKY28 and WRKY46 as possible regulators of ICS1 and PBS3. Results Expression studies with ICS1 promoter::?-glucuronidase (GUS) genes in Arabidopsis thaliana protoplasts cotransfected with 35S::WRKY28 showed that over expression of WRKY28 resulted in a strong increase in GUS expression. Moreover, qRT-PCR analyses indicated that the endogenous ICS1 and PBS3 genes were highly expressed in protoplasts overexpressing WRKY28 or WRKY46, respectively. Electrophoretic mobility shift assays indentified potential WRKY28 binding sites in the ICS1 promoter, positioned -445 and -460 base pairs upstream of the transcription start site. Mutation of these sites in protoplast transactivation assays showed that these binding sites are functionally important for activation of the ICS1 promoter. Chromatin immunoprecipitation assays with haemagglutinin-epitope-tagged WRKY28 showed that the region of the ICS1 promoter containing the binding sites at -445 and -460 was highly enriched in the immunoprecipitated DNA. Conclusions The results obtained here confirm results from our multiple microarray co-expression analyses indicating that WRKY28 and WRKY46 are transcriptional activators of ICS1 and PBS3, respectively, and support this in silico screening as a powerful tool for identifying new components of stress signaling pathways.

2011-01-01

210

Two Proteins with Ornithine Acetyltransferase Activity Show Different Functions in Streptomyces clavuligerus: Oat2 Modulates Clavulanic Acid Biosynthesis in Response to Arginine  

Microsoft Academic Search

The oat2 gene, located in the clavulanic acid gene cluster in Streptomyces clavuligerus, is similar to argJ, which encodes N-acetylornithine:glutamic acid acetyltransferase activity. Purified proteins obtained by expression in Escherichia coli of the argJ and oat2 genes of S. clavuligerus posses N-acetyltransferase activity. The kinetics and substrate specificities of both proteins are very similar. Deletion of the oat2 gene did

A. de la Fuente; J. F. Martin; A. Rodriguez-Garcia; P. Liras

2004-01-01

211

The CXCR3(+)CD56Bright Phenotype Characterizes a Distinct NK Cell Subset with Anti-Fibrotic Potential That Shows Dys-Regulated Activity in Hepatitis C  

Microsoft Academic Search

BackgroundIn mouse models, natural killer (NK) cells have been shown to exert anti-fibrotic activity via killing of activated hepatic stellate cells (HSC). Chemokines and chemokine receptors critically modulate hepatic recruitment of NK cells. In hepatitis C, the chemokine receptor CXCR3 and its ligands have been shown to be associated with stage of fibrosis suggesting a role of these chemokines in

Marianne Eisenhardt; Andreas Glässner; Benjamin Krämer; Christian Körner; Bernhard Sibbing; Pavlos Kokordelis; Hans Dieter Nischalke; Tilman Sauerbruch; Ulrich Spengler; Jacob Nattermann

2012-01-01

212

Allosteric mutants show that PrfA activation is dispensable for vacuole escape but required for efficient spread and Listeria survival in vivo  

PubMed Central

The transcriptional regulator PrfA controls key virulence determinants of the facultative intracellular pathogen Listeria monocytogenes. PrfA-dependent gene expression is strongly induced within host cells. While the basis of this activation is unknown, the structural homology of PrfA with the cAMP receptor protein (Crp) and the finding of constitutively activated PrfA* mutants suggests it may involve ligand-induced allostery. Here, we report the identification of a solvent-accessible cavity within the PrfA N-terminal domain that may accommodate an activating ligand. The pocket occupies a similar position to the cAMP binding site in Crp but lacks the cyclic nucleotide-anchoring motif and has its entrance on the opposite side of the ?-barrel. Site-directed mutations in this pocket impaired intracellular PrfA-dependent gene activation without causing extensive structural/functional alterations to PrfA. Two substitutions, L48F and Y63W, almost completely abolished intracellular virulence gene induction and thus displayed the expected phenotype for allosteric activation-deficient PrfA mutations. Neither PrfAallo substitution affected vacuole escape and initial intracellular growth of L. monocytogenes in epithelial cells and macrophages but caused defective cell-to-cell spread and strong attenuation in mice. Our data support the hypothesis that PrfA is allosterically activated during intracellular infection and identify the probable binding site for the effector ligand. They also indicate that PrfA allosteric activation is not required for early intracellular survival but is essential for full Listeria virulence and colonization of host tissues.

Deshayes, Caroline; Bielecka, Magdalena K; Cain, Robert J; Scortti, Mariela; de las Heras, Aitor; Pietras, Zbigniew; Luisi, Ben F; Nunez Miguel, Ricardo; Vazquez-Boland, Jose A

2012-01-01

213

KSC Wildlife Show  

NASA Technical Reports Server (NTRS)

This video highlights footage of the many forms of animal and plant life that inhabit the environs surrounding KSC. Shown are birds, alligators, butterflies, and plants as they react to shuttle launches and other activities eminating from KSC.

1994-01-01

214

Macrophages in resistance to rickettsial infection: strains of mice susceptible to the lethal effects of Rickettsia akari show defective macrophage Rickettsicidal activity in vitro.  

PubMed Central

Activation of macrophages was assessed in strains of mice inoculated intraperitoneally with 1,000 times the 50% lethal dose of Rickettsia akari. Macrophages from mice resistant to R. akari infection (C3H/HeN, C57BL/10J, and BALB/cN) were nonspecifically tumoricidal 2 to 4 days after rickettsial inoculation. Moreover, these macrophages were microbial for R. akari in vitro; cells were resistant to infection with the bacterium and were capable of killing intracellular rickettsiae. In contrast, macrophages from strains of mice susceptible to R. akari (C3H/HeJ, C57BL/10SnCR, and A/J) failed to develop nonspecific tumoricidal activity over the course of lethal disease and became infected with R. akari in vivo within 2 days of rickettsial inoculation. Macrophages from uninfected mice of strains susceptible to R. akari also could not be activated for rickettsicidal or tumoricidal activities by treatment with macrophage-activating agents (Mycobacterium bovis BCG) in vivo or by treatment with lymphokines in vitro.

Nacy, C A; Meltzer, M S

1982-01-01

215

Quantitative genome-wide enhancer activity maps for five Drosophila species show functional enhancer conservation and turnover during cis-regulatory evolution.  

PubMed

Phenotypic differences between closely related species are thought to arise primarily from changes in gene expression due to mutations in cis-regulatory sequences (enhancers). However, it has remained unclear how frequently mutations alter enhancer activity or create functional enhancers de novo. Here we use STARR-seq, a recently developed quantitative enhancer assay, to determine genome-wide enhancer activity profiles for five Drosophila species in the constant trans-regulatory environment of Drosophila melanogaster S2 cells. We find that the functions of a large fraction of D. melanogaster enhancers are conserved for their orthologous sequences owing to selection and stabilizing turnover of transcription factor motifs. Moreover, hundreds of enhancers have been gained since the D. melanogaster-Drosophila yakuba split about 11 million years ago without apparent adaptive selection and can contribute to changes in gene expression in vivo. Our finding that enhancer activity is often deeply conserved and frequently gained provides functional insights into regulatory evolution. PMID:24908250

Arnold, Cosmas D; Gerlach, Daniel; Spies, Daniel; Matts, Jessica A; Sytnikova, Yuliya A; Pagani, Michaela; Lau, Nelson C; Stark, Alexander

2014-07-01

216

Supplementation with sodium selenite and selenium-enriched microalgae biomass show varying effects on blood enzymes activities, antioxidant response, and accumulation in common barbel (Barbus barbus).  

PubMed

Yearling common barbel (Barbus barbus L.) were fed four purified casein-based diets for 6 weeks in outdoor cages. Besides control diet, these were supplemented with 0.3 mg kg(-1) dw selenium (Se) from sodium selenite, or 0.3 and 1.0 mg kg(-1) from Se-enriched microalgae biomass (Chlorella), a previously untested Se source for fish. Fish mortality, growth, Se accumulation in muscle and liver, and activity of selected enzymes in blood plasma, muscle, liver, and intestine were evaluated. There was no mortality, and no differences in fish growth, among groups. Se concentrations in muscle and liver, activity of alanine aminotransferase and creatine kinase in blood plasma, glutathione reductase (GR) in muscle, and GR and catalase in muscle and liver suggested that selenium from Se-enriched Chlorella is more readily accumulated and biologically active while being less toxic than sodium selenite. PMID:24772422

Kouba, Antonín; Velíšek, Josef; Stará, Alžb?ta; Masojídek, Ji?í; Kozák, Pavel

2014-01-01

217

A GPBAR1 (TGR5) Small Molecule Agonist Shows Specific Inhibitory Effects on Myeloid Cell Activation In Vitro and Reduces Experimental Autoimmune Encephalitis (EAE) In Vivo.  

PubMed

GPBAR1 is a G protein-coupled receptor that is activated by certain bile acids and plays an important role in the regulation of bile acid synthesis, lipid metabolism, and energy homeostasis. Recent evidence suggests that GPBAR1 may also have important effects in reducing the inflammatory response through its expression on monocytes and macrophages. To further understand the role of GPBAR1 in inflammation, we generated a novel, selective, proprietary GPBAR1 agonist and tested its effectiveness at reducing monocyte and macrophage activation in vitro and in vivo. We have used this agonist, together with previously described agonists to study agonism of GPBAR1, and shown that they can all induce cAMP and reduce TLR activation-induced cytokine production in human monocytes and monocyte-derived macrophages in vitro. Additionally, through the usage of RNA sequencing (RNA-Seq), we identified a select set of genes that are regulated by GPBAR1 agonism during LPS activation. To further define the in vivo role of GPBAR1 in inflammation, we assessed GPBAR1 expression and found high levels on circulating mouse monocytes. Agonism of GPBAR1 reduced LPS-induced cytokine production in mouse monocytes ex vivo and serum cytokine levels in vivo. Agonism of GPBAR1 also had profound effects in the experimental autoimmune encephalomyelitis (EAE) mouse model of multiple sclerosis, where monocytes play an important role. Mice treated with the GPBAR1 agonist exhibited a significant reduction in the EAE clinical score which correlated with reduced monocyte and microglial activation and reduced trafficking of monocytes and T cells into the CNS. These data confirm the importance of GPBAR1 in controlling monocyte and macrophage activation in vivo and support the rationale for selective agonists of GPBAR1 in the treatment of inflammatory diseases. PMID:24967665

Lewis, Nuruddeen D; Patnaude, Lori A; Pelletier, Josephine; Souza, Donald J; Lukas, Susan M; King, F James; Hill, Jonathan D; Stefanopoulos, Dimitria E; Ryan, Kelli; Desai, Sudha; Skow, Donna; Kauschke, Stefan G; Broermann, Andre; Kuzmich, Daniel; Harcken, Christian; Hickey, Eugene R; Modis, Louise K

2014-01-01

218

A GPBAR1 (TGR5) Small Molecule Agonist Shows Specific Inhibitory Effects on Myeloid Cell Activation In Vitro and Reduces Experimental Autoimmune Encephalitis (EAE) In Vivo  

PubMed Central

GPBAR1 is a G protein-coupled receptor that is activated by certain bile acids and plays an important role in the regulation of bile acid synthesis, lipid metabolism, and energy homeostasis. Recent evidence suggests that GPBAR1 may also have important effects in reducing the inflammatory response through its expression on monocytes and macrophages. To further understand the role of GPBAR1 in inflammation, we generated a novel, selective, proprietary GPBAR1 agonist and tested its effectiveness at reducing monocyte and macrophage activation in vitro and in vivo. We have used this agonist, together with previously described agonists to study agonism of GPBAR1, and shown that they can all induce cAMP and reduce TLR activation-induced cytokine production in human monocytes and monocyte-derived macrophages in vitro. Additionally, through the usage of RNA sequencing (RNA-Seq), we identified a select set of genes that are regulated by GPBAR1 agonism during LPS activation. To further define the in vivo role of GPBAR1 in inflammation, we assessed GPBAR1 expression and found high levels on circulating mouse monocytes. Agonism of GPBAR1 reduced LPS-induced cytokine production in mouse monocytes ex vivo and serum cytokine levels in vivo. Agonism of GPBAR1 also had profound effects in the experimental autoimmune encephalomyelitis (EAE) mouse model of multiple sclerosis, where monocytes play an important role. Mice treated with the GPBAR1 agonist exhibited a significant reduction in the EAE clinical score which correlated with reduced monocyte and microglial activation and reduced trafficking of monocytes and T cells into the CNS. These data confirm the importance of GPBAR1 in controlling monocyte and macrophage activation in vivo and support the rationale for selective agonists of GPBAR1 in the treatment of inflammatory diseases.

Lewis, Nuruddeen D.; Patnaude, Lori A.; Pelletier, Josephine; Souza, Donald J.; Lukas, Susan M.; King, F. James; Hill, Jonathan D.; Stefanopoulos, Dimitria E.; Ryan, Kelli; Desai, Sudha; Skow, Donna; Kauschke, Stefan G.; Broermann, Andre; Kuzmich, Daniel; Harcken, Christian; Hickey, Eugene R.; Modis, Louise K.

2014-01-01

219

The dipeptide H-Trp-Glu-OH (WE) shows agonistic activity to peroxisome proliferator-activated protein-? and reduces hepatic lipid accumulation in lipid-loaded H4IIE cells.  

PubMed

Dipeptides digested from dietary proteins can be directly absorbed by the intestine and delivered to the circulatory system. However, the dipeptides' metabolic roles and biological activities are largely unknown. Lipid-loaded HII4E cells stimulated with H-Trp-Glu-OH (WE) exhibited reduced lipid accumulation, of which the effect was abolished by peroxisome proliferator-activated receptor (PPAR) ? gene knock down. A luciferase assay showed that the WE dipeptide induced PPAR? transactivation in a dose-dependent manner. Surface plasmon resonance and time-resolved fluorescence resonance energy transfer analyses demonstrated that WE interacts directly with the PPAR? ligand binding domain (KD, 120?M; EC50, 83?M). Cells stimulated with WE induced PPAR? and its responsive genes and increased cellular fatty acid uptake. In conclusion, WE reduces hepatic lipid accumulation in lipid-loaded hepatocytes via the activation of PPAR? by a direct interaction. PMID:24821375

Jia, Yaoyao; Kim, Jong-Ho; Nam, Bora; Kim, Jiyoung; Lee, Ji Hae; Hwang, Kwang-Yeon; Lee, Sung-Joon

2014-07-01

220

Turkish freshwater and marine macrophyte extracts show in vitro antiprotozoal activity and inhibit FabI, a key enzyme of Plasmodium falciparum fatty acid biosynthesis  

Microsoft Academic Search

The ethanolic extracts of a number of Turkish freshwater macrophytes (Potamogeton perfoliatus, Ranunculus tricophyllus and Cladophora glomerata) and marine macroalgae (Dictyota dichotoma, Halopteris scoparia, Posidonia oceanica, Scinaia furcellata, Sargassum natans and Ulva lactuca) were assayed for their in vitro antiprotozoal activity. Trypanosoma brucei rhodesiense, Trypanosoma cruzi, Leishmania donovani and Plasmodium falciparum were used as test organisms. The cytotoxicity of the

I. Orhan; B. Sener; T. At?c?; R. Brun; R. Perozzo; D. Tasdemir

2006-01-01

221

A Holographic Road Show.  

ERIC Educational Resources Information Center

Describes the viewing sessions and the holograms of a holographic road show. The traveling exhibits, believed to stimulate interest in physics, include a wide variety of holograms and demonstrate several physical principles. (GA)

Kirkpatrick, Larry D.; Rugheimer, Mac

1979-01-01

222

Trade Show Managers  

Microsoft Academic Search

Trade show management is a multi-faceted field, requiring a breadth of skills on the part of those engaged in the craft. Whether they go by the title of Show Manager, Director of Marketing, Vice President of Meetings\\/Conventions, or Director of Meetings\\/Conventions, these professionals work with exhibitors, attendees, and service providers to produce their events. The managers of the 200 largest

Susan Gregory; Deborah Breiter

2001-01-01

223

Ni(II), Cu(II), and Zn(II) Diethyldithiocarbamate Complexes Show Various Activities Against the Proteasome in Breast Cancer Cells  

PubMed Central

A series of three complexes with diethyldithiocarbamate ligand and three different metals (Ni, Cu, Zn) was prepared, confirmed by X-ray crystallography, and tested in human breast cancer MDA-MB-231 cells. Zinc and copper complexes, but not nickel complex, were found to be more active against cellular 26S proteasome than against purified 20S proteasome core particle. One of the possible explanations is inhibition of JAMM domain in the 19S proteasome lid.

Cvek, Boris; Milacic, Vesna; Taraba, Jan; Dou, Q. Ping

2008-01-01

224

The triple combination of tenofovir, emtricitabine and efavirenz shows synergistic anti-HIV-1 activity in vitro: a mechanism of action study  

Microsoft Academic Search

BACKGROUND: Tenofovir disoproxil fumarate (TDF), emtricitabine (FTC), and efavirenz (EFV) are the three components of the once-daily, single tablet regimen (Atripla) for treatment of HIV-1 infection. Previous cell culture studies have demonstrated that the double combination of tenofovir (TFV), the parent drug of TDF, and FTC were additive to synergistic in their anti-HIV activity, which correlated with increased levels of

Joy Y Feng; John K Ly; Florence Myrick; Derrick Goodman; Kirsten L White; Evguenia S Svarovskaia; Katyna Borroto-Esoda; Michael D Miller

2009-01-01

225

HaCaT Keratinocytes Overexpressing the S100 Proteins S100A8 and S100A9 Show Increased NADPH Oxidase and NF-?B Activities  

Microsoft Academic Search

The calcium- and arachidonic acid (AA)-binding proteins S100A8 and S100A9 are involved in nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activation in phagocytes. They are specifically expressed in myeloid cells, and are also found in epithelial cells in various (patho)physiological conditions. We have investigated the consequences of S100A8\\/A9 overexpression in epithelial cell lines on reactive oxygen species (ROS) generation and downstream

Malgorzata Benedyk; Claudia Sopalla; Wolfgang Nacken; Günther Bode; Harut Melkonyan; Botond Banfi; Claus Kerkhoff

2007-01-01

226

Plasma renin-angiotensin system-regulating aminopeptidase activities are modified in early stage Alzheimer's disease and show gender differences but are not related to apolipoprotein E genotype.  

PubMed

Alterations in blood pressure and components of the renin-angiotensin system (RAS) contribute to the development and progression of Alzheimer's disease (AD), resulting in changes that can lead or contribute to cognitive decline. Aspartyl aminopeptidase (ASAP), aminopeptidase A (APA), aminopeptidase N (APN) and aminopeptidase B (APB) catabolise circulating angiotensins, whereas insulin-regulated aminopeptidase (IRAP) has been described as the AT4 receptor. We have found in AD patients a significant decrease of APA activity in men but not in women, and of APN, APB and IRAP in both genders, when compared with control subjects. No changes were found in ASAP activity. Also, APN, APB and IRAP but not APA correlated with the Mini-Mental test, but no relationship with APOE genotype was found. We conclude that several components of the RAS are modified in AD patients, with gender differences. Furthermore, ROC analysis indicates that APN, APB and IRAP activities could be useful non-invasive biomarkers of AD from the earliest stages. PMID:23500679

Puertas, María Del Carmen; Martínez-Martos, José Manuel; Cobo, Manuela; Lorite, Pedro; Sandalio, Rosa María; Palomeque, Teresa; Torres, María Isabel; Carrera-González, María Pilar; Mayas, María Dolores; Ramírez-Expósito, María Jesús

2013-06-01

227

Type 2 diabetic patients and their offspring show altered parameters of iron status, oxidative stress and genes related to mitochondrial activity.  

PubMed

Type 2 diabetes (T2D) is directly related to alterations in iron status, oxidative stress and decreased mitochondrial activity, but the possible interaction of these parameters among T2D patients and their offspring is unclear. The whole study included 301 subjects: 77 T2D patients and one of their offspring and 51 control subjects with one of their offspring. The offspring were older than 20 years old. We measured parameters of iron status (serum iron, ferritin and transferrin receptor), diabetes (pre and post-prandial glucose, insulin, lipids), oxidative stress (Heme oxygenase activity, TBARS, SOD, GSH, Vitamin E), as well as the expression of genes in blood leukocytes related to mitochondrial apopotosis (mitofusin and Bcl/Bax ratios). The offspring of T2D patients had increased levels of serum ferritin (P < 0.01) and lower transferrin receptor (P < 0.008); higher insulin (P < 0.03) and total and LDL cholesterol; higher heme oxygenase and SOD activities increased TBARS and lower GSH; decreased mitofusin and Bcl/Bax expression ratios compared to offspring of normal subjects. These results suggest that the offspring of T2D patients could have an increased metabolic risk of develop a cardiovascular disease mediated by oxidative stress and iron status. PMID:22450556

Le Blanc, Solange; Villarroel, Pia; Candia, Valeria; Gavilán, Natalia; Soto, Néstor; Pérez-Bravo, Francisco; Arredondo, Miguel

2012-08-01

228

Demonstration Road Show  

NSDL National Science Digital Library

The Idaho State University Department of Physics conducts science demonstration shows at S. E. Idaho schools. Four different presentations are currently available; "Forces and Motion", "States of Matter", "Electricity and Magnetism", and "Sound and Waves". Information provided includes descriptions of the material and links to other resources.

Shropshire, Steven

2009-04-06

229

Blue Ribbon Art Show.  

ERIC Educational Resources Information Center

Describes the process of selecting judges for a Blue Ribbon Art Show (Springfield, Missouri). Used adults (teachers, custodians, professional artists, parents, and principals) chosen by the Willard South Elementary School art teacher to judge student artwork. States that nominated students received blue ribbons. (CMK)

Bowen, Judy Domeny

2002-01-01

230

Convolvulus galaticus, Crocus antalyensis, and Lilium candidum extracts show their antitumor activity through induction of p53-mediated apoptosis on human breast cancer cell line MCF-7 cells.  

PubMed

Conventional and newly emerging treatment procedures such as chemotherapy, catalytic therapy, photodynamic therapy, and radiotherapy have not succeeded in reversing the outcome of cancer diseases to any drastic extent, which has led researchers to investigate alternative treatment options. The extensive repertoire of traditional medicinal knowledge systems from various parts of the world are being re-investigated for their healing properties. It has been reported that several members of the Convolvulaceae, Iridaceae, and Liliaceae families have antitumor activity against some tumor cell lines. Here we first report that Convolvulus galaticus, Crocus antalyensis, and Lilium candidum species have cytotoxic activity on human breast cancer cell line MCF-7 cells. Plant samples were collected and identified, and their cytotoxic effects on the MCF-7 cell line were examined at different concentrations of methanol extracts. We found that all three plants have cytotoxic effects on MCF-7 cells but that C. galaticus has the strongest cytotoxic effect even in the lowest extract concentration tested (0.32??g/mL). Our results indicate that these plant extracts have cytotoxic effects on human breast carcinoma cell line MCF-7 cells and that this cytotoxic effect comes from p53-mediated stimulation of apoptosis. PMID:22897477

Tokgun, Onur; Akca, Hakan; Mammadov, Ramazan; Aykurt, Candan; Deniz, Gökhan

2012-11-01

231

What Do Maps Show?  

NSDL National Science Digital Library

The United States Geological Survey's Learning Web site Working with Maps contains a complete Web-based lesson for students between grades 5 and 8. The site introduces the basic concepts related to cartography, including the definition of a map, various map types, and information that you can find on them. Printable activity sheets are available for download, covering four lessons: Introduction to Maps, Some Things You Need to Know to Read a Map, What You Can Learn From a Map, and How to Read a Topographic Map. The tradition of quality material from the USGS continues with this well done, attractive site that gives educators premium content and ease of use.

232

Show Me the Money  

NSDL National Science Digital Library

In this math lesson, learners make wallets or purses and then participate in a variety of money-related activities. First, learners make their own paper money and paper wallets or purses. Next, learners identify coins while singing a money-themed song. Then, learners consider how they would spend various denominations of coins and record their thoughts in a Money Journal. Finally, learners rotate through various Money Centers, where they record coin flips (heads vs. tails), sort coins, make a money cube graph, create a book of coin rubbings, and play store.

Lessonplans, Utah

2012-09-26

233

Show-Me Center  

NSDL National Science Digital Library

The Show-Me Center is a partnership of four NSF-sponsored middle grades mathematics curriculum development Satellite Centers (University of Wisconsin, Michigan State University, University of Montana, and the Educational Development Center). The group's website provides "information and resources needed to support selection and implementation of standards-based middle grades mathematics curricula." The Video Showcase includes segments on Number, Algebra, Geometry, Measure, and Data Analysis, with information on ways to obtain the complete video set. The Curricula Showcase provides general information, unit goals, sample lessons and teacher pages spanning four projects: the Connected Mathematics Project (CMP), Mathematics in Context (MiC), MathScape: Seeing and Thinking Mathematically, and Middle Grades Math Thematics. The website also posts Show-Me Center newsletters, information on upcoming conferences and workshops, and links to resources including published articles and unpublished commentary on mathematics school reform.

234

The Graphing Game Show  

NSDL National Science Digital Library

This lesson plan assesses student interpretation of graphs utilizing cooperative learning to further students understanding. Types of graphs used are horizontal and vertical bar graphs, picture graphs, and pictographs. In the lesson students play a game called the Graphing Game Show, in which they must work as a team to answer questions about specific graphs. The lesson includes four student resource worksheets and suggestions for extension and differentiation.

2011-01-01

235

Synthesis and structural characterization of silver(I), aluminium(III) and cobalt(II) complexes with 4-isopropyltropolone (hinokitiol) showing noteworthy biological activities. Action of silver(I)-oxygen bonding complexes on the antimicrobial activities  

Microsoft Academic Search

Through two unequivalent oxygen donor atoms of the hinokitiol (Hhino; C10H12O2; 4-isopropyltropolone) ligand that showed noteworthy biological activities, the dimeric, silver(I)-oxygen bonding complex [Ag(hino)]21, the monomeric aluminium(III) complex [Al(hino)3]·0.5H2O 4 and the cobalt(II) complex “[Co(hino)2]2 ·H2O” 6 were synthesized and characterized with elemental analysis, thermogravimetric and differential thermal analysis (TG\\/DTA), FTIR and solution (1H and 13C) NMR spectroscopy. The crystal

Kenji Nomiya; Akira Yoshizawa; Ken Tsukagoshi; Noriko Chikaraishi Kasuga; Shoko Hirakawa; Jun Watanabe

2004-01-01

236

Education Statistics Slide Show  

NSDL National Science Digital Library

Created by Grace York, coordinator of the University of Michigan's Documents Center, the Education Statistics Slide Show is an online presentation demonstrating how to locate, obtain, and manipulate educational data on the Web. The presentation consists of 72 slides and offers instruction on the use of several Websites including the US Census Bureau's American Factfinder site (see the April 2, 1999 Scout Report), the Center for International Earth Science Information Network (CIESIN) Census Mapping site, the National Center for Education Statistics (NCES) site, the FEDSTATS site (see the May 30, 1997 Scout Report), and many more. The tutorial presentation also provides ten practice questions and a detailed Webliography.

237

The Q175 Mouse Model of Huntington's Disease Shows Gene Dosage- and Age-Related Decline in Circadian Rhythms of Activity and Sleep  

PubMed Central

Sleep and circadian disruptions are commonly reported by patients with neurodegenerative diseases, suggesting these may be an endophenotype of the disorders. Several mouse models of Huntington’s disease (HD) that recapitulate the disease progression and motor dysfunction of HD also exhibit sleep and circadian rhythm disruption. Of these, the strongest effects are observed in the transgenic models with multiple copies of mutant huntingtin gene. For developing treatments of the human disease, knock-in (KI) models offer advantages of genetic precision of the insertion and control of mutation copy number. Therefore, we assayed locomotor activity and immobility-defined sleep in a new model of HD with an expansion of the KI repeats (Q175). We found evidence for gene dose- and age-dependent circadian disruption in the behavior of the Q175 line. We did not see evidence for loss of cells or disruption of the molecular oscillator in the master pacemaker, the suprachiasmatic nucleus (SCN). The combination of the precise genetic targeting in the Q175 model and the observed sleep and circadian disruptions make it tractable to study the interaction of the underlying pathology of HD and the mechanisms by which the disruptions occur.

Loh, Dawn H.; Kudo, Takashi; Truong, Danny; Wu, Yingfei; Colwell, Christopher S.

2013-01-01

238

A membrane-associated movement protein of Pelargonium flower break virus shows RNA-binding activity and contains a biologically relevant leucine zipper-like motif.  

PubMed

Two small viral proteins (DGBp1 and DGBp2) have been proposed to act in a concerted manner to aid intra- and intercellular trafficking of carmoviruses though the distribution of functions and mode of action of each protein partner are not yet clear. Here we have confirmed the requirement of the DGBps of Pelargonium flower break virus (PFBV), p7 and p12, for pathogen movement. Studies focused on p12 have shown that it associates to cellular membranes, which is in accordance to its hydrophobic profile and to that reported for several homologs. However, peculiarities that distinguish p12 from other DGBps2 have been found. Firstly, it contains a leucine zipper-like motif which is essential for virus infectivity in plants. Secondly, it has an unusually long and basic N-terminal region that confers RNA binding activity. The results suggest that PFBV p12 may differ mechanistically from related proteins and possible roles of PFBV DGBps are discussed. PMID:21444100

Martínez-Turiño, Sandra; Hernández, Carmen

2011-05-10

239

A Monoclonal Antibody to O-Acetyl-GD2 Ganglioside and Not to GD2 Shows Potent Anti-Tumor Activity without Peripheral Nervous System Cross-Reactivity  

PubMed Central

Background Monoclonal antibodies (mAb) against GD2 ganglioside have been shown to be effective for the treatment of neuroblastoma. Beneficial actions are, however, associated with generalized pain due to the binding of anti- GD2 mAbs to peripheral nerve fibers followed by complement activation. Neuroblastoma cells that express GD2 also express its O-acetyl derivative, O-acetyl- GD2 ganglioside (OAcGD2). Hence, we investigated the distribution of OAcGD2 in human tissues using mAb 8B6 to study the cross-reactivity of mAb 8B6 with human tissues. Methodology/Principal Findings The distribution of OAcGD2 was performed in normal and malignant tissues using an immunoperoxydase technique. Anti-tumor properties of mAb 8B6 were studied in vitro and in vivo in a transplanted tumor model in mice. We found that OAcGD2 is not expressed by peripheral nerve fibers. Furthermore, we demonstrated that mAb 8B6 was very effective in the in vitro and in vivo suppression of the growth of tumor cells. Importantly, mAb 8B6 anti-tumor efficacy was comparable to that of mAb 14G2a specific to GD2. Conclusion/Significance Development of therapeutic antibodies specific to OAcGD2 may offer treatment options with reduced adverse side effects, thereby allowing dose escalation of antibodies.

Cochonneau, Denis; Chaumette, Tanguy; Clemenceau, Beatrice; Leprieur, Stephanie; Bougras, Gwenola; Supiot, Stephane; Mussini, Jean-Marie; Barbet, Jacques; Saba, Julie; Paris, Francois; Aubry, Jacques; Birkle, Stephane

2011-01-01

240

Acute ethanol intoxication shows no effect on Ca sup 2+ -uptake of Ca sup 2+ -dependent ATPase activity in myocardial sarcoplasmic reticulum vesicles  

SciTech Connect

We have previously shown that acute ethanolism impairs left ventricular (LV) function. We hypothesized that cardiac dysfunction may be related to altered Ca{sup 2+} pump function by the sarcoplasmic reticulum (SR). In this study, LV function (in isolated perfused hearts) was compared to Ca{sup 2+} transport in SR vesicles isolated from nonperfused hearts in control (C) and acutely intoxicated (ETOH, 2.5 ml/kg IV) guinea pigs. Compared to control hearts, ETOH hearts had significantly lower LV systolic pressure maximal rate of LV pressure rise and fall. Ca{sup 2+}-ATPase activity was not significantly different in either group of animals. Although maximum Ca{sup 2+} uptake tended to be slightly lower in ETOH compared to control hearts coupling ratios (mol Ca{sup 2+} transported/mol ATP hydrolyzed) were not significantly different. We conclude that changes in SR Ca{sup 2+} pump function are not responsible for the depressed LV function seen in acute ethanolism.

McAllister, K.P.; Horton, J.W.; Kaufman, T.M.; White, D.J. (Univ. of Texas Southwestern Medical Center, Dallas (USA))

1989-02-09

241

Performance on a pattern separation task by Alzheimer's patients shows possible links between disrupted dentate gyrus activity and apolipoprotein E ?4 status and cerebrospinal fluid amyloid-?42 levels  

PubMed Central

Introduction Emerging evidence suggests that decreased adult hippocampal neurogenesis represents an early critical event in the course of Alzheimer’s disease (AD). In mice, adult neurogenesis is reduced by knock-in alleles for human apolipoprotein E (ApoE) ?4. Decreased dentate gyrus (DG) neural progenitor cells proliferation has been observed in the triple-transgenic mouse model of AD (3xTg-AD); this reduction being directly associated with the presence of amyloid-? (A?) plaques and an increase in the number of A?-containing neurons in the hippocampus. Cognitive tasks involving difficult pattern separations have been shown to reflect DG activity and thus potentially neurogenesis in both animals and man. This study involved the administration of a pattern separation paradigm to Alzheimer’s patients to investigate relationships between task performance and both ApoE status and cerebrospinal fluid (CSF) A?42 levels. Methods The CDR System pattern separation task involves the presentation of pictures that must later be discriminated from closely similar pictures. This paper presents pattern separation data from 66 mild to moderate AD patients, of which 50 were genotyped and 65 in whom CSF A?42 was measured. Results ApoE ?4 homozygotes were not compromised on the easy pattern separations compared with the other patients, but they were statistically significantly poorer at the difficult separations. In all patients CSF A?42 correlated significantly with the ability to make the difficult discriminations, but not easier discriminations. Pattern separation speed correlated negatively with CSF A?42, and thus the association was not due to increased impulsivity. Conclusions These are, to our knowledge, the first human pattern separation data to suggest a possible genetic link to poor hippocampal neurogenesis in AD, as well as a relationship to A?42. Therapies which target neurogenesis may thus be useful in preventing the early stages of AD, notably in ApoE ?4 homocygotes.

2014-01-01

242

Organometallic cyclic polyphenols derived from 1,2-(alpha-keto tri or tetra methylene) ferrocene show strong antiproliferative activity on hormone-independent breast cancer cells.  

PubMed

We have previously shown that achiral ferrocenyl diphenol butene derivatives are strong antitumor agents against both hormone-dependent and -independent breast cancer cell lines. We report now examples of a new series of two planar chiral diphenol derivatives, namely 1,2-[1-[1,1-bis(4-hydroxyphenyl)methylidene]trimethylene] ferrocene (4), and 1,2-[1-[1,1-bis(4-hydroxyphenyl)methylidene]tetramethylene]ferrocene (5). They were prepared under racemic form from a McMurry coupling reaction with 30% and 16% yields, respectively. Compound 5 gave crystals suitable for X-ray structural analysis. Compounds 4 and 5 were tested for ERalpha and ERbeta affinity, lipophilicity, and proliferative/antiproliferative effects against the hormone-dependent breast cancer cell line MCF-7, and the hormone-independent breast cancer cell line MDA-MB-231. Both compounds exhibit better affinity for ERbeta (16.4 +/- 0.1, and 7.0 +/- 0.4, respectively) than for ERalpha (6.4 +/- 0.2, and 6.6 +/- 0.2). The test on hormone-independent breast cancer cell line MDA-MB-231 showed that 4 with a 5-membered ring gives an IC50 value of 2.7 microM while with 5 in which the ring has 6 carbons, the value is reduced to IC50 = 1.23 microM. PMID:20614088

Plazuk, Damian; Top, Siden; Vessières, Anne; Plamont, Marie-Aude; Huché, Michel; Zakrzewski, Janusz; Makal, Anna; Wo?niak, Krzysztof; Jaouen, Gérard

2010-08-28

243

Show-Me Center  

NSDL National Science Digital Library

The Show-Me Center, located at the University of Missouri, is a math education project of the National Science Foundation. The center's Web site "provides information and resources needed to support selection and implementation of standards-based middle grades mathematic curricula." There are some sample lesson plans offered, but most of the material is solely for use by teachers. Five different middle grade math curriculums were started in 1992, and now, the implementation and results of each curriculum are presented on this site. Teachers can examine each one, view video clips, and read case studies and other reports to choose which parts of the curriculums would fit best into their own classes.

244

Synthesis and photochemical transformation of 3?,21-dihydroxypregna-5,7-dien-20-one to novel secosteroids that show anti-melanoma activity  

PubMed Central

We have synthesized 3?,21-dihydroxypregna-5,7-dien-20-one (21(OH) 7DHP) and used UVB radiation to induce its photoconversion to analogues of vitamin D (pD), lumisterol (pL) and tachysterol (pT). The number and character of the products and the dynamics of the process were dependent on the UVB dose. The main products: pD and pT compounds were characterized by UV absorption, MS and NMR spectroscopy after RP-HPLC chromatography. In addition, formation of multiple oxidized derivatives of the primary products was detected and one of these derivatives was characterized as oxidized 21-hydroxyisotachysterol compound (21(OH)oxy-piT). These newly synthesized compounds inhibited growth of human melanoma cells in a dose dependent manner, with greater or equal potency to calcitriol. 3?,21-Dihydroxy-9?,10?-pregna-5,7-dien-20-one (21(OH)pL) and 21(OH)oxy-piT had higher potency against pigmented melanoma cells, while the EC50 for compounds 21(OH)7DHP and (5Z,7E)-3?,21-dihydroxy-9,10-secopregna-5,7,10(19)-trien-20-one (21(OH)pD) were similar in both pigmented and non-pigmented cells. Moreover, 21(OH)7DHP and its derivatives inhibited proliferation of human epidermal HaCaT keratinocytes, albeit at a lower activity compared to melanoma cells. Importantly, 21(OH)7DHP derivatives strongly inhibited the colony formation of human melanoma cells with 21(OH)pD being the most potent. The potential mechanism of action of newly synthesized compounds was similar to that mediated by 1,25(OH)2D3 and involved ligand-induced translocation of vitamin D receptor into the nucleus. In summary, we have characterized for the first time products of UVB-induced conversion of 21(OH)7DHP and documented that these compounds have selective, inhibitory effects on melanoma cells.

Zmijewski, Michal A.; Li, Wei; Chen, Jianjun; Kim, Tae-Kang; Zjawiony, Jordan K.; Sweatman, Trevor W.; Miller, Duane D.; Slominski, Andrzej T.

2010-01-01

245

Synthesis and photochemical transformation of 3?,21-dihydroxypregna-5,7-dien-20-one to novel secosteroids that show anti-melanoma activity.  

PubMed

We have synthesized 3?,21-dihydroxypregna-5,7-dien-20-one (21(OH) 7DHP) and used UVB radiation to induce its photoconversion to analogues of vitamin D (pD), lumisterol (pL) and tachysterol (pT). The number and character of the products and the dynamics of the process were dependent on the UVB dose. The main products: pD and pT compounds were characterized by UV absorption, MS and NMR spectroscopy after RP-HPLC chromatography. In addition, formation of multiple oxidized derivatives of the primary products was detected and one of these derivatives was characterized as oxidized 21-hydroxyisotachysterol compound (21(OH)oxy-piT). These newly synthesized compounds inhibited growth of human melanoma cells in a dose dependent manner, with greater or equal potency to calcitriol. 3?,21-Dihydroxy-9?,10?-pregna-5,7-dien-20-one (21(OH)pL) and 21(OH)oxy-piT had higher potency against pigmented melanoma cells, while the EC(50) for compounds 21(OH)7DHP and (5Z,7E)-3?,21-dihydroxy-9,10-secopregna-5,7,10(19)-trien-20-one (21(OH)pD) were similar in both pigmented and non-pigmented cells. Moreover, 21(OH)7DHP and its derivatives inhibited proliferation of human epidermal HaCaT keratinocytes, albeit at a lower activity compared to melanoma cells. Importantly, 21(OH)7DHP derivatives strongly inhibited the colony formation of human melanoma cells with 21(OH)pD being the most potent. The potential mechanism of action of newly synthesized compounds was similar to that mediated by 1,25(OH)(2)D(3) and involved ligand-induced translocation of vitamin D receptor into the nucleus. In summary, we have characterized for the first time products of UVB-induced conversion of 21(OH)7DHP and documented that these compounds have selective, inhibitory effects on melanoma cells. PMID:21070794

Zmijewski, Michal A; Li, Wei; Chen, Jianjun; Kim, Tae-Kang; Zjawiony, Jordan K; Sweatman, Trevor W; Miller, Duane D; Slominski, Andrzej T

2011-01-01

246

Different tumours, transduced with different cytokine genes as G-CSF and IL-2, show inhibition of tumour take through neutrophil activation but differ in T cell functions.  

PubMed

The immune effectors and the cellular mechanisms responsible for tumour rejection of two different tumours transduced with different cytokines have been characterized by immunocytochemistry and in situ hybridization. A colon (C-26) and a mammary (TS/A) adenocarcinoma engineered to release, respectively, 90 pg/ml of G-CSF (C-26/G-CSF) and 30 U or 6000 U of IL-2 (low B1.30 and high B4.6000 level of IL-2, respectively) were compared for the type of infiltrating leucocytes and for the repertoire of secondary cytokines produced by the leucocytes recruited at the tumour site. The results indicate that in both systems tumour rejection is associated with prominent infiltration of CD45+/RB6-8C5+ polymorphonuclear (PMN) leucocytes expressing mRNA for IL-1 alpha, IL-1 beta and TNF alpha. TS/A B1.30 and B4.6000 also showed a small proportion of infiltrating T lymphocytes, expressing IFN gamma and IL-4 mRNA, which were virtually absent in the C-26/G-CSF tumour. In mice injected with C-26/G-CSF cells after 600 rad irradiation, the tumours grew to about 1.5 cm and then regressed completely. During the regression phase, T lymphocytes were recruited within C-26/G-CSF, and the infiltrating leucocytes were similar, in terms of PMN/macrophages/T lymphocytes ratio, to those found during the memory response elicited by injection of a challenging dose of parental TS/A into mice pre-immunized with B1.30 IL-2-producing cells. The memory response was characterized by a CD4/CD8 ratio of 0.4 and by IFN-gamma and IL-4 mRNA expression, whereas the T lymphocytes present within regressing C-26/G-CSF were mostly CD4 (CD4/CD8 ratio of 2.1) and expressed IFN gamma mRNA only. The gene transfers of cytokines as different as G-CSF and IL-2 are able to inhibit tumour take through a similar anti-tumour immune response mostly due to non-specific effectors (PMN), thus resembling acute inflammation phenomena. Regression of C-26/G-CSF initially established in irradiated mice as well as rejection of TS/A tumour injected into B1.30 immunized mice are similar to a chronic infection but the immune reaction elicited by C-26/G-CSF has an impaired T-lymphocyte function. PMID:7525363

Stoppacciaro, A; Forni, G; Colombo, M P

1994-01-01

247

Heat shock, copper sulfate and oxidative stress activate the retrotransposon MAGGY resident in the plant pathogenic fungus Magnaporthe grisea  

Microsoft Academic Search

MAGGY is a gypsy-like retrotransposon isolated from the plant pathogenic fungus Magnaporthe grisea. The ability of various stresses to activate MAGGY was tested in the original and in a heterologous host (Colletotrichum lagenarium), using #-glucuronidase (GUS) as a reporter. The MAGGY promoter was activated in M. grisea by either heat shock, copper sulfate, or oxidative stress, but not by the

K. Ikeda; H. Nakayashiki; M. Takagi; Y. Tosa; S. Mayama

2001-01-01

248

Arabidopsis DNA methyltransferase AtDNMT2 associates with histone deacetylase AtHD2s activity  

SciTech Connect

DNA methyltransferase2 (DNMT2) is always deemed to be enigmatic, because it contains highly conserved DNA methyltransferase motifs but lacks the DNA methylation catalytic capability. Here we show that Arabidopsis DNA methyltransferase2 (AtDNMT2) is localized in nucleus and associates with histone deacetylation. Bimolecular fluorescence complementation and pull-down assays show AtDNMT2 interacts with type-2 histone deacetylases (AtHD2s), a unique type of histone deacetylase family in plants. Through analyzing the expression of AtDNMT2: ss-glucuronidase (GUS) fusion protein, we demonstrate that AtDNMT2 has the ability to repress gene expression at transcription level. Meanwhile, the expression of AtDNMT2 gene is altered in athd2c mutant plants. We propose that AtDNMT2 possibly involves in the activity of histone deacetylation and plant epigenetic regulatory network.

Song, Yuan [Key Laboratory of Arid and Grassland Agroecology, Ministry of Education, School of Life Science, Lanzhou University, Lanzhou 730000 (China) [Key Laboratory of Arid and Grassland Agroecology, Ministry of Education, School of Life Science, Lanzhou University, Lanzhou 730000 (China); Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, 1391 Sandford Street, London, ON, Canada N5V4T3 (Canada); Wu, Keqiang [Institute of Plant Biology, National Taiwan University, Taipei 106, Taiwan (China)] [Institute of Plant Biology, National Taiwan University, Taipei 106, Taiwan (China); Dhaubhadel, Sangeeta [Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, 1391 Sandford Street, London, ON, Canada N5V4T3 (Canada)] [Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, 1391 Sandford Street, London, ON, Canada N5V4T3 (Canada); An, Lizhe, E-mail: lizhean@lzu.edu.cn [Key Laboratory of Arid and Grassland Agroecology, Ministry of Education, School of Life Science, Lanzhou University, Lanzhou 730000 (China)] [Key Laboratory of Arid and Grassland Agroecology, Ministry of Education, School of Life Science, Lanzhou University, Lanzhou 730000 (China); Tian, Lining, E-mail: tianl@agr.gc.ca [Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, 1391 Sandford Street, London, ON, Canada N5V4T3 (Canada)] [Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, 1391 Sandford Street, London, ON, Canada N5V4T3 (Canada)

2010-05-28

249

Differential expression of rice calmodulin promoters in response to stimuli and developmental tissue in transgenic tobacco plants.  

PubMed

The promoters of OsCaM1 and OsCaM3 were characterized after sequencing and fused to the reporter gene, GUS. The constructs were then transformed into the tobacco plant. Histochemical analysis of GUS showed different expression patterns in pOsCaM1::GUS and pOsCaM3:: GUS transgenic plants. The expression of pOsCaM1::GUS in 4- to 15-day-old seedlings in particular was observed only in the root, while the expression of pOsCaM3::GUS was detected in both the cotyledons and root. Also, pRCaM1::GUS was detected in all the tissues surrounding the root system, while the presence of pOsCaM3::GUS was observed in the root, except in the root meristem. However, in mature transgenic plants, the expression of pOsCaM1::GUS and OsRCaM3::GUS was scarcely detected. Under wounding stress, the GUS activity of pOsCaM1 and pOsCaM3 was strongly induced, and the activity of pOsCaM3 especially, was retained for long periods. In the phloem, pOsCaM3 activity induced by hormone treatments and abiotic stresses was also identified. [BMB reports 2010; 43(1): 9-16]. PMID:20132729

Jung, Kim Yu; Kyung, Cho Eun; In, Lee Soo; Lim, Chae Oh; Ju, Choi Young

2010-01-01

250

Marine Bacteria from Danish Coastal Waters Show Antifouling Activity against the Marine Fouling Bacterium Pseudoalteromonas sp. Strain S91 and Zoospores of the Green Alga Ulva australis Independent of Bacteriocidal Activity?†  

PubMed Central

The aims of this study were to determine if marine bacteria from Danish coastal waters produce antifouling compounds and if antifouling bacteria could be ascribed to specific niches or seasons. We further assess if antibacterial effect is a good proxy for antifouling activity. We isolated 110 bacteria with anti-Vibrio activity from different sample types and locations during a 1-year sampling from Danish coastal waters. The strains were identified as Pseudoalteromonas, Phaeobacter, and Vibrionaceae based on phenotypic tests and partial 16S rRNA gene sequence similarity. The numbers of bioactive bacteria were significantly higher in warmer than in colder months. While some species were isolated at all sampling locations, others were niche specific. We repeatedly isolated Phaeobacter gallaeciensis at surfaces from one site and Pseudoalteromonas tunicata at two others. Twenty-two strains, representing the major taxonomic groups, different seasons, and isolation strategies, were tested for antiadhesive effect against the marine biofilm-forming bacterium Pseudoalteromonas sp. strain S91 and zoospores of the green alga Ulva australis. The antiadhesive effects were assessed by quantifying the number of strain S91 or Ulva spores attaching to a preformed biofilm of each of the 22 strains. The strongest antifouling activity was found in Pseudoalteromonas strains. Biofilms of Pseudoalteromonas piscicida, Pseudoalteromonas tunicata, and Pseudoalteromonas ulvae prevented Pseudoalteromonas S91 from attaching to steel surfaces. P. piscicida killed S91 bacteria in the suspension cultures, whereas P. tunicata and P. ulvae did not; however, they did prevent adhesion by nonbactericidal mechanism(s). Seven Pseudoalteromonas species, including P. piscicida and P. tunicata, reduced the number of settling Ulva zoospores to less than 10% of the number settling on control surfaces. The antifouling alpP gene was detected only in P. tunicata strains (with purple and yellow pigmentation), so other compounds/mechanisms must be present in the other Pseudoalteromonas strains with antifouling activity.

Bernbom, Nete; Ng, Yoke Yin; Kjelleberg, Staffan; Harder, Tilmann; Gram, Lone

2011-01-01

251

Schwann Cells Express NDF and SMDF\\/n-ARIA mRNAs, Secrete Neuregulin, and Show Constitutive Activation of erbB3 Receptors: Evidence for a Neuregulin Autocrine Loop  

Microsoft Academic Search

Cultured Schwann cells secreted low levels (30 pg\\/ml\\/1.5×106cells) of a 45-kDa neuregulin protein and showed constitutive activation of a neuregulin receptor, Erb-B3, suggesting the existence of an autocrine loop involving neuregulins in Schwann cells. RT-PCR analyses indicated that Schwann cells and fibroblasts in culture produced SMDF\\/n-ARIA and NDF but not GGF neuregulin messages. Schwann cell and fibroblast neuregulin messages encoded

Claudia Rosenbaum; Saikumar Karyala; Mark A. Marchionni; Haesun A. Kim; Alexei L. Krasnoselsky; Beth Happel; Indu Isaacs; Robert Brackenbury; Nancy Ratner

1997-01-01

252

HaCaT keratinocytes overexpressing the S100 proteins S100A8 and S100A9 show increased NADPH oxidase and NF-kappaB activities.  

PubMed

The calcium- and arachidonic acid (AA)-binding proteins S100A8 and S100A9 are involved in nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activation in phagocytes. They are specifically expressed in myeloid cells, and are also found in epithelial cells in various (patho)physiological conditions. We have investigated the consequences of S100A8/A9 overexpression in epithelial cell lines on reactive oxygen species (ROS) generation and downstream signaling. Epithelial carcinoma HeLa cells, which exclusively express Nox2, showed dramatically increased activation of NADPH oxidase by phorbol 12-myristate 13-acetate after S100A8/A9 gene transfection. HaCaT keratinocytes overexpressing S100A8/A9 showed enhanced, transient ROS generation in response to the calcium ionophore A23187 compared to mock-transfected cells. Polymerase chain reaction analysis revealed mRNA transcripts for Nox1, Nox2, and Nox5 in HaCaT keratinocytes. Detailed transfection studies confirmed that NADPH oxidase activities in Nox1- and Nox5-transfected HeLa cells were enhanced after S100A8/A9 gene complementation. Furthermore, mutational analysis revealed that AA binding and Thr113 phosphorylation are important for S100A8/A9-enhanced activation of NADPH oxidase. Nuclear factor-kappaB (NF-kappaB) activation and interleukin-8 mRNA levels were increased in S100A8/A9-HaCaT keratinocytes, consistent with the view that NF-kappaB is a redox-sensitive transcription factor. Because they are expressed in epithelia under specific conditions, S100A8 and S100A9 might be involved in skin pathogenesis by modulating aspects of downstream signaling. PMID:17429438

Benedyk, Malgorzata; Sopalla, Claudia; Nacken, Wolfgang; Bode, Günther; Melkonyan, Harut; Banfi, Botond; Kerkhoff, Claus

2007-08-01

253

Mice deficient for the synaptic vesicle protein Rab3a show impaired spatial reversal learning and increased explorative activity but none of the behavioral changes shown by mice deficient for the Rab3a regulator Gdi1.  

PubMed

Rab proteins are small GTPases involved in intracellular trafficking. Among the 60 different Rab proteins described in mammals, Rab3a is the most abundant in brain, where it is involved in synaptic vesicle fusion and neurotransmitter release. Rab3a constitutive knockout mice (Rab3a(-/-)) are characterized by deficient short- and long-term synaptic plasticity in the mossy fiber pathway and altered circadian motor activity, while no effects on spatial learning have been reported so far for these mice. The goals of this study were to analyse possible behavioral consequences of the lack of synaptic plasticity in the mossy fiber pathway using a broad battery of sensitive behavioral measures that has been used previously to analyse the behavior of Gdi1 mice lacking a protein thought to regulate Rab3a. Rab3a(-/-) mice showed normal acquisition but moderately impaired platform reversal learning in the water maze including reference memory and episodic-like memory tasks. A mild deficit in spatial working memory was also observed when tested in the radial maze. Analysis of explorative behavior revealed increased locomotor activity and enhanced exploratory activity in open field, O-maze, dark/light box and novel object tests. Spontaneous activity in normal home cage settings was unaffected but Rab3a(-/-) mice showed increased motor activity when the home cage was equipped with a wheel. No differences were found for delayed and trace fear conditioning or for conditioned taste aversion learning. Congruent with earlier data, these results suggest that Rab3a-dependent synaptic plasticity might play a specific role in the reactivity to novel stimuli and behavioral stability rather than being involved in memory processing. On the other hand, the phenotypic changes in the Rab3a(-/-) mice bore no relation to the behavioral changes as observed in the Gdi1 mice. Such divergence in phenotypes implies that the putative synaptic interaction between Gdi1 and Rab3a should be reconsidered and re-analysed. PMID:15078563

D'Adamo, Patrizia; Wolfer, David P; Kopp, Caroline; Tobler, Irene; Toniolo, Daniela; Lipp, Hans-Peter

2004-04-01

254

Gall formation in clubroot-infected Arabidopsis results from an increase in existing meristematic activities of the host but is not essential for the completion of the pathogen life cycle.  

PubMed

Plasmodiophora brassicae (clubroot) infection leads to reprogramming of host development resulting in the formation of characteristic galls. In this work we explored the cellular events that underly gall formation in Arabidopsis thaliana with the help of molecular markers of cell division (CYCB1:GUS) and meristematic activity (ANT:GUS). Our results show that gall development involved the amplification of existing meristematic activities within the vascular cambium (VC) and phloem parenchyma (PP) cells in the region of the hypocotyl. Additionally we found that the increase in VC activity and prolonged maintenance of cambial-derived cells in a meristematic state was crucial for gall formation; disruption of the VC activity significantly decreased the gall size. Gall formation also perturbed vascular development with a significant reduction in xylem and increase in PP in infected plants. This situation was reflected in a decrease in transcripts of key factors promoting xylogenesis (VND6, VND7 and MYB46) and an increase in those promoting phloem formation and function (APL, SUC2). Finally we show, using the cell cycle inhibitor ICK1/KRP1 and a cle41 mutant with altered regulation of cambial stem cell maintenance and differentiation, that a decrease in gall formation did not prevent pathogen development. This finding demonstrates that although gall formation is a typical symptom of the disease and influences numbers of spores produced, it is not required for completion of the pathogen life cycle. Together, these results provide an insight into the relationship of the cellular events that accompany Plasmodiophora infection and their role in disease progression. PMID:22394393

Malinowski, Robert; Smith, Jody A; Fleming, Andrew J; Scholes, Julie D; Rolfe, Stephen A

2012-07-01

255

A Mouse with a Loss-of-function Mutation in the c-Cbl TKB Domain Shows Perturbed Thymocyte Signaling without Enhancing the Activity of the ZAP-70 Tyrosine Kinase  

PubMed Central

The unique tyrosine kinase binding (TKB) domain of Cbl targets phosphorylated tyrosines on activated protein tyrosine kinases (PTKs); this targeting is considered essential for Cbl proteins to negatively regulate PTKs. Here, a loss-of-function mutation (G304E) in the c-Cbl TKB domain, first identified in Caenorhabditis elegans, was introduced into a mouse and its effects in thymocytes and T cells were studied. In marked contrast to the c-Cbl knockout mouse, we found no evidence of enhanced activity of the ZAP-70 PTK in thymocytes from the TKB domain mutant mouse. This finding contradicts the accepted mechanism of c-Cbl–mediated negative regulation, which requires TKB domain targeting of phosphotyrosine 292 in ZAP-70. However, the TKB domain mutant mouse does show aspects of enhanced signaling that parallel those of the c-Cbl knockout mouse, but these involve the constitutive activation of Rac and not enhanced PTK activity. Furthermore, the enhanced signaling in CD4+CD8+ double positive thymocytes appears to be compensated by the selective down-regulation of CD3 on mature thymocytes and peripheral T cells from both strains of mutant c-Cbl mice.

Thien, Christine B.F.; Scaife, Robin M.; Papadimitriou, John M.; Murphy, Maria A.; Bowtell, David D.L.; Langdon, Wallace Y.

2003-01-01

256

Ecology- and bioassay-guided drug discovery for treatments of tropical parasitic disease: 5alpha,8alpha-epidioxycholest-6-en-3beta-ol isolated from the mollusk Dolabrifera dolabrifera shows significant activity against Leishmania donovani.  

PubMed

An ecology- and bioassay-guided search employed to discover compounds with activity against tropical parasitic diseases and cancer from the opisthobranch mollusk, Dolabrifera dolabrifera, led to the discovery of antileishmanial properties in the known compound, 5alpha,8alpha-epidioxycholest-6-en-3beta-ol (1). Compound 1 was identified through nuclear magnetic resonance spectroscopy (1H, 13C) and mass spectrometry. The compound was concentrated in the digestive gland of D. dolabrifera, but was not detected in other body parts, fecal matter or mucus. Compound 1 showed an IC50 of 4.9 microM towards the amastigote form of Leishmania donovani compared with an IC50 of 281 microM towards the control Vero cell line, a 57.3-fold difference, and demonstrated no measurable activity against Plasmodium falciparum, Trypanosoma cruzi, and the breast cancer cell line, MCF-7. PMID:24427935

Clark, Kathryn E; Capper, Angela; Della Togna, Gina; Paul, Valerie J; Romero, Luz I; Johns, Timothy; Cubilla-Rios, Luis; Capson, Todd L

2013-11-01

257

Glionitrin A, a new diketopiperazine disulfide, activates ATM-ATR-Chk1/2 via 53BP1 phosphorylation in DU145 cells and shows antitumor effect in xenograft model.  

PubMed

In a recent study, we isolated the diketopiperazine disulfide glionitrin A from the co-culture broth of a mine drainage-derived fungus (Aspergillus fumigatus KMC901) and bacterium (Sphingomonas KMK001). Here, we investigated the antitumor activity of glionitrin A and its underlying molecular mechanisms in human prostate cancer DU145 cells. Glionitrin A showed significant cytotoxicity, promoting cell cycle arrest and apoptosis. Glionitrin A-treated cells exhibited elevated levels of phospho-histone 2AX (Ser139), a marker of DNA damage, and accumulated in both S phase and G2/M phase due to the activation of checkpoints associated with the ataxia-telangiectasia-mutated and ataxia-telangiectasia-mutated-Rad3-related Chk1/2 pathway downstream of p53-binding protein 1 phosphorylation at Ser1778. In addition, glionitrin A induced apoptosis through both caspase-dependent and -independent pathways. Glionitrin A activated caspase-8, -9 and -3 and also released endonuclease G from the mitochondria to the nucleus in a dose-dependent manner. Our in vivo study performed in nude mice bearing xenografts of DU145 cells showed that glionitrin A dramatically reduced the tumor volume by an average of 38.2% (5?mg/kg, per os (p.o.)) and 71.3% (10?mg/kg, p.o.) at 27?d after the beginning of treatment. Taken together, these findings provide a detailed description of the mechanism underlying the biological activity of the new natural product glionitrin A, which has the potential to be developed as an anti-prostate cancer agent. PMID:24583858

Kim, Young-Joo; Park, Hyun Bong; Yoo, Ji-Hye; Kwon, Hak Cheol; Kim, Joonki; Yang, Hyun Ok

2014-01-01

258

Asia: Showing the Changing Seasons  

NSDL National Science Digital Library

SeaWiFS false color data showing seasonal change in the oceans and on land for Asia. The data is seasonally averaged, and shows the sequence: fall, winter, spring, summer, fall, winter, spring (for the Northern Hemisphere).

Allen, Jesse; Newcombe, Marte; Feldman, Gene

1998-09-09

259

What Do Blood Tests Show?  

MedlinePLUS

... page from the NHLBI on Twitter. What Do Blood Tests Show? Blood tests show whether the levels ... changes may work best. Result Ranges for Common Blood Tests This section presents the result ranges for ...

260

13,14-bis(cis-3,5-dimethyl-1-piperazinyl)-?-elemene, a novel ?-elemene derivative, shows potent antitumor activities via inhibition of mTOR in human breast cancer cells.  

PubMed

Elemene has been approved for the treatment of advanced cancer in China, however, it inhibits cell growth only at high concentrations and is an essential oil with poor water solubility and stability. The discovery of new ?-elemene derivatives is of increasing interest. We recently reported that the compound 13,14-bis(cis-3,5-dimethyl-1-piperazinyl)-?-elemene (IIi), a novel ?-elemene derivative with a cis-2,6-dimethylpiperazine substitution, is a potent agent for inhibiting the proliferation of SGC-7901 and HeLa cells. In the present study, we further verified that IIi is cytotoxic to a wide spectrum of human cancer cells in culture, including those of breast, ovarian, lung, gastric, hepatocellular and colon cancer, as well as leukemia cell lines, with an average IC50 of 3.44 ?mol/l. Notably, IIi showed significant cytotoxicity in two multidrug-resistant (MDR) cell lines, with an average resistance factor (RF) of 1.66. Moreover, in mice with S-180 sarcoma xenografts, the intraperitoneal administration of IIi inhibited tumor growth. The immunoblotting study showed that treatment with IIi decreases phosphorylated p70S6K1 and 4EBP1 levels in the human breast cancer MCF-7 and MDA-MB-468 cells. In the MCF-7 cells, IIi also significantly increased the expression of cleaved LC3. This indicated that IIi inhibits mTOR activity and induces autophagy. The mTOR inhibitory function and the potent antitumor activity, taken together with the appreciable anti-multidrug resistance action, shows IIi to be a novel potential antitumor agent, which merits further research and development. PMID:23761818

Ding, Xiao-Fei; Shen, Mao; Xu, Li-Ying; Dong, Jin-Hua; Chen, Guang

2013-05-01

261

Optical and SPION-enhanced MR imaging shows that trans-stilbene inhibitors of NF-?B concomitantly lower Alzheimer's disease plaque formation and microglial activation in A?PP/PS-1 transgenic mouse brain.  

PubMed

Alzheimer's disease (AD) is associated with a microglia-dependent neuroinflammatory response against plaques containing the fibrous protein amyloid-? (A?). Activation of microglia, which closely associate with A? plaques, engenders the release of pro-inflammatory cytokines and the internalization of A? fibrils. Since the pro-inflammatory transcription factor NF-?B is one of the major regulators of A?-induced inflammation, we treated transgenic amyloid-? protein protein/presenilin-1 (A?PP/PS1) mice for one year with a low dose (0.01% by weight in the diet) of either of two trans-stilbene NF-?B inhibitors, resveratrol or a synthetic analog LD55. The 3D distribution of A? plaques was measured ex vivo in intact brains at 60 ?m resolution by quantitative magnetic resonance imaging (MRI) using blood-brain barrier-permeable, anti-A?PP-conjugated superparamagentic iron oxide nanoparticles (SPIONs). The MRI measurements were confirmed by optical microscopy of thioflavin-stained brain tissue sections and indicated that supplementation with either of the two trans-stilbenes lowered A? plaque density in the cortex, caudoputamen, and hippocampus by 1.4 to 2-fold. The optical measurements also included the hippocampus and indicated that resveratrol and LD55 reduced average A? plaque density by 2.3-fold and 3.1-fold, respectively. Ex vivo measurements of the regional distribution of microglial activation by Iba-1 immunofluorescence of brain tissue sections showed that resveratrol and LD55 reduced average microglial activation by 4.2- fold and 3.5-fold, respectively. Since LD55 lacked hydroxyl groups but both resveratrol and LD55 concomitantly reduced both A? plaque burden and neuroinflammation to a similar extent, it appears that the antioxidant potential of resveratrol is not an important factor in plaque reduction. PMID:24413613

Solberg, Nathan O; Chamberlin, Ryan; Vigil, Jenette R; Deck, Lorraine M; Heidrich, John E; Brown, David C; Brady, Christina I; Vander Jagt, Thomas A; Garwood, Michael; Bisoffi, Marco; Severns, Virginia; Vander Jagt, David L; Sillerud, Laurel O

2014-01-01

262

Planning a Successful Tech Show  

ERIC Educational Resources Information Center

Tech shows are a great way to introduce prospective students, parents, and local business and industry to a technology and engineering or career and technical education program. In addition to showcasing instructional programs, a tech show allows students to demonstrate their professionalism and skills, practice public presentations, and interact…

Nikirk, Martin

2011-01-01

263

The promoter of the nematode resistance gene Hs1pro-1 activates a nematode-responsive and feeding site-specific gene expression in sugar beet (Beta vulgaris L.) and Arabidopsis thaliana.  

PubMed

The Hs1pro-1 gene confers resistance to the beet cyst nematode Heterodera schachtii in sugar beet (Beta vulgaris L.) on the basis of a gene-for-gene relationship. RNA-gel blot analysis revealed that the transcript of Hs1pro-1 was present in uninfected roots of resistant beet at low levels but increased by about fourfold one day after nematode infection. Treatments of plants with external stimuli including salicylic acid, jasmonic acid, gibberellic acid and abscisic acid as well as wounding or salt stress did not result in changes in the gene transcription, indicating de novo transcription of Hs1pro-1 upon nematode infection specifically. To study transcriptional regulation of Hs1pro-1 expression at the cellular level, a 3082 bp genomic fragment representing the Hs1pro-1 promoter, isolated from the YAC-DNA housing the Hs1pro-1 gene, was fused to the beta-glucuronidase reporter gene (1832prm1::GUS) and transformed into susceptible beet roots and Arabidopsis plants, respectively. Fluorometric and histochemical GUS assays on transgenic beet roots and Arabidopsis plants carrying the 1832prm1::GUS construct demonstrated that the Hs1pro-1 promoter is functional in both species and drives a nematode responsive and feeding site-specific GUS-expression. GUS activity was detected as early as at initiation of the nematode feeding sites and GUS staining was restricted to the nematode feeding sites. To delineate the regulatory domains of the Hs1pro-1 promoter, fusion genes with various 5' deletions of the Hs1pro-1 promoter and the GUS gene were constructed and analysed in transgenic beet roots as well. Cis elements responsible for feeding site-specific gene expression reside between -355 and +247 from the transcriptional initiation site of Hs1pro-1 whereas an enhancer region necessary for higher gene expression is located between -1199 and -705 of the promoter. The Hs1pro-1 promoter drives a nematode feeding site-specific GUS expression in both sugar beet and Arabidopsis suggesting a conserved mechanism of regulation of Hs1pro-1 expression in these two species. PMID:12956533

Thurau, Tim; Kifle, Sirak; Jung, Christian; Cai, Daguang

2003-06-01

264

A reconstituted HDL containing V156K or R173C apoA-I exhibited anti-inflammatory activity in apo-E deficient mice and showed resistance to myeloperoxidase-mediated oxidation  

PubMed Central

It has been hypothesized that blood infusion of reconstituted HDL (rHDL) is a possible therapeutic strategy for the treatment of coronary artery disese. To compare short-term anti-inflammatory activity of wildtype (WT) apoA-I and point mutants, each rHDL containing WT, V156K, or R173C was infused into apo-E deficient atherosclerotic mice. Each rHDL was injected via the tail vein at a dosage of 120 mg/kg of body weight in 0.4 ml of tris-buffered saline (TBS), and blood was then collected at 24 and 48 h post-injection. Although regression activity was observed in each of the rHDL infused groups, a 30% reduction in the lipid-stained area of the aortic sinus was observed in the V156K and R173C-rHDL groups when compared to that of the WT-rHDL group, and this reduction was well correlated with an approximately 60% reduction in the accumulation of macrophages in the lesion area. Additionally, the groups that received the V156K and R173C-rHDL treatments showed smaller increases in the GOT, GPT, interleukin-6, myeloperoxidase (MPO) and lipid hydroperoxide (LPO) serum levels than those that received the WT-rHDL treatment. In addition, the strongest serum paraoxonase and ferric reducing ability was observed in the V156K and R173C-rHDL groups. In vitro nitration and chlorination of apoA-I by MPO treatment revealed that V156K-rHDL and R173C-rHDL were less susceptible to chlorination. Furthermore, rHDL treatment inhibited cellular uptake of oxidized LDL by macrophage cells and the production of proatherogenic species in culture media. In conclusion, blood infusions of the rHDLs exerted in vivo regression activity with anti-inflammatory and antioxidant activity in apo-E deficient mice and THP-1 cells, especially in those that were treated with V156K and R173C apoA-I.

Kim, Jae-Ryong

2009-01-01

265

Comparisons show construction management's benefits.  

PubMed

Analysis of competitive bid, design-build, and construction management approaches shows that construction management offers owners substantial advantages regarding project cost and quality and substantial involvement and control throughout the process. PMID:110667

Payette, T M

1979-09-01

266

Using Graphs to Show Connections  

NSDL National Science Digital Library

The purpose of this resource is to show how graphs of GLOBE data over time show the interconnectedness of Earth's system components at the local level. Students visit a study site, where they observe and recall their existing knowledge of air, water, soil, and living things to make a list of interconnections among the four Earth system components. They make predictions about the effects of a change in a system, inferring ways these changes affect the characteristics of other related components.

The GLOBE Program, University Corporation for Atmospheric Research (UCAR)

2003-08-01

267

Show-and-Tell Physics.  

ERIC Educational Resources Information Center

Describes a program where teams of fifth graders and high school junior and senior physics students work together to explore physical science principles and create a student-constructed exploratorium. Incorporates and synthesizes scientific inquiry, hands-on activities, and creative and critical problem-solving approaches in a motivating and fun…

Jones, T. Griffith; And Others

1996-01-01

268

International Plowing Match & Farm Machinery Show  

NSDL National Science Digital Library

The 1995 International Plowing Match & Farm Machinery Show in Ontario, Canada has a site of the Web. The IPM is a non-profit organization of volunteers which annually organizes Canada's largest farm machinery show. The event is commercial and educational. Thousands of school children and educators attend and participate in organized educational activities.

1995-01-01

269

What Do These Images Show?  

NSDL National Science Digital Library

In this activity, students, working in small groups, will examine images of surface features on Mars and attempt to identify them, speculate as to their origins, and determine their size. They will share their ideas in a class discussion and attempt to reach a consensus for the interpretation of each image. Most of the images are from the Viking missions; they are accompanied by sample questions for the class.

270

The Maths File Game Show  

NSDL National Science Digital Library

The British Broadcasting Corporation offers this great collection of fun, educational games that help children learn basic concepts of mathematics. Twelve interactive games are available in all, illustrating principles of data handling, numbers, algebra, and measurement. Children can practice interpreting Cartesian coordinates by guiding a space ship across a grid, or compete with an animated character in a test of fractions and percentages. In addition to the online games, several other activities are presented in one-page documents for easy printing.

2007-12-12

271

The OOPSLA trivia show (TOOTS)  

Microsoft Academic Search

OOPSLA has a longstanding tradition of being a forum for discussing the cutting edge of technology in a fun and participatory environment. The type of events sponsored by OOPSLA sometimes border on the unconventional. This event represents an atypical panel that conforms to the concept of a game show that is focused on questions and answers related to OOPSLA themes.

Jeff Gray; Douglas C. Schmidt

2009-01-01

272

Schwann cells express NDF and SMDF/n-ARIA mRNAs, secrete neuregulin, and show constitutive activation of erbB3 receptors: evidence for a neuregulin autocrine loop.  

PubMed

Cultured Schwann cells secreted low levels (30 pg/ml/1.5 x 10(6) cells) of a 45-kDa neuregulin protein and showed constitutive activation of a neuregulin receptor, Erb-B3, suggesting the existence of an autocrine loop involving neuregulins in Schwann cells. RT-PCR analyses indicated that Schwann cells and fibroblasts in culture produced SMDF/n-ARIA and NDF but not GGF neuregulin messages. Schwann cell and fibroblast neuregulin messages encoded both beta and alpha domains; Schwann cell transcripts encoded only transmembrane neuregulin forms while fibroblast messages encoded transmembrane and secreted forms. SMDF/n-ARIA and NDF messages were also expressed in early postnatal rat sciatic nerve, suggesting a role for neuregulins in peripheral nerve development. An anti-neuregulin antibody inhibited the mitogenic response of Schwann cells to cultured neurons and to extracts of cultured neurons or embryonic brain, consistent with the accepted paracrine role of neuregulins on Schwann cells. Surprisingly, the same antibody inhibited Schwann cell proliferation stimulated by several unrelated mitogens including bFGF, HGF, and TGF-beta1. These data implicate both paracrine and autocrine pathways involving neuregulin form(s) in Schwann cell mitogenic responses. PMID:9417836

Rosenbaum, C; Karyala, S; Marchionni, M A; Kim, H A; Krasnoselsky, A L; Happel, B; Isaacs, I; Brackenbury, R; Ratner, N

1997-12-01

273

Casimir experiments showing saturation effects  

NASA Astrophysics Data System (ADS)

We address several different Casimir experiments where theory and experiment disagree. First out is the classical Casimir force measurement between two metal half spaces; here both in the form of the torsion pendulum experiment by Lamoreaux and in the form of the Casimir pressure measurement between a gold sphere and a gold plate as performed by Decca ; theory predicts a large negative thermal correction, absent in the high precision experiments. The third experiment is the measurement of the Casimir force between a metal plate and a laser irradiated semiconductor membrane as performed by Chen ; the change in force with laser intensity is larger than predicted by theory. The fourth experiment is the measurement of the Casimir force between an atom and a wall in the form of the measurement by Obrecht of the change in oscillation frequency of a R87b Bose-Einstein condensate trapped to a fused silica wall; the change is smaller than predicted by theory. We show that saturation effects can explain the discrepancies between theory and experiment observed in all these cases.

Sernelius, Bo E.

2009-10-01

274

Casimir experiments showing saturation effects  

SciTech Connect

We address several different Casimir experiments where theory and experiment disagree. First out is the classical Casimir force measurement between two metal half spaces; here both in the form of the torsion pendulum experiment by Lamoreaux and in the form of the Casimir pressure measurement between a gold sphere and a gold plate as performed by Decca et al.; theory predicts a large negative thermal correction, absent in the high precision experiments. The third experiment is the measurement of the Casimir force between a metal plate and a laser irradiated semiconductor membrane as performed by Chen et al.; the change in force with laser intensity is larger than predicted by theory. The fourth experiment is the measurement of the Casimir force between an atom and a wall in the form of the measurement by Obrecht et al. of the change in oscillation frequency of a {sup 87}Rb Bose-Einstein condensate trapped to a fused silica wall; the change is smaller than predicted by theory. We show that saturation effects can explain the discrepancies between theory and experiment observed in all these cases.

Sernelius, Bo E. [Division of Theory and Modeling, Department of Physics, Chemistry and Biology, Linkoeping University, SE-581 83 Linkoeping (Sweden)

2009-10-15

275

Mimas Showing False Colors #1  

NASA Technical Reports Server (NTRS)

False color images of Saturn's moon, Mimas, reveal variation in either the composition or texture across its surface.

During its approach to Mimas on Aug. 2, 2005, the Cassini spacecraft narrow-angle camera obtained multi-spectral views of the moon from a range of 228,000 kilometers (142,500 miles).

The image at the left is a narrow angle clear-filter image, which was separately processed to enhance the contrast in brightness and sharpness of visible features. The image at the right is a color composite of narrow-angle ultraviolet, green, infrared and clear filter images, which have been specially processed to accentuate subtle changes in the spectral properties of Mimas' surface materials. To create this view, three color images (ultraviolet, green and infrared) were combined into a single black and white picture that isolates and maps regional color differences. This 'color map' was then superimposed over the clear-filter image at the left.

The combination of color map and brightness image shows how the color differences across the Mimas surface materials are tied to geological features. Shades of blue and violet in the image at the right are used to identify surface materials that are bluer in color and have a weaker infrared brightness than average Mimas materials, which are represented by green.

Herschel crater, a 140-kilometer-wide (88-mile) impact feature with a prominent central peak, is visible in the upper right of each image. The unusual bluer materials are seen to broadly surround Herschel crater. However, the bluer material is not uniformly distributed in and around the crater. Instead, it appears to be concentrated on the outside of the crater and more to the west than to the north or south. The origin of the color differences is not yet understood. It may represent ejecta material that was excavated from inside Mimas when the Herschel impact occurred. The bluer color of these materials may be caused by subtle differences in the surface composition or the sizes of grains making up the icy soil.

The images were obtained when the Cassini spacecraft was above 25 degrees south, 134 degrees west latitude and longitude. The Sun-Mimas-spacecraft angle was 45 degrees and north is at the top.

The Cassini-Huygens mission is a cooperative project of NASA, the European Space Agency and the Italian Space Agency. The Jet Propulsion Laboratory, a division of the California Institute of Technology in Pasadena, manages the mission for NASA's Science Mission Directorate, Washington, D.C. The Cassini orbiter and its two onboard cameras were designed, developed and assembled at JPL. The imaging operations center is based at the Space Science Institute in Boulder, Colo.

For more information about the Cassini-Huygens mission visit http://saturn.jpl.nasa.gov . The Cassini imaging team homepage is at http://ciclops.org .

2005-01-01

276

Transgenic rice plants expressing the ferredoxin-like protein (AP1) from sweet pepper show enhanced resistance to Xanthomonas oryzae pv. oryzae.  

PubMed

We used particle bombardment to cotransform mature seed-derived rice callus (Oryza sativa L., ssp. japonica, cv. Eyi 105) with plasmids containing the linked marker genes gusA and hpt, and the ap1 gene encoding an amphipathic protein previously shown to delay the hypersensitive response induced in non-host plants by the pathogen Pseudomonas syringae pv. syringae (Pss). Thirty-two independent lines of transgenic rice plants were regenerated, and 27 of these lines carried all three transgenes as shown by molecular analysis. A bacterial blight inoculation test was carried out on ten lines. In each case, plants carrying the ap1 gene showed enhanced resistance to Xanthomonas oryzae pv. oryzae (Xoo) race 6 at various levels. This suggests the ap1 gene could be a useful candidate for genetic engineering strategies in rice to provide bacterial blight resistance. PMID:11297801

Tang, K; Sun, X; Hu, Q; Wu, A; Lin, C -H.; Lin, H -J.; Twyman, R M.; Christou, P; Feng, T

2001-04-01

277

Deep-Ultraviolet Transparent Phosphates RbBa2(PO3)5 and Rb2Ba3(P2O7)2 Show Nonlinear Optical Activity from Condensation of [PO4](3-) Units.  

PubMed

It is challenging to explore deep-ultraviolet (deep-UV) nonlinear optical (NLO) materials that can achieve a subtle balance between deep-UV transparency and high NLO activity. Known deep-UV NLO materials are almost exclusively limited to borates, except few newly discovered phosphates despite their small NLO activities. Here we report two asymmetric phosphates, RbBa2(PO3)5 (I) and Rb2Ba3(P2O7)2 (II), which feature [PO3]? chains and [P2O7](4-) dimers formed by condensation of [PO4](3-) units, respectively. Remarkably, I achieves the desired balance, with the shortest deep-UV absorption edge at 163 nm and the largest NLO activity of 1.4 × KDP (KH2PO4) in deep-UV NLO phosphates. According to first-principles calculations, the enhanced macroscopic SHG response of I can be attributed to the [PO3]? chains which exhibit significantly larger microscopic SHG coefficients as compared with the [P2O7](4-) dimers. PMID:24906077

Zhao, Sangen; Gong, Pifu; Luo, Siyang; Bai, Lei; Lin, Zheshuai; Ji, Chengmin; Chen, Tianliang; Hong, Maochun; Luo, Junhua

2014-06-18

278

Factors affecting transient gene expression in protoplasts isolated from very slowly growing embryogenic callus cultures of wheat (Triticum aestivum L.).  

PubMed

Protoplasts isolated from embryogenic ('Mustang' and 'Chinese Spring') and non-embryogenic ('Mit') calli of wheat (Triticum aestivum L.) genotypes transiently expressed ?-glucuronidase (GUS) activity when electroporated with a plasmid containing the GUS gene and driven by an enhanced 35S promoter and a TMV leader sequence. Conditions for the maximum expression of GUS activity were: electroporation of the freshly isolated protoplasts at 250 Vcm(-1) and 250 ?F for 2 s using 50 ?g/ml of plasmid DNA; incubation of the protoplasts with the plasmid before the pulse for 2 h; and a 15-min recovery period on ice after the pulse. In general, a higher GUS activity was obtained in protoplasts of non-embryogenic (NE) callus origin than in those of embryogenic (E) callus origin. Only GUS constructs containing a duplicate 35S promoter derivative resulted in a significant level of GUS expression. The presence of the TMV viral leader sequence in the pAGUS1-TN2 plasmid construct resulted in a significant increase of GUS activity in the electroporated protoplasts of both callus types. On the other hand, protoplasts electroporated with the Adh1 promoter and intron showed a threefold less GUS activity than those electroporated with pAGUS1-TN2. Optimized conditions for DNA uptake and expression were very similar for protoplasts of both callus types. The importance of these findings for the successful regeneration of transgenic and fertile wheat plants is discussed. PMID:24193782

Zaghmout, O M; Trolinder, N L

1993-07-01

279

The cauliflower mosaic virus (CaMV) 35S promoter sequence alters the level and patterns of activity of adjacent tissue- and organ-specific gene promoters  

Microsoft Academic Search

Here we report the effect of the 35S promoter sequence on activities of the tissue- and organ-specific gene promoters in tobacco\\u000a plants. In the absence of the 35S promoter sequence the AAP2 promoter is active only in vascular tissues as indicated by expression of the AAP2:GUS gene. With the 35S promoter sequence in the same T-plasmid, transgenic plants exhibit twofold

Xuelian Zheng; Wei Deng; Keming Luo; Hui Duan; Yongqin Chen; Richard McAvoy; Shuiqing Song; Yan Pei; Yi Li

2007-01-01

280

Stable transformation of the food yam Dioscorea alata L. by particle bombardment.  

PubMed

A biolistic particle gun was used to deliver genetic material into intact yam cells. Cultured suspension cells of D. alata were bombarded with microprojectiles coated with pBI221.2 DNA and histochemical assays were carried out to show transient GUS expression in bombarded cells. Stably transformed D. alata cells were recovered from cultured cells after bombardment with microprojectiles coated with pRT99gus harbouring both the nptII and uidA genes. Bombarded cells were selected on a medium containing geneticin (G418). Two months after bombardment, calli resistant to G418 were assayed for GUS expression. There was a 100% correlation between resistance to G418 and GUS expression. From these calli, four cell lines were established and GUS activity in each line was determined fluorometrically. The use of a specific GUS inhibitor showed that the GUS activity was due to the introduced uidA gene rather than to any intrinsic GUS-like activity originating from the plant. Incorporation of the introduced DNA into the plant genomic DNA was confirmed by Southern analysis. PMID:24197354

Tör, M; Ainsworth, C; Mantell, S H

1993-05-01

281

Two negative cis-regulatory regions involved in fruit-specific promoter activity from watermelon (Citrullus vulgaris S.)  

PubMed Central

A 1.8?kb 5?-flanking region of the large subunit of ADP-glucose pyrophosphorylase, isolated from watermelon (Citrullus vulgaris S.), has fruit-specific promoter activity in transgenic tomato plants. Two negative regulatory regions, from –986 to –959 and from –472 to –424, were identified in this promoter region by fine deletion analyses. Removal of both regions led to constitutive expression in epidermal cells. Gain-of-function experiments showed that these two regions were sufficient to inhibit RFP (red fluorescent protein) expression in transformed epidermal cells when fused to the cauliflower mosaic virus (CaMV) 35S minimal promoter. Gel mobility shift experiments demonstrated the presence of leaf nuclear factors that interact with these two elements. A TCCAAAA motif was identified in these two regions, as well as one in the reverse orientation, which was confirmed to be a novel specific cis-element. A quantitative ?-glucuronidase (GUS) activity assay of stable transgenic tomato plants showed that the activities of chimeric promoters harbouring only one of the two cis-elements, or both, were ?10-fold higher in fruits than in leaves. These data confirm that the TCCAAAA motif functions as a fruit-specific element by inhibiting gene expression in leaves.

Yin, Tao; Wu, Hanying; Zhang, Shanglong; Liu, Jingmei; Lu, Hongyu; Zhang, Lingxiao; Xu, Yong; Chen, Daming

2009-01-01

282

Activity of the Arabidopsis RD29A and RD29B promoter elements in soybean under water stress.  

PubMed

The constitutive and drought-induced activities of the Arabidopsis thaliana RD29A and RD29B promoters were monitored in soybean (Glycine max (L.) Merr.] via fusions with the visual marker gene ?-glucuronidase (GUS). Physiological responses of soybean plants were monitored over 9 days of water deprivation under greenhouse conditions. Data were used to select appropriate time points to monitor the activities of the respective promoter elements. Qualitative and quantitative assays for GUS expression were conducted in root and leaf tissues, from plants under well-watered and dry-down conditions. Both RD29A and RD29B promoters were significantly activated in soybean plants subjected to dry-down conditions. However, a low level of constitutive promoter activity was also observed in both root and leaves of plants under well-watered conditions. GUS expression was notably higher in roots than in leaves. These observations suggest that the respective drought-responsive regulatory elements present in the RD29X promoters may be useful in controlling targeted transgenes to mitigate abiotic stress in soybean, provided the transgene under control of these promoters does not invoke agronomic penalties with leaky expression when no abiotic stress is imposed. PMID:22983672

Bihmidine, Saadia; Lin, Jiusheng; Stone, Julie M; Awada, Tala; Specht, James E; Clemente, Tom E

2013-01-01

283

In Plant Activation: An Inducible, Hyperexpression Platform for Recombinant Protein Production in Plants[W][OPEN  

PubMed Central

In this study, we describe a novel protein production platform that provides both activation and amplification of transgene expression in planta. The In Plant Activation (INPACT) system is based on the replication machinery of tobacco yellow dwarf mastrevirus (TYDV) and is essentially transient gene expression from a stably transformed plant, thus combining the advantages of both means of expression. The INPACT cassette is uniquely arranged such that the gene of interest is split and only reconstituted in the presence of the TYDV-encoded Rep/RepA proteins. Rep/RepA expression is placed under the control of the AlcA:AlcR gene switch, which is responsive to trace levels of ethanol. Transgenic tobacco (Nicotiana tabacum cv Samsun) plants containing an INPACT cassette encoding the ?-glucuronidase (GUS) reporter had negligible background expression but accumulated very high GUS levels (up to 10% total soluble protein) throughout the plant, within 3 d of a 1% ethanol application. The GUS reporter was replaced with a gene encoding a lethal ribonuclease, barnase, demonstrating that the INPACT system provides exquisite control of transgene expression and can be adapted to potentially toxic or inhibitory compounds. The INPACT gene expression platform is scalable, not host-limited, and has been used to express both a therapeutic and an industrial protein.

Dugdale, Benjamin; Mortimer, Cara L.; Kato, Maiko; James, Tess A.; Harding, Robert M.; Dale, James L.

2013-01-01

284

huBC1IL12, an immunocytokine which targets EDB-containing oncofetal fibronectin in tumors and tumor vasculature, shows potent anti-tumor activity in human tumor models  

Microsoft Academic Search

IL-12 is a cytokine which showed anti-tumor effects in clinical trials, but also produced serious toxicity. We describe a\\u000a fusion protein, huBC1-IL12, designed to achieve an improved therapeutic index by specifically targeting IL-12 to tumor and\\u000a tumor vasculature. huBC-1 is a humanized antibody that targets a cryptic sequence of the human ED-B-containing fibronectin\\u000a isoform, B-FN, present in the subendothelial extracellular

Kin-Ming Lo; Yan Lan; Scott Lauder; Jinyang Zhang; Beatrice Brunkhorst; Guozhong Qin; Rakesh Verma; Nigel Courtenay-Luck; Stephen D. Gillies

2007-01-01

285

Enhancement of foreign gene expression by a dicot intron in rice but not in tobacco is correlated with an increased level of mRNA and an efficient splicing of the intron.  

PubMed

The first intron of castor bean catalase gene, cat-1 was placed in the N-terminal region of the coding sequence of the beta-glucuronidase gene (gusA) and the intron-containing gusA was used with the cauliflower mosaic virus (CaMV) 35S promoter. Using this plasmid, pIG221, the effect of the intron on expression of beta-glucuronidase (GUS) activity was examined in transgenic rice calli and plants (a monocotyledon), and transgenic tobacco plants (a dicotyledon). The intron-containing plasmid increased the level of GUS enzyme activity 10 to 40-fold and 80 to 90-fold compared with the intronless plasmid, pBI221, in transgenic rice protoplasts and transgenic rice tissues, respectively. In contrast, the presence of the intron hardly influenced the expression of the GUS activity in transgenic tobacco plants. Northern blot analysis showed that the catalase intron was efficiently spliced in rice cells while transgenic tobacco plants contained both spliced and unspliced gusA transcripts in equal amounts. Furthermore, the level of the mature gusA transcript in transformed rice calli was greatly increased in the presence of the intron. The catalase intron was removed at the same splice junctions in transgenic rice and tobacco plants. These findings indicate that the stimulating effect of the intron on GUS expression is correlated with an efficient splicing of pre-mRNA and an increased level of mature mRNA. PMID:2263444

Tanaka, A; Mita, S; Ohta, S; Kyozuka, J; Shimamoto, K; Nakamura, K

1990-12-11

286

Enhancement of foreign gene expression by a dicot intron in rice but not in tobacco is correlated with an increased level of mRNA and an efficient splicing of the intron.  

PubMed Central

The first intron of castor bean catalase gene, cat-1 was placed in the N-terminal region of the coding sequence of the beta-glucuronidase gene (gusA) and the intron-containing gusA was used with the cauliflower mosaic virus (CaMV) 35S promoter. Using this plasmid, pIG221, the effect of the intron on expression of beta-glucuronidase (GUS) activity was examined in transgenic rice calli and plants (a monocotyledon), and transgenic tobacco plants (a dicotyledon). The intron-containing plasmid increased the level of GUS enzyme activity 10 to 40-fold and 80 to 90-fold compared with the intronless plasmid, pBI221, in transgenic rice protoplasts and transgenic rice tissues, respectively. In contrast, the presence of the intron hardly influenced the expression of the GUS activity in transgenic tobacco plants. Northern blot analysis showed that the catalase intron was efficiently spliced in rice cells while transgenic tobacco plants contained both spliced and unspliced gusA transcripts in equal amounts. Furthermore, the level of the mature gusA transcript in transformed rice calli was greatly increased in the presence of the intron. The catalase intron was removed at the same splice junctions in transgenic rice and tobacco plants. These findings indicate that the stimulating effect of the intron on GUS expression is correlated with an efficient splicing of pre-mRNA and an increased level of mature mRNA. Images

Tanaka, A; Mita, S; Ohta, S; Kyozuka, J; Shimamoto, K; Nakamura, K

1990-01-01

287

Promoter elements of rice susceptibility genes are bound and activated by specific TAL effectors from the bacterial blight pathogen, Xanthomonas oryzae pv. oryzae.  

PubMed

*Plant pathogenic bacteria of the genus Xanthomonas inject transcription activator-like effector (TALe) proteins that bind to and activate host promoters, thereby promoting disease or inducing plant defense. TALes bind to corresponding UPT (up-regulated by TALe) promoter boxes via tandemly arranged 34/35-amino acid repeats. Recent studies uncovered the TALe code in which two amino acid residues of each repeat define specific pairing to UPT boxes. *Here we employed the TALe code to predict potential UPT boxes in TALe-induced host promoters and analyzed these via beta-glucuronidase (GUS) reporter and electrophoretic mobility shift assays (EMSA). *We demonstrate that the Xa13, OsTFX1 and Os11N3 promoters from rice are induced directly by the Xanthomonas oryzae pv. oryzae TALes PthXo1, PthXo6 and AvrXa7, respectively. We identified and functionally validated a UPT box in the corresponding rice target promoter for each TALe and show that box mutations suppress TALe-mediated promoter activation. Finally, EMSA demonstrate that code-predicted UPT boxes interact specifically with corresponding TALes. *Our findings show that variations in the UPT boxes of different rice accessions correlate with susceptibility or resistance of these accessions to the bacterial blight pathogen. PMID:20345643

Römer, Patrick; Recht, Sabine; Strauss, Tina; Elsaesser, Janett; Schornack, Sebastian; Boch, Jens; Wang, Shiping; Lahaye, Thomas

2010-09-01

288

Aranciamycin analogs generated by combinatorial biosynthesis show improved antitumor activity  

Microsoft Academic Search

Expression of the aranciamycin biosynthetic gene cluster in Streptomyces diastatochromogenes Tü6028 resulted in production of four novel compounds, aranciamycins E, F, G, and H with different decorations in the tetracyclic\\u000a backbone. Two derivatives contain a d-amicetose moiety at C7 (aranciamycins F and G), two are hydroxylated at position C1 (aranciamycins E and G), and one is hydroxylated at C13 (aranciamycin

Andriy Luzhetskyy; Jens Hoffmann; Stefan Pelzer; Sven-Eric Wohlert; Andreas Vente; Andreas Bechthold

2008-01-01

289

ADA plaintiff must show AIDS limits major life activities.  

PubMed

In a rare case, a Federal court ruled that AIDS does not automatically qualify a plaintiff for legal protection under the Americans with Disabilities Act (ADA). [Name removed], an Illinois Wal-Mart stock clerk, was fired weeks after telling the store's general manager of his HIV status. [Name removed] alleges that the firing was due solely to his disease. Wal-Mart contends that [name removed] was fired for sexually harassing a co-worker, and says that since [name removed] was asymptomatic and asked for no accommodations, he does not qualify for ADA protection. Magistrate Morton Denlow agreed, saying that [name removed] raised no genuine issues about whether the ADA should protect him. A trial is scheduled for May. PMID:11365337

1998-05-15

290

Cotransformation of Trichoderma harzianum with beta-glucuronidase and green fluorescent protein genes provides a useful tool for monitoring fungal growth and activity in natural soils.  

PubMed

Trichoderma harzianum was cotransformed with genes encoding green fluorescent protein (GFP), beta-glucuronidase (GUS), and hygromycin B (hygB) resistance, using polyethylene glycol-mediated transformation. One cotransformant (ThzID1-M3) was mitotically stable for 6 months despite successive subculturing without selection pressure. ThzID1-M3 morphology was similar to that of the wild type; however, the mycelial growth rate on agar was reduced. ThzID1-M3 was formed into calcium alginate pellets and placed onto buried glass slides in a nonsterile soil, and its ability to grow, sporulate, and colonize sclerotia of Sclerotinia sclerotiorum was compared with that of the wild-type strain. Wild-type and transformant strains both colonized sclerotia at levels above those of indigenous Trichoderma spp. in untreated controls. There were no significant differences in colonization levels between wild-type and cotransformant strains; however, the presence of the GFP and GUS marker genes permitted differentiation of introduced Trichoderma from indigenous strains. GFP activity was a useful tool for nondestructive monitoring of the hyphal growth of the transformant in a natural soil. The green color of cotransformant hyphae was clearly visible with a UV epifluorescence microscope, while indigenous fungi in the same samples were barely visible. Green-fluorescing conidiophores and conidia were observed within the first 3 days of incubation in soil, and this was followed by the formation of terminal and intercalary chlamydospores and subsequent disintegration of older hyphal segments. Addition of 5-bromo-4-chloro-3-indolyl-beta-D-glucuronic acid (X-Gluc) substrate to recovered glass slides confirmed the activity of GUS as well as GFP in soil. Our results suggest that cotransformation with GFP and GUS can provide a valuable tool for the detection and monitoring of specific strains of T. harzianum released into the soil. PMID:10653755

Bae, Y S; Knudsen, G R

2000-02-01

291

Women showing off: notes on female exhibitionism.  

PubMed

The limitations of the phallocentric cast of earlier psychoanalytic formulations of "female exhibitionism" linger into the present. In part this connects to certain historical expectations for women's social behavior, and to the vicissitudes of Freud's insufficient knowledge of women in his libidinal psychosexual phasing used as a basis for analytic understanding. The contemporary fade of libido theory contributes to the neglect of such topics as they relate to the biological body. Yet ease and conflict regarding conscious and unconscious female body image representations related to that stepchild of theory-pregnancy and childbirth in particular-play a major role in female body display. Recognition of such body fantasies and female body meanings from early childhood into maturity tends to be marginalized within all of the psychoanalytic theories current today. The focus here on female exhibitionism suggests a normative spectrum for pleasurably active sex seeking and pleasurable procreative desire and fantasy that is present in a female's use of her body and which (of course, but secondarily) can become caught up in conflict. Two cases accenting analyses of female "showing off" behavior are included. PMID:18430704

Balsam, Rosemary H

2008-03-01

292

Lhcb transcription is coordinated with cell size and chlorophyll accumulation. Studies on fluorescence-activated, cell-sorter-purified single cells from wild-type and immutans Arabidopsis thaliana  

SciTech Connect

To study the mechanisms that integrate pigment and chlorophyll a/b-binding apoprotein biosynthesis during light-harvesting complex II assembly, we have examined {beta}-glucuronidase (GUS) enzyme activities, cell-sorting-separated single cells sizes in fluorescence activated, cell-sorting-separated single cells from transgenic Arabidopsis thaliana wild-type and immutans variegation mutant plants that express an Lhcb (photosystem II chlorophyll a/b-binding polypeptide gene)/GUS promoter fusion. We found that GUS activities are positively correlated with chlorophyll content and cell size in green cells from the control and immutans plants, indicating that Lhcb gene transcription is coordinated with cell size in this species. Compared with the control plants, however, chlorophyll production is enhanced in the green cells of immutans; this may represent part of a strategy to maximize photosynthesis in the white sectors of the mutant. Lhcb transcription is significantly higher in pure-white cells of the transgenic immutans plants than in pure-white cells from norflurazon-treated, photooxidized A. thaliana leaves. This suggests that immutans partially uncouples Lhcb transcription from its normal dependence on chlorophyll accumulation and chloroplast development. We conclude that immutans may play a role in regulating Lhcb transcription, and may be a key component in the signal transduction pathways that control chloroplast biogenesis. 58 refs., 5 figs., 2 tabs.

Meehan, L.; Harkins, K.; Rodermel, S. [Iowa State Univ., Ames, IA (United States)] [and others

1996-11-01

293

Expression and parent-of-origin effects for FIS2, MEA, and FIE in the endosperm and embryo of developing Arabidopsis seeds  

PubMed Central

The promoters of MEA (FIS1), FIS2, and FIE (FIS3), genes that repress seed development in the absence of pollination, were fused to ?-glucuronidase (GUS) to study their activity pattern. The FIS2?GUS product is found in the embryo sac, in each of the polar cell nuclei, and in the central cell nucleus. After pollination, the maternally derived FIS2?GUS protein occurs in the nuclei of the cenocytic endosperm. Before cellularization of the endosperm, activity is terminated in the micropylar and central nuclei of the endosperm and subsequently in the nuclei of the chalazal cyst. MEA?GUS has a pattern of activity similar to that of FIS2?GUS, but FIE?GUS protein is found in many tissues, including the prepollination embryo sac, and in embryo and endosperm postpollination. The similarity in mutant phenotypes; the activity of FIE, MEA, and FIS2 in the same cells in the embryo sac; and the fact that MEA and FIE proteins interact in a yeast two-hybrid system suggest that these proteins operate in the same system of control of seed development. Maternal and not paternal FIS2?GUS, MEA?GUS, and FIE?GUS show activity in early endosperm, so these genes may be imprinted. When fis2, mea, and fie mutants are pollinated, seed development is arrested at the heart embryo stage. The seed arrest of mea and fis2 is avoided when they are fertilized by a low methylation parent. The wild-type alleles of MEA or FIS2 are not required. The parent-of-origin-determined differential activity of MEA, FIS2, and FIE is not dependent on DNA methylation, but methylation does control some gene(s) that have key roles in seed development.

Luo, Ming; Bilodeau, Pierre; Dennis, Elizabeth S.; Peacock, W. James; Chaudhury, Abed

2000-01-01

294

The cauliflower mosaic virus (CaMV) 35S promoter sequence alters the level and patterns of activity of adjacent tissue- and organ-specific gene promoters.  

PubMed

Here we report the effect of the 35S promoter sequence on activities of the tissue- and organ-specific gene promoters in tobacco plants. In the absence of the 35S promoter sequence the AAP2 promoter is active only in vascular tissues as indicated by expression of the AAP2:GUS gene. With the 35S promoter sequence in the same T-plasmid, transgenic plants exhibit twofold to fivefold increase in AAP2 promoter activity and the promoter becomes active in all tissue types. Transgenic plants hosting the ovary-specific AGL5:iaaM gene (iaaM coding an auxin biosynthetic gene) showed a wild-type phenotype except production of seedless fruits, whereas plants hosting the AGL5:iaaM gene along with the 35S promoter sequence showed drastic morphological alterations. RT-PCR analysis confirms that the phenotype was caused by activation of the AGL5:iaaM gene in non-ovary organs including roots, stems and flowers. When the pollen-, ovule- and early embryo-specific PAB5:barnase gene (barnase coding a RNase gene) was transformed, the presence of 35S promoter sequence drastically reduced transformation efficiencies. However, the transformation efficiencies were restored in the absence of 35S promoter, indicating that the 35S promoter might activate the expression of PAB5:barnase in non-reproductive organs such as calli and shoot primordia. Furthermore, if the 35S promoter sequence was replaced with the NOS promoter sequence, no alteration in AAP2, AGL5 or PAB5 promoter activities was observed. Our results demonstrate that the 35S promoter sequence can convert an adjacent tissue- and organ-specific gene promoter into a globally active promoter. PMID:17340093

Zheng, Xuelian; Deng, Wei; Luo, Keming; Duan, Hui; Chen, Yongqin; McAvoy, Richard; Song, Shuiqing; Pei, Yan; Li, Yi

2007-08-01

295

Flat Globe: Showing the Changing Seasons  

NSDL National Science Digital Library

SeaWiFS false color data showing seasonal change in the oceans and on land for the entire globe. The data is seasonally averaged, and shows the sequence: fall, winter, spring, summer, fall, winter, spring (for the Northern Hemisphere).

Allen, Jesse; Newcombe, Marte; Feldman, Gene

1998-09-09

296

The Franklin Institute's Traveling Science Shows  

NSDL National Science Digital Library

The Franklin Institute's team of science educators are available for shows on a variety of science topics. Traveling Science shows are aligned with National Science Education Standards, and focus on Physics, Biology and Chemistry.

Shows, Traveling S.

2004-04-05

297

A Learning Model of Trade Show Attendance  

Microsoft Academic Search

This study examines trade show attendees' show choice behavior from a learning perspective. The purpose is to create a model that can be useful in attracting attendees, either to a show or even to a specific booth. Reasons for attending were found to be three-dimensional: shopping, career development, and general industry awareness. These dimensions were then used to develop clusters,

John F. Tanner Jr; Lawrence B. Chonko; Thomas V. Ponzurick

2001-01-01

298

The Wonders of Physics Traveling Show  

NSDL National Science Digital Library

The Wonders of Physics is a live physics show designed to stimulate interest in science in people of all ages and backgrounds. The program's fast-paced presentation is supplemented by a variety of media tools. In addition, a smaller traveling show is based in Madison, WI, but does shows all over the United States and Canada.

Physics, The W.

2004-06-02

299

Colonization by the arbuscular mycorrhizal fungus Glomus versiforme induces a defense response against the root-knot nematode Meloidogyne incognita in the grapevine (Vitis amurensis Rupr.), which includes transcriptional activation of the class III chitinase gene VCH3.  

PubMed

Inoculation of the grapevine (Vitis amurensis Rupr.) with the arbuscular mycorrhizal (AM) fungus Glomus versiforme significantly increased resistance against the root-knot nematode (RKN) Meloidogyne incognita. Studies using relative quantitative reverse transcription-PCR (RQRT-PCR) analysis of grapevine root inoculation with the AM fungus revealed an up-regulation of VCH3 transcripts. This increase was greater than that observed following infection with RKN. However, inoculation of the mycorrhizal grapevine roots with RKN was able to enhance VCH3 transcript expression further. Moreover, the increase in VCH3 transcripts appeared to result in a higher level of resistance against subsequent RKN infection. Constitutive expression of VCH3 cDNA in transgenic tobacco under the control of the cauliflower mosaic virus 35S promoter also conferred resistance against RKN, but had no significant effect on the growth of the AM fungus. We analyzed beta-glucuronidase (GUS) activity directed by a 1,216 bp VCH3 promoter in transgenic tobacco following inoculation with both the AM fungus and RKN. GUS activity was negligible in the root tissues before inoculation, and was more effectively induced after inoculation with the AM fungus than with RKN. Moreover, GUS staining in the mycorrhizal transgenic tobacco roots was enhanced by subsequent RKN infection, and was found ubiquitously throughout the whole root tissue. Together, these results suggest that AM fungus induced a defense response against RKN in the mycorrhizal grapevine roots, which appeared to involve transcriptional control of VCH3 expression throughout the whole root tissue. PMID:16326755

Li, Hai-Yan; Yang, Guo-Dong; Shu, Huai-Rui; Yang, Yu-Tao; Ye, Bao-Xing; Nishida, Ikuo; Zheng, Cheng-Chao

2006-01-01

300

Solar System Odyssey - Fulldome Digital Planetarium Show  

NSDL National Science Digital Library

This is a Fulldome Digital Planetarium Show. Learners go on a futuristic journey through our Solar System. They explore the inner and outer planets, then the moons: Titan, Europa, and Callisto as possible places to establish a human colony. A full-length preview of the show is available on the website, you need to scroll down about 3/4 of the page - under section on children's shows, direct link not available.

301

Production of bialaphos-resistant Nierembergia repens by electroporation  

Microsoft Academic Search

Transgenic plants with the herbicide-resistance gene (bar gene) were obtained via organogenesis from isolated mesophyll protoplasts of Nierembergia repens after applying electroporation. Transient ?-glucuronidase (GUS) activity of electroporated protoplasts assayed 2 days after\\u000a applying an electric pulse showed that optimum condition (transient GUS activity 319 pmol 4 MU\\/mg per min and plating efficiency\\u000a 2.43%) for electroporation was 0.5 kV\\/cm in field strength

Yoshiaki Shizukawa; Masahiro Mii

2008-01-01

302

25 CFR 141.56 - Show cause procedures.  

Code of Federal Regulations, 2011 CFR

...AFFAIRS, DEPARTMENT OF THE INTERIOR FINANCIAL ACTIVITIES BUSINESS PRACTICES ON THE NAVAJO, HOPI AND ZUNI RESERVATIONS Enforcement Powers, Procedures and Remedies § 141.56 Show cause procedures. (a) When the Commissioner believes...

2011-04-01

303

The Language of Show Biz: A Dictionary.  

ERIC Educational Resources Information Center

This dictionary of the language of show biz provides the layman with definitions and essays on terms and expressions often used in show business. The overall pattern of selection was intended to be more rather than less inclusive, though radio, television, and film terms were deliberately omitted. Lengthy explanations are sometimes used to express…

Sergel, Sherman Louis, Ed.

304

Show Me: Automatic Presentation for Visual Analysis  

Microsoft Academic Search

This paper describes Show Me, an integrated set of user interface commands and defaults that incorporate automatic presentation into a commercial visual analysis system called Tableau. A key aspect of Tableau is VizQL, a language for specifying views, which is used by Show Me to extend automatic presentation to the generation of tables of views (commonly called small multiple displays).

Jock D. Mackinlay; Pat Hanrahan; Chris Stolte

2007-01-01

305

Virus infection elevates transcriptional activity of miR164a promoter in plants  

PubMed Central

Background Micro RNAs (miRs) constitute a large group of endogenous small RNAs that have crucial roles in many important plant functions. Virus infection and transgenic expression of viral proteins alter accumulation and activity of miRs and so far, most of the published evidence involves post-transcriptional regulations. Results Using transgenic plants expressing a reporter gene under the promoter region of a characterized miR (P-miR164a), we monitored the reporter gene expression in different tissues and during Arabidopsis development. Strong expression was detected in both vascular tissues and hydathodes. P-miR164a activity was developmentally regulated in plants with a maximum expression at stages 1.12 to 5.1 (according to Boyes, 2001) along the transition from vegetative to reproductive growth. Upon quantification of P-miR164a-derived GUS activity after Tobacco mosaic virus Cg or Oilseed rape mosaic virus (ORMV) infection and after hormone treatments, we demonstrated that ORMV and gibberellic acid elevated P-miR164a activity. Accordingly, total mature miR164, precursor of miR164a and CUC1 mRNA (a miR164 target) levels increased after virus infection and interestingly the most severe virus (ORMV) produced the strongest promoter induction. Conclusion This work shows for the first time that the alteration of miR pathways produced by viral infections possesses a transcriptional component. In addition, the degree of miR alteration correlates with virus severity since a more severe virus produces a stronger P-miR164a induction.

2009-01-01

306

KGB- agents (Part 2: Music and show -bussines)  

NASA Astrophysics Data System (ADS)

The paper represents a continuation of the previous one 2008stuk.rept.....G and reffers to activity of KGB in the Show Busyness in the former USSR. During 1970-th the activity of KGB was concerned to control the market of musical disks, the disco clubs, the programs and the personal componence of the musical groups and orchestra.

Gaina, Alex

2008-10-01

307

Comparison of Weather Shows in Eastern Europe  

NASA Astrophysics Data System (ADS)

Comparison of Weather Shows in Eastern Europe Television weather shows in Eastern Europe have in most cases in the high graphical standard. There is though a wast difference in duration and information content in the weather shows. There are few signs and regularities by which we can see the character of the weather show. The main differences are mainly caused by the income structure of the TV station. Either it is a fully privately funded TV relying on the TV commercials income. Or it is a public service TV station funded mainly by the national budget or fixed fee structure/tax. There are wast differences in duration and even a graphical presentation of the weather. Next important aspect is a supplier of the weather information and /or the processor. Shortly we can say, that when the TV show is produced by the national met office, the TV show consists of more scientific terms, synoptic maps, satellite imagery, etc. If the supplier is the private meteorological company, the weather show is more user-friendly, laical with less scientific terms. We are experiencing a massive shift in public weather knowledge and demand for information. In the past, weather shows consisted only of maps with weather icons. In todaýs world, even the laic weather shows consist partly of numerical weather model outputs - they are of course designed to be understandable and graphically attractive. Outputs of the numerical weather models used to be only a part of daily life of a professional meteorologist, today they are common part of life of regular people. Video samples are a part of this presentation.

Najman, M.

2009-09-01

308

Melanoma Drug Trials Show Significant Promise  

MedlinePLUS

... sharing features on this page, please enable JavaScript. Melanoma Drug Trials Show Significant Promise By targeting immune ... Monday, June 2, 2014 Related MedlinePlus Pages Medicines Melanoma MONDAY, June 2, 2014 (HealthDay News) -- A relatively ...

309

Spacecraft Image Mashup Shows Galactic Collision  

NASA Video Gallery

This new composite image from the Chandra X-ray Observatory, the Hubble Space Telescope, and the Spitzer Space Telescope shows two colliding galaxies more than a 100 million years after they first ...

310

Adult Stem Cell Research Shows Promise  

MedlinePLUS

... Tobacco Products Vaccines, Blood & Biologics Articulos en Espanol Adult Stem Cell Research Shows Promise Search the Consumer Updates Section ... re looking at a particular kind of multipotent adult stem cell—the MSC—which is being used in a ...

311

TRMM Satellite Shows Heavy Rainfall in Cristina  

NASA Video Gallery

NASA's TRMM satellite rainfall data was overlaid on an enhanced visible/infrared image from NOAA's GOES-East satellite showing cloud and rainfall extent. Green areas indicate rainfall at over 20 mm...

312

GOES Satellite Data Shows Tornado Development  

NASA Video Gallery

This animation of NOAA's GOES-East satellite data shows the development and movement of the weather system that spawned tornadoes affecting the southern and eastern U.S. states on April 27-29, 2014...

313

New Psoriasis Drug Shows Promise in Trials  

MedlinePLUS

... features on this page, please enable JavaScript. New Psoriasis Drug Shows Promise in Trials Secukinumab appears more ... Wednesday, July 9, 2014 Related MedlinePlus Pages Medicines Psoriasis WEDNESDAY, July 9, 2014 (HealthDay News) -- A new ...

314

map showing predicted habitat potentional for tortoise  

USGS Multimedia Gallery

This map shows the spatial representation of the predicted habitat potential index values for desert tortoise in the Mojave and parts of the Sonoran Deserts of California, Nevada, Utah, and Arizona. Map: USGS. ...

2009-05-21

315

Learning to Show You're Listening  

ERIC Educational Resources Information Center

Good listeners generally produce back-channel feedback, that is, short utterances such as "uh-huh" which signal active listening. As the rules governing back-channeling vary from language to language, second-language learners may need help acquiring this skill. This paper is an initial exploration of how to provide this. It presents a training…

Ward, Nigel G.; Escalante, Rafael; Al Bayyari, Yaffa; Solorio, Thamar

2007-01-01

316

Purification and Characterization of a CkTLP Protein from Cynanchum komarovii Seeds that Confers Antifungal Activity  

PubMed Central

Background Cynanchum komarovii Al Iljinski is a desert plant that has been used as analgesic, anthelminthic and antidiarrheal, but also as a herbal medicine to treat cholecystitis in people. We have found that the protein extractions from C. komarovii seeds have strong antifungal activity. There is strong interest to develop protein medication and antifungal pesticides from C. komarovii for pharmacological or other uses. Methodology/Principal Findings An antifungal protein with sequence homology to thaumatin-like proteins (TLPs) was isolated from C. komarovii seeds and named CkTLP. The three-dimensional structure prediction of CkTLP indicated the protein has an acid cleft and a hydrophobic patch. The protein showed antifungal activity against fungal growth of Verticillium dahliae, Fusarium oxysporum, Rhizoctonia solani, Botrytis cinerea and Valsa mali. The full-length cDNA was cloned by RT-PCR and RACE-PCR according to the partial protein sequences obtained by nanoESI-MS/MS. The real-time PCR showed the transcription level of CkTLP had a significant increase under the stress of abscisic acid (ABA), salicylic acid (SA), methyl jasmonate (MeJA), NaCl and drought, which indicates that CkTLP may play an important role in response to abiotic stresses. Histochemical staining showed GUS activity in almost the whole plant, especially in cotyledons, trichomes and vascular tissues of primary root and inflorescences. The CkTLP protein was located in the extracellular space/cell wall by CkTLP::GFP fusion protein in transgenic Arabidopsis. Furthermore, over-expression of CkTLP significantly enhanced the resistance of Arabidopsis against V. dahliae. Conclusions/Significance The results suggest that the CkTLP is a good candidate protein or gene for contributing to the development of disease-resistant crops.

Wang, Qinghua; Li, Fuguang; Zhang, Xue; Zhang, Yongan; Hou, Yuxia; Zhang, Shengrui; Wu, Zhixia

2011-01-01

317

Educational Outreach: The Space Science Road Show  

NASA Astrophysics Data System (ADS)

The poster presented will give an overview of a study towards a "Space Road Show". The topic of this show is space science. The target group is adolescents, aged 12 to 15, at Dutch high schools. The show and its accompanying experiments would be supported with suitable educational material. Science teachers at schools can decide for themselves if they want to use this material in advance, afterwards or not at all. The aims of this outreach effort are: to motivate students for space science and engineering, to help them understand the importance of (space) research, to give them a positive feeling about the possibilities offered by space and in the process give them useful knowledge on space basics. The show revolves around three main themes: applications, science and society. First the students will get some historical background on the importance of space/astronomy to civilization. Secondly they will learn more about novel uses of space. On the one hand they will learn of "Views on Earth" involving technologies like Remote Sensing (or Spying), Communication, Broadcasting, GPS and Telemedicine. On the other hand they will experience "Views on Space" illustrated by past, present and future space research missions, like the space exploration missions (Cassini/Huygens, Mars Express and Rosetta) and the astronomy missions (Soho and XMM). Meanwhile, the students will learn more about the technology of launchers and satellites needed to accomplish these space missions. Throughout the show and especially towards the end attention will be paid to the third theme "Why go to space"? Other reasons for people to get into space will be explored. An important question in this is the commercial (manned) exploration of space. Thus, the questions of benefit of space to society are integrated in the entire show. It raises some fundamental questions about the effects of space travel on our environment, poverty and other moral issues. The show attempts to connect scientific with community thought. The difficulty with a show this elaborate and intricate is communicating on a level understandable for teenagers, whilst not treating them like children. Professional space scientists know how easy it is to lose oneself in technical specifics. This would, of course, only confuse young people. The author would like to discuss the ideas for this show with a knowledgeable audience and hopefully get some (constructive) feedback.

Cox, N. L. J.

2002-01-01

318

Sequence analysis of a "true" chalcone synthase (chs_H1) oligofamily from hop (Humulus lupulus L.) and PAP1 activation of chs_H1 in heterologous systems.  

PubMed

Screening of a cDNA library of the hop cv. Osvald's 72 and genomic cloning were used to isolate members of an oligofamily of chs_H1 genes that codetermine the biosynthesis of prenylated chalcones known to be valuable medicinal compounds present in hop (Humulus lupulus L.). chs_H1 oligofamily members showed more than 99% and 98% identity on nucleotide and amino acid levels, respectively, and retained all conserved amino acids that form the catalytic center characteristic for "true" chalcone synthases. The chs_H1 promoter exhibited low sequence variability in addition to conservation of all predicted cis-regulatory elements. Possible transactivation of the chs_H1 gene with the transcription factor PAP1 from Arabidopsis thaliana was assayed using Agrobacterium tumefaciens infiltrations of Nicotiana benthamiana and Petunia hybrida plants. Infiltration of N. benthamiana leaves with chs_H1 promoter/GUS chimeras led to a 24.8-fold increase of the GUS activity when coinfiltrated with the pap1 gene. Coinfiltration of the "native" chs_H1 gene with pap1 led to an increased accumulation of chs_H1 mRNA as observed by semiquantitative reverse transcription-polymerase chain reaction. Transgenic lines of P. hybrida expressing the pap1 gene showed unusual patterns of UV-A-inducible pigmentation and anthocyanin accumulation in parenchymatic and medulla cells. Infiltration of transgenic leaves of P. hybrida with chs_H1 and pap1 genes arranged as a tandem led to quick pigmentation within 12 h after UV-A irradiation. It is indicated that the chs_H1 promoter contains functional element(s) mediating an efficient response to PAP1 expression and UV-A irradiation. UV-A also induced chs_H1 mRNA and accumulation of flavonol glycosides in hop leaves. It can be expected that the PAP1 factor could significantly influence the expression of the chs_H1 oligofamily in transgenic hop and modify the hop metabolome. PMID:17002429

Matousek, Jaroslav; Vrba, Lukás; Skopek, Josef; Orctova, Lidmila; Pesina, Karel; Heyerick, Arne; Baulcombe, David; De Keukeleire, Denis

2006-10-01

319

Liquid Crystal Research Shows Deformation By Drying  

NASA Technical Reports Server (NTRS)

These images, from David Weitz's liquid crystal research, show ordered uniform sized droplets (upper left) before they are dried from their solution. After the droplets are dried (upper right), they are viewed with crossed polarizers that show the deformation caused by drying, a process that orients the bipolar structure of the liquid crystal within the droplets. When an electric field is applied to the dried droplets (lower left), and then increased (lower right), the liquid crystal within the droplets switches its alignment, thereby reducing the amount of light that can be scattered by the droplets when a beam is shone through them.

2003-01-01

320

The object-oriented trivia show (TOOTS)  

Microsoft Academic Search

OOPSLA has a longstanding tradition of being a forum for discussing the cutting edge of technology in a fun and participatory environment. The type of events sponsored by OOPSLA sometimes border on the unconventional. This event represents an atypical panel that conforms to the concept of a game show that is focused on questions and answers related to SPLASH, OOPSLA,

Jeff Gray; Jules White

2010-01-01

321

Creating Computer Slide Shows the Easy Way.  

ERIC Educational Resources Information Center

Explains how to create slide shows via computer using common software programs such as ClarisWorks. Highlights include identifying the audience, organizing the information, becoming familiar with technical specifications of the equipment, the use of text, style consistency, the use of graphics, and the use of color. (LRW)

Anderson, Mary Alice

1996-01-01

322

Video showing use of circuit construction kit  

NSDL National Science Digital Library

The link below shows "Mr. Anderson" using the circuit construction kit in order to explore Ohm's Law. It allows the students to predict either general patterns or specific variables when changes are made to a circuit. http://www.bozemanscience.com/science-videos/2011/6/26/voltage-current-and-resistance.html

2012-02-28

323

Showing Enantiomorphous Crystals of Tartaric Acid  

ERIC Educational Resources Information Center

Most of the articles and textbooks that show drawings of enantiomorphous crystals use an inadequate view to appreciate the fact that they are non-superimposable mirror images of one another. If a graphical presentation of crystal chirality is not evident, the main attribute of crystal enantiomorphism can not be recognized by students. The classic…

Andrade-Gamboa, Julio

2007-01-01

324

Analysis shows process industry accident losses rising  

Microsoft Academic Search

An analysis of the 150 largest losses caused by accidents and natural phenomena in the hydrocarbon processing and chemical industries during a period of 30 years ending Jan. 1, 1989, shows that the cost and number of losses is increasing. The catastrophic losses analyzed were used to develop statistical trends from the losses in a data base. The trended data

J. A. Krembs; J. M. Connolly

1990-01-01

325

Olaparib shows promise in multiple tumor types.  

PubMed

A phase II study of the PARP inhibitor olaparib (AstraZeneca) for cancer patients with inherited BRCA1 and BRCA2 gene mutations confirmed earlier results showing clinical benefit for advanced breast and ovarian cancers, and demonstrated evidence of effectiveness against pancreatic and prostate cancers. PMID:23847380

2013-07-01

326

Where the Boys Are: Show Chorus.  

ERIC Educational Resources Information Center

Boys are given a chance to experience choral music through the aid of a system that lessens peer pressure. Through auditions, the teacher can evaluate musical ability, coordination level, interaction with peers, and ability to learn rapidly. Show chorus has brought a unique interest to the music program. (AU/CS)

Mancuso, Sandra L.

1983-01-01

327

Food safety at shows and fairs  

Microsoft Academic Search

Food events, such as food festivals, agricultural shows and village fetes take place throughout the UK, usually in outdoor locations. Consumers’ overall satisfaction with the food purchased at such events is high and they have few or no concerns about the food safety on sale. A wide variety of foods, including some high-risk products are offered for sale to the

Denise Worsfold

2003-01-01

328

Laser entertainment and light shows in education  

NASA Astrophysics Data System (ADS)

Laser shows and beam effects have been a source of entertainment since its first public performance May 9, 1969, at Mills College in Oakland, California. Since 1997, the Photonics Center, NgeeAnn Polytechnic, Singapore, has been using laser shows as a teaching tool. Students are able to exhibit their creative skills and learn at the same time how lasers are used in the entertainment industry. Students will acquire a number of skills including handling three- phase power supply, operation of cooling system, and laser alignment. Students also acquire an appreciation of the arts, learning about shapes and contours as they develop graphics for the shows. After holography, laser show animation provides a combination of the arts and technology. This paper aims to briefly describe how a krypton-argon laser, galvanometer scanners, a polychromatic acousto-optic modulator and related electronics are put together to develop a laser projector. The paper also describes how students are trained to make their own laser animation and beam effects with music, and at the same time have an appreciation of the operation of a Class IV laser and the handling of optical components.

Sabaratnam, Andrew T.; Symons, Charles

2002-05-01

329

Slide Show Maker 3.53  

NSDL National Science Digital Library

Slide Show Maker 3.53 is a small application that allows users to create an .avi file from a number of .bmp or .jpg images. After creating these files, users have the ability to add professional looking effects, such as text overlays and fade-in or fade-out effects. The interface used to implement these effects is well laid out, and users can begin to use the program after a few minutes of trying out the various commands and tools. Additionally, the homepage contains a FAQ section and several screen shots of the program in use. Slide Show Maker 3.53 is compatible with all systems running Windows 95 and higher.

330

Learning helicopter control through “teaching by showing  

Microsoft Academic Search

A model-free “teaching by showing” methodology is developed to train a fuzzy-neural controller for an autonomous robot helicopter. The controller is generated and tuned using training data gathered while a teacher operates the helicopter. A hierarchical behavior-based control architecture is used, with each behavior implemented as a hybrid fuzzy logic controller (FLC) and general regression neural network controller (GRNNC). The

James F. Montgomery; George A. Bekey

1998-01-01

331

Ge Crystals on Si Show Their Light  

NASA Astrophysics Data System (ADS)

We report micron-sized Ge crystal arrays grown on deeply patterned Si substrates that yield a surge of the interband photoluminescence intensity by more than 2 orders of magnitude with respect to that typical for epitaxial layers directly grown on planar substrates. This finding is ascribed to the strongly modified internal quantum efficiency induced by controlling the nonradiative recombination at dislocations and to the improved light extraction offered by the array architecture. By spectrally resolving the interband and the dislocation-related luminescence, we address the parasitic activity of extended defects and its impact on the optical properties of the heterosystem. Such results are then exploited along with band gap engineering to design SiGe reflectors and Ge quantum wells that are effective in further amplifying the emission yield.

Pezzoli, F.; Isa, F.; Isella, G.; Falub, C. V.; Kreiliger, T.; Salvalaglio, M.; Bergamaschini, R.; Grilli, E.; Guzzi, M.; von Känel, H.; Miglio, L.

2014-05-01

332

Groks Science Show for February 12, 2003  

NSDL National Science Digital Library

Chemistry has applications in many diverse fields from astronomy to zoology. With new applications emerging, it is the job of Chemical and Engineering News (CEN), a publication of the American Chemical Society, to report the breadth of activity in chemistry. This radio broadcast features an interview with the editor of CEN, Madeleine Jacobs, who discusses the CEN magazine and its handling of potentially controversial issues such as global warming; how chemistry has become increasingly interdisciplinary; the frontiers of chemistry, such as in nanotechnology; and the often negative public image of chemistry as solely toxic. The interview with Jacobs begins at 11 minutes and 50 seconds into the audio file; the interview is 14 minutes and 30 seconds in length.

2007-05-17

333

Normalization of gas shows improves evaluation  

SciTech Connect

A normalization scheme has been developed that allows mud-log gas curves to be correlated with each other and with other well logs. The normalized mud logs may also be used to enhance formation and geopressure evaluation. The method, which requires relatively simple calculations and uses data already available in the mud logging unit, overcomes a major weakness of traditional mud logging methods: too many factors, other than reservoir content, affected gas show magnitude. As a result, mud log gas analyses could not be numerically integrated with other well logs. Often, even mud logs from nearby wells might not be reliably correlated with each other.

Whittaker, A.; Sellens, G.

1987-04-20

334

Latest European coelacanth shows Gondwanan affinities  

PubMed Central

The last European fossil occurrence of a coelacanth is from the Mid-Cretaceous of the English Chalk (Turonian, 90 million years ago). Here, we report the discovery of a coelacanth from Late Cretaceous non-marine rocks in southern France. It consists of a left angular bone showing structures that imply close phylogenetic affinities with some extinct Mawsoniidae. The closest relatives are otherwise known from Cretaceous continental deposits of southern continents and suggest that the dispersal of freshwater organisms from Africa to Europe occurred in the Late Cretaceous.

Cavin, Lionel; Forey, Peter L; Buffetaut, Eric; Tong, Haiyan

2005-01-01

335

Asteroid Ida - 6 Views Showing Rotation  

NASA Technical Reports Server (NTRS)

This composite image shows the asteroid 243 Ida as seen from the Galileo spacecraft during its approach on August 28, 1993. The six views were shuttered through the camera's green filter and show Ida's rotation over a period of about 3 hours 18 minutes. The asteroid makes a complete rotation every 4 hours 38 minutes; therefore, this set of images spans about 3/4 of Ida's rotation period and shows most of Ida's surface. By combining the information in these views with that from the highest resolution images returned from the spacecraft in September 1993, the size and shape of this irregular body can now be determined accurately The asteroid appears to be about 58 kilometers (36 miles) long and about 23 kilometers wide, with a very irregular shape and volume of some 16,000 cubic kilometers. The images are arranged in chronological order from a time 3 hours 51 minutes before closest approach (upper left), through upper right, middle left, middle right lower left and lower right (33 minutes before closest approach). The six images show Ida at the same scale throughout. Ida's rotation axis is roughly vertical in these images, and the rotation causes the right-hand end of Ida to move toward the viewer as time progresses. The first image was taken from a range of about 171,000 km (106,000 miles) and provides an image resolution of about 1,700 meters per pixel (the highest resolution achieved for Ida is about 25 meters per pixel). The second, taken 70 minutes later, is from 119,000 kilometers, followed by 102,000 kilometers, 85,000 kilometers, 50,000 kilometers, and 25,000 kilometers. The features on Ida are less sharp in the earlier views because of the greater distances. Prominent in the middle three views is a deep depression across the short axis of the Asteroid. This feature tends to support the idea that Ida may have originally been formed from two or more separate large objects that collided softly and stuck together. Also visible in the lower left view is an apparent linear albedo or reflectance boundary. Color images yet to be returned from the Galileo spacecraft may help resolve the question of whether or not the two ends of Ida are made of different materials.

1994-01-01

336

VLA Shows "Boiling" in Atmosphere of Betelgeuse  

NASA Astrophysics Data System (ADS)

A team of astronomers says that observations with the National Science Foundation's Very Large Array (VLA) radio telescope show that a neighboring bloated star has giant convective plumes propelling gas from its surface (photosphere) up into the star's atmosphere. This new information contradicts long-held ideas that such stellar atmospheres are more uniform, and may resolve questions about how the star's atmosphere attains its enormous size as well as how dust and gas is driven away from the star. Jeremy Lim of the Academia Sinica Institute of Astronomy & Astrophysics in Taiwan; Chris Carilli, Anthony Beasley, and Ralph Marson of the National Radio Astronomy Observatory (NRAO) in Socorro, NM; and Stephen White of the University of Maryland studied the red-supergiant star Betelgeuse, about 430 light-years away in the constellation Orion. They reported their findings in the April 9 issue of the scientific journal Nature. "These radio-telescope images confirm that Betelgeuse -- already more than 600 times larger than our Sun -- has a dense atmosphere that extends to many times larger still than the star itself," said Lim. "The highest-resolution image shows the star's atmosphere to have a remarkably complex structure." "To our surprise," added White, "the images also show that most of the gas in the atmosphere is only about as hot as that on the surface. Previously, all of it was thought to be very much hotter." The astronomers used the VLA to make images of Betelgeuse at a variety of radio frequencies. The series of radio observations measured the temperature of the star's atmosphere at different heights. Previous observations with the Hubble Space Telescope (HST) at ultraviolet wavelengths showed that the star's atmosphere contains very hot gas at about twice the surface temperature. The VLA images showed that there also is lower-temperature gas throughout the atmosphere. This gas is near the surface temperature at low heights and decreases in temperature progressively outwards. Although its existence was not previously suspected, this lower-temperature gas turns out to be the most abundant constituent of Betelgeuse's atmosphere. "This alters our basic understanding of red-supergiant star atmospheres," explains Lim. "Instead of the star's atmosphere expanding uniformly because of gas heated to very high temperatures near its surface, it now appears that several giant convection cells propel gas from the star's surface into its atmosphere. This creates the complex structure we observe for Betelgeuse's atmosphere." Betelgeuse can be likened to an enormous "boiling" ball of gas heated by the release of energy from nuclear fusion in its core. The circulating boiling pattern -- convection -- appears as large regions of hot upwelling gas on the star's surface. "The idea that red-supergiant stars have enormous convection cells is not new," noted Marson. "This was suggested by Martin Schwarzschild more than 20 years ago, and was seen in optical images of Betelgeuse's surface in 1990." The new picture of Betelgeuse's atmosphere also helps resolve the mystery of how massive amounts of dust and gas are expelled from red supergiant stars, an important source of enrichment for the interstellar medium. If their atmospheres were entirely very hot at lower levels, dust grains would not be able to condense there. Dust grains could possibly condense at higher levels, but there they would not get enough "push" from the star's radiation to explain their outward movement. In the new picture, the relatively cool environment at lower levels allows dust grains to condense effectively; here they can be strongly propelled by the more-intense starlight, carrying gas with them. Indeed, dust has previously been inferred to form sporadically near Betelgeuse's surface, but its presence there was difficult to reconcile with the old picture. "This method for propelling the mass outflows of red giant and supergiant stars was proposed by Sun Kwok i

1998-04-01

337

Star Shows It Has The Right Stuff  

NASA Astrophysics Data System (ADS)

Astronomers have used an observation by NASA's Chandra X-ray Observatory to make the best case yet that a star can be engulfed by its companion star and survive. This discovery will help astronomers better understand how closely coupled stars, and perhaps even stars and planets, evolve when one of the stars expands enormously in its red giant phase. The binary star system known as V471 Tauri comprises a white dwarf star (the primary) in a close orbit -- one thirtieth of the distance between Mercury and the Sun -- with a normal Sun-like star (the secondary). Chandra's data showed that the hot upper atmosphere of the secondary star has a deficit of carbon atoms relative to nitrogen atoms. "This deficit of carbon atoms is the first clear observational evidence that the normal star was engulfed by its companion in the past," according to Jeremy Drake of the Smithsonian Astrophysical Observatory in Cambridge, MA, who coauthored an article on V471 in The Astrophysical Journal Letters with Marek Sarna of the N. Copernicus Astronomical Center in Poland. The white dwarf star was once a star several times as massive as the Sun. Nuclear fusion reactions in the core of such a star convert carbon into nitrogen over a period of about a billion years. When the fuel in the core of the star is exhausted, the core collapses, triggering more energetic nuclear reactions that cause the star to expand and transform into a red giant before eventually collapsing to become a white dwarf. The carbon-poor material in the core of the red giant is mixed with outer part of the star, so its atmosphere shows a deficit of carbon, as compared with Sun-like stars. The X-ray spectra of a red giant star (top panel) and a Sun-like star (bottom panel) show the large difference in the peaks due to carbon atoms in the two stars. Theoretical calculations indicate that a red giant in a binary system can completely envelop its companion star and dramatically affect its evolution. During this common envelope phase, friction causes the companion star to spiral inward rapidly where it will either be destroyed by the red giant, or it will survive when much of the envelope is spun away. If the companion star manages to survive, it will bear the marks of its ordeal in the form of contamination by carbon-poor material that it accreted while it was inside the red giant envelope. The X-ray spectrum of V471 Tauri in the middle panel shows just this effect - the carbon peak is intermediate between that of a Sun-like star and an isolated red giant star. The data indicate that about 10 percent of the star's mass has been accreted from the red giant. In the future the companion star can return the favor when it expands and dumps material back onto the white dwarf. If enough material is dumped on the white dwarf, it could cause the white dwarf to explode as a supernova. "It's a dog-eat-dog world out there," observed Drake. V471 Tau was observed for approximately one day by Chandra using the Low Energy Transmission Grating and High Resolution Camera on January 24-25, 2002. NASA's Marshall Space Flight Center, Huntsville, Ala., manages the Chandra program for the Office of Space Science, NASA Headquarters, Washington. Northrop Grumman of Redondo Beach, Calif., formerly TRW, Inc., was the prime development contractor for the observatory. The Smithsonian Astrophysical Observatory controls science and flight operations from the Chandra X-ray Center in Cambridge, Mass.

2004-01-01

338

Lemurs and macaques show similar numerical sensitivity.  

PubMed

We investigated the precision of the approximate number system (ANS) in three lemur species (Lemur catta, Eulemur mongoz, and Eulemur macaco flavifrons), one Old World monkey species (Macaca mulatta) and humans (Homo sapiens). In Experiment 1, four individuals of each nonhuman primate species were trained to select the numerically larger of two visual arrays on a touchscreen. We estimated numerical acuity by modeling Weber fractions (w) and found quantitatively equivalent performance among all four nonhuman primate species. In Experiment 2, we tested adult humans in a similar procedure, and they outperformed the four nonhuman species but showed qualitatively similar performance. These results indicate that the ANS is conserved over the primate order. PMID:24068469

Jones, Sarah M; Pearson, John; DeWind, Nicholas K; Paulsen, David; Tenekedjieva, Ana-Maria; Brannon, Elizabeth M

2014-05-01

339

Microbiological and environmental issues in show caves.  

PubMed

Cultural tourism expanded in the last half of the twentieth century, and the interest of visitors has come to include caves containing archaeological remains. Some show caves attracted mass tourism, and economical interests prevailed over conservation, which led to a deterioration of the subterranean environment and the rock art. The presence and the role of microorganisms in caves is a topic that is often ignored in cave management. Knowledge of the colonisation patterns, the dispersion mechanisms, and the effect on human health and, when present, over rock art paintings of these microorganisms is of the utmost importance. In this review the most recent advances in the study of microorganisms in caves are presented, together with the environmental implications of the findings. PMID:22806150

Saiz-Jimenez, Cesareo

2012-07-01

340

ArtBots: The Robot Talent Show  

NSDL National Science Digital Library

Sponsored by the Columbia University Computer Music Center and the Digital Media Center of Columbia University's School of the Arts, ArtBots is "an international art exhibition for robotic art and art-making robots." The annual event invites artists from around the world to enter their robots based on the broad guidelines: "if you think it's a robot and you think it's art, we encourage you to submit." The Participants section includes a photo, description (including some interpretive analysis) and website link for each of the ArtBot 2004 participants. Some sample entries include a robot that draws, one that picks up balls and a "robotic sculpture built around a section of tree trunk." The website even offers a sample of the Theme Song from the show.

341

832 Karin Shows No Rotational Spectral Variations  

NASA Astrophysics Data System (ADS)

Sasaki et al. (2004, 2005) claimed that 832 Karin, the brightest member of the very young (5.75 Myr) Karin cluster of the Koronis family, shows dramatically different colors as a function of rotational phase. It was interpreted that Karin is a fragment of the recently broken-up asteroid, showing the reddish space-weathered exterior surface of the precursor asteroid as well as an interior face, which has not had time to become space-weathered. On five nights during UT 7-14 January 2006, we observed Karin with the SpeX instrument, 0.8-2.5 microns, on the IRTF. We sampled its spectrum well throughout its rotation. We analyzed the data in 50 deg. intervals of rotational longitude; some longitudes were sampled during two different nights. We find that Karin exhibits minimal spectral variations with rotation, certainly nothing of the magnitude reported by Sasaki et al. Since our data resemble Sasaki et al.'s "blue" and "green" sets, we suggest that their "red" set is spurious. Indeed, it is difficult to understand how the reported color change could have occurred during such a modest interval ( 4%) of rotational longitude. (Note that we have not determined Karin's pole position nor the phase of the Sasaki et al. data within our own coverage, so the refutation of dramatic color change is not absolutely secure.) Karin and its family members are not quite as red as typical S-types, yet have shallow absorption bands. Perhaps the space-weathering process affecting these young asteroids has had time to reduce spectral contrast, but has not operated long enough to redden them -- an intermediate case of space weathering, which has gone to completion for older main-belt asteroids of these sizes. Supported by the NASA Planetary Astronomy Program. T. Sasaki et al. 2004. ApJ 615, L161-L164; T. Sasaki et al. 2005. LPSC XXXVI, 1590.pdf.

Chapman, Clark R.; Enke, B.; Merline, W. J.; Nesvorny, D.; Tamblyn, P.; Young, E. F.

2006-09-01

342

Genotypically defined lissencephalies show distinct pathologies.  

PubMed

Lissencephaly is traditionally divided into 2 distinct pathologic forms: classic (type I) and cobblestone (type II). To date, mutations in 4 genes, LIS1, DCX, RELN, and ARX, have been associated with distinct type I lissencephaly syndromes. Each of these genes has been shown to play a role in normal cell migration, consistent with the presumed pathogenesis of type I lissencephaly. Based on these data, we hypothesized that all forms of radiographically defined type I lissencephaly independent of genotype would be pathologically similar. To test this hypothesis, we examined brains from 16 patients, including 15 lissencephalic patients and one patient with subcortical band heterotopia. Of these 16 patients, 6 had LIS1 deletions, 2 had DCX mutations, and 2 had ARX mutations. In addition, 6 patients had no defined genetic defect, although the patient with subcortical band heterotopia exhibited the same pattern of malformation expected with an XLIS mutation. In all cases, the cortex was thickened; however, the topographic distribution of the cortical pathology varied, ranging from frontal- to occipital-biased pathology to diffuse involvement of the neocortex. Although brains with LIS1 deletions exhibited the classic 4-layer lissencephalic architecture, patients with DCX and ARX mutations each had unique cytoarchitectural findings distinct from LIS1. Furthermore, 2 of the 5 patients with no known genetic defect showed a fourth type of histopathology characterized by a 2-layered cortex. Interestingly, the 2 brains with the fourth type of lissencephaly showed profound brainstem and cerebellar abnormalities. In summary, we identified at least 4 distinct histopathologic subtypes of lissencephaly that stratify with the underlying genetic defect. Based on these data, a new classification for lissencephaly is proposed that incorporates both pathologic and genetic findings. PMID:16215456

Forman, Mark S; Squier, Waney; Dobyns, William B; Golden, Jeffrey A

2005-10-01

343

Plant species descriptions show signs of disease.  

PubMed

It is well known that diseases can greatly influence the morphology of plants, but often the incidence of disease is either too rare or the symptoms too obvious for the 'abnormalities' to cause confusion in systematics. However, we have recently come across several misinterpretations of disease-induced traits that may have been perpetuated into modern species inventories. Anther-smut disease (caused by the fungus Microbotryum violaceum) is common in many members of the Caryophyllaceae and related plant families. This disease causes anthers of infected plants to be filled with dark-violet fungal spores rather than pollen. Otherwise, their vegetative morphology is within the normal range of healthy plants. Here, we present the results of a herbarium survey showing that a number of type specimens (on which the species name and original description are based) in the genus Silene from Asia are diseased with anther smut. The primary visible disease symptom, namely the dark-violet anthers, is incorporated into the original species descriptions and some of these descriptions have persisted unchanged into modern floras. This raises the question of whether diseased type specimens have erroneously been given unique species names. PMID:14667368

Hood, Michael E; Antonovics, Janis

2003-11-01

344

Fading Supernova Creates Spectacular Light Show  

NASA Technical Reports Server (NTRS)

This image of SN 1987A, taken November 28, 2003 by the Advanced Camera for Surveys aboard NASA's Hubble Space Telescope (HST), shows many bright spots along a ring of gas, like pearls on a necklace. These cosmic pearls are being produced as superior shock waves unleashed during an explosion slam into the ring at more than a million miles per hour. The collision is heating the gas ring, causing its irnermost regions to glow. Astronomers detected the first of these hot spots in 1996, but now they see dozens of them all around the ring. With temperatures surging from a few thousand degrees to a million degrees, the flares are increasing in number. In the next few years, the entire ring will be ablaze as it absorbs the full force of the crash and is expected to become bright enough to illuminate the star's surroundings. Astronomers will then be able to obtain information on how the star ejected material before the explosion. The elongated and expanding object in the center of the ring is debris form the supernova blast which is being heated by radioactive elements, principally titanium 44, that were created in the explosion. This explosion was first observed by astronomers seventeen years ago in 1987, although the explosion took place about 160,000 years ago.

2003-01-01

345

Activation of silent gal genes in the lac-gal regulon of Streptococcus thermophilus.  

PubMed

Streptococcus thermophilus strain CNRZ 302 is unable to ferment galactose, neither that generated intracellularly by lactose hydrolysis nor the free sugar. Nevertheless, sequence analysis and complementation studies with Escherichia coli demonstrated that strain CNRZ 302 contained structurally intact genes for the Leloir pathway enzymes. These were organized into an operon in the order galKTE, which was preceded by a divergently transcribed regulator gene, galR, and followed by a galM gene and the lactose operon lacSZ. Results of Northern blot analysis showed that the structural gal genes were transcribed weakly, and only in medium containing lactose, by strain CNRZ 302. However, in a spontaneous galactose-fermenting mutant, designated NZ302G, the galKTE genes were well expressed in cells grown on lactose or galactose. In both CNRZ 302 and the Gal(+) mutant NZ302G, the transcription of the galR gene was induced by growth on lactose. Disruption of galR indicated that it functioned as a transcriptional activator of both the gal and lac operons while negatively regulating its own expression. Sequence analysis of the gal promoter regions of NZ302G and nine other independently isolated Gal(+) mutants of CNRZ 302 revealed mutations at three positions in the galK promoter region, which included substitutions at positions -9 and -15 as well as a single-base-pair insertion at position -37 with respect to the main transcription initiation point. Galactokinase activity measurements and analysis of gusA reporter gene fusions in strains containing the mutated promoters suggested that they were gal promoter-up mutations. We propose that poor expression of the gal genes in the galactose-negative S. thermophilus CNRZ 302 is caused by naturally occurring mutations in the galK promoter. PMID:11157930

Vaughan, E E; van den Bogaard, P T; Catzeddu, P; Kuipers, O P; de Vos, W M

2001-02-01

346

Webcams, TV Shows and Mobile phones: Empowering Exhibitionism  

Microsoft Academic Search

The roles of visual representations have been multiplied. In contrast of being targets of the ever-increasing surveillance, people seek to play an active role in the production of images, thus, reclaiming the copyright of their own lives. In this article, three examples of this development are examined. 'Reality shows' in TV aim to create an impression of the viewer participating

Hille Koskela

2004-01-01

347

69. (Credit JTL) View beneath marble meter bench showing hydraulic ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

69. (Credit JTL) View beneath marble meter bench showing hydraulic lines leading to water valve hydraulic control cylinders from control handles in bench; strings and pulleys activate meters. - McNeil Street Pumping Station, McNeil Street & Cross Bayou, Shreveport, Caddo Parish, LA

348

Interior of control house showing remains of controller. Moving the ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Interior of control house showing remains of controller. Moving the handle rotated the vertical shaft and porcelain cams to engage various electrical switches and activate the lift mechanism. All electrical components have been removed. - Potomac Edison Company, Chesapeake & Ohio Canal Bridge, Spanning C & O Canal South of U.S. 11, Williamsport, Washington County, MD

349

11. Historic view of Building 100 control room, showing personnel ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

11. Historic view of Building 100 control room, showing personnel operating rocket engine test controls and observer watching activity from observation room. May 27, 1957. On file at NASA Plumbrook Research Center, Sandusky, Ohio. NASA photo number C-45020. - Rocket Engine Testing Facility, GRC Building No. 100, NASA Glenn Research Center, Cleveland, Cuyahoga County, OH

350

Dissection of the wheat transcription factor HBP1a(17) reveals a modular structure for the activation domain  

Microsoft Academic Search

The wheat bZIP protein HBP-1a(17) is a putative transcription factor regulating histone gene expression. To delineate the\\u000a functional domain(s) of this factor, we made a series of effector constructs expressing fusion proteins, in which various\\u000a portions of HBP-1a(17) are fused to the DNA-binding domain of the yeast transcriptional activator GAL4, in plant cells. When\\u000a the ?-glucuronidase (GUS) reporter gene, driven

T. Nakayama; M. Okanami; T. Meshi; M. Iwabuchi

1997-01-01

351

Sudan-?-d-glucuronides and their use for the histochemical localization of ?-glucuronidase activity in transgenic plants  

Microsoft Academic Search

Synthesis of five different Sudan-?-d-glucuronides (I, II, III, IV, and RedB) was performed by condensation of a set of red Sudan diazo dyes with methyl (1-deoxy-2,3,4-tri-O-acetyl-1-trichloroacetimidoyl-?-d-glucopyran)uronate. After the acid and alcohol groups had been deprotected, the resulting compounds were used for histochemical\\u000a localization of ?-glucuronidase (GUS) activity in transgenic plants (Petunia hybrida, Arabidopsis thaliana, and Nicotiana tabacum) that contained the

E. Van der Eycken; N. Terryn; J. L. Goeman; G. Carlens; W. Nerinckx; M. Claeyssens; J. Van der Eycken; M. Van Montagu; M. Brito-Arias; G. Engler

2000-01-01

352

Activation of camalexin biosynthesis in Arabidopsis thaliana in response to perception of bacterial lipopolysaccharides: a gene-to-metabolite study.  

PubMed

Lipopolysaccharides (LPS), as lipoglycan microbe-associated molecular pattern molecules, trigger activation of signal transduction pathways involved in defence that generate an enhanced defensive capacity in plants. The transcriptional regulation of the genes for tryptophan synthase B, TSB1, and the cytochrome P450 monooxygenases CYP79B2 and CYP71B15, involved in the camalexin biosynthetic pathway, were investigated in response to LPS treatment. GUS-reporter assays for CYP71B15 and CYP79B2 gene promoter activation were performed on transgenic plants and showed positive histochemical staining in response to LPS treatment, indicating activation of the promoters. Quantitative PCR revealed that transcripts of TSB1, CYP79B2 and CYP71B15 exhibited differential, transient up-regulation. TSB1 transcript levels were up-regulated between 6 and 9 h after LPS-induction, while CYP71B15 and CYP79B2 both exhibited maxima at 12 h. To obtain information on the gene-to-metabolite network, the effect of the transcriptome changes on the metabolome was correlated to camalexin production. Increases in camalexin concentration were quantified by ultra pressure liquid chromatography-mass spectrometry and both absorbance spectra and elemental composition confirmed its identity. The concentrations increased from 0.03 to 3.7 ?g g(-1) fresh weight over a 24-h time period, thus indicating that the up-regulation of the biosynthetic pathway in response to LPS was accompanied by a time-dependent increase in camalexin concentration. Metabolomic analysis through principal component analysis-derived scores plots revealed clusters of sample replicates for 0, 6, 12, 18 and 24 h while loadings plots for LPS data identified camalexin as a biomarker that clearly demonstrated the variability between samples. PMID:22350766

Beets, Caryn Ann; Huang, Ju-Chi; Madala, Ntakadzeni Edwin; Dubery, Ian

2012-07-01

353

Use of the VvMybA1 gene for non-destructive quantification of promoter activity via color histogram analysis in grapevine (Vitis vinifera) and tobacco.  

PubMed

We report the development of a convenient plant-based reporter system to analyze promoters and facilitate selection of genetically engineered plants. The VvMybA1 gene of grapevine (Vitis vinifera L.) regulates the last metabolic step of anthocyanin biosynthesis and its ectopic expression leads to anthocyanin production in otherwise non-pigmented cells. To develop an anthocyanin-based quantitative reporter system, the VvMybA1 gene was isolated from V. vinifera 'Merlot' and placed under control of three promoters to test its ability to distinguish different activity levels. Promoters included a double enhanced CaMV35S (d35S) promoter, a double enhanced CsVMV (dCsVMV) promoter or a bi-directional dual promoter (BDDP), resulting in transformation vectors DAT, CAT and DEAT, respectively. These vectors were introduced into grapevine and tobacco via Agrobacterium-mediated transformation for transient and stable expression analysis. A linear relationship between the mean red brightness (MRB) and optical density (OD) values with a 0.99 regression coefficient was identified in a dilution series of anthocyanin, thus allowing the use of histogram data for non-destructive and real-time assessment of transcriptional activity. Results of histogram-based analysis of color images from transformed grapevine somatic embryos (SE) and various tissues of transgenic tobacco showed a consistent six to sevenfold promoter activity increase of DEAT over DAT. This expression increase was verified by spectroscopic measurement of anthocyanin concentrations in sepal tissue of transgenic tobacco plants. These results were congruent with previously findings of promoter activity derived from GUS fluorometric assay, thus demonstrating for the first time that the VvMybA1 gene could offer a simple, versatile and reliable plant-based alternative for quantitative promoter analysis in plants. PMID:21229312

Li, Zhijian T; Dhekney, Sadanand A; Gray, Dennis J

2011-10-01

354

Characterization of a baculovirus lacking the alkaline nuclease gene.  

PubMed

The Autographa californica multiple nucleocapsid nucleopolyhedrovirus (AcMNPV) alkaline nuclease (AN) associates with the baculovirus single-stranded DNA binding protein LEF-3 and possesses both a 5'-->3' exonuclease and an endonuclease activity. These activities are thought to be involved in DNA recombination and replication. To investigate the role of AN in AcMNPV replication, the lambda Red system was used to replace the an open reading frame with a chloramphenicol acetyltransferase gene (cat) and a bacmid containing the AcMNPV genome in Escherichia coli. The AcMNPV an knockout bacmid (vAcAN-KO/GUS) was unable to propagate in Sf9 cells, although an an-rescued bacmid (vAcAN-KO/GUS-Res) propagated normally. In addition, the mutant did not appear to produce budded virions. These data indicated that an is an essential baculovirus gene. Slot blot and DpnI assays of DNA replication in Sf9 cells transfected with vAcAN-KO/GUS, vAcAN-KO/GUS-Res, and a wild-type bacmid showed that the vAcAN-KO/GUS bacmid was able to replicate to levels similar to those seen with the vAcAN-KO/GUS-Res and wild-type bacmids at early stages posttransfection. However, at later time points DNA did not accumulate to the levels seen with the repaired or wild-type bacmids. Northern analysis of Sf9 cells transfected with bacmid vAcAN-KO/GUS showed that transcription of late and very late genes was lower at later times posttransfection relative to the results seen with wild-type and vAcAN-KO/GUS-Res bacmids. These data suggest that the an gene might be involved in the maturation of viral DNA or packaging of the DNA into virions. PMID:15367632

Okano, Kazuhiro; Vanarsdall, Adam L; Rohrmann, George F

2004-10-01

355

Do men and women show love differently in marriage?  

PubMed

In Western societies, women are considered more adept than men at expressing love in romantic relationships. Although scholars have argued that this view of love gives short shrift to men's ways of showing love (e.g., Cancian, 1986; Noller, 1996), the widely embraced premise that men and women "love differently" has rarely been examined empirically. Using data collected at four time points over 13 years of marriage, the authors examined whether love is associated with different behaviors for husbands and wives. Multilevel analyses revealed that, counter to theoretical expectations, both genders were equally likely to show love through affection. But whereas wives expressed love by enacting fewer negative or antagonistic behaviors, husbands showed love by initiating sex, sharing leisure activities, and doing household work together with their wives. Overall, the findings indicate that men and women show their love in more nuanced ways than cultural stereotypes suggest. PMID:22711739

Schoenfeld, Elizabeth A; Bredow, Carrie A; Huston, Ted L

2012-11-01

356

The Physics Circus -- Engaging students through a demo show  

NASA Astrophysics Data System (ADS)

The Physics Circus is a fun and exciting demo show presented by the UT Austin Physics Department at elementary, middle, and high schools in central Texas with the aim of engaging students. I will discuss the goals of this program, its facilitation, and the impact it has on the students who see the presentation as well as the university students who actively do the demonstrations. I will also demonstrate some highlights from the show, including the electric pickle, the flaming dollar, and the exploding trash can.

Hinko, Kathleen

2010-03-01

357

Hormonal regulation of tomato gibberellin 20-oxidase1 expressed in Arabidopsis.  

PubMed

Gibberellin 20-oxidases, enzymes of gibberellin (GA) biosynthesis, play an important role in (GA) homeostasis. To investigate the regulation of tomato SlGA20ox1 expression, a genomic clone was isolated, its promoter transcriptionally fused to the GUS reporter gene, and the construct used to transform Arabidopsis. Expression was found in diverse vegetative (leaves and roots) and reproductive (flowers) organs. GUS staining was also localized in the columella of secondary roots. GA negative feed-back regulation of SlGA20ox1:GUS was shown to be active both in tomato and in transformed Arabidopsis. Auxin (indol-3-acetic acid, 2,4-dichlorophenoxyacetic acid and naphtaleneacetic acid), triiodobenzoic acid (an inhibitor of auxin transport) and benzyladenine (a cytokinin) treatment induced SlGA20ox1:GUS expression associated with increased auxin content and/or signaling, detected using DR5:GUS expression as a marker. Interestingly, SlGA20ox:GUS expression was induced by auxin and root excision in the hypocotyl, an organ not showing GUS staining in control seedlings. In etiolated seedlings, SlGA20ox1:GUS expression occurred in the elongating hypocotyl region of etiolated seedlings and was down-regulated upon transfer to light associated with decrease of growth rate elongation. Our results show that feed-back, auxin and light regulation of SlGA20ox1 expression depends on DNA elements contained within the first 834bp of the 5' upstream promoter region. Putative DNA regulatory sequences involved in negative feed-back regulation and auxin response were identified in that promoter. PMID:20570010

Martí, Esmeralda; Carrera, Esther; Ruiz-Rivero, Omar; García-Martínez, José Luis

2010-09-15

358

Showing and Telling Farming: Agricultural Shows and Re-Imaging British Agriculture  

ERIC Educational Resources Information Center

Some actors in the ''mainstream'' agricultural sector are beginning to engage in strategies of influencing public perceptions of farming, responding to public anxieties over industrialised agriculture and to a supposed separation of non-farming publics from food production. This paper focuses on agricultural shows as sites and events central to…

Holloway, Lewis

2004-01-01

359

Salt tolerance and activity of antioxidative enzymes of transgenic finger millet overexpressing a vacuolar H(+)-pyrophosphatase gene (SbVPPase) from Sorghum bicolor.  

PubMed

A vacuolar proton pyrophosphatase cDNA clone was isolated from Sorghum bicolor (SbVPPase) using end-to-end gene-specific primer amplification. It showed 80-90% homology at the nucleotide and 85-95% homology at the amino acid level with other VPPases. The gene was introduced into expression vector pCAMBIA1301 under the control of the cauliflower mosaic virus 35S (CaMV35S) promoter and transformed into Agrobacterium tumifaciens strain LBA4404 to infect embryogenic calli of finger millet (Eleusine coracana). Successful transfer of SbVPPase was confirmed by a GUS histochemical assay and PCR analysis. Both, controls and transgenic plants were subjected to 100 and 200mM NaCl and certain biochemical and physiological parameters were studied. Relative water content (RWC), plant height, leaf expansion, finger length and width and grain weight were severely reduced (50-70%), and the flowering period was delayed by 20% in control plants compared to transgenic plants under salinity stress. With increasing salt stress, the proline and chlorophyll contents as well as the enzyme activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and glutathione reductase (GR) increased by 25-100% in transgenics, while malondialdehyde (MDA) showed a 2-4-fold decrease. The increased activities of antioxidant enzymes and the reduction in the MDA content suggest efficient scavenging of reactive oxygen species (ROS) in transgenics and, as a consequence, probably alleviation of salt stress. Also, the leaf tissues of the transgenics accumulated 1.5-2.5-fold higher Na(+) and 0.4-0.8-fold higher K(+) levels. Together, these results clearly demonstrate that overexpression of SbVPPase in transgenic finger millet enhances the plant's performance under salt stress. PMID:24877670

Anjaneyulu, Ediga; Reddy, Palle Surender; Sunita, Merla Srilakshmi; Kishor, Polavarapu B Kavi; Meriga, Balaji

2014-06-15

360

Cloning and expression of beta-glucuronidase from Lactobacillus brevis in E. coli and application in the bioconversion of baicalin and wogonoside.  

PubMed

The beta-glucuronidase (GUS) gene from Lactobacillus brevis RO1 was cloned and expressed in Escherichia coli GMS407. The GUS gene was composed of 1812 bp, encoding a 603-amino-acid protein belonging to the glycosyl hydrolase family 2 with three conserved domains. The amino acid similarity was higher than 70% with the beta-glucuronidases of various microorganisms, yet less than 58% with the beta-glucuronidase of L. gasseri ADH. Overexpression and purification of the GUS was performed in beta-glucuronidase-deficient E. coli GMS407. The purified GUS protein was 71 kDa and showed 1284 U/mg of specific activity at optimum condition of pH 5.0 and 37 degrees C. At 37 degrees C, the GUS remained stable for 80 min at pH values ranging from 5.0 to 8.0. The purified enzyme exhibited a half-life of 1 h at 60 degrees C and more than 2 h at 50 degrees C. When the purified GUS was applied to transform baicalin and wogonoside into their corresponding aglycones, 150 microM of baicalin and 125 microM of wogonoside were completely transformed into baicalein and wogonin, respectively, within 3 h. PMID:20075633

Kim, Hyun Sung; Kim, Jin Yong; Park, Myeong Soo; Zheng, Hua; Ji, Geun Eog

2009-12-01

361

DIME Students Show Off their Lego(TM) Challenge Creation  

NASA Technical Reports Server (NTRS)

Two students show the Lego (TM) Challenge device they designed and built to operate in the portable drop tower demonstrator as part of the second Dropping in a Microgravity Environment (DIME) competition held April 23-25, 2002, at NASA's Glenn Research Center. Competitors included two teams from Sycamore High School, Cincinnati, OH, and one each from Bay High School, Bay Village, OH, and COSI Academy, Columbus, OH. DIME is part of NASA's education and outreach activities. Details are on line at http://microgravity.grc.nasa.gov/DIME_2002.html.

2002-01-01

362

Children with Autism Show Reduced Somatosensory Response: An MEG Study  

PubMed Central

Lay Abstract Autism spectrum disorders are reported to affect nearly one out of every one hundred children, with over 90% of these children showing behavioral disturbances related to the processing of basic sensory information. Behavioral sensitivity to light touch, such as profound discomfort with clothing tags and physical contact, is a ubiquitous finding in children on the autism spectrum. In this study, we investigate the strength and timing of brain activity in response to simple, light taps to the fingertip. Our results suggest that children with autism show a diminished early response in the primary somatosensory cortex (S1). This finding is most evident in the left hemisphere. In exploratory analysis, we also show that tactile sensory behavior, as measured by the Sensory Profile, may be a better predictor of the intensity and timing of brain activity related to touch than a clinical autism diagnosis. We report that children with atypical tactile behavior have significantly lower amplitude somatosensory cortical responses in both hemispheres. Thus sensory behavioral phenotype appears to be a more powerful strategy for investigating neural activity in this cohort. This study provides evidence for atypical brain activity during sensory processing in autistic children and suggests that our sensory behavior based methodology may be an important approach to investigating brain activity in people with autism and neurodevelopmental disorders. Scientific Abstract The neural underpinnings of sensory processing differences in autism remain poorly understood. This prospective magnetoencephalography (MEG) study investigates whether children with autism show atypical cortical activity in the primary somatosensory cortex (S1) in comparison to matched controls. Tactile stimuli were clearly detectable, painless taps applied to the distal phalanx of the second (D2) and third (D3) fingers of the right and left hands. Three tactile paradigms were administered: an oddball paradigm (standard taps to D3 at an inter-stimulus interval (ISI) of 0.33 and deviant taps to D2 with ISI ranging from 1.32–1.64s); a slow-rate paradigm (D2) with an ISI matching the deviant taps in the oddball paradigm; and a fast-rate paradigm (D2) with an ISI matching the standard taps in the oddball. Study subjects were boys (age 7–11 years) with and without autism disorder. Sensory behavior was quantified using the Sensory Profile questionnaire. Boys with autism exhibited smaller amplitude left hemisphere S1 response to slow and deviant stimuli during the right hand paradigms. In post-hoc analysis, tactile behavior directly correlated with the amplitude of cortical response. Consequently, the children were re-categorized by degree of parent-report tactile sensitivity. This regrouping created a more robust distinction between the groups with amplitude diminution in the left and right hemispheres and latency prolongation in the right hemisphere in the deviant and slow-rate paradigms for the affected children. This study suggests that children with autism have early differences in somatosensory processing, which likely influence later stages of cortical activity from integration to motor response.

Marco, Elysa J.; Khatibi, Kasra; Hill, Susanna S.; Siegel, Bryna; Arroyo, Monica S.; Dowling, Anne F.; Neuhaus, John M.; Sherr, Elliott H.; Hinkley, Leighton N. B.; Nagarajan, Srikantan S.

2012-01-01

363

Activities.  

ERIC Educational Resources Information Center

An activity is described as an example of an application that draws on the behavior of current in electrical circuits to create a symbolic algebra. Five worksheets are included, with discussion of their use. (MNS)

Hirsch, Christian R., Ed.; And Others

1987-01-01

364

VASA localization requires the SPRY-domain and SOCS-box containing protein, GUSTAVUS.  

PubMed

VASA (VAS), a key protein in establishing the specialized translational activity of the Drosophila pole plasm, accumulates at the posterior pole of the developing oocyte. We identified a gene, gustavus (gus), that encodes a protein that interacts with VAS. A gus mutation blocks posterior localization of VAS, as does deletion of a segment of VAS containing the GUS binding site. Like VAS, GUS is present in cytoplasmic ribonucleoprotein particles. Heterozygotes for gus or a deletion including gus produce embryos with fewer pole cells and posterior patterning defects. Therefore, GUS is essential for the posterior localization of VAS. However, gus is not required for the posterior localization of oskar (osk). Apparent gus orthologs are present in mammalian genomes. PMID:12479811

Styhler, Sylvia; Nakamura, Akira; Lasko, Paul

2002-12-01

365

Functional characterization of the Ginkgo biloba chalcone synthase gene promoter in transgenic tobacco.  

PubMed

The regulative sequence (2273 bp) of the chalcone synthase gene promoter of biloba was cloned by genomic walking. A 2273-bp promoter 5' upstream translation start site of GbCHS was cloned and designated as GbCHSP. pBI121+CHSP:GUS and pBI121-35S:GUS were constructed and transformed into tobacco by LBA4404. We found that GbCHSP could drive transient expression of GUS in tobacco and differentially expressed in root, stem and leaf tissues of this plant. GUS activity regulated by the CHSP promoter were located in tissues (apical meristems) at the growing points of roots and stems. pBI121+CHSP:GUS could be induced by wounding, copper, UV-B, abscisic acid, and ethephon treatments of transgenic seedlings. This activity was weakly inhibited by gibberellin. Deletion analysis of the CHSP promoter in transgenic tobacco showed that CHSP1 complete promoter conferred a GUS expression and activity similar to that of 35 S(CaMV). GUS activity dropped dramatically when there were CHSP4, CHSP5 constructs and was almost totally absent when the CHSP6 construct was present. We conclude that the upstream sequence -1548 to -306 of GbCHSP is the main region for transcriptional regulation of the CHS gene and that it is activated by hormone and stress factors in G. biloba. These results will help us to understand the transcriptional regulatory mechanisms involved in GbCHS expression and flavonoid accumulation in G. biloba. PMID:24841790

Li, L L; Cheng, H; Yuan, H H; Xu, F; Cheng, S Y; Cao, F L

2014-01-01

366

Activities.  

ERIC Educational Resources Information Center

The material presented is designed to help students explore geometric patterns involving Fibonnaci numbers and the golden ratio, and to aid in review of basic geometry skills. Worksheet masters intended for duplication are provided. Suggestions are made of possible classroom extensions to the initial activities. (MP)

Mathematics Teacher, 1982

1982-01-01

367

Eccentric muscle challenge shows osteopontin polymorphism modulation of muscle damage.  

PubMed

A promoter polymorphism of the osteopontin (OPN) gene (rs28357094) has been associated with multiple inflammatory states, severity of Duchenne muscular dystrophy (DMD) and muscle size in healthy young adults. We sought to define the mechanism of action of the polymorphism, using allele-specific in vitro reporter assays in muscle cells, and a genotype-stratified intervention in healthy controls. In vitro reporter constructs showed the G allele to respond to estrogen treatment, whereas the T allele showed no transcriptional response. Young adult volunteers (n = 187) were enrolled into a baseline study, and subjects with specific rs28357094 genotypes enrolled into an eccentric muscle challenge intervention [n = 3 TT; n = 3 GG/GT (dominant inheritance model)]. Female volunteers carrying the G allele showed significantly greater inflammation and increased muscle volume change as determined by magnetic resonance imaging T1- and T2-weighted images after eccentric challenge, as well as greater decrement in biceps muscle force. Our data suggest a model where the G allele enables enhanced activities of upstream enhancer elements due to loss of Sp1 binding at the polymorphic site. This results in significantly greater expression of the pro-inflammatory OPN cytokine during tissue remodeling in response to challenge in G allele carriers, promoting muscle hypertrophy in normal females, but increased damage in DMD patients. PMID:24626632

Barfield, Whitney L; Uaesoontrachoon, Kitipong; Wu, Chung-Sheih; Lin, Stephen; Chen, Yue; Wang, Paul C; Kanaan, Yasmine; Bond, Vernon; Hoffman, Eric P

2014-08-01

368

Eccentric muscle challenge shows osteopontin polymorphism modulation of muscle damage  

PubMed Central

A promoter polymorphism of the osteopontin (OPN) gene (rs28357094) has been associated with multiple inflammatory states, severity of Duchenne muscular dystrophy (DMD) and muscle size in healthy young adults. We sought to define the mechanism of action of the polymorphism, using allele-specific in vitro reporter assays in muscle cells, and a genotype-stratified intervention in healthy controls. In vitro reporter constructs showed the G allele to respond to estrogen treatment, whereas the T allele showed no transcriptional response. Young adult volunteers (n = 187) were enrolled into a baseline study, and subjects with specific rs28357094 genotypes enrolled into an eccentric muscle challenge intervention [n = 3 TT; n = 3 GG/GT (dominant inheritance model)]. Female volunteers carrying the G allele showed significantly greater inflammation and increased muscle volume change as determined by magnetic resonance imaging T1- and T2-weighted images after eccentric challenge, as well as greater decrement in biceps muscle force. Our data suggest a model where the G allele enables enhanced activities of upstream enhancer elements due to loss of Sp1 binding at the polymorphic site. This results in significantly greater expression of the pro-inflammatory OPN cytokine during tissue remodeling in response to challenge in G allele carriers, promoting muscle hypertrophy in normal females, but increased damage in DMD patients.

Barfield, Whitney L.; Uaesoontrachoon, Kitipong; Wu, Chung-Sheih; Lin, Stephen; Chen, Yue; Wang, Paul C.; Kanaan, Yasmine; Bond, Vernon; Hoffman, Eric P.

2014-01-01

369

Cotton leaf curl Multan betasatellite as a plant gene delivery vector trans-activated by taxonomically diverse geminiviruses.  

PubMed

Cotton leaf curl Multan betasatellite (CLCuMB) replicates in tobacco, tomato and datura plants in the presence of the helper viruses tomato leaf curl virus-Australia, Iranian isolates of tomato yellow leaf curl virus, tomato leaf curl Karnataka virus, and beet severe curly top virus (BSCTV). Infectious recombinant CLCuMB constructs were made in which segments of either the CaMV 35S or the petunia ChsA promoter replaced the CLCuMB ?C1 ORF, and these were designated pBin??C1-35S and pBin??C1-ChsA, respectively. Inoculation of tobacco plants containing a functional 35S-GUS transgene with pBin??C1-35S, and normal petunia plants with pBin??C1-ChsA, in the presence of helper viruses resulted in silencing of GUS and ChsA activities in transgenic tobacco and non-transgenic petunia plants, respectively. Replication of CLCuMB with different geminiviruses, especially BSCTV, a curtovirus with a broad host range, makes it a valuable gene delivery vector to the large number of host plant species of geminiviruses that support CLCuMB. PMID:22476203

Kharazmi, S; Behjatnia, S A A; Hamzehzarghani, H; Niazi, A

2012-07-01

370

Mixtures of thermostable enzymes show high performance in biomass saccharification.  

PubMed

Optimal enzyme mixtures of six Trichoderma reesei enzymes and five thermostable enzyme components were developed for the hydrolysis of hydrothermally pretreated wheat straw, alkaline oxidised sugar cane bagasse and steam-exploded bagasse by statistically designed experiments. Preliminary studies to narrow down the optimization parameters showed that a cellobiohydrolase/endoglucanase (CBH/EG) ratio of 4:1 or higher of thermostable enzymes gave the maximal CBH-EG synergy in the hydrolysis of hydrothermally pretreated wheat straw. The composition of optimal enzyme mixtures depended clearly on the substrate and on the enzyme system studied. The optimal enzyme mixture of thermostable enzymes was dominated by Cel7A and required a relatively high amount of xylanase, whereas with T. reesei enzymes, the high proportion of Cel7B appeared to provide the required xylanase activity. The main effect of the pretreatment method was that the required proportion of xylanase was higher and the proportion of Cel7A lower in the optimized mixture for hydrolysis of alkaline oxidised bagasse than steam-exploded bagasse. In prolonged hydrolyses, less Cel7A was generally required in the optimal mixture. Five-component mixtures of thermostable enzymes showed comparable hydrolysis yields to those of commercial enzyme mixtures. PMID:24752938

Kallioinen, Anne; Puranen, Terhi; Siika-Aho, Matti

2014-07-01

371

[How children show positive and negative relationships on their drawings].  

PubMed

This study analyses, whether pictures of children showing a positive relationship are significantly different from those showing a negative one with respect to several criteria. The study involved a random selection of 45 children aged 4;6 to 11;6 years. The children painted a picture with themselves and a person they liked and a picture of themselves with someone they disliked. For the most part, the children drew pictures of themselves with peers both with respect to positive as well as negative images. In an interview afterwards, the children specified the criteria in their drawings by which the quality of the particular relationship can be identified. Positive and negative relationship paintings differ in the character of activity described. The sun as an element in children's paintings is painted not more frequent on positive compared to negative pictures. The colour black is used more often in the drawings signifying negative relationships. While girls used more colour in negative relationship drawings, boys used more colour in the positive ones. There was no significant difference in the use of favourite colours and decorative elements between the two groups. Only in negative relationship drawings people were looking away from each other. Smiling individuals were more common in the positive relationship pictures and in pictures painted by the 6 to 8 year olds. A greater distance between the individuals emerged on negative relationship drawings of the girls. PMID:15730148

Gramel, Sabine

2005-01-01

372

Ligand-exchangeability of 2-coordinate phosphinegold(I) complexes with AuSP and AuNP cores showing selective antimicrobial activities against Gram-positive bacteria. Crystal structures of [Au(2-Hmpa)(PPh 3)] and [Au(6-Hmna)(PPh 3)] (2-H 2mpa=2-mercaptopropionic acid, 6-H 2mna=6-mercaptonicotinic acid)  

Microsoft Academic Search

Selective and effective antimicrobial activities against Gram-positive bacteria (B. subtilis and\\/or S. aureus) were found in 2-coordinate gold(I)–PPh3 complexes with AuSP and AuNP cores, i.e. [Au(L)(PPh3)] (HL=2-H2mna [H2mna=mercaptonicotinic acid] 3, d-H2pen [H2pen=penicillamine] 4, d,l-H2pen 5, 4-H2mba [H2mba=mercaptobenzoic acid] 8, Hpz [Hpz=pyrazole] 9, Him [Him=imidazole] 10, 1,2,3-Htriz [Htriz=triazole] 11, 1,2,4-Htriz 12, Htetz [Htetz=tetrazole] 13), whereas no activity was observed in 2-coordinate

Kenji Nomiya; Satoshi Yamamoto; Ryusuke Noguchi; Hironari Yokoyama; Noriko Chikaraishi Kasuga; Kei Ohyama; Chieko Kato

2003-01-01

373

Assays of dioxins and dioxin-like compounds in actually contaminated soils using transgenic tobacco plants carrying a recombinant mouse aryl hydrocarbon receptor-mediated ?-glucuronidase reporter gene expression system.  

PubMed

The transgenic tobacco plant XD4V-26 carrying the recombinant mouse aryl hydrocarbon receptor XD4V-mediated ?-glucuronidase (GUS) reporter gene expression system was used for assay of dioxins and dioxin-like compounds consisting of polychlorinated dibenzeno-p-dioxins, polychlorinated dibenzofurans, and coplanar polychlorinated biphenyls (Co-PCBs) in actually contaminated soils. The transgenic tobacco plant XD4V-26 showed a significant dose-dependent induced GUS activity when cultured on MS medium containing PCB126 [toxic equivalency factor (TEF) = 0.1]. In contrast, PCB169 and PCB180, which have 0.03 of TEF and unassigned TEF values, respectively, did not significantly induce GUS activity under the same conditions as with PCB126. When the tobacco plants were cultivated for up to 5 weeks on actually contaminated soils with dioxins and dioxin-like compounds collected from the periphery of an incinerator used for disposal of residential and industrial wastes, GUS activity in the leaves was dose-dependently increased. The plants clearly detected 360 pg-TEQ g(-1) of dioxins and dioxin-like compounds in this assay. There was a positive correlation between GUS activity and TEQ value of dioxins and dioxin-like compounds in the plants. This assay does not require any extraction and purification processes for the actually contaminated soil samples. PMID:22022789

Inui, Hideyuki; Gion, Keiko; Utani, Yasushi; Wakai, Taketo; Kodama, Susumu; Eun, Heesoo; Kim, Yun-Seok; Ohkawa, Hideo

2012-01-01

374

Assays of dioxins and dioxin-like compounds in actually contaminated soils using transgenic tobacco plants carrying a recombinant mouse aryl hydrocarbon receptor-mediated ?-glucuronidase reporter gene expression system.  

PubMed

The transgenic tobacco plant XD4V-26 carrying the recombinant mouse aryl hydrocarbon receptor XD4V-mediated ?-glucuronidase (GUS) reporter gene expression system was used for assay of dioxins and dioxin-like compounds consisting of polychlorodibenzo-p-dioxins, polychlorinated dibenzofurans, and coplanar polychlorinated biphenyls (Co-PCBs) in actually contaminated soils. The transgenic tobacco plant XD4V-26 showed a significant dose-dependent induced GUS activity when cultured on MS medium containing PCB126 [toxic equivalency factor (TEF) = 0.1]. In contrast, PCB169 and PCB180, which have 0.03 of TEF and unassigned TEF values, respectively, did not significantly induce GUS activity under the same conditions as with PCB126. When the tobacco plants were cultivated for up to 5 weeks on actually contaminated soils with dioxins and dioxin-like compounds collected from the periphery of an incinerator used for disposal of life and industrial wastes, GUS activity in the leaves was dose-dependently increased. The plants clearly detected 360 pg-TEQ g(-1) of dioxins and dioxin-like compounds in this assay. There was a positive correlation between GUS activity and TEQ value of dioxins and dioxin-like compounds in the plants. This assay does not require any extraction and purification processes for the actually contaminated soil samples. PMID:22428884

Inui, Hideyuki; Gion, Keiko; Utani, Yasushi; Wakai, Taketo; Kodama, Susumu; Eun, Heesoo; Kim, Yun-Seok; Ohkawa, Hideo

2012-01-01

375

Spatial and temporal patterns of transcription of a wound-induced gene in potato.  

PubMed Central

Transcriptional fusions between the gene encoding win2 from potato and the reporter gene encoding beta-glucuronidase (GUS) have been used to study the spatial and temporal patterns of wound induced gene activity in transgenic potato and tobacco plants. Gene fusions containing a full length win2 promoter were found to be correctly regulated in response to mechanical wounding in transgenic potato, but not in the heterologous host, tobacco. Sequences greater than 560 bp upstream of the transcription start site of win2 were shown to be important for wound inducibility. The dramatic induction of GUS activity detected using fluorometric assays of extracts of wounded and aged leaves of several independent win2--GUS transformants was consistent with the kinetics of win2 mRNA accumulation. Histochemical analysis of wounded leaves showed that transcription first occurred in cells immediately adjacent to the wound, and was then progressively induced in cells associated with the vascular system at a distance from the wound site. In tubers, a localized response to wounding was observed, and this only spread to other parts of the tuber if it had started to sprout. It was concluded that active vascular transport was necessary for the spread of wound response. Win2--GUS fusions were also expressed as part of normal plant development, as GUS activity was detected in the developing buds and in a layer of cells associated with the lenticels of unwounded tubers. Images Fig. 2. Fig. 3. Fig. 4. Fig. 5.

Stanford, A C; Northcote, D H; Bevan, M W

1990-01-01

376

Assays of dioxins and dioxin-like compounds in actually contaminated soils using transgenic tobacco plants carrying a recombinant mouse aryl hydrocarbon receptor-mediated ?-glucuronidase reporter gene expression system  

PubMed Central

The transgenic tobacco plant XD4V-26 carrying the recombinant mouse aryl hydrocarbon receptor XD4V-mediated ?-glucuronidase (GUS) reporter gene expression system was used for assay of dioxins and dioxin-like compounds consisting of polychlorodibenzo-p-dioxins, polychlorinated dibenzofurans, and coplanar polychlorinated biphenyls (Co-PCBs) in actually contaminated soils. The transgenic tobacco plant XD4V-26 showed a significant dose-dependent induced GUS activity when cultured on MS medium containing PCB126 [toxic equivalency factor (TEF) = 0.1]. In contrast, PCB169 and PCB180, which have 0.03 of TEF and unassigned TEF values, respectively, did not significantly induce GUS activity under the same conditions as with PCB126. When the tobacco plants were cultivated for up to 5 weeks on actually contaminated soils with dioxins and dioxin-like compounds collected from the periphery of an incinerator used for disposal of life and industrial wastes, GUS activity in the leaves was dose-dependently increased. The plants clearly detected 360 pg-TEQ g?1 of dioxins and dioxin-like compounds in this assay. There was a positive correlation between GUS activity and TEQ value of dioxins and dioxin-like compounds in the plants. This assay does not require any extraction and purification processes for the actually contaminated soil samples.

Inui, Hideyuki; Gion, Keiko; Utani, Yasushi; Wakai, Taketo; Kodama, Susumu; Eun, Heesoo; Kim, Yun-Seok; Ohkawa, Hideo

2012-01-01

377

Sediment score shows mountain off beat with climate change  

NASA Astrophysics Data System (ADS)

Neodymium (Nd) isotopes recorded in marine sediments are usually used in paleoceanography as tracers of changes of past climate. The Indian Ocean raises the interest because of its connections with the Atlantic and the Pacific Oceans, and because of the proximity of the Himalayans drained by the Ganga-Brahmaputra-Meghna (GBM) rivers system. The origin of the variations of seawater Nd isotopic data observed over the last Glacial/Interglacial cycles remains unclear. They are either interpreted as changes in the global oceanic circulation or as changes in the continental input of Nd related to changes in the continental rainfalls. Here we present a simple technique to discriminate these two interpretations at a given site, based on the correlation of ?18O and ?Nd seawater signals. We show in-phase records at site SK129-CR2 (Arabian Sea) and out-of phase records at site ODP-758 (Bay of Bengal), suggesting that the two sites have recorded different phenomena through time. Arabian Site fluctuations were interpreted as changes of the thermohaline circulation and Bay of Bengal Site fluctuations as changes on the Himalayan input. As Himalayan rivers input is linked to the Earth's climate variability, we filtered the time series of ?18O and ?Nd at Site ODP 758 around the three periods related to the three main orbital parameters. We show that the time lag between ?18O and ?Nd increases from 1000 to 2000 and then to 7000 years for the 23 ky, 41 ky, and 100 ky filtered signals. To explain the delays between temperature changes recorded by ?18O and ?Nd, two models were proposed: diffusive and erosion models of Himalayans. For the first time, we demonstrate that a geochemical dataset can record and thus constrain the time lag in the erosional response of an active mountain belt to climate change.

Gourlan, Alexandra T.; Voisin, Christophe; Chauvel, Catherine; Braun, Jean

2013-04-01

378

Coagulation tests show significant differences in patients with breast cancer.  

PubMed

Activated coagulation and fibrinolytic system in cancer patients is associated with tumor stroma formation and metastasis in different cancer types. The aim of this study is to explore the correlation of blood coagulation assays for various clinicopathologic factors in breast cancer patients. A total of 123 female breast cancer patients were enrolled into the study. All the patients were treatment naïve. Pretreatment blood coagulation tests including PT, APTT, PTA, INR, D-dimer, fibrinogen levels, and platelet counts were evaluated. Median age of diagnosis was 51 years old (range 26-82). Twenty-two percent of the group consisted of metastatic breast cancer patients. The plasma level of all coagulation tests revealed statistically significant difference between patient and control group except for PT (p?50 years) was associated with higher D-dimer levels (p?=?0.003). Metastatic patients exhibited significantly higher D-dimer values when compared with early breast cancer patients (p?=?0.049). Advanced tumor stage (T3 and T4) was associated with higher INR (p?=?0.05) and lower PTA (p?=?0.025). In conclusion, coagulation tests show significant differences in patients with breast cancer. PMID:24596033

Tas, Faruk; Kilic, Leyla; Duranyildiz, Derya

2014-06-01

379

Chemokine analogues show suitable stability for development as microbicides.  

PubMed

New prevention strategies are urgently needed to slow the spread of the HIV/AIDS pandemic, and in the absence of an effective vaccine, there is hope that "microbicides"-HIV inhibitors applied to mucosal surfaces before sexual intercourse-may be able to make an impact. Because developing countries are at the center of the epidemic, affordability and stability during storage are key criteria for candidate microbicides. Furthermore, because formulation strategies that provide long-duration protection after a single dose may enhance acceptability and compliance, stability in the vaginal environment and in the presence of semen should also be considered. PSC-RANTES, a human chemokine analog, has shown promise as a candidate microbicide, but because it contains nonnatural structures that necessitate chemical synthesis steps, it is not suitable for production at a feasible cost and scale for general distribution in developing countries. We have recently developed 2 new fully recombinant chemokine analogs, 5P12-RANTES and 6P4-RANTES, which show equivalent anti-HIV activity to PSC-RANTES. In this study, we tested the stability of these molecules under conditions related to use as microbicides. Our results suggest that stability issues will not present a major obstacle to the further development of these promising molecules as microbicides. PMID:18989226

Cerini, Fabrice; Landay, Alan; Gichinga, Carolyne; Lederman, Michael M; Flyckt, Rebecca; Starks, David; Offord, Robin E; Le Gal, François; Hartley, Oliver

2008-12-15

380

Resetting of FLOWERING LOCUS C expression after epigenetic repression by vernalization  

PubMed Central

The epigenetic repression of FLOWERING LOCUS C (FLC) in winter-annual ecotypes of Arabidopsis by prolonged cold ensures that plants flower in spring and not during winter. Resetting of the FLC expression level in progeny is an important step in the life cycle of the plant. We show that both the paternally derived and the maternally derived FLC:GUS genes are reset to activity but that the timing of their first expression differs. The paternal FLC:GUS gene in vernalized plants is expressed in the male reproductive organs, the anthers, in both somatic tissue and in the sporogenous pollen mother cells, but there is no expression in mature pollen. In the progeny generation, the paternally derived FLC:GUS gene is expressed in the single-celled zygote (fertilized egg cell) and through embryo development, but not in the fertilized central cell, which generates the endosperm of the progeny seed. FLC:GUS is not expressed during female gametogenesis, with the maternally derived FLC:GUS being first expressed in the early multicellular embryo. We show that FLC activity during late embryo development is a prerequisite for the repressive action of FLC on flowering.

Sheldon, Candice C.; Hills, Melissa J.; Lister, Clare; Dean, Caroline; Dennis, Elizabeth S.; Peacock, W. James

2008-01-01

381

Tennis players show a lower coactivation of the elbow antagonist muscles during isokinetic exercises  

Microsoft Academic Search

PurposePrevious studies have suggested that muscle coactivation could be reduced by a recurrent activity (training, daily activities). If this was correct, skilled athletes should show a specific muscle activation pattern with a low level of coactivation of muscles which are typically involved in their discipline. In particular, the aim of this study was to verify the hypothesis that the amount

Ilenia Bazzucchi; Maria Elena Riccio; Francesco Felici

2008-01-01

382

Map Showing Lava Inundation Zones for Mauna Loa, Hawaii  

USGS Publications Warehouse

Introduction The Island of Hawaii is composed of five coalesced basaltic volcanoes. Lava flows constitute the greatest volcanic hazard from these volcanoes. This report is concerned with lava flow hazards on Mauna Loa, the largest of the island shield volcanoes. Hilo lies 58 km from the summit of Mauna Loa, the Kona coast 33 km, and the southernmost point of the island 61 km. Hawaiian volcanoes erupt two morphologically distinct types of lava, aa and pahoehoe. The surfaces of pahoehoe flows are rather smooth and undulating. Pahoehoe flows are commonly fed by lava tubes, which are well insulated, lava-filled conduits contained within the flows. The surfaces of aa flows are extremely rough and composed of lava fragments. Aa flows usually form lava channels rather than lava tubes. In Hawaii, lava flows are known to reach distances of 50 km or more. The flows usually advance slowly enough that people can escape from their paths. Anything overwhelmed by a flow will be damaged or destroyed by burial, crushing, or ignition. Mauna Loa makes up 51 percent of the surface area of the Island of Hawaii. Geologic mapping shows that lava flows have covered more than 40 percent of the surface every 1,000 years. Since written descriptions of its activity began in A.D. 1832, Mauna Loa has erupted 33 times. Some eruptions begin with only brief seismic unrest, whereas others start several months to a year following increased seismic activity. Once underway, the eruptions can produce lava flows that reach the sea in less than 24 hours, severing roads and utilities. For example, the 1950 flows from the southwest rift zone reached the ocean in approximately three hours. The two longest flows of Mauna Loa are pahoehoe flows from the 50-kilometer-long 1859 and the 48-kilometer-long 1880-81 eruptions. Mauna Loa will undoubtedly erupt again. When it does, the first critical question that must be answered is: Which areas are threatened with inundation? Once the threatened areas are established, we can address the second critical question: What people, property, and facilities are at risk? These questions can be answered by estimating the areas most likely to be affected by eruptions originating on various parts of the volcano. This report contains such estimates, based on the known source vents and areas affected by past eruptions. We have divided the volcano into potential lava inundation zones and prepared maps of these zones, which are presented here on the accompanying map sheets.

Trusdell, F. A.; Graves, P.; Tincher, C. R.

2002-01-01

383

Activation and repression of transcription by auxin-response factors.  

PubMed

Auxin-response factors (ARFs) bind with specificity to TGTCTC auxin-response elements (AuxREs), which are found in promoters of primary/early auxin-response genes. Nine different ARFs have been analyzed for their capacity to activate or repress transcription in transient expression assays employing auxin-responsive GUS reporter genes. One ARF appears to act as a repressor. Four ARFs function as activators and contain glutamine-rich activation domains. To achieve transcriptional activation on TGTCTC AuxREs in transient expression assays, ARFs require a conserved dimerization domain found in both ARF and Aux/IAA proteins, but they do not absolutely require their DNA-binding domains. Our results suggest that ARFs can activate or repress transcription by binding to AuxREs directly and that selected ARFs, when overexpressed, may potentiate activation further by associating with an endogenous transcription factor(s) (e.g., an ARF) that is bound to AuxREs. Transfection experiments suggest that TGTCTC AuxREs are occupied regardless of the auxin status in cells and that these occupied AuxREs are activated when exogenous auxin is applied to cells or when ARF activators are overexpressed. The results provide new insight into mechanisms involved with auxin regulation of primary/early-response genes. PMID:10318972

Ulmasov, T; Hagen, G; Guilfoyle, T J

1999-05-11

384

Reducing appointment no-shows: going from theory to practice.  

PubMed

Addiction appointment no-shows adversely impact clinical outcomes and healthcare productivity. During 2007-2010, 67 treatment organizations in the Strengthening Treatment Access and Retention program were asked to reduce their no-show rates by using practices taken from no-show research and theory. These treatment organizations reduced outpatient no-show rates from 37.4% to 19.9% (p = .000), demonstrated which practices they preferred to implement, and which practices were most effective in reducing no-show rates. This study provides an applied synthesis of addiction treatment no-show research and suggests future directions for no-show research and practice. PMID:23607670

Molfenter, Todd

2013-06-01

385

Robot innovations highlight numerous recent shows in Chicago  

Microsoft Academic Search

Purpose – The purpose of this paper is to review the robots at the 2007 ATExpo Show and related Electronics Assembly Show, Quality, PlasTec and National Manufacturing Week Shows, as well as at the FABTECH Show and the Rockwell Automation Show held in Chicago. Design\\/methodology\\/approach – The paper incorporates in-depth interviews with robot exhibitors, as well as suppliers of accessories

Richard Bloss

2008-01-01

386

Strong seed-specific protein expression from the Vigna radiata storage protein 8SG? promoter in transgenic Arabidopsis seeds.  

PubMed

Vigna radiata (mung bean) is an important crop plant and is a major protein source in developing countries. Mung bean 8S globulins constitute nearly 90% of total seed storage protein and consist of three subunits designated as 8SG?, 8SG?' and 8SG?. The 5'-flanking sequences of 8SG?' has been reported to confer high expression in transgenic Arabidopsis seeds. In this study, a 472-bp 5'-flanking sequence of 8SG? was identified by genome walking. Computational analysis subsequently revealed the presence of numerous putative seed-specific cis-elements within. The 8SG? promoter was then fused to the gene encoding ?-glucuronidase (GUS) to create a reporter construct for Arabidopsis thaliana transformation. The spatial and temporal expression of 8SG??GUS, as investigated using GUS histochemical assays, showed GUS expression exclusively in transgenic Arabidopsis seeds. Quantitative GUS assays revealed that the 8SG? promoter showed 2- to 4-fold higher activity than the Cauliflower Mosaic Virus (CaMV) 35S promoter. This study has identified a seed-specific promoter of high promoter strength, which is potentially useful for directing foreign protein expression in seed bioreactors. PMID:24503210

Chen, Mo-Xian; Zheng, Shu-Xiao; Yang, Yue-Ning; Xu, Chao; Liu, Jie-Sheng; Yang, Wei-Dong; Chye, Mee-Len; Li, Hong-Ye

2014-03-20

387

Auxin regulates the promoter of the root-inducing rolB gene of Agrobacterium rhizogenes in transgenic tobacco  

Microsoft Academic Search

The regulation in tobacco of the rolB and rolC promoters of Agrobacterium rhizogenes pRi 1855 TL-DNA was studied by using the ß-glucuronidase (GUS) reporter system in transgenic plants. A 20- to 100-fold increase of GUS activity was selectively induced by auxin in rolB-GUS transformed mesophyll protoplasts, whereas this auxin-dependent increase was only 5-fold in rolC-GUS protoplasts. Moreover, both gene fusions

Christophe Maurel; Jean Brevet; Hélène Barbier-Brygoo; Jean Guern; Jacques Tempé

1990-01-01

388

78. VIEW OF UNCOMPLETED RESERVOIR, SHOWING FOREBAY AND FLUME; ALSO ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

78. VIEW OF UNCOMPLETED RESERVOIR, SHOWING FOREBAY AND FLUME; ALSO SHOWING POOL ARRANGEMENT FOR TEMPORARILY UTILIZING WATER WITHOUT FILLING THE RESERVOIR, Print No. 232, April 1904 - Electron Hydroelectric Project, Along Puyallup River, Electron, Pierce County, WA

389

North Pacific and North America: Showing the Changing Seasons  

NSDL National Science Digital Library

SeaWiFS false color data showing seasonal change in the oceans and on land for North America and the North Pacific. The data is seasonally averaged, and shows fall, winter, spring, summer, fall, winter, spring, and summer.

Allen, Jesse; Newcombe, Marte; Feldman, Gene

1998-09-09

390

16. SECOND FLOOR, EAST ROOM, SHOWING DOORWAYS TO BOXED WINDER ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

16. SECOND FLOOR, EAST ROOM, SHOWING DOORWAYS TO BOXED WINDER STAIRWAYS-- UP AND DOWN-- AND WALL CABINET WITH DOORS OPEN TO SHOW STAIRWAY - High Farm, House, Creek Road, 1 mile West of Easton Road, Pipersville, Bucks County, PA

391

Journey to the Stars Educator's Guide: Teaching with the Show  

NSDL National Science Digital Library

This is an educators guide to accompany the Journey to the Stars planetarium show. The materials include a section, titled Teaching With the Show, containing guiding questions to encourage class discussions about the life cycle of stars.

392

22. DETAIL EXTERIOR VIEW LOOKING NORTHWEST, SHOWING FISH LADDER AT ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

22. DETAIL EXTERIOR VIEW LOOKING NORTHWEST, SHOWING FISH LADDER AT NORTH END OF DAM/SPILLWAY; VIEW SHOWS SECTION OF FISH LADDER NEAR WHERE IT ENTERS THE COLUMBIA RIVER. - Bonneville Project, Bonneville Dam, Columbia River, Bonneville, Multnomah County, OR

393

1. AERIAL VIEW, LOOKING NORTH, SHOWING COVERED BARGE (VESSEL 37) ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

1. AERIAL VIEW, LOOKING NORTH, SHOWING COVERED BARGE (VESSEL 37) IN CENTER OF PICTURE WITH FOUR HATCHES SHOWING IN SUPERSTRUCTURE Charles Wisniewski, photographer, January 1985 - Shooters Island, Ships Graveyard, Vessel No. 37, Newark Bay, Staten Island (subdivision), Richmond County, NY

394

Payload specialist Reinhard Furrer show evidence of previous blood sampling  

NASA Technical Reports Server (NTRS)

Payload specialist Reinhard Furrer shows evidence of previous blood sampling while Wubbo J. Ockels, Dutch payload specialist (only partially visible), extends his right arm after a sample has been taken. Both men show bruises on their arms.

1985-01-01

395

42 CFR 456.655 - Validation of showings.  

Code of Federal Regulations, 2013 CFR

...2013-10-01 2013-10-01 false Validation of showings. 456.655 Section...Utilization Control Program § 456.655 Validation of showings. (a) The Administrator...showings submitted under § 456.654. Validation procedures will include on-site...

2013-10-01

396

77. GENERAL VIEW SHOWING RIVERFRONT SIDE OF THE PROPERTY, FROM ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

77. GENERAL VIEW SHOWING RIVERFRONT SIDE OF THE PROPERTY, FROM WEST 77. GENERAL VIEW SHOWING RIVERFRONT SIDE OF THE PROPERTY, FROM WEST 77. GENERAL VIEW SHOWING RIVERFRONT SIDE OF THE PROPERTY, FROM WEST - Westover, State Route 633, Westover, Charles City, VA

397

When Oprah Intervenes: Political Correlates of Daytime Talk Show Viewing  

Microsoft Academic Search

This study examined the influence of daytime talk shows on opinion formation. Using agenda-setting and cultivation perspectives, it was hypothesized that both exposure to daytime talk shows and the apparent reality of these shows would be positively related to support for government involvement in social issues. In addition to exposure and apparent reality being positively related to levels of support,

Carroll J. Glynn; Michael Huge; Jason B. Reineke; Bruce W. Hardy; James Shanahan

2007-01-01

398

Best in Show: Teaching Old Dogs to Use New Rubrics  

Microsoft Academic Search

This paper discusses an instructional approach designed to help preservice teachers understand how assessments can be influenced by personal biases. In order to achieve this objective, we developed an analogy- based activity called \\

Austin M. Hitt; Emory C. Helms

2009-01-01

399

INTERIOR VIEW OF ENTRANCE TO LABORATORY, SHOWING HANDHAMMERED ALUMINUM DOORS ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

INTERIOR VIEW OF ENTRANCE TO LABORATORY, SHOWING HAND-HAMMERED ALUMINUM DOORS AND MARBLE. NOTE ALUMINUM LIGHT FIXTURE - Alcoa Research Laboratory, Freeport Road, New Kensington, Westmoreland County, PA

400

Interior view, detail of the staircase to show the burnished ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Interior view, detail of the staircase to show the burnished aluminum and brass balustrade - Departmental Auditorium, Constitution Avenue between Twelfth and Fourteenth Streets, Washington, District of Columbia, DC

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